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Vol. 49(4), December 2012

PAKISTAN ACADEMY OF SCIENCES
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C O N T E N T S
Volume 49, No. 4, December 2012
Page
Research Articles
Engineering Sciences
Electrochemical Behavior of Yttria Stabilized Thermal Barrier Coating on
— M. Farooq, K. M. Deen, S. Naseem, S. Alam, R. Ahmad and A. Imran
Life Sciences
— M. Jawed Iqbal, Zaeem Uddin Ali and S. Shahid Ali
— Aneela Yasmin, Akhtar A. Jalbani and Abdul Samad Chandio
Comparative Sequence Analysis of Some Rdr1 Resistance Genes from Rosa
— Aneela Yasmin, Akhtar A. Jalbani and Thomas Debener
Medical Sciences
— Naveed Akhtar, Rashida Parveen, Barkat Ali Khan, Muhammad Jamshaid and Haroon Khan
Review Articles
— Muhammad Hassan Khalil, Jia Dong Xu and Tsolmon Tumenjargal
— Wasam Liaqat Tarar, Sundus Sonia Butt, Fatima Amin and Mariam Zaka Butt
Physical Sciences
— M. Ayub Khan Yousuf Zai and Khusro Mian
— Muhammad Iqbal, Muhammad Iqbal Bhatti and Silvestru Sever Dragomir
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Proceedings of the Pakistan Academy of Sciences 49 (4): 235–240 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Research Article
Electrochemical Behavior of Yttria Stabilized Thermal Barrier
M. Farooq 1 , K. M. Deen 2* , S. Naseem 3 , S. Alam 4 , R. Ahmad 2 and A. Imran 4
1
Institute of Chemical Engineering & Technology,
University of the Punjab, Lahore 54590, Pakistan
2
Department of Metallurgy and Materials Engineering, CEET,
University of the Punjab, Lahore 54590, Pakistan
3
Center of Solid State Physics, University of the Punjab, Lahore 54590, Pakistan
4
Abstract: The plasma sprayed Yttria stabilized thermal barrier coating electrochemical behavior was
Electrochemical Impedance Spectroscopy (EIS) methods at room temperature. Thermal properties of applied
coating after 600 hours exposure in 3.5% NaCl solution demonstrated 0.5% weight loss at about constant
analysis revealed that the open circuit potential became positive after 600 hours compared to 0 hour with
decrease in polarization resistance. The 28% decrease in charge density after 600 hours exposure in 3.5%
NaCl solution was in support to decrease in polarization resistance. The decrease in charge transfer resistance
was attributed to the ingress of chloride ions at the coating metal interface and localized corrosion reactions
Keywords: Thermal barrier coating, plasma spray, corrosion resistance, electrochemical
1. INTRODUCTION
Thermal-barrier coatings (TBCs) have great
importance for insulation of components operating
at higher temperatures rather than protecting base
from oxidation. The TBCs have markedly increased
elevated temperatures [1]. The TBC is usually
a multilayer composite coating system consists
on metallic bond coat and ceramic top coat. The
metallic bond coat is multifunctional interlayer
which provides oxidation resistance to substrate
and promotes strong bonding between top coat and
base alloy [2–4].
A typical TBCs top coat is yttria-stabilized
zirconia (YSZ) with a metallic bond coat. The bond
————————————————
Received, August 2012; Accepted, November 2012
*Corresponding author: K. M. Deen; Email: kashif_mairaj83@yahoo.com
coat may have different recipes such as Ni-Cr-Al-Y
super alloy, Ni-Al, and Cr-Ni-Al, etc. YSZ top coat
is considered best as it has low thermal conductivity
and high temperature stability [5]. During operation
at high temperature the hot gases diffuse through
the pores in the top coat and oxidize the bond coat.
Long term thermal exposure results in the growth
of oxide layer between the bond and top coat and
hence cracking of TBCs.
At room temperature the diffusion of electrolyte
through micro-pores within the top coat will
facilitate growth of oxide layer at bond and top coat
interface and, eventually, failure of coating would
become certain. To evaluate degradation mechanism

236 M. Farooq et al
impedance spectroscopy (EIS) is a reliable method.
The porosity within topcoat and diffusion of
electrolyte can be evaluated by Equivalent Electrical
Circuit Model (ECM) simulated to the physical
system at room temperature [6–9].
The objective of this study was to assess the
corrosion tendency of TBCs when exposed to sea
water at room temperature. The physical absorption
of electrolyte in the coating and structural changes
were estimated by thermal analysis. Additionally,
the mechanism of electrochemical reactions at
the coating/metal interface was investigated by
electrochemical impedance spectroscopy.
2. MATERIALS AND METHODS
2.1. Substrate Coating
Coupons of carbon steel (0.08% C, 2.0% Mn, 1.09
% Si, 0.045% P and 0.03% S) of 12.5x 25mm
size were grit blasted with steel grit # 29 to attain
approximate surface roughness of 9µm. The
coupons were cleaned in distilled water and then
by acetone followed by ultrasonic cleaning at 60 o C.
Coupons were again rinsed in distilled water and
dried with compressed air to ensure removal of
aqueous medium.
Table 1.
coating.
—————————————————————————
Parameter
Value
—————————————————————————
Torch current intensity
500 Amp
Pressure & composition of gas 50psi (Ar/H 2
)
Pressure of carrier gas (Ar)
Powder feed rate
60 psi
6 lbs/hr
Spray distance & angle 6 inches at 90 0
Preheating temperature
150 0 C
Substrate cooling temperature
Normalizing
—————————————————————————
The Sulzer Metco Air Plasma Spray (APS)
coating system was used for TBC to deposit over
the surface. Before coating the powder was heated
at 105 ° C in an oven for about 30 minutes to remove
moisture. Ni 5
Al (Metco 450 NS powder) was
deposited at the substrate by APS as a bond coat
2
–8 wt% Y 2
3
) top coat was applied
over bond coat similarly. The operating parameters
of APS are given in Table 1. The thermal analysis
such as Differential Scanning Calorimeter (DSC)
and Thermo-gravimetric Analysis (TGA) of the
top coat after 600 hours immersion in 3.5% NaCl
solution was carried out to evaluate its physical
absorption of electrolyte characteristics.
2.2 Electrochemical Testing
The electrochemical characteristics of applied
Thermal Barrier Coating (TBC) were evaluated by
Polarization Method (LPR) and Electrochemical
Impedance Spectroscopy (EIS).The coated
surface was exposed to 3.5% NaCl solution in
a three electrode cell system while other sides of
the samples were covered with polyester resin. In
this cell Saturated Calomel Electrode (SCE) was
a reference, graphite as an auxiliary and coated
sample acted as a working electrode. The electrical
contact was made with the sample by soldering a
potential range -200 to +200mV Vs. SCE) were
taken after every 72hours upto 600 hours providing
an initial stabilization delay of 24hours. Similarly
EIS spectrums were obtained with 10mV AC
perturbation at the open circuit potential and
frequency range of 100 KHz–10 mHz. The whole
electrochemical investigation was exercised by
using Gamry Potentiostat (PC14-750).
3. RESULTS AND DISCUSSION
3.1 Thermal Analysis of Thermal Barrier
Coating (TBC)
The results of differential scanning calorimetry
(DSC) and thermogravimetric analysis (TGA) of
YSZ top coat when exposed to 3.5% NaCl solution
after 600 hours are shown in Fig. 1a & b, respectively.
It was evaluated that loss in mass of coating when

Electrochemical Behavior of Yttria Stabilized Thermal Barrier Coating 237
Fig. 1. Thermal analysis of TBC (a); DSC (b); TGA.
behavior with continuous decrease in 0.5% weight
in TGA was attributed to the stabilization of YSZ.
was due to the dehydration process with about 0.2%
weight loss which demonstrated the actual ability
of physical absorption of water in the coating upto
this temperature. The endothermic behavior of YSZ
diffusion of anions (oxygen, chlorides) within the
2
after stabilization with Y 2
3
from the electrolyte. The addition of ‘Y ions’ in
Zirconia occupies lattice positions which produce
oxygen vacancies. With the addition of Yttria in
Zirconia promote oxygen ions to ingress within the
YSZ lattice (oxidation) with increase in temperature
[10].
coated sample. The potential at 0 hour was -594 mV
and gradually increased to noble potential (-345.2
mV) after 600 hours (Fig. 2). The tendency to shift
potential in noble direction (positive direction) was
coat surface but still active (-345.2 mV) potential
was related to the presence of chloride ions in
The shift of potential at the rate of 0.297 mV/hour
towards noble direction indicated development of
3.2 Open Circuit Potential (OCP)
The open circuit potential (E ocp
) is a predictive
parameter to estimate corrosion tendency of a
system. The E ocp
of TBC samples with respect
to Saturated Calomel Electrode (SCE) was
determined for 600 hours when immersed in 3.5%
NaCl solution. The values of E ocp
were determined
after every 72 hours after initial stabilization for
24 hours. The potential trend was extrapolated at 0
hour to evaluate corrosion tendency of unexposed
Fig. 2.
time.
3.3 Linear Polarization Resistance (LPR)
corrosion rates and to understand kinetically

238 M. Farooq et al
Fig. 3. Variation in Polarization Resistance (R p
).
controlled electrochemical reactions. The LPR
and polarization resistance (R p
) and current
density (i d
) values after each 72 hour interval were
determined. The trend of change in R p
value with
time (Fig. 3). The increase in R p
and decrease in i d
values in 600 hours of exposure was observed and
this tendency was considered due to the formation
and accumulation of corrosion product within the
pores of coating after 600 hours at the interface
of top ceramic coat and bond coat. The initial R p
and i d
values after 24hours exposure were 21.80
with respect to initial values reached to 46.31
independence of Yittria stabalized Ziconia (YSZ)
Fig. 4. Decrease in charge density measured by LPR.
top coat to hinder the diffusion of electrolyte. The
charge density of YSZ coating when immersed in
3.5% NaCl solution was calculated by integrating
the current potential values of LPR curves. It was
deduced that initial charge density (160.6 µC/
cm 2 ) of coating was deteriorated to 114.6 µC/cm 2
as presented in Fig. 4. This decrease of 46 µC/
cm 2 in charge density and increase in polarization
resistance suggested the formation of corrosion
product and accumulation within the pores in the
coating after 600 hours exposure. This was also
responsible for the reduction in the active sites for
electrochemical reactions.
3.4 Electrochemical Impedance Spectroscopy
(EIS) of TBC
The EIS Nyquist plots for coated samples were
obtained at room temperature as shown in Fig.
5. These plots were modeled and simulated with
equivalent electrical circuit which was investigated
95%. It was apparent from the EIS investigation that
there were three time constants in the spectrums.
The solution resistance and YSZ coating resistance
at higher frequency and in the second time constant
depicted kinetically controlled reactions at the
interface of top coat and bond coat by developing
hour. This decrease in R ct
was due chloride ingress
at the interface and localized reactions with oxide
corrosion products. YSZ top coat resistance was very
low due to free penetration of electrolyte through
inherent micro-pores in the YSZ. The EIS study
evaluated kinetic and mass transport controlled
behavior of YSZ top coat and capacitative behavior
of bond coat at low frequency in a third time constant
was related with the formation of unstablized oxide
interface of bond coat and top coat. This result for
YSZ demonstrates typical behavior of TBC that
mass transport controlled reactions transformed to
kinetically controlled reactions at low frequency
regime of impedance spectrum.

Electrochemical Behavior of Yttria Stabilized Thermal Barrier Coating 239
Fig. 5. Electrochemical impedance spectroscopy (EIS) Nyquist plots.
In TBC the kinetics of electrochemical reactions
depend on the ionic resistance of electrolyte,
thickness, porosity and intrinsic defects within the
coating [7].
4. CONCLUSIONS
The thermal analysis of coating after exposure
to 3.5% NaCl solution revealed the absorption
of electrolyte within the coating and exothermic
regime and a sharp dip for endothermic reactions
corresponded to the stabilization of YSZ and
dehydration of electrolyte, respectively. The initial
loss in weight as revealed by thermal analysis was
related to desorption of electrolyte while further
increase in temperature and exothermic behavior
of top coat and heat evolution corresponded to the
stabilization of top coat. The shift of open circuit
potential in positive direction and increase in
polarization resistance with exposure time were
of top and bond coat. But the decrease in charge
transfer resistance (R ct
) as determined by EIS was
due chloride ingress at the interface and localized
since their greater capability of hydrolyzing the
corrosion product.
5. REFERENCES
1. Birks, N., G. H. Meier, & F. S. Pettit. Introduction
to the High-Temperature Oxidation of Metals, 2nd
ed. Cambridge University Press, Cambridge, UK
(2006).
2. Yu, Z., D.D. Hass, & H.N.G. Wadley. NiAl bond
coats made by a directed vapor deposition approach.
Materials Science and Engineering A 394: 43–52
(2005).
3. Zhou, Z. F., E. Chalkova, S.N. Lvov, P. Chou, & R.
Pathania. Development of hydrothermal deposition
process for applying Zirconia coating on BWR
materials for IGSCC mitigation. Corrosion Science
49 (2): 830–843 (2007).
4. Podor, R., N. David, C. Rapin, M. Vilasi, & P.
Berthod. Mechnism of corrosion layer formation
during Zirconium immersion in (Fe) bearing glass
melt. Corrosion Science 49 (8): 3226–3240 (2007).
5. Wang, Z., A. Kulkarni, S. Deshpande, T. Nakamura,
& H. Herman. Effect of pores and interfaces on
effective properties of plasma sprayed Zirconia
coatings. Acta Materialia 51: 5319–5334 (2003).
6. Heung, R., X. Wang, & P. Xiao. Characterization of
impedance spectroscopy. Electrochimica Acta 5:
1789-1793 (2006).
7. Zhang, J., & V. Desai. Evaluation of thickness,
porosity and pore shape of plasma sprayed TBC by
electrochemical impedance spectroscopy. Surface
Coatings Technology 190: 98-109 (2005).
8. Byeon, J. W., B. Jayaraj, S. Vishweswaraiah, S.
Rhee, V.H. Desai, & Y.H. Sohn. Non-destructive
evaluation of degradation in multi-layered thermal
barrier coatings by electrochemical impedance
spectroscopy. Material Science and Engineering: A

240 M. Farooq et al
407 (1-2): 213-225 (2005).
9. Jayaraj, B., S.Vishweswaraiah,V.H.Desai, & Y.H.
Sohn. Electrochemical impedance spectroscopy of
thermal barrier coatings as a function of isothermal
and cyclic thermal exposure. Surface Coatings
Technology 177–178: 140–151 (2004).
10. American Ceramic Society. Progress in Thermal
Barrier Coatings. John Wiley and Sons (2009).

Proceedings of the Pakistan Academy of Sciences 49 (4): 241–249 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Research Article
Agroclimatic Modelling for Estimation of Wheat Production
in the Punjab Province, Pakistan
M. Jawed Iqbal 1,2* , Zaeem Uddin Ali 1 and S. Shahid Ali 3
1
Institute of Space and Planetary Astrophysics, University of Karachi, Karachi, Pakistan
2
Department of Mathematics, University of Karachi, Karachi, Pakistan
3
Department of Geography, University of Karachi, Karachi, Pakistan
Abstract: Pakistan’s economy hinges on agriculture and the most important agricultural commodity of the
country is wheat. The province of Punjab has the predominant share in wheat production of the country.
As agriculture sector is highly vulnerable to the climate change phenomena, the current global climatic
article reports on an attempted agroclimatic model for the estimation of wheat production in Punjab using
meteorological parameters.
Keywords: Agroclimatic model, prediction of wheat yield, agrometeorological variables
1. INTRODUCTION
Agriculture is the linchpin of our national economy.
According to economic survey of Pakistan 2011–
2012 it has a share of 21% in the national GDP
and employs 45% of the national labour force. The
agricultural production system of Pakistan is based
on irrigation. In Pakistan 84% of the total cultivated
area (22.05 million ha) is under irrigation and the
remaining is entirely rainfed or barani [1]. More
than 90% of the available fresh-water resources in
the country are utilized for irrigated agriculture [2].
Punjab is Pakistan’s second largest province with
205,344 km 2 (79,284 mi 2 ), after Balochistan, and
is located at the northwestern edge of the
geologic Indian plate in South Asia. The Fig. 1
depicts the distribution of average yield of wheat
in various districts across the country. According to
McCarty’s measure, 28 out of 35 districts of Punjab
have higher yield than the country’s average yield,
i.e., 2.05 ton per hectare. The northern districts of
the Punjab have lower productivity per unit land
area, primarily due to the topographical features
and more relative humidity [3]. Several studies have
highlighted agro-ecological zones for the growth
————————————————
Received, April 2012; Accepted, November 2012
*Corresponding author, M. Jawed Iqbal; Email: javiqbal@uok.edu.pk
and development of crops across the country [3, 4].
The cultivate areas of the entire country are divided
into 10 agro-ecological zones (Fig. 2). Out of which
four agro-ecological zones, i.e., irrigated plains,
Barani (rain fed) region, Thal region and Marginal
Lands occur in the Punjab province.
The agricultural sector of Pakistan is confronted
with many problems. The provision of livelihood
and food security for the fast growing population,
without affecting the fragile ecosystem, are the
main challenges to the agriculture sector. Being
open to vagaries of nature, agriculture sector is
highly susceptible to climate change phenomena.
The scholars opine that human activities have
reached a level where these are adversely affecting
global environment. Anthropogenic perturbations
are causing the global climate change at a much
faster rate compared to natural pace. The current
on sustainable agriculture production and food
security as described by Adams et al. [5]. Increasing
change in ambient temperature and rainfall and
the resultant increased frequency and intensity of

242 M. Jawed Iqbal et al
Fig. 1. Wheat growing areas in Pakistan, 2004-05 (courtesy: Dr. S.S. Ali).
Fig. 2. Agro-ecological zones of Punjab (courtesy: Dr. S.S. Ali ).

Agroclimatic Modelling for Estimation of Wheat Production 243
the crop-lands, pastures and forests. Thus, it was
thought important to investigate the impact of
current climate change on agriculture of the country.
Estimates of evapotranspiration offer an outlook of
soil-water balance in association with the amount
and irrigation scheduling [6]. Also, these estimates
can help determine the nature of agro-climate,
agro-climatic potential of the region and suitability
of various crops or varieties which can be grown
for obtaining the best economic returns [7].
A recent study employed Growing Degree Days
(GDD) for expressing the relationship between
period of every plant growth phonological stage
and the temperature degree [8]. Growing Degree
Days of all Andhra Pradesh were correlated with
variability of rice yield of this Indian state [9].
Satellite-derived product of normalized difference
vegetation index (NDVI) were used by Chakraborty
et al. [10] for the estimation of rice yield. In this
study as well, we employed NDVI for estimating
the wheat yield of Punjab province by examining
the relationship between wheat productivity per unit
area and integrated NDVI. Finally, we constructed
a multiple regression model of wheat in the Punjab
using agro-meteorological variables derived from
nearest stations, i.e., the remotely sensed satellite
data (e.g. NDVI, GDD, and SOI).
2. DATA AND METHODOLOGY
2.1. Data
Yield of wheat crop has been collected from website
of Agriculture Marketing Information Service
Directorate of Agriculture (Economics& Marketing)
Punjab, Lahore. The Pakistan Meteorological
Department provided data of daily maximum and
minimum temperature of Lahore, Jhelum, Khanpur,
Multan, Faisalabad, Islamabad, Bahawalpur, and
annual data of rainfall of Islamabad, Jhelum, Lahore,
Sargodha, Faisalabad, Multan, Bahawalpur.
The following formula was used to compute the
Growing Degree Days (GDD):
where T max
and T min
are daily maximum and minimum
temperature, T b
is the threshold temperature below
which growth of wheat crop is not possible, and
T b
of monthly GDD temperature for each mentioned
stations using T max
and T min
. Next, the monthly data
of GDD temperature of Punjab was computed by
taking average of monthly GDD temperature for
each mentioned stations.
Normalized Difference Vegetation Index (NDVI)
is the satellite-derived product which represents the
green vegetation in area. It works on the phenomena
that growing green plants absorb radiations in
visible region and emit infra red radiation. Data of
NDVI can be downloaded from the website www.
jisao.washington.edu. NVDI was computed using
the formula:
Where NIR
R
This spectral pattern is described by a value
ranging from -1 (usually water) to +1 (strongly
vegetative growth). We computed spatial average
of monthly NVDI over the region of Punjab
province.
Other parameters which were used in this study
is South Oscillation Index (SOI). SOI and Nino 3
were obtained from the website www.cdc.noaa.
gov. SOI was used to indicate El Nino and La Nina
events. It is calculated by using the Mean Sea Level
Pressure (MSLP) difference between Tahiti and
Darwin. It can be calculated as:
where P diff
= (Average Tahiti MSLP for the
month) - (Average Darwin
MSLP for the month)
P diffav
= Long term average of P diff
for
the month
SD (P diff
) = Long term standard deviation
of P diff
for the month

244 M. Jawed Iqbal et al
2.2. Methodology
To examine the impact of agro-climatic parameters
season mean of crops from November to April. It
is important to understand the role of agro-climatic
parameters in modelling for wheat crop. Albeit
techniques for identifying parameters range from
simple correlation analysis to advanced procedures
such as canonical correlation and neural network
[13, 14], we used the Pearson correlation analysis
calculated between crop yields and agro-climatic
parameters (e.g. rainfall, GDD, NVDI, SOI, etc.).
Secondly, multiple linear regression equation
was constructed between wheat yields of Punjab
variance associated with each parameter designates
the relative importance of the index in modulating
the crop variability. Moreover, the total variance
inter-annual variations. The linear regression model
Y = b 0 + b i X i
, (i {0, 1, 2, 3, ... ,p}),
where Y is the estimated wheat yield in standardised
unit, b 0 is the intercept, X i
is a value of the i th
predictor (agro-climatic parameter) in the model,
b i is the model parameters representing the linear
relationship between the predictor and predicted, p
the number of the predictors variable in the model.
3. RESULTS AND DISCUSSION
Percentage departure of wheat yield in Punjab
province from median 599.2 is plotted in Fig. 3.
Percentage departure started from 65.5% and
there is a fall initially up to minimum value of
132.2% (in 1952). After that it rose linearly with
some ups and downs. It had the maximum value
of 46.6% in the year 2006. Wheat yield was below
the median till year 1977. Since 1978 the yield was
above the median, except in 1983. In our study we
also observed that during La Nina years (i.e., 1947,
1948, 1949, 1954, 1955, 1956, 1964, 1967, 1970,
1971, 1973, 1975, 1988, 1998, 2000, and 2007) the
average yield of wheat was 558.32 kg/acre. During
El-Nino Years (i.e., 1951, 1957, 1963, 1965, 1969,
1972, 1976, 1982, 1986, 1987, 1991, 1997, 2002
and 2009) the average yield was 621.68 kg/acre,
which clearly indicated that warm phase of ENSO
had positive effect on wheat yield. . Sarma et al [9]
have also obtained similar results for rice production
in Andhra Paradesh, a province of India.
3.1 Variation in Wheat Yield with Meteorological
Parameters
increasing trend (Fig. 4). The annual rainfall and
wheat crop yields are plotted in Fig. 5. It shows that
the wheat yield increased linearly and its trend is
linear from 1987 to 1998. But overall its trend did
not described the variation in wheat yield. The
highest wheat yield was recorded in the year 1999
when the rainfall was observed 453 mm and the
minimum wheat yield was recorded in the year 1983
when the rainfall was 766 mm. Pearson correlation
0.16,
wheat area is irrigated by tube wells, this might be
relationship of crop yield with the rainfall was also
observed by Sarma et al [9].
Fig. 6 depicts the plot of wheat yield with Nino
Table 1.
Parameters/Months Nov Dec Jan Feb Mar Apr Nov-Apr Selected Months
SOI 0.38 * 0.36 * 0.12 0.30 0.25 0.14 0.28 0.38 * Nov-Dec
GDD 0.45 ** 0.42 * -0.12 0.24 0.09 0.37 * 0.41 * 0.50 ** Nov-Dec
NDVI -0.08 -0.29 0.40 * 0.58 *** 0.46 ** 0.03 0.22 0.52 ** Jan-Mar

Agroclimatic Modelling for Estimation of Wheat Production 245
Fig. 3. Percentage departure of wheat yield in the Punjab.
Fig. 4. Five-year moving average of wheat yield in the Punjab.
Fig. 5. Variability of wheat yield with rainfall in the Punjab.

246 M. Jawed Iqbal et al
3 SST. It was observed that there was no correlation
between these variables; the Pearson correlation
0.1, which was
relation with Kharif crops whereas for Rabi crops
it was not meaningful. There are different stages in
growth of wheat plant. SOI has been tested month
wise and its correlation with wheat yield which
correlation (with p-value less than 0.1) during the
months of November and December, we took the
average of SOI for these two months for further
analysis. This showed that the average SOI of the
phonological stages of wheat plant growth. Fig. 7
illustrates the variability of wheat yield with the
Southern Oscillation index (during Nov-Dec).
Southern Oscillation index varied from 2.69 in the
ENSO year 1982 for wheat yield of 684 kg to 1.59
in the ENSO year 1988 for wheat yield of 761.6
kg.
relationship with wheat yield in the months of
November and December. So we understand that
GDD explain the initial development of wheat plant
Fig. 6. Variability of wheat yield with Nino 3 SST in the Punjab.
Fig. 7. Variability of wheat yield with SOI in the Punjab.

Agroclimatic Modelling for Estimation of Wheat Production 247
growth. Wheat yield had linear dependency on GDD
(Fig. 8). GDD had the range of 135 units with the
lowest value of 646 units in the ENSO year 1997
and highest value of 781 units in the year 1999 for
the maximum yield of 1079 kg. It exhibited positive
the period of January to March. Fig. 9 represents the
proportionality of NDVI (for the months January
to March) with wheat yield. Pearson correlation
among all the other parameters. Its value was 0.52
of NDVI was 0.79 in the ENSO year 1982 for wheat
yield of 684 kg and the maximum value of NDVI
was recorded in the year 1993, which was declared
this year the wheat yield was 786.6 kg. It is relevant
the model developed by Bazgeer et al [16] for the
prediction of wheat yield of Hoshiarpur district of
Punjab, India.
3.2 Statistical Model for Prediction of Wheat
Yield
meteorological parameters SOI (Nov-Dec), NDVI
(Jan-Mar) and Growing Degree Day Temperature
(Jan-Mar) for the Punjab province which (Table
2). It shows that NDVI and GDD were mutually
Fig. 9. Variability of wheat yield with NDVI in the Punjab.
Fig. 10. A comparison of actual wheat yield with estimated wheat yield.

248 M. Jawed Iqbal et al
Fig. 8. Variability of wheat yield with GDD in the Punjab.
independent. The statistical model of wheat yield
in the Punjab province for was constructed with
two independent parameters (i.e. NDVI and GDD)
by using multi-regression analysis. The regression
equation was:
Y = 587.54*X 1
+1.34*X 2
717.06
where Y = Wheat Yield in kg/acre
X 1
= Seasonal average of NDVI of
Punjab for the months from
January to March
X 2
= Seasonal average of Growing
Degree Day Temperature of
Punjab for the months from
November to December
Table 2.
Parameter NDVI GDD SOI
NDVI 1.00 0.22
GDD 0.22 -
SOI 0.20 0.41 * -
model was +0.65. Thus, our model explained
43% variability of wheat yield for the period from
1980 to 2000. Fig. 10 shows the comparison of
actual wheat yield and estimated wheat yield. The
maximum difference was approximately 190.82 kg
in the year 2000 which was less than actual wheat
yield.
4. CONCLUSIONS
This study demonstrated the impact of regional
meteorological parameters (i.e., NDVI and SOI)
wheat yield in Punjab province of Pakistan. The
wheat yield was not dependant on rainfall; also
Nino 3 SST had no explanation with regard to
wheat productivity in the Punjab. However, South
oscillation exhibited some relationship with wheat
yield, but it was not included in the model because
GDD temperature had greater impact on wheat
yield. NDVI had a stronger correlation with wheat
yield, compared to other parameters. Thus, we
construct a multi-linear model for wheat yield by
employing NDVI and GDD. This model explains
the 43% variability of wheat yield for the 1980–
2000 period.
5. REFERENCES
1. Iqbal, M. M., M. A. Goheer & A. M. Khan. Climate-
Change Aspersion on Food Security of Pakistan.
Science Vision 15(1): 15-23 (2009).
2. Bhatti, M.A., and Akhtar. Increasing irrigated
agricultural productivity for Poverty reduction
in Pakistan. In: Second South Asia Water Forum,
Pakistan Water Partnership, Vol. 2. 14-16 December
2002, Islamabad, p. 600 (2002).
3. Ali, S. S. Structure and Spatial Patterns of
Agriculture in Pakistan: A Study in Regionalization.
PhD thesis, University of Karachi, Karachi, Pakistan
(2010).

Agroclimatic Modelling for Estimation of Wheat Production 249
4. PARC. Agro-ecological Regions of Pakistan.
Pakistan Agricultural Research Council, Islamabad
(1980).
5. Adams, R.M., B. H. S. Hurd, Lenhart, & N. Leary.
Effects of global climate change on agriculture: an
interpretative review. Climate Research 11: 19–30
(1998).
6. Rasul, G. Water requirement of wheat crop in
Pakistan. J. of Engg. & App. Sci. 3(1): 65-79
(1992).
7. Rasul, G. & A. B. Farooqi. Water requirement of
cotton crop in Pakistan. J. of Engg. & App. Sci.
4(2): 154-165 (1993).
8. Esfandiary, F., G. Aghaie, & A. D. Mehr. Wheat
Yield Prediction through Agro Meteorological
Indices for Ardebil District. World Academy of
Science, Engineering and Technology 49: 32-35
(2009).
9. Sarma. A.A.L.N., T V. L. Kumar & K. Koteswararao.
Development of an agroclimatic model for the
estimation of rice yield. J. Ind. Geophys. Union 12
(2): 89-96 (2008).
10. Chakraborty, M., Panigraohy & J.S. Parihar. Use of
NOAA AVHRR data in cloud cover prone areas for
rice yield and production estimation: A case study
in Orissa state. Proc. Nat, Symp on Remote Sensing
Applications for Resource Management with Special
Emphasis on North East Region, Guwahati, Nov
25-27, 1993, p. 285-291 (1993).
11. Dubey, R. P., M. H. Kalubarme, O. P. Jhorar, & S.
S. Cheema. Wheat yield models and production
estimates for Patiala and Ludhiana districts based
on Landsat – MSS and agro-meteorological data.
Applications Center, Ahmadabad, p. 1-34 (1987).
12. Sharma, A. R. K. Sood, & M. H.Kalubarme.
Agrometeorological wheat yield forecast in
Himachal Pradesh. J. Agromet. 6: 153-160 (2004).

Proceedings of the Pakistan Academy of Sciences 49 (4): 251–257 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Research Article
MATLAB-based Sequence Analysis of muRdr1H, a Functionally
Characterized Resistance Gene of Roses
Aneela Yasmin 1 , Akhtar A. Jalbani 2 * and Abdul Samad Chandio 3
1
Department of Biotechnology, Faculty of Crop Production,
Sindh Agriculture University, Tandojam-70060, Pakistan
2
Information Technology Center, Sindh Agriculture University,
Tandojam-70060, Pakistan
3
Department of Irrigation and Drainage, Faculty of Agricultural Engineering,
Sindh Agriculture University, Tandojam-70060, Pakistan
Abstract: One of the practical applications of Bioinformatics is sequence analysis through sequence
softwares available for the biological sequence alignment; in this paper some basic alignment algorithms are
discussed and implemented in MATLAB Bioinformatics tool box. Two sequences analyzed by this tool box
were muRdr1H and resistance protein of Populus trichocarpa (ACCESSION XP_002329162). muRdr1H is
a functionally characterized member of Rdr1 resistance gene family of roses, active against black spot. The
global sequence alignment. Our proposed work provides useful application of MATLAB which can help in
interpretation and visualization of the data in molecular biology.
Keywords: Sequence alignment, MATLAB, Bioinformatics, protein sequence
1. INTRODUCTION
Computer science in combination with biological
science is a relatively new multidisciplinary area of
science, known as Bioinformatics. In this research
area, Informatics involves the technology that
uses computer for different purposes; for example,
storage of biological data and for performing
manipulation and distribution. As the data contains
biological information, hence the distribution of
information is related to biological aspects, such
as DNA, RNA and proteins [1]. In this domain,
research focuses on the usage of computers because
most of the tasks in genomics data analysis are
highly repetitive. Moreover, the common activities
performed in Bioinformatics research include
mapping and analyzing or aligning DNA and protein
sequences, comparing and creating 3-D models of
drug design and discovery.
In computational biology the main aspects
of using Informatics is to develop innovative
algorithms to compute biological sequencerelated
problems. In handling biological
sequences, sequence alignment is a key process of
Bioinformatics and computational biology. During
alignment, sequences are compared by identifying
similar patterns and establishing residue-residue
correspondence among related sequences [2, 3].
Hence sequence alignment is a way of arranging
primary sequences of DNA, RNA and proteins to
detect similar regions that may be consequences of
————————————————
Received, May 2012; Accepted, November 2012
*Corresponding author, Akhtar A. Jalbani; Email: akjalbani@sau.edu.pk

252 Aneela Yasmin et al
functional, structural or evolutionary relationship
between the sequences. The resulting alignment
produces revise transcript of mismatches, i.e.,
insertions and deletions, where mismatches can
be inferred as point mutations. As a result, we can
infer how sequences with the identical origin would
deviate from one another.
Nowadays, new biological sequences are being
generated at an exponential rate; hence sequence
comparison is widely used in biological research to
identify new proteins and can search for existing
protein for drug or diseases discovery. In any
genome project, newly determined sequences are
in the genomic databases, such as NCBI, in order
to discover similarities. As a result of comparison,
one or more sequence alignments can be produced.
Similarity score is one factor that can be associated
with the sequence alignment. If new sequences are
to the biological databases, like NCBI, ENTREZ
(which integrates GenBank) [4]. These databanks
are remotely accessible. Researchers take full
advantages of these databases to query and compare
their sequences using different Bioinformatics
tools.
expose the structural or functional importance of
unknown sequences. Here in this paper, two very
important types of DNA alignments are discussed
namely Global and Local alignment in MATLAB.
MATLAB is a powerful tool for the modeling and
simulation of various domains. It contains different
toolbox for the different domain for example
communication systems, image compression,
Bioinformatics, etc. MATLAB provides tight
integration of compiler and its simulating
environment. In this paper, we focused on the
Bioinformatics tool box, which contains different
solutions for the biological research; sequence
alignment is one of them. In local alignment, the
assumption is made based on that the two sequences
are not similar over entire length [5]. Hence it only
similarities and the aligned two sequences show
region based alignment without considering the
alignment of the rest of the regions. It is possible
that the two sequences to be aligned can be of
different lengths [6, 7] whereas global alignment
uses dynamic programming to obtain global
alignment between the two selected sequences.
These two algorithms were applied in MATLAB.
The two sequences selected to analyze in
MATLAB were muRdr1H and TIR-NBS-
LRR-resistance protein of Populus trichocarpa
(ACCESSION XP_002329162). muRdr1H is a
member of Rdr1 resistance gene family of roses
active against black spot, a fungal disease. This gene
active resistance genes against Dort E4, race 6 of
diplocarpon rosae [8, 9] whereas TIR-NBS-LRRresistance
protein of Populus trichocarpa shares
the highest identity (41%) to muRdr1H protein.
It is important to state that the TIR-NBS-LRRresistance
protein of Populus trichocarpa was not
functionally characterized.
2. MATERIALS AND METHODS
To identify the homologues of muRdr1H gene
BLASTx searches were carried out against the
GeneBank non-redundant database (http://blast.
ncbi.nlm.nih.gov). The selected two sequences were
tested using MATLAB Bioinformatics software
tools (www.mathworks.com). Those two sequences
are assumed to be identical in length. The sequence
alignment is carried out from beginning to the end
alignment. The two types of alignment methods
considered for the practical assignment were local
alignment and global alignment.
2.1 Algorithms for the DNA Sequence
Alignment
DNA sequence strings consist of four alphabet
letters (A, C, G, T) called nucleotides. The length
of sequence is variable; hence algorithm should
produce high quality sequence alignment from
these four letters. In local alignment, we need to
identify the isolated regions of high similarity from
the entire DNA sequence that makes better choice
in some situation for this type of alignment method
but it is more complex in general [10].
2.1.1. Local Alignment
The local alignment is performed through very

MATLAB-based Sequence Analysis of muRdr1H 253
well-known algorithm known as Smith-Waterman
algorithm. It is used for determining identical regions
between two nucleotides or proteins. The algorithm
does not look for total sequence but it compares
segments of all possible lengths and optimizes the
similarity measures. Following steps should be
considered for Smith-Waterman algorithm:
Step-1: Fill all dynamic matrix
max value and trace of max value for that patch
which leads to the max value or score.
The Smith-Waterman algorithm compares
two DNA sequences based individual pair-wise
comparison between the characteristics.
2.1.2. Global Alignment
Global alignment method uses dynamic
programming, for this Needleman-Wunsch
algorithm is the best algorithm for this type of
sequence alignment. From two sequences, this
algorithm helps to identify the global alignment.
This uses all elements of the two sequences for the
alignment procedure. This is also called end-to-end
alignment.
All elements are considered in global sequence
alignment method, therefore the scoring matrix will
also become m*n (where m is the longer sequence
and n is the shorter sequence). The optimal score
can be calculated at each matrix position by adding
current match score to last scored position and
subtracting the gap penalties. Hence each matrix
position may have +ve or –ve or even zero value.
2.2. Sequence Alignment with MATLAB
The algorithms discussed above can easily be
best possible sequence alignment for nucleotide
or protein. MATLAB contains different built-in
functions to access the already stored sequencing
data on the gene databanks. Using MATLAB we
can apply global and local alignment method to
entered by accession numbers of the sequences,
sequences were retrieved in its Open Reading
Frames (ORF). After bringing the information in
MATLAB environment, we applied algorithms for
comparison of sequences using global and local
alignment with the score that determines the degree
of similarity. At the end, Monte Carlo Techniques
were applied in MATLAB environment to assess
sequences were generated and their scores were
statistics toolbox, the parameters of bar distribution
were estimated and the probability density function
of the estimated distribution was plotted in red line
(Fig. 1).
3. RESULTS AND DISCUSSION
3.1. Selection of Sequences to Study Alignment
Algorithms
According to BLASTx searches muRdr1H protein
shares the maximum, i.e., 41%, identity to TIR-NBS-
LRR-resistance proteins of Populus trichocarpa
(ACCESSION XP_002329162), both genes belong
to the same class of TIR-NBS-LRR resistance
genes, followed by 39-44% to hypothetical proteins
of Vitis vinifera, 40% to TIR of Medicago truncatula
(ACCESSION ABD28703), 40% to CMR1 of
Phaseolus vulgaris (ACCESSION ABH07384)
and 39% identity to N-like protein of N. tabacum
(ACCESSION BAF95888) for resistance to the
Tobacco Mosaic Virus. Out of these sequences
TIR-NBS-LRR-resistance proteins of Populus
trichocarpa was selected for the alignment with
muRdr1H using MATLAB to demonstrate the
utility of this software for searching differences in
their protein sequences.
3.2. Open Reading Frame of Selected Sequences
in MATLAB Environment
The protein sequences of both genes were in text
format. We used the same sequence by using
seqshoworfs MATLAB method to open it in open
reading frame work reader (ORF) as shown in Fig.
2.
3.3. Dot Plot based Comparison in MATLAB
Environment
The dot plot representation of selected sequences
is shown in Fig. 3. This basic sequence alignment
method compares two sequences in graphical
method by comparing two dimensional matrix; two
sequences are written in vertical and horizontal

254 Aneela Yasmin et al
Fig. 1. muRdr1H and TIR-NBS-LRR resistance
protein of Populus trichocarpa are assessed using Monte Carlo Techniques.
Fig. 2. Open Reading Framework (ORF) options for muRdr1H Sequence in MATLAB.

MATLAB-based Sequence Analysis of muRdr1H 255
Fig. 3. Dot plot of muRdr1H and TIR-NBS-LRR resistance protein of Populus trichocarpa in MATLAB
environment.
Fig. 4. Global Alignment of muRdr1H and TIR-NBS-LRR resistance protein of Populus
trichocarpa.

256 Aneela Yasmin et al
Fig. 5. Local Alignment of of muRdr1H and TIR-NBS-LRR resistance protein of Populus trichocarpa.
axes of the matrix and compared [10]. The selected
sequences exhibited substantial regions of similarity
(Fig. 3). Many dots are lined up in a diagonal line
revealing some sequence alignment that will be
and global alignment. It is obvious from the line that
TIR and NBS region of protein has more similarity
as compared to the end region that represents LRR
both proteins, which was 41% as described above.
3.4. Global and Local Sequence Alignments
Both sequences are compared using global and local
alignment methods. For global alignment we used
Needle-Wunsch algorithm in the MATLAB and the
resultant aligned global sequence is shown in Fig
3. The scoring of global alignment is 749.33 with
522/ 1232 (42%) identities and 822/ 1232 (67%)
positives. The same sequences were used for local
alignment. In which we used Smith-Waterman
algorithm, where pair wise comparison is shown
is 766.66 with 43% identities and 67% positives.
Sequences with good similarity did not show much
difference between local and global alignments.
conserved regions of a gene.
The number of identities and positives in global
and local alignments are shown in Fig. 4 and Fig.
5, respectively. Actually this did not represent the
Techniques in MATLAB environment to assess
methods. The random sequences were generated in
MATLAB and their scores were plotted as bars (Fig
gave the scores of 49.28 and 9.5. The probability
density function of the estimated distribution was
plotted which clearly showed that global alignment

MATLAB-based Sequence Analysis of muRdr1H 257
In this paper, global and local alignment
algorithms were developed and simulated using
MATLAB. For global alignment, Needleman-
Wunsch algorithm and for local alignment Smith-
Waterman algorithm were used in MATLAB. This
demonstration of data mining, visualization and of
course interpretation in MATLAB can facilitate the
sequence data handling in molecular biology.
4. REFERENCES
1. Mathkour, H. & M. Ahmad. A comprehensive
survey on genome sequence analysis. In: IEEE
International Conference on Bioinformatics and
Biomedical Technology, p. 14-18 (2010).
2. Benkrid, K., Y. Liu & A. Benkrid. A highly
for pairwise biological sequence alignment. IEEE
Transactions on Very Large Scale Integration
Systems 17(4): 561-570 (2009).
3. Boukerche, A., J. M. Correa, A. Cristina M.A de
Melo & R. P. Jacob. A hardware accelerator for
the fast retrieval of DIALIGN biological sequence
alignments in linear space. IEEE Transactions on
Computers 59(6): 808-821 (2010).
sequence alignment. IEEE/ACM Transactions on
Computational Biology and Bioinformatics 6(4):
570-562 (2009).
5. Nguyen, V., A. Cornu & D. Lavenier. Implementing
protein seed-based comparison algorithm on the
SGI RASC-100 platform. In: IEEE Symposium on
Parallel and Distributed Processing. p. 1-7 (2009).
6. Bandyopadhayay, S. & R. Mitra. A parallel
pairwise local sequence alignment algorithm. IEEE
Transactions on Nano Bioscience 8(2): 139-146
(2009).
7. Nahar, N., M. Popstova & J. Gogarten. GPX: A tool
for the exploration and visualization of genome
evolution. In: 7th IEEE International Conference on
Bioinformatics and Bioengineering, p. 1338-1342
(2007).
8. Yasmin, A.
Characterization of Rdr1 Resistance Gene from
Roses. PhD thesis, University of Hannover, Germany
(2010).
9. Terefe-Ayana, D., A. Yasmin, T. L. Le, H. Kaufmann,
A. Biber, A. Kuehr, M. Linde & T. Debener. Mining
disease resistance genes in roses: functional and
molecular characterization of the Rdr1 locus.
Frontiors of Plant Science 2:35, doi: 10.3389/
fpls.2011.00035 (2011).
10. Mai, S. M., M. Hamdy, M. Aboelfotoh & Y. M.
Kadah. BIOINFTool: Bioinformatics and sequence
data analysis in molecular biology using MATLAB.
In: Cairo International Biomedical Engineering
Conference, p. 1-9 (2006).

Proceedings of the Pakistan Academy of Sciences 49 (4): 259–267 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Original Article
Comparative Sequence Analysis of Some Rdr1 Resistance Genes
from
Aneela Yasmin 1 , Akhtar A. Jalbani 2 * and Thomas Debener
1
Department of Biotechnology, Sindh Agriculture University, Tandojam, Pakistan
2
Information Technology Center, Sindh Agriculture University, Tandojam, Pakistan
3
Institute of Plant Genetics, University of Hannover, D-3419, Hannover, Germany
Abstract: Rdr1
active against a fungal disease, black spot, of roses. Rdr1 locus consists of nine genes named as muRdr1A
to muRdr1I. Functional and molecular characterization of this locus, revealed the presence of one gene,
muRdr1H, active against Dort E4, race 6 of Diplocarpon rosae. In the present study we aimed to analyze
the protein sequences of three members of this Rdr1 gene family for their homologies, important domains,
conserved regions and selection pressure to get insights of their functionality based on protein sequence.
The protein sequences of one active gene, muRdr1H and two inactive genes, muRdr1C and muRdr1G, were
deduced and dissected using internet based bioinformatics tools. All three proteins belong to the TIR-NBS-
LRR family of resistance genes. Although these proteins carry all necessary domains and conserved regions
necessary for the functionality of TIR-NBS-LRR proteins, LRR region of the proteins, found under selection
Keywords: Rdr1, rose, black spot, TIR-NBS-LRR proteins, resistance genes
1. INTRODUCTION
The most devastating fungal diseases of roses are
powdery mildew, black spot and downy mildew.
Powdery mildew usually infects roses grown in
greenhouses whereas the black spot is a problem
moist conditions throughout the world [1]. The
present control of this disease is fungicidal sprays.
On the other hand use of resistant varieties with
durable genetic resistance is the safest option for
the sustainable environment. Most of the cultivated
roses lack natural resistance against black spot
thus it has to be introgressed from wild roses [2].
Nevertheless the exploitation of natural genetic
resistance requires understanding of the resistance
genes in terms of diversity, genomic organization
and functionality.
Rdr1
resistance gene locus found active against black
spot (Diplocarpon rosae) of roses through
phytopathological methods [3]. This Rdr1 locus
was later mapped to a telomeric position of linkage
group 1 in the diploid population 94/1 [4]. The
construction of two large insert BAC libraries [5,
location of Rdr1 gene within a 220 kb region [6].
The 220 kb region contains 9 copies of “resistancegene-analogues”
sequences (RGA) of the T1R-
NBS-LRR type of resistance genes [7]. Out of these
9 genes one gene muRdr1H was found as major
active gene against Dort E4, race-6 of Diplocarpon
rosae by functional characterization [8, 9].
In TIR-NBS-LRR resistance genes, TIR motif
is similar to toll/interleukin-1-receptor (TIR),
the NBS is a part of a nucleotide binding (NB)-
ARC domain that belongs to the STAND (signal
transduction ATPases with numerous domains)
family of NTPases. These proteins are proposed
————————————————
Received, May 2012; Accepted November 2012
*Corresponding author: Akhtar A. Jalbani; Email: akjalbani@sau.edu.pk

260 Aneela Yasmin et al
to regulate signal transduction as NB domain
hydrolyzes NTP and changes its conformational
states [10]. The NB-ARC domain has three subdomains
conserved in NBS-LRR proteins: a P-loop
bundle, and an ARC2 adopting a winged-helix fold
that is connected to the LRR domain by a short
linker [10]. LRR domains contain various numbers
of tandemly repeated leucine-rich motifs with a
conserved core consensus of L-X-X-L-X-L-X-X-N
structure of the LRR domain suggests its role in
different intra and intermolecular interactions of
direct recognition of pathogen effectors, regulating
protein activation and signal transduction [12].
The current research was aimed to analyze protein
sequences of three paralogs of Rdr1 locus namely
muRdr1H (AEE43932), muRdr1C (AEE43927) and
muRdr1G (AEE43931). These paralogs are unique
in a sense that these are the only members of this
family for which 3’ and 5’ RACE products are
available and are functionally characterized against
Dort.E4. In this study we tried to get an insight in
protein functionality based on their sequences and
evaluated the selection pressure exerted on the genes.
For this sequence variability, nucleotide substitution
rates and amino acid homology check were carried
out using different bioinformatics tools.
2. MATERIAL AND METHODS
technique was used to obtain the 5’ and 3’ ends of
RNA transcripts of muRdr1C, G and H transiently
expressed in tobacco [8, 9]. Isolated and sequenced
RACE products were used to get cDNA sequence
from previously sequenced BAC clones [6, 9].
Protein sequences of three proteins were obtained
using full length cDNA [8] in Soft-ware Bioedit
Version 7.0.9 [13].The same software was used for
the protein sequence editing, alignment, multiple
alignment (ClustalW) and local BLAST searches to
analyze and obtain conserved regions. Nucleotide
sequence of muRdr1H (cDNA) was used for Blastx
searches (http://www.ncbi.nlm.nih.gov/blast/
proteins to muRdr1H. For Neignbor-joining (NJ)
analysis the homologous protein sequences were
downloaded from NCBI database and phylogenetic
tree was constructed in MEGA4 [14, 15]. The gene
prediction and protein analysis were performed
on different free internet sources as http://www.
expasy.ch/, http://swissmodel.expasy.org/SWISS-
MODEL. html, http://www.ebi.ac.uk/interpro/,http://
linux1.softberry.com/berry.phtml whereas the
ratio of synonymous (K s
) and non-synonymous
(K a
) substitutions rates per synonymous/ nonsynonymous
site were calculated using software
DnaSP v 5 [16].
3. RESULTS AND DISCUSSION
Rdr1 resistance locus in consists of
a cluster of nine paralogs disease resistance genes
that play important roles in innate immunity against
black spot. The protein sequences of three members,
muRdr1C, muRdr1H and muRdr1G, were deduced
and subjected to sequence variability, nucleotide
substitution rates and amino acid homology check.
According to previous studies, among the nine R
genes in this locus muRdr1H confers partial resistance
to race 6 of the fungal pathogen Diplocarpon rosae
[8, 9, 17]. The muRdr1H transcript resulted in the
Open Reading Frame (ORF) of 1122 amino acids
muRdr1C has the predicted
protein of 1139 aa whereas the deduced protein of
muRdr1G has 944 amino acids. All three proteins
show the presence of TIR, NBS and LRR domains
with all conserved motifs as suggested by Lukasik
and Takken [18] and Meyers and colleagues [19]
(Fig. 1).
The homologues of muRdr1H protein were
out against the GenBank non-redundant database
(http://blast.ncbi.nlm.nih.gov). The muRdr1H
protein shares the highest identity (41%) to TIR-
NBS-LRR-resistance protein of Populus trichocarpa
(ACCESSION XP_002329162). It also shows
identity to hypothetical proteins of Vitis vinifera
(39-44%), to TIR of Medicago truncatula (40%;
ACCESSION ABD28703), to CMR1 of Phaseolus
vulgaris (40%; ACCESSION ABH07384) and to
N-like protein of N. tabacum (39%; ACCESSION
BAF95888 for resistance to Tobacco Mosaic Virus).
Fig. 2 shows phylogenetic tree of different amino

261

262 Aneela Yasmin et al
Fig. 1. Alignment of the proteins of muRdr1H, C and G of roses, TIR-NBS-LRR resistance protein
of Populus trichocarpa and N like protein of N. tabacum. The muRdr1H protein and its predicted
TIR, NBS and LRR domains with conserved motifs are under-lined. Stars on muRdr1H protein sequence
represent unique amino acids of this protein when compared to proteins of muRdr1C and G.
Within muRdr1H protein sequence the end amino acid of each domain is marked bold. a- Numbers in
muRdr1H (1), muRdr1C (2), muRdr1G (3), resistance protein of Populus
trichocarpa (4) and N like protein of N. tabacum (5); b and c- sequence positions; dashes indicate gaps
inserted to maintain optimal alignment.
acid sequences showing considerable identity to
muRdr1H protein, these full length aligned protein
sequences were downloaded from NCBI and tree
was constructed based on the bootstrap neignborjoining
(NJ) method with the Kimura2-parameter
model by MEGA4 [14, 15]. It is interesting to code
here that the CMR1 is a viral resistance gene from
common bean that functions across plant families
[20]. This protein sequence shows 40% identity
to the muRdr1H protein that found to be active
against black spot fungus, which suggests that the
predicted NB-LRR structures and recognition of
certain pathogen type lack correlation.
The kind of selection pressure exerted on different
domains of the genes was estimated by calculating
ratio of synonymous (K s
) and non-synonymous
(K a
) rates of substitutions for each synonymous/
non-synonymous site of different parts of the ORF
of three paralogs (muRdr1H, G and C). When K a
/ K s
ratio is 1 it represents random mutation or no
selection pressure operating on sequences may be
K a
/ K s
> 1) or conservation of
sequence (K a
/ K s
< 1) [21]. Estimation of the number
of synonyms and non-synonyms nucleotide per site

263
Fig. 2. NJ analysis of protein sequence showing considerable similarities to
muRdr1H. The full length aligned protein sequences were downloaded from NCBI
and analyzed in MEGA 4.0.
showed that the three genes are under conservation
selection and all domains (TIR, NBS and LRR
domain) are not under positive selection compared
motifs which showed K a
/ K s
ratio > 1 (Table 1). The
comparison of complete LRR domains of muRdr1H
and muRdr1C resulted in higher K a
values than K s
actually the result of lower K s
for the N- and C-
shows a K a
/ K s
>1 (Table 1). Although three genes
show conserved type of selection, LRR domains
show the highest frequency of non-synonymous
substitutions followed by NBS domain. The K a
/ K s
ratios were calculated for C-terminal region
in this region. This supported the hypothesis that
the parts of proteins which participate in pathogen
recognition are under diversifying selection. This
pattern is in agreement to previous studies that show
diversifying selection acts on the LRR encoding
domain of various plant disease resistance genes
[11].
Comparison of muRdr1H protein sequence with
other two (muRdr1C and muRdr1G) members
of the same Rdr1 family revealed 119 muRdr1H
which are part of TIR (2 aa; 0.002%), NBS (40 aa;
3.6%), and LRR (77 aa; 6.8%) domains (Fig. 3).
For RAG8, as the majority of unique amino acids
are located in LRR region followed by NBS region
suggesting some important role of these domains
in functionality of muRdr1H. The deduced Gro1-4
protein (resistance gene) showed only 16 nonconserved
differences in amino acids sequence

264 Aneela Yasmin et al
Fig. 3. Comparison of protein sequences encoded by 3-members of Rdr1 family. Protein sequences
muRdr1H (1), muRdr1C (2) and muRdr1G (3) were aligned to check amino acid similarities
and divergence. Stars represent consensus sequence and dots show differences in amino acids. a-
muRdr1H, G & C proteins sequences; b and c- sequence positions; dashes indicate gaps inserted
to maintain optimal alignment.

265
Table 1. Sequence variability and nucleotide diversity in different regions of three paralogs of Rdr1
family.
aa 2 (%) K a
K s
K a
/ K s
R-genes compared Analyzed regions 1 Nucleotide substitutions 3
muRdr1H vs.
muRdr1C
Complete CDS
80 0.0977 0.1041 0.92
TIR domain
90 0.0484 0.0836 0.58
NBS domain
76
0.1168
0.1196
0.98
Complete LRR domain
79
0.1034
0.0983*
1.05
-
0.1146
0.1108
1.03
xxLxLxx motif
-
0.0722
0.0935
0.77
muRdr1H vs.
muRdr1G
Complete CDS
TIR domain
-
75
98
0.1679
0.0991
0.0079
0.1156
0.1354
0.0356
1.45
0.73
0.22
NBS domain
78
0.1184
0.1853
0.64
Complete LRR domain
66
0.1209
0.1429
0.85
-
0.1148
0.0530
2.17
xxLxLxx motif
-
0.0717
0.1142
0.63
muRdr1G vs.
muRdr1C
Complete CDS
TIR domain
-
64
90
0.5067
0.1180
0.0512
0.5738
0.1497
0.1134
0.88
0.79
0.45
NBS domain
71
0.1439
0.1737
0.83
Complete LRR domain
52
0.1267
0.1508
0.84
-
0.0740
0.1132
0.65
xxLxLxx motif
-
0.0793
0.1249
0.64
muRdr1H vs.
muRdr1G &C
Complete CDS
TIR domain
-
58
89
0.5657
0.09228
0.02736
0.5029
0.11032
0.05688
1.13
0.83
0.47
NBS domain
67
0.10896
0.13689
0.78
Complete LRR domain
49
0.10421
0.11089
0.94
-
0.10613
0.07759
1.40
xxLxLxx motif
-
0.06861
0.09678
0.69
-
0.25922
0.25760
1.01
1
Different region of R-genes is analyzed for nucleotide variability.
2
Amino acid (aa) homology in percentage.
3
The ratio of synonymous (K s
) and non-synonymous (K a
) substitutions rates per synonymous/ non-synonymous site were calculated using software
DnaSP v 5.
*
Under-lined numbers represent the exceeded value of synonymous (K s
) compared to non-synonymous (K a

266 Aneela Yasmin et al
when compared to non-functional members of the
Gro1 gene family [22]. Mutagenesis analysis of
the muRdr1H protein or comparison of functional
and non-functional orthologs could determine
the essential residues necessary for pathogen
recognition and/ or downstream signaling. The Rdr1
D. rosae [23]. It is possible that the other members
of this family are also functionally active against
other races of D. rosae. Although the amino acid
sequence identity of these three paralogs of Rdr1
resistance gene cluster ranges between 58% and
80% and muRdr1H shares the highest overall amino
acid sequence homology to muRdr1C (80%; Table
1) the functionality of muRdr1C and muRdr1G
should be explored against different isolates of D.
rosae and/ or some other taxonomically different
pathogens.
Alignment of deduced amino acids of muRdr1H,
C and G show that the higher degree of sequence
similarity is present in N-terminal halves of
proteins that harbour putative effector domains
when compared to the C-terminal halves of
proteins (Fig. 3) suggesting the LRR domain under
selection as demonstrated for other closely related
NBS-LRR proteins [24]. This kind of selection in
LRR domain is exerted by single base changes,
insertions, deletions and unequal exchange of
meiotic recombination events for the evolution
between closely linked R-genes in a cluster [25].
4. CONCLUSION
The functionally characterized muRdr1H which
was found active against Dort E4 and other two
sequences has all conserved regions of TIR-NBS-
LRR genes. muRdr1G and C were inactive when
challenged by Dort E4. Due to the homology of two
paralogs to muRdr1H, we suggest their functional
characterization against other races of D. rosae to
assess their functionality on molecular level. We
observed less selection pressure exerted on these
genes as the pathogen D. rosae has low mobility
and races.
5. REFERENCES
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5. Kaufmann, H., L. Mattiesch, H. Lörz & T. Debener.
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6. Biber, A., H. Kaufmann, M. Linde, M. Spiller, D.
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marker-assisted selection in roses. Theoretical and
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(2009).
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susceptible rose genotypes. Acta Horticulture
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8. Yasmin, A.
Characterization of Rdr1 Resistance Gene from
Roses. PhD thesis, Univercity. of Hannover,
Germany (2010).
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A. Biber, A. Kuehr, M. Linde & T. Debener. Mining
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15. Kimura, M. A simple method for estimating
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& R. Rozas. DnaSP, DNA polymorphism analyses
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19. Meyers, B. C., A. W. Dickerman, R. W. Michelmore,
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Jeon, P. Ronald, W. J. Lucas & R. L. Gilbertson. A
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M. C. Menendez, F. Salamini, A. Ballvora & C.
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Proceedings of the Pakistan Academy of Sciences 49 (4): 269–278 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Research Article
Development of Skin-friendly Dermatological Water-in-Oil
Emulsion of Pomegranate Juice
Naveed Akhtar 1 , Rashida Parveen 1 , Barkat Ali Khan 1, 2* ,
Muhammad Jamshaid 3 and Haroon Khan 4
1
Department of Pharmacy, Islamia University, Bahawalpur, Pakistan
2
Department of Pharmaceutics, Faculty of Pharmacy, Gomal University, D.I Khan, Pakistan
3
Faculty of Pharmacy, University of Central Punjab, Lahore, Pakistan
4
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Gomal University,
D.I. Khan, Pakistan
Abstract: This study was designed to develop a topical skin-care cream (w/o emulsion) of 4% pomegranate
extracts versus its vehicle (the base) as control and evaluate its effects on skin-melanin, skin erythema,
skin moisture, skin sebum and TEWL. Concentrated pomegranate (Punica granatum) juice was entrapped
in the inner aqueous phase of w/o emulsion. The base containing no extract and a the formulation containing
4% concentrated juice of pomegranate was formulated. The odour was adjusted with few drops of lemon oil.
Both the base and the formulation were stored at different storage conditions for a period of four weeks to
predict their stability. The stability parameters, i.e., physical stability, centrifugation and pH, were monitored
at different time intervals. Both the base and the formulation were applied to the cheeks of 25 healthy human
volunteers for a period of 8 weeks. The pharmaceutical stability of creams was achiewed from 4 weeks
in-vitro study period. Odour disapperaed with the passage of time due to volatilization of lemon oil. The
and the formulation tended to increase sebum content The results showed a good stability over 4 weeks invitro
observation period of both the base and the formulation and the formulation exhibited bleaching, anti
respect to sensory evaluation.
Keywords: Antioxidant, emulsion, Punica granatum extract, skin, TEWL
1. INTRODUCTION
Punica granatum (Punicaceae) is a symbol of life,
longevity, health, feminity, fecundity, knowledge,
morality, immortality and spirituality. In Ayurvedic
medicine Punica granatum is considered “A
Pharmacy unto Itself ” [1]. This herb was used in
skin, mucosa and joints. The major constituents
are tannins (25–28%) including punicalagin;
polyphenols such as ellagic acid, ascorbic acid,
niacin, potassium and piperidine alkaloids.
Punica granatum functions as an astringent. It
has documented antimicrobial activity for gramnegative
bacteria, Saccharomyces fungus, parasites
and viruses [2].
Pomegrante juice comprises of 85% water, 10%
antioxidants and coloring material [3]. Pomegrante
is the potent source of polyphenolic compounds,
pedunculagin, punicalagin, gallic acid and ellagic
acid) which have 92% radical scavenging activity
[4, 5]. It is a rich source of polyphenolics, which
anticarcinogenic activity in numerous in vivo and
in vitro studies [6]. This application relates to
————————————————
Received, October 2012; Accepted, November 2012
*Corresponding author: Barkat Ali Khan; Email: barki.gold@gmail.com

270 Naveed Akhtar et al
dermatological agents for treating dermatological
disorders. The dermatological agents include a
therapeutically effective amount of at least one
to neutralize free radicals, a moisturizing agent in
skin, and a pharmaceutically acceptable carrier
[7]. Pomegrante extract is water miscible and is
incorporated in the aqueous phase of water-in-oil
emulsion. The skin lightening effects of ellagic
acid may due to chelating copper at the active site
of tyrosinase. Skin whitening, moisturizing, sun
protection and ant wrinkle, prevent and treat the
acne, anti-stain and anti-freckles effects are mainly
attributed to ellagic acid [8, 9]. An emulsion is
a biphasic system consisting of two immiscible
and uniformly dispersed as globules throughout the
other phase (the continuous phase) [10]. The main
advantage of emulsions is that they increase the
solubility and bioavailability of therapeutic drugs
as well as the ability to favor the topical transport
of hydrophilic solute. It has been shown that
microemulsions can be form spontaneously and
are thermodynamically stable, on one hand they
improve drug solubilization and bioavailability
and on the other hand they act as potential drug
delivery systems by integrating a wide range of drug
molecules. The main advantage of using the topical
emulsions is to avoid gastrointestinal environment
surfactant which is chemically cetyl dimethicone
formulation of water-in-oil emulsions, w/o/w and
o/w/o multiple emulsions. Preparations with ABIL
skin has 10-20% water content and its hydration
level can be enhanced by topical applied hydrating
cosmetics. Skin is viscoelastic in nature due to
contents improve skin elasticity [14].
2. MATERIALS AND METHODS
2.1 Ingredients
Pomegrantefruit was used as a plant material.
Extract of Punica granatum (purchased locally)
was prepared in lab of Pharmacy Department, The
Islamia University of Bahawalpur, Pakistan. ABIL-
KGaA Darmstadt (Germany), while Distilled Water
was prepared in labs of Pharmacy Department, The
Islamia University of Bahawalpur, Pakistan.
Punica granatum (Family:
Punicaceae) was performed in Cholistan Institute
of Desert Studies (CIDS) at The Islamia University
of Bahawalpur, Pakistan. The specimen was
deposited in the herbarium; the Voucher number
is: P. granatum 7072/21-04-1925 Rawal Chand
Herbarium of Department of Botany, University of
Agriculture, Faislabad, Pakistan.
The free radical scavenging activity of Punica
granatum was determined in accordance to
DPPH (1, 1-diphenil-2-picrylhydrazyl) which is
a stable free radical [15]. Fresh extract of Punica
(9 mL) buffer with a pH of 7.4.One mL of DPPH
obtained mixture was kept at room temperature for
20 minutes. Then, the absorption of the mixture at
517 nm was taken, in comparison with the control
solution (mixture without DPPH). The activity
of free radicals was calculated in % inhibition
according to the following relation:
% Inhibition = (A control- A test) x100
A control
The antioxidant activity P. granatum was
observed to be 78%. The apparatus used were:
Germany
Germany
Taiwan

Development of Skin Friendly Dermatological Water-in-Oil Emulsion 271
Homogenizer, Euro-Star, IKA D 230, Germany
Refrigerator, Dawlance, Pakistan
12.0
2.2. Preparation of Extract
Extract of Punica granatum was obtained by
concentrating the aqueous juice of pomegranate.
The fruits were thoroughly rinsed with water
before subjecting them to juice extraction. Thickskinned
pomegranate fruits were cut in halves prior
using a mechanical juice extractor. Extracted juice,
as came from the extractor was cloudy, containing
under reduced pressure; for the complete removal
of the seeds and pulp from the extract. Then, the
quantity of extracted juice was measured. In order
to maintain the quality of the extract, the container
of juice was covered with aluminum foil and kept
immediately in the refrigerator after it was removed
from the extractor and screened. After screening the
extract was then concentrated, bottled and frozen.
pomegranate extract was concentrated to half of
its original extracted amount; by just removing
maximum part of its water. To prevent the loss of
important compounds by heating, the extract was
providing low heat treatment, up to 45 o C. Finally, the
concentrated extract was packaged in glass tubes,
sealed and frozen. Later, whenever required, the
frozen extract of pomegranate was removed from
the freezer, kept for sometime at room temperature
to liquefy (i.e. freeze thawing) and used as an active
ingredient in the aqueous phase of w/o emulsion
[16].
2.3. Preparation of Emulsions
The W/O emulsions (base and formulation) were
prepared by the addition of aqueous phase to the oily
phase with continuous agitation. For the preparation
751 0 C. At the same time, aqueous phase consisting
of water was heated to the same temperature and
then the Punica granatum extract was added in it.
After that, aqueous phase was added to the oil phase
drop by drop. Stirring was continued at 2000 rpm
by the mechanical mixer for about 15 minutes until
complete aqueous phase was added, 2 to 3 drops
of lemon oil were added during this stirring time
to give good fragrance to the formulation. After
the complete addition of the aqueous phase, the
speed of the mixer was reduced to 1000 rpm for
homogenization, for a period of 5 minutes and then
the speed of the mixer was further reduced to 500
rpm for 5 minutes for complete homogenization;
until the emulsion cooled to room temperature.
The base was also prepared by the above stated
method using the same ingredients, excluding
pomegranate extract (i.e., the active ingredient)
[16].
Composition of the Base
Oily Phase:
4%.
Aqueous phase: Distilled water (q.s.), 100%.
Composition of the Formulation
Oily Phase:
4%.
Aqueous phase: Pomegranate extract, 4%
(concentrated); and Distilled water (q.s.),
100%.
Both the creams were analyzed to assure the
formulation of desired type. Emulsion was analyzed
organoleptically (color, thickness, look, feel,
liquefaction) and physically (creaming and phase
separation).
Type of emulsion was analyzed by diluting the
emulsion with oil and water separately.
pH value of freshly prepared emulsion and
emulsions kept at different conditions were
cosmetic laboratory.
Electrical conductivity of freshly prepared

272 Naveed Akhtar et al
emulsion and emulsions kept at different conditions
were monitored by a digital conductivity-meter.
Centrifugal tests were performed for emulsions
immediately after preparation. The centrifugal tests
were repeated for emulsions after 24hrs, 7, 14, 21
and 28 days of preparation. The centrifugal tests
were performed at 25 0 C and at 5000 rpm for 10
minutes by placing the 5g of sample in disposable
stoppered centrifugal tubes. Stability tests were
performed at different conditions for emulsions to
note the effect of these conditions on the storage of
emulsions. These tests were performed on samples
kept at 8 0 C 0.1 0 C (in refrigerator), 25 0 C0.1 0 C
(in incubator), 40 0 C0.1 0 C (in incubator) and
40 0 C0.1 0 C (in incubator) with 75% RH. Physical
characteristic of simple emulsions, i.e. color,
creaming and liquefaction, were noted at various
intervals for 28 days.
One-sided blind study was designed with placebo
control in the month of August to September (the
moderate season in Pakistan). 25 healthy human
volunteers who signed the informed consent, with
were included in this work as they were easily
available with regular under control observations.
Prior to the tests, the volunteers were examined by
a cosmetic expert for any serious skin disease or
damage especially on cheeks and forearms. All the
skin tests were performed at 21±01 0 C and 40±2%
relative humidity conditions [14].
The experiments were carried out on the cheeks
of volunteers as cheeks are uniformly and more
(Burchard test) was performed on the forearms of
each volunteer to determine any possible reactions
to the emulsions. On the second day, each volunteer
was provided with two creams. One cream was base
and the other one was formulation containing the
active ingredients. Each cream was marked with
“right” or “left” indicating application of that cream
to the respective cheek. The creams were applied by
the volunteers themselves as instructed for 60 days.
Every individual was instructed to come on 1 st , 2 nd ,
3 rd , 4 th , 6 th and 8 th week for the skin measurements.
2.4. Patch Tests (Burchard Tests)
performed on the both forearms of each volunteer.
A 5cm X 4cm region was marked on the forearms.
The patch (Bandage disc) for the right forearm was
saturated with 1.0 g of base while the patch for left
forearm was saturated with 1.0 g of formulation.
Each were applied to the 5cm X 4cm marked regions
separately on each forearm. The regions were
covered with the surgical dressing after application.
The patches were removed after 48 hours and the
forearms were washed with physiological saline
[16]. After 48 hours, scores were recorded for the
presence of erythema (skin redness) using a scale
with 4 points from 0 to 3.0 stands for absence of
erythema,1 for mild erythema,2 for moderate
erythema while 3 stands for severe erythema.
Each volunteer was asked to note their irritation/
itching towards the patches and then assign a score
from the same scale. Average score with respect to
volunteers is given in Fig. 1.
Fig. 1.
of the base and the formulation after 24 hours (Patch
melanin on the cheeks of human volunteers, on
and then on 1 st , 2 nd , 3 rd , 4 th , 6 th and 8 th week.With
the help of a Corneometer, stratum corneum (SC)
capacitance was measured before the application
of any cream and then on 1 st , 2 nd , 3 rd , 4 th , 6 th and
8 th week. Sebumeter was used to measure sebum
content of the skin before application of any cream
and then on 1 st , 2 nd , 3 rd , 4 th , 6 th and 8 th week.
Every individual was provided with a form
prepared previously to test the sensory values of
creams. This form consisted of seven parameters
to be evaluated and every parameter was assigned
very good, respectively. This form was asked to be

Development of Skin Friendly Dermatological Water-in-Oil Emulsion 273
completed independently by each individual on day
60.
The percentage changes for individual values of
different parameters, taken every week, of volunteers
were calculated by the following formula:
Percentage Change = [(A – B) / B]*100
where;
A = Individual value of any parameter (from 1 st to
8 th week)
B = Zero hour value of that parameter
The measured values obtained for different
parameters (skin moisture, sebum, melanin,
erythema, elasticity and pH) were analyzed using
SPSS 12.0 on the PC computer (paired samples
t-test for variation between the two preparations;
two-way ANOVA for variation between different
the base and formulation on the cheeks of human
volunteers have been demonstrated in Fig. 3, 4, 5, 6
and 7, respectively.
Sensory evaluation of both the base and
formulation by the volunteers has been presented in
Fig. 8.
Both the base and the formulation were divided
in to four samples separately and these samples
were kept at different storage conditions i.e. at 8 ° C
in refrigerator, at 25 ° C, 40 ° C and at 40 °
relative humidity (RH) in stability chambers. These
3. RESULTS AND DISCUSSION
3.1 Stability and Properties during Storage
Stability of the base and of the formulation kept
at different storage conditions was studied and
physical characteristics regarding the stability of
the base and formulation have been discussed.
Centrifugation tests for the base and the
formulation of freshly prepared creams and for
samples kept at different storage conditions were
performed and phase separation in these samples
was observed for 28 days at different time intervals.
No phase separation was observed in any sample
of the base or the formulation throughout the study
period. Electrical conductivity values of the base
and formulation of fresh creams and samples kept
at different storage conditions for 28 were also been
determined. No electrical conductivity was noticed
in any sample of the base and formulation throughout
the study period. pH values of the base and the
formulation of fresh creams and samples kept at
different storage conditions up to 28 days have been
determined and reported in Fig. 2. The percentages
of changes in the amounts of erythema, melanin,
skin moisture, skin sebum and transepidermal
Water loss (TEWL) following the applications of
Fig. 2. pH of the base and the formulation kept at 8 o C,
25 o C, 40 o C and 40 o
Fig. 3. Percentage change in skin melanin content after
application of the base and formulation.
Fig. 4. Percentage change in skin erythema content after
application of the base and the formulation.

274 Naveed Akhtar et al
samples of the base and formulation at different
storage conditions were observed for a period of 28
days at different intervals with respect to change in
color, liquefaction and phase separation.
Fig. 5. Percentage change in skin moisture content after
application of the base and formulation.
Fig. 6. Percentage of change in skin sebum after
application of the base and formulation.
Fig .7. Percentage change in trans-epidermal water
loss (TEWL) after application of the base and the
formulation.
Fig. 8. Average results for panel test.
(1= Ease of application, 2= Spreadability, 3= Sense just
after application
4= Sense in long term, 5= Irritation, 6= Shine on skin, 7=
Sense of softness)
The freshly prepared the base and formulation
were creamy white in color. There was no change
in color of any sample of the base or formulation
at different storage conditions i.e., 8 0 C, 25 0 C, 40
0
C and the at 40 0
the observation period of 28 days. Thus both the
base and formulation were stable at the studied
storage conditions up to 28 days.
No liquefaction was observed in any of the
sample of the base and formulation kept at 8 0 C and
25 0 C during whole observation period of 28 days.
A Slight liquefaction was observed in the sample
of the base and formulation kept at 40 0 C and 40 0 C
st and 28 th day of observation The
samples of the base and formulation were stable at
8 0 C,25 0 C, but slight phase separation in the sample
of the base occurred at 40 0 C and 40 0
on 28 th day of observation but the formulation was
stable at 40 0 C and 40 0 th day of
observation. The centrifugation test was performed
on the samples of the base and formulation kept at
different storage conditions up to a period of 28 days
centrifugation was seen in any of the samples kept
at different storage conditions i.e. 8 0 C, 25 0 C, 40 0 C
and 40 0 th day of observation.
This indicated that the emulsions were stable at all
the storage conditions for 28 days. Conductivity
test was performed for all the samples of the
base and the formulation kept at different storage
intervals. No electrical conductivity was seen in
any of the samples of the base and formulation kept
at different storage conditions i.e. 8 0 C, 25 0 C, 40 0 C
and 40 0
This indicated that creams were stable at different
storage conditions.
pH of freshly prepared the base and formulation
was 5.35 and 5.65 respectively, which is within the
range of skin pH. The pH values of the samples of
the base kept at different storage conditions i.e. 8 0 C,
25 0 C, 40 0 C and 40 0
increasing gradually in the 3 days and then it started

Development of Skin Friendly Dermatological Water-in-Oil Emulsion 275
to decline continuously till 28 th day with some
variations. At the end of study pH of the samples
of the base at 8 0 C, 25 0 C, 40 0 C and 40 0
was 4.52, 4.60, 4.35 and 4.08 respectively. Whereas
pH of the samples of formulation kept at 8 0 C, 25 0 C,
40 0 C and 40 0
in pH values with slight variations with time. The
pH values of samples of formulation kept at 8 0 C,
25 0 C, 40 0 C and 40 0
3.39 and 4.01 at 28 th day respectively. By using two-
it was found that the change in pH of different
samples of the base as well as formulation was
levels of time and temperature. The decrease in pH
of the formulation at different storage conditions
might be due to the production of any acidic
metabolite.
3.2. Dermatological Tests
The mel
event can enter the dermis where it is engulfed by
macrophages, producing ‘‘melanophages’’. These
cells are often retained in the upper dermis for
prolonged periods, as removal of dermal melanin
apparently is a very slow process [17]. The major
source of color in human skin derives from the
presence within the epidermis of specialized melanin
melanocytes are acquired by keratinocytes and
transported within them to the epidermal surface
[16]. The base produced no effects on skin melanin
with respect to volunteers. The formulation
respect to volunteers and time. The decrease in skin
melanin content after application of formulation
may be attributed to the antioxidant activity of
pomegranate extract which is rich in ellagic acid,
punicagalin and vitamins C [7]. Ellagic acid causes
inhibition of tyrosinase and melanin synthesis thus
inhibiting melanogenesis [19]. Vitamin C promotes
collagen growth to maintain and improve tone,
through several mechanisms. Among them is
mediators such as Interleukin- 1-alpha, endothelin-1,
and stem cell factor. Reactive oxygen species,
such as superoxide and nitric oxide, generated in
damaged skin (for example, from UV exposure) or
the safety of cosmetics, the important point is that
cosmetics must not cause any contact dermatitis
when applied to the skin. Skin irritation is caused
by the direct toxicity of chemicals on cells or blood
vessels in the skin and is different from contact
allergy which is caused by immune response [20].
The base produced no effects on skin erythema
at different time intervals, whereas application of
on skin erythema at time and volunteer level. With
the help of paired sample t-test it was evident that
respect to the base and formulation throughout
the study period. As pomegranate extract is rich
in ellagic acid, vitamin C and contain potent
on skin and therefore reduced erythema [17].
and anti carcinogenic potential [6, 21].
The moisturizing treatment involves repairing
the skin barrier, retaining/increasing water content,
reducing TEWL, restoring the lipid barriers’
ability to attract, hold and redistribute water, and
maintaining skin integrity and appearance. The
base affected no change in moisture content with
with respect to volunteers throughout the study
period of 60 days whereas the formulation showed
study period of 60 days with respect to time and
content was observed throughout study period.
values were observed after application of the base
and the formulation throughout the study period,
except after 4 th , 6 th and 8 th
increase in moisture retaining capacity on skin
after application of formulation may be because of
vitamin C because collagen synthesis increases and
hydration level improves [9].

276 Naveed Akhtar et al
and epidermal lipid. Of the two, sebum, which
is secreted by the sebaceous glands, is the major
vary individually according to age, sex, inherited
traits, and topographical variations of the skin
[22]. Sebum, the product of sebaceous glands, is a
complex of various lipids that are thought to act as
an epidermal and/or follicular lubricant [23]. The
probe is then placed into the device which radiates a
and then into an instrument called a photomultiplier,
which measures the amount of light in the beam. The
more sebum on the skin, the more transparent is the
In this study, the effects of the base and
formulation on the sebum contents of human cheeks
were investigated. Sebum was measured every week
in all the individuals. The base increased the sebum
contents throughout the study period of 60 days.
base and the formulation on skin sebum throughout
the study period. The increase in sebum content
after application of the base and the formulation
may be attributed to the oily nature of w/o emulsion
oil [24].
3.3. Trans Epidermal Water Loss (TEWL)
Skin barrier capacity is measured by an increased
transepidermal water loss. TEWL measures the
difference in vapor pressure at two points situated
on a line perpendicular to the skin surface inside a
tunnel pressed toward the skin. In the absence of
skin irritation, TEWL may be considered an indirect
measure of stratum corneum water permeability
[19].
The changes in TEWL produced by the base and
retaining properties; the formulation and the base
enhanced the stratum corneum ability to attract, hold
and redistribute water thus reducing the TEWL. Of
the oils, a mineral oil is frequently used ingredient
but can reduce trans epidermal water loss only by
about 30% [20].
3.4. Panel Test
A questionnaire containing seven questions was
prepared and the two copies of this form were
given to each volunteer for sensory evaluation of
the two creams. Average points were calculated
from the points assigned by each volunteer for each
question for both of the creams. Average points for
were found to be 4.02 and 4.18 for the base and
formulation, respectively. This indicated that the
base and formulation can be easily applied on the
skin. Average points regarding spreadability were
4.36 for the base and 4.37 for formulation which
meant that the formulation spread on skin better
than the base. Average points for feel on application
were 3.84 for the base and 3.68 for formulation.
This indicated that the base and formulation were
felt well on the skin. Average points for the sense
in long-term application of creams were 4.08 and
4.00 for the base and formulation respectively. This
showed that formulation and the base produced
pleasant feeling on application to skin. There was
no irritation on the skin in both cases i.e. the base
and formulation, as these were assigned 0.00 point
for irritation by all the volunteers. Shine on skin
was 4.00 for the base and 4.04 for formulation.
This was expected since the base and formulation
the formulation led to more softness of the skin
than the base as the average point was 4.41 for the
base and 4.43 for formulation.
The paired sample t-test results revealed no
difference between the average points of sensitivity
for the base and the formulation. Thus, both of the
creams behaved similarly from the sensory point of
view.
4. CONCLUSIONS
From this investigations we concluded that a stable
W/O emulsion containing extract of pomegranate
melanin content of the volunteers after application
of the formulation indicated that the formulation
possessed skin depigmenting effects. The skin
erythema content of the skin decreased after
application of the formulation, and did not cause

Development of Skin Friendly Dermatological Water-in-Oil Emulsion 277
skin irritation throughout the study period. The
formulation produced a pronounced increase in
moisture content of the skin at the end of study
that the formulation had anti-wrinkle effects. Thus,
this investigation revealed a promising future use
of pomegranate extract in skincare formulations
without any adverse dermal toxic effects, cosmetics
and/or pharmaceutical preparations.
5. ACKNOWLEDGEMENTS
Financial support for this study was provided by
Department of Pharmacy, The Islamia University of
Bahawalpur, Pakistan. The authors thank to the Chairman,
Department of Pharmacy, The Islamia University of
Bahawalpur for his support to accomplish this task.
6. REFERENCES
1
(Pomegranate) and Its Potential for Prevention
J.
Ethnopharmacol. 109: 177–206 (2007).
2. Carl, T.T. Cosmeceuticals Containing Herbs: Fact,
Fiction, and Future. Dermatol. Surg. 31: 873–880
(2005).
Platelet Aggregation: Studies in Humans and In
Am. J. Clin. Nutr. 71: 1062-1076 (2005).
Derived Products for Cancer Chemoprevention.
Sem. Can. Biol. 17: 377–385 (2007).
Its Relationship with Phenolic Composition and
Processing. J. Agric. Food. Chem. 48: 4581-4589
(2000).
Effects of Standardized
Pomegranate (Punica granatum L.) Polyphenolic
Extract in Ultraviolet-Irradiated Human Skin
Fibroblasts. J. Agric. Food. Chem. 56 (18): 8434–
8441 (2008).
for Treating Dermatological Disorders. United
States Patent 6800292.
Ellagic Acid Rich Pomegrante Extract on Tyrosinase
Activiy and Ultra-Violet Induced Pigmenation.
Biosci. Biotechnol. Biochem. 12: 2368-2373
(2008).
vitamin C on collagen biosynthesis and degree
of birefringence in polarization sensitive optical
coherence tomography (PS-OCT). Afr. J. Biotechnol.
7: 2049-54 (2008).
10. Agarwal, S.P. Physical Pharmacy. 2 nd ed. CBS
Publishers. New Delhi. p. 177-184 (2007).
Health care applications-An Overview. J. Disp. Sci.
Technol. 23: 419-439 (2002).
swelling-breakdown of w/o/w multiple emulsions:
possible mechanisms for the lipophillic surfactant
effect. J. Cont. Release. 52: 99-107 (1998).
13. Evonik. ABIL ®
of cosmetic w/o creams and lotions, 2008: http://
www.evonik.com/personal-care
solid lipid Nano-particles on skin hydration and
viscoelasticity-in vivo study. Eur. J. Pharm.
Biopharm. 56: 67-72 (2003).
and antioxidant activity in pomegranate arils during
fruit development. Food. Chem. 93: 319-324
(2005).
two moisturizers on skin barrier damage in allergic
contact dermatitis. Eur. J. Dermatol. 12: 136-138
(2002).
Hyper pigmentation. J. Invest. Dermatol. 13: 10–14
(2008).
Antioxidants on UV-Induced Erythema Formation
when Administered after Exposure. J. Dermatol.
198: 52-55 (1999).
Administration of Ellagic Acid Rich Pomegranate
Extract on Ultra-Violet Induced Pigmentation in
the Human Skin. J. Nutr. Sci. Vitamin. 52: 383-388
(2006).
Pomegranate as a Cosmeceutical Source:
Pomegranate Fractions Promote Proliferation
Cells. J. Ethnopharmacol. 103: 311–318 (2006).
of nutrition on canine sebum secretion: a preliminary
report. Vet. Dermatol. 11: 2-3 (2000).
Visualization of Lipid Domains in Human Skin

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and Temperature. Biophys. J. 93: 3142–55 (2007).
23. Zoe, D.D. Dermatological Aspects of Cosmetics:
Dermatol. Clin. 18: 597-
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melanin. Cosmet. Toil. 123: 55-68 (2008).

Proceedings of the Pakistan Academy of Sciences 49 (4): 279–288 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Review Article
Microwave Imaging: Potential for Early Breast Cancer Detection
Muhammad Hassan Khalil*, Jia Dong Xu and Tsolmon Tumenjargal
Department of Electronics and Information Technology,
Northwestern Polytechnical University, Xi’an, NWPU Xi’an, China 710072
Abstract: Early breast cancer detection can save the women infected by malignant tumors. Microwave
imaging has recently been proposed for detecting small malignant breast tumors at early stages. This type of
cancer is the top-most cause of death among women due to malignant tumors. The detection of early-stage
tumors in the breast by microwave imaging is challenged by both the moderate endogenous dielectric contrast
between healthy and malignant glandular tissues and the spatial resolution available from illumination at
to be imaged and the low level of signal scattered from a tumor relative to the clutter scattered by normal
of the dielectric properties within the breast, have the potential to reconstruct both normal and cancerous
tissue structures. However, the ill-posedness of the associated inverse problem often limits the frequency
of microwave illumination to the ultra high frequency (UHF) band within which early-stage cancers have
sub-wavelength dimensions. This review presents the research status of microwave imaging for malignant
tumor detection. Many methods have been used, i.e., active, passive, and hybrid. However, it is important to
remember that, in addition to microwave imaging, several alternative breast cancer detection modalities are
actively being pursued, including optical imaging methods.
Keywords: Breast cancer detection, microwave imaging, inverse scattering
1. INTRODUCTION
Detecting breast cancer in its earliest stages is looked
upon as the best hope for successful treatment of the
disease [2]. The limitations of conventional X-ray
mammograms are well-recognized [2, 3] and, in
response to these limitations, several complementary
modalities for breast cancer are under investigation.
Currently, X-ray mammography represents the
gold standard method of breast imaging. Other
methods, including magnetic resonance imaging
(MRI) and ultrasound approaches are as yet either
less effective or too expensive for mass-screening
purposes. X-ray mammography is based on the
which affect mammogram formation. Mammogram
is 2-D map that represents the intensity of X-ray
radiation passed through the previously compressed
breast (in order to reduce image blurring and to
reach uniformity of tissue). However, relatively
small contrast between affected and normal tissues
(4–34%) and false-positive (70%) rates [4]. Besides
this, very early stage tumors do not necessarily
radiation is accumulated over repeated scans.
As a supplement to X-ray mammography,
microwave imaging is a new and promising
technique for breast cancer detection. It has a lot
of advantages, including real time monitoring, noninvasive
and can function over wide ranges of time
and size scales involved in biomedical processes.
As is well known, microwave imaging is a
technique aimed at sensing a given scene by means
of interrogating microwaves. This active technique
————————————————
Received, April 2012; Accepted, November 2012
*Corresponding author: Muhammad Hassan Khalil; Email: hassan_janjua86@yahoo.com

280 Muhammad Hassan Khalil et al
was considered for a long time as an emerging
technique. Microwave imaging has recently
proven capable of providing excellent diagnostic
capabilities in several areas, including civil and
industrial engineering, geophysical prospecting
and biomedical engineering. To date, the primary
motivation for developing microwave systems has
been the need for improved detection of malignant
breast tumors [1].
Microwave imaging approaches to early breast
cancer detection offer an attractive alternative
to conventional mammography. The attraction
is motivated by dielectric property contrast
between normal and malignant breast tissue at
microwave frequencies (between 2:1 and 10:1)
[5, 6]. Endogenous contrast and spatial resolution
are two important considerations in the detection
and location of tumors by microwave imaging
techniques. Recent studies of the histopathology
and microwave-frequency dielectric properties
of excised breast tissues suggested a much lower
contrast between healthy and cancerous tissues
than was previously understood [16]. The ex vivo
microwave dielectric properties of malignant
glandular tissues were observed to be about ten
times those of adipose tissue, but only one-tenth
higher than the properties of normal glandular
tissues. Furthermore, the electrical dimensions of
early- stage cancers are at or below the nominal
half-wavelength resolution limit of the UHF-band
frequencies (0.3–3.0 GHz) typically employed in
frequency-domain microwave inverse scattering.
Although super-resolution has been observed
and attributed to evanescent waves in near-
environments [17, 18], microwave detection of
early-stage malignancies is nevertheless challenged
by the moderate endogenous dielectric contrast, the
small scattering area of these malignancies, and the
heterogeneous scattering environment of healthy
glandular tissue in which tumors often form.
Ultra-wideband (UWB) radar technique was
introduced by S.C. Hagness in 1998 [5–13]. The
technique, based on the ideas of ground penetrating
chosen inside the breast, without reconstruction
The key point of the technique is time shifting
and summing (synthetic focusing) of scattered
waveforms. Later, Hagness and colleagues
presented further development of UWB radar
approach which improved the process of scattered
energy estimation, namely Microwave Imaging via
Space Time (MIST) Beam Forming method in both
time and frequency domains [14, 15]. In contrast
to the image recovery goal of tomography, the
proposed UWB radar approach solves a simpler
computational problem by seeking only to identify
such as malignant breast tumors. In the UWB
radar approach, high bandwidths and large antenna
apertures are used to improve spatial resolution at
microwave frequencies.
Another class of numerical technique in radar
terminology is inverse scattering method. The
inverse scattering problem using iterative methods.
In addition to obtaining the shape of the object, a
quantitative description of the dielectric constant
valuable diagnostic information.
2. TUMOR DETECTION AND
IDENTIFICATION SCHEMES
Currently, three directions in microwave breast
imaging can be distinguished: hybrid microwaveinduced
acoustic imaging [7–8], microwave
tomography [9–10], and UWB radar technique
to heat (and, thus, expand) tumors; the pressure
waves so generated are then detected by ultrasound
transducers. Microwave tomography relies on
basis of measurements of narrow-band microwave
signals transmitted through the breast. This type
requires solving both forward scattering problem
and nonlinear ill-conditioned inverse scattering
problem iteratively; until the computed and the
measured data are close enough [12–10]. From
the basic concepts of each approach, researchers
have investigated different techniques to achieve
a suitable system for breast cancer detection; the
investigated techniques are illustrated in Fig.1.
Currently, tomographic and radar are the two
major approaches [19]. Both passive and active

Microwave Imaging: Potential for Early Breast Cancer Detection 281
Fig.1. Schematic diagram of microwave imaging.
microwave imaging methods are being researched
for breast cancer detection [30].
2.1. Passive Method
Passive microwave radiometry [31, 32] exploits
temperature differences between malignant and
normal breast tissue due to elevated metabolism in
fast-growing malignant tumors.
The principle of operation refers to measurement
of temperature of the breast by radiometric
techniques and comparison of the formed
thermical map “images” of the suspicious area with
corresponding healthy breast area obtained at the
microwave frequencies. The malignant indication
are made by recognizing the temperature difference
in the asymmetry between two images [33, 34].
Microwave radiometry (also called thermography)
has been explored for many years especially as
adjuvant to mammography.
2.2. Active Methods
Three types of active microwave breast imaging
techniques have been proposed: hybrid-induced
acoustic imaging, microwave tomography, ultra
wideband microwave radar techniques.
In general, active microwave imaging falls
under the broad category of inverse problems. The
objective of MWI is to reconstruct the unknown
distribution of the complex permittivity inside the
mammary tissue given a set of data measured along
the perimeter of the volume. There are several
groups currently performing research in active
microwave imaging [25]. While these systems
acquisition and image reconstruction, they share
the common potential advantages that microwave
imaging offers over traditional mammography.
First, active microwave imaging systems do not
require the use of ionizing radiation. In addition,
MWI systems would eliminate the need for
uncomfortable breast compression. Furthermore,
microwave imaging systems are expected to be
less expensive than x-ray systems and much less
costly than MRI. All of these characteristics would
allow for earlier and more frequent examinations.
Finally, microwave imaging has the potential for a
much higher sensitivity in detecting tumors, as well
and benign tumors. This is because the electrical
contrast is likely to be much higher than the density
contrast, particularly for malignant lesions. Active
methods can be divided into further two groups,
i.e., tomography and radar-based.
2.3. Hybrid Method
Hybrid methods could include, for example, twostep
procedures in which fast qualitative algorithms
are combined with quantitative methods in order to

282 Muhammad Hassan Khalil et al
and successively retrieve the distributions of their
dielectric parameters. The principle of operation
is to illuminate the breast by using microwave
frequency and measure the signal by the ultrasound
transducer, since more energy is deposited in tumors
due to its higher conductivity, as a result the tumor
expanding and generates pressure waves, which are
detected by the ultrasound transducer.
Two approaches of hybrid microwave acoustic
imaging system has been proposed, namely,
computed thermo-acoustic tomography (CTT) and
scanning thermo-acoustic tomography (STT).
2.3.1. Computed Thermo-acoustic Tomography
(CTT)
The breast is placed in a water-bath and illuminated
with 0.5 µs pluses of 434 MHz signals using
waveguide; 0.5 µs waves are used to generate
ultrasound waves in the medical region [20,
21]. Ultrasound transducers are arranged on a
hemisphere, and data are recorded as the transducers
Filtered back projection algorithms adapted
from X-ray computed tomography has been used
to form the images. The clinical results have been
obtained with this method to demonstrate images,
which show internal tissues structure in the breast.
2.3.2. Scanning Thermo-acoustic Tomography
(STT)
L. Wang and his colleagues developed the
Scanning Thermo-acoustic Tomography (STT)
technique; good experimental results were obtained
with various properties and thickness phantoms
containing block of fat and muscles [21, 22].
Numerous improvements on imaging algorithms
for the STT approach were derived for planner
and cylindrical systems [23, 24]. For the spherical
proposed [23]. The resolution of the obtained
images was approximately 0.5 mm, which shows a
good detection capability.
Although research groups have taken a variety
of approaches to microwave imaging, these
approaches can be divided into two main categories:
tomographic methods that utilize traditional inverse
scattering approach both sequential 2D slice
methods and full 3D methods and methods that
use beamsteering approaches confocal imaging,
MIST, TSAR (including UWB). The fundamental
difference between these two methods is the way
in which a voxel of material data is localized. In
the case of inverse scattering approaches, the
electrical properties of all locations in the volume
are determined simultaneously using an inversion
algorithm that operates on the aggregate of the
measured data. Alternatively, beamsteering
approaches isolate a particular location in the
volume to be measured, and data is taken for each
individual voxel. Statistical methods are then
employed to determine a given voxel’s electrical
properties.
2.4. Tomographic Imaging Systems
In tomographic microwave imaging systems, a set
of measurement data is collected using antennas on
the surface of a chamber, and the unknown complex
permittivity distribution of the material inside can
then be found using one of a variety of methods
that have been developed to address this ill-posed
inverse problem. These systems can generally be
categorized into two main types based on whether
the inverse problem is solved for a series of twodimensional
slices using data collected from a
planar array of antennas (2D imaging) or for the
entire volume using data collected from antennas
positioned throughout the perimeter of the chamber
(3D imaging).
2.4.1. Sequential 2D Slice Approach
In the case of the 2D slice approach, the volume to be
scanned is surrounded by the 2D array of antennas,
typically arranged in a ring, and sequential 2D
images are generated by translating the entire array
vertically through a series of positions. There are
several advantages to using such an approach. First,
a relatively small number of simple antennas can be
the complexity of the system to be modeled in
the reconstruction algorithm. Furthermore, the
the inversion process. In addition, a translating
2D array reduces the complexity of the hardware
system design and data-acquisition process.

Microwave Imaging: Potential for Early Breast Cancer Detection 283
eliminated, and isolating sensitive electronics from
accomplish. Finally, the positioning of the antennas
are determined by the CNC motor. This approach
is not without its disadvantages, however. Due
to the nature of the motorized translation system,
data-acquisition time will be much slower than an
electronically switched 3D imaging system. This
prolonged acquisition time, combined with the
motion united with the translating antenna array,
will likely lead to increased noise. Furthermore, a
2D approach to data collection reduces the number
of possible antenna combinations and does not have
the ability to utilize data from out of plane antenna
combinations. Finally, such a 2D approach is likely
to result in reduced resolution and errors since the
translation in the transverse direction is on the order
of a wavelength. For a further discussion of the
trade offs associated with 2D imaging, the reader is
referred to [26].
2.4.2. Full 3D Inversion Approach
The full 3D inverse scattering approach offers many
potential advantages over the other two approaches
detailed in this paper, often at the cost of added
complexity. First, a full 3D array of densely packed
antennas that are switched electronically offers
effcient data collection and highly precise antenna
positioning. A complete 3D array also provides
the maximum combination of transmit and receive
antennas, and enables the collection of out-of-plane
transmission data. Finally, a fully 3D system is able
to take advantage of the optimizations and advances
in 3D inversion techniques that have recently been
made. All of these contribute to the potential of
3D inversion methods to offer greater resolution,
increased SNR, and more rapid data collection [27,
28, 29].
2.5. Beam Steering Approaches
Beamforming systems have their roots in optics,
confocal microscopy, and RADAR. The general
approach in such systems is to focus an illuminating
microwave signal at a particular point in the scanning
volume and then to refocus the scattered signal
back to the point of illumination. By systematically
scanning the focal point within a set of preselected
voxels throughout the breast volume, a 3D image
can be constructed. The ability of these systems to
detect tumors relies on the increased backscatter
caused by malignant tissue. The primary advantages
of this approach are two-fold: there is no need for
complex inversion techniques and simple timegating
methods can be used to reduce clutter and
multiple scattering effects.
2.5.1. Confocal Imaging: Simple Delay-and-sum
Beamforming
In a typical confocal breast imaging system, ultra
wideband (UWB) pulses are generated by an
antenna located on or near the surface of the breast.
The backscattered waveform at that particular
antenna location is then collected and stored in a
computer.
Using both electronic switching and mechanical
scanning, this procedure is repeated for each
antenna element in an N element antenna array. The
set of N backscattered signals are then time-shifted
is then scanned throughout the breast by adjusting
the relative amount of time-shift applied to each
backscattered signal. Statistical analysis is then
used to predict the absence or presence of a tumor
at each scanned voxel. Finally, all of the statistical
decisions for each voxel are aggregated to form an
image. It should be noted that this method does not
seek to generate a map of the dielectric properties
of the breast. Instead, it only seeks to identify and
locate the presence of strong scatterers within the
breast [5].
2.5.2. Microwave Imaging via Space-time
Beamforming (MIST)
At the University of Wisconsin, Hagnees and
colleagues have proposed a planar system termed
Microwave Imaging via Space-time (MIST) system
[36], the patient to be scanned lies in a supine
position. Either one antenna or an array is placed
scanned to multiple locations. Special liquid has
been used to approximate the dielectric properties
of the medium to those of the object.
studies [5], with using a simple monopole antenna

284 Muhammad Hassan Khalil et al
and planer breast phantom, data simulated using the
were obtained at 17 antenna locations and spherical
tumor with 5 mm diameter was located 3 cm beneath
the array was detected successfully. Resistively
loaded bowtie antenna, 8 cm long was used as a
sensor to perform -D study [12], spherical tumor
1.76 cm diameter located 5 cm below the antenna
was detected.
Successful detection was performed of small
below the surface of the 2 cm thick skin layer in
a complex phantom with planer system and more
realistic MR based 2-D models [37]. Multi-static
approaches were introduced [38], and initial
results using data from a realistic FDTD model
demonstrated successful detection of a small tumor
2mm in lossy, inhomogeneous human breast.
2.6. Tissue Sensing Adaptive Radar (TSAR)
As an alternative to MIST, Fear and Sill developed
a similar system that has been termed Tissue
Sensing Adaptive Radar. Like MIST, it seeks to
address the shortcomings of the simple confocal
system; however, there are a few fundamental
difference between the two. For example, in the
Tissue Sensing Adaptive Radar (TSAR) system,
the patient lies prone and the pendulous breast is
scanned from surrounding locations. Additionally,
the TSAR system uses less complicated clutter
reduction methods than MIST.
Simple time shifting and adding algorithm were
used to reconstruct images in TSAR system, the
steps of this algorithm were:
i) First, tissue sensing carried out to locate
the breast in the tank; second, the skin
were formed into an image.
ii) The results of this technique showed the
ability for detecting image, for detecting
and localizing tumors of greater than 4 mm
diameter [36].
iii) The feasibility of the new technique,
maximum-a-posteriori (MAP), for
estimating the tumor response contained
investigated and good results were obtained
[39].
2.7. Indirect Holographic Method
Many of the radar methods are based on direct
microwave holographic method to diagnose
tumors. The obtained results of this method were
encouraging. However, this method uses vector
images, therefore is slow and expensive [43].
Another method which uses Indirect Holographic
method as an alternative to detect tumors was also
investigated [44].
Indirect holographic method uses the application
and direct back transformation onto the measured
interference pattern, the use of continuous wave
signal for imaging avoids the problems associated
with pulsed systems.
This method comprises of two stages. Stage-one
is the recording of a holographic intensity pattern,
by combing the signal scattered from the object,
with a phase coherent reference plane wave signal.
The second stage is the image reconstruction from
the 2D holographic intensity pattern produced
[45].
3. NUMERICAL METHODS FOR
DETECTION
As time-domain microwave breast imaging
algorithms are usually based on numerical
simulations with the Finite-difference Time-domain
(FDTD) method [40], accurately modeling all the
different aspects of the problem is very important
for the evaluation of these systems. Biological tissue
in general is quite dispersive in the microwave
frequency range. Modeling of the frequency
dependence for the various types of tissue is an
important challenge for time-domain analysis of
microwave breast cancer detection systems. FDTD
is a useful method for the simultaneous acquisition
bandwidth of interest. The sample spacing of the
forward solution grid must be dense enough to limit
the numerical dispersion in the FDTD simulation,
but sparse enough to limit the computational cost of

Microwave Imaging: Potential for Early Breast Cancer Detection 285
Fig. 2. Simple delay and sum beam former [35].
Fig. 3.
Fig. 4. Multiple scattering effects inside the object.
the simulations. The choice of the FDTD method is
motivated by the fact that it has a distinct advantage
domain. An understanding of these interactions
imaging is moving towards the search of better
signal to noise ratio (SNR) and image resolution.
A uniform 2.0 mm sample spacing was selected
for the forward FDTD solutions to balance this
trade off. Since multiple FDTD simulations of the
numerical phantoms are still computationally costly
on a 2.0 mm grid, the simulations on a GPU based
hardware accelerator were run to achieve a feasible
imaging time. Alternative numerical techniques
for reducing the computation time of the forward
solution have been presented elsewhere [41, 28,
42]. For further explanation, consult Jocob et al
[46].
4. CONCEPT OF INVERSE SCATTERING
IN MICROWAVE IMAGING FOR
TUMOR DETECTION
object is inferred from the measurement data
collected at a distance from the scatterer. In addition
to obtaining the shape of the object, a quantitative
obtainable from the inverse scattering experiment.
The inverse scattering depends on the multiple
scattering effects that take place within the object.

286 Muhammad Hassan Khalil et al
linearly relate to the object function; which is
a function that describes the velocity, relative
permittivity and conductivity distribution of the
object. A solution to the inverse scattering is
A solution to the inverse problem is non-unique,
due to the generation of evanescent waves by high
spatial frequency portions of the object. These
waves are exponentially small at the receiver
locations and in practice not measurable unless the
receivers are very close to the object. This gives rise
tomography.
the scattering object is a non-linear one. This nonlinearity
arises from the multiple scattering effects
within the object as shown in Fig. 4.
When only scatterer 1 is present, the scattered
1s
and when only scatterer 2 is present
2s
. However when both the
scatterers are simultaneously present, the scattered
1s
+ Eff 2s
+ Eff ms
where Eff ms
is a result of
multiple scattering between the scatterers.
5. CONCLUSIONS
Motivated by the clinical desire, microwave breast
imaging has been an active research area over the
past two decades. Most of the works reported were
simulations, and a few were tested experimentally
on phantoms. Reconstruction of 2-D tomographic
images from experimentally collected scattered
cancerous inclusion) in the presence of a matching
coupling medium was not tried. The analyses for
distorted Born iterative method to solve the inverse
scattering problem using experimentally collected
scattered data on breast tissues and on breast
phantoms. This method was adopted as it is simple,
easy to implement and could develop images
with reasonable accuracy, for strong detection of
malignant tumor by microwave technology.
Though these efforts have generated an expectation
for something close to an ideal breast cancer
detection system, a number of fundamental and
contemporary issues deserve further research
consideration about this microwave modality.
Also, it is important to remember that in addition
to microwave imaging, several other alternative
breast cancer detection modalities are actively
being pursued, including optical imaging methods.
6. ACKNOWLEDGMENTS
The authors are thankful to China Scholarship Council
and Northwestern Polytechnical University Xi’an,
China for the research support.
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Proceedings of the Pakistan Academy of Sciences 49 (4): 289-294 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Review Article
Pain Management in Cancer Patients: A Review
Wasam Liaqat Tarar*, Sundus Sonia Butt, Fatima Amin and Mariam Zaka Butt
Department of Pharmacy, Lahore College for Women University, Lahore, Pakistan
Abstract: Pain management in cancer patients is a dire need which is often overlooked in the overall healthcare
provided by oncologists. This review focuses on revealing the crucial trends and issues on effective pain
management. The meta-analyses of previous research studies demonstrate that pain management in cancer
patients is often suboptimal and there are many barriers which lead to poor pain management. According
to the WHO guidelines there should be scheduled administration of drugs orally on the occurrence of pain,
starting with non-opioid drugs such as acetaminophen, NSAIDs or COX-2 inhibitors following mild opioids
and later strong opioids. Pain management in cancer patients is often inadequate due to the reasons as poor
diagnosis of pain and reluctance of patient to take opioids or report pain. On physician’s part reasons are
managing pain in cancer patients as the guidelines are available. Effective alliance with pain experts and
experts on palliative care can also be helpful. There is a need for collaboration in the form of clinical teams
including nurses, doctors and pharmacists rather than individual efforts to be involved in managing cancer
pain.
Keywords: Pain management, cancer patients, oncologists, opioids
1. INTRODUCTION
A disagreeable sensory and emotional experience
which is related with tissue damage either actual
or potential, or expressed in terms of such damage
cancer may by the tumor itself or by some medical
interventions performed while diagnosing and
treating the disease. Pain is divided into two sub-
to the central nervous system (CNS) which results
the meaning or interpretation of the pain which is
(e.g., more fear may be induced by “crushing” chest
pain than by the same pain intensity in any other
part of human body). The experiences, behaviours
The reactive component has wide variability from
one individual to another [1].
2. PREVALENCE OF CANCER PAIN
Pain in cancer is widespread and its prevalence
varies according to the stages of illness. Patients
with early disease (48%), patients undergoing
cancer treatment (59%) and 64-74% with advanced
disease have cancer pain. Most of the time of cancer
patients with pain is spent in the community settings
until the last days of their lives. Overall, cancer pain
pervasiveness in Europe is 72%. Elderly patients
and those in care homes are particularly more prone
to under-treatment of pain. Primary care teams
along with palliative care teams are more likely to
initiate and treat cancer pain, but the education of
patients, caregivers and healthcare providers is of
prime importance for improved patient outcomes
[2, 3].
2.1. Age and Cancer Pain
Age is not found to have any correlation with the
pain intensity but older patients are found to respond
adversely to lower doses of drugs or potency of
analgesic drugs in secondary care settings (out-
————————————————
Received, September 2012; Accepted, November 2012
*Corresponding author: Wasam Liaqat Tarar; E-mail: wasam_tarar@hotmail.com

290 Wasam Liaqat Tarar et al
patient clinics and hospice inpatient units) [4, 5,
6]. A scantiness of research on community based
patients having cancer pain is found. Cognitive
impairment in older people is associated with greater
intensity of cancer pain [7]. Their experience does
not differ from younger people as they do not suffer
from more adverse effects, and do not need increase
in dose or the need for changing opioid [5, 8].
2.2. Pain Assessment Scale
Different categories of pain severity i.e. mild,
moderate and severe pain, in terms of their
0-10 point numerical scale aforesaid three distinct
correspond to mild pain, 5-6 to moderate pain, and
7-10 to severe pain based on extent of interference
with cancer patient’s performance and activity. It
demonstrates that the pain severity-interference
relationship is non-linear. There were only slight
pain affects, whereas these cut-points were same
pain levels are found to be useful in clinical trials,
clinical evaluation and also epidemiology [9].
The hospitalized cancer patients report more pain
intensity and in many it is under-treated. Patients at
community settings have even greater intensity of
pain than those in secondary care hence effective
strategies for pain management in primary care
must be implemented [10, 11] .
2.3. Intensity of Cancer Pain
The patients having moderate pain occurring
several times weekly often suffer from moderateto-severe
pain in a month. Many patients receive
prescription analgesics and others receive strong
opioids either alone or as combinations with milder
opioids or NSAIDs. The patients undergoing
prescription analgesics therapy still experience
breakthrough pain. Many of the cancer patients
life routine activities. Whereas many patients hold
that the quality of life is not considered as a priority
by their physicians in overall course of therapy. The
treatment of pain in cancer is suboptimal as patients
are not pain free. The management of pain should
be given a prime importance and the pain should
not be considered as part and parcel of cancer. The
pain treatment guidelines should be revised by
healthcare teams for effective pain control in cancer
patients [12].
2.4. Concurrent Symptoms
Cancer patients suffer 3.3 symptoms as an average
along with pain. These symptoms include anorexia,
insomnia, constipation, sweating, nausea, vomiting,
diarrhea, dysphagia, dyspnea, neuropsychiatric
symptoms, urinary symptoms, dyspepsia, paresis,
pruritus, and dermatological symptoms. The
occurrence of these symptoms is associated with
the site of tumor, intensity of pain, and opioid
therapy. The association between symptoms and
These symptoms must be addressed to improve
patient’s quality of life. Only highlighting the pain
while treating cancer patients with pain is not going
to serve the purpose for patients rather a more
global and erstwhile approach is the need of hour
for associated symptoms management [13].
3. TREATMENT GUIDELINES
Chronic pain in cancer is treated by opioid
analgesics as they are drugs of choice. The dose
equianalgesic to the previous opioid should be
a patient who has already been treated with opioids.
Approximate equianalgesic dose for codeine is
120 mg and for hydromorphine is 2 mg. Patients
taking appropriate dose of acetaminophen i.e., 1gm
every four hours and still their pain is not controlled
then another drug should be started rather than
increasing acetaminophen dose. Hepatotoxicity,
is often reported while treating cancer pain with
acetaminophen. The dose of morphine varies.
Continuous infusion is found to be helpful in
patients who require frequent dosing or those who
are unable to take medicine orally. Due regard
must be given to infusion rates. Many guidelines
have come to the forefront for continuous opioid
infusions. Methadone should be avoided in elderly
patients, patients suffering from severe liver
dysfunction and in patients with retarded pulmonary
function. Meperidine should be avoided when
chronic frequent use is required and in patients

Pain Management in Cancer Patients 291
with renal dysfunction. Meperidine can lead to
seizures as it’s metabolism forms normeperidine
which possesses CNS stimulatory activity. Orally
administered heroin is same as morphine as it is
metabolized to morphine. Parenteral heroin can
b given in small volume as it has better solubility
than morphine therefore more drug quantity can be
delivered. Propoxyphene results in the same effects
as all other opioid analgesics. However it is less
potent as compared to other opioids, which makes
it a suitable candidate to be administered along with
non-opioid analgesics [1].
3.1. Effectiveness of Opioids when given
Parenterally
It is established that it is easier to control pain by
preventing it from recurring rather than treating it
when it has recurred as we relate with the words
‘precaution is better than cure’. Hence medication
on frequent schedule basis is preferred rather than
PRN for patients with persistent chronic pain.
Also the anxiety level in patient increases as he
knows the effect of last dose is to subside and he
has to experience pain before the next dose. This
The parenteral route is more effective for all opoids
given by oral route. The ratio of oral:parenteral
effectiveness of morphine is found to be 1:6. Thus,
when drug switching from oral to parenteral route
is to be done, doses can be made half or reduced to
one-third, and when switching from parenteral to
oral route, doses can be made double or triple [1].
3.2. Oncologists’s Ability
When oncologists are interviewed about their cancer
pain management ability they rate themselves high
on a numeric scale of 0-10 {median, 7; interquartile
range [IQR], 6 to 8}. They rate their peers and
fellows as more conservative prescribers compared
to themselves as they still follow long-established
guidelines. The quality of pain management
training conducted in medical school and during
residency is rated as 3 and 5 respectively which is
3.3. Inadequate Analgesia
There exists a discrepancy in treating patients as
the patients at centers treating minorities more
frequently suffer from under treatment of pain as
compared to other hospital settings. The physician’s
poor management of cancer pain. Other factor
that contribute to inadequate pain management
include physicians do not consider the pain to be
associated with cancer, better performance status
of patients, elderly patients and female patients.
Thus, many patients report severe pain even after
taking the analgesic therapy. This is pain is severe
enough to hinder their daily life activities. The pain
management in cancer patients is inadequate till
date despite availability of cancer pain management
guidelines [16].
3.4. Controlled Use of Opioids
Opioids have been used for the treatment of
pain since ages but it has been recently (around
past 60 years) brought under controlled use.
The legitimate use of opioids is only under the
supervision of licensed practitioners. According
to current available guidelines the dose escalation
and discontinuation of opioid drugs if treatment
plan is not being accomplished, is to be carefully
monitored. Unfortunately in busy practice settings
this is not the scenario. Sometimes higher doses of
opioids are being prescribed to cancer patients with
chronic pain which is not the result of advanced
cancer stage. Earlier there was a misconception
that unlimited increase in dose of opioids is safe
but now it has been established that unnecessary,
prolonged and high dose opioid therapy is neither
effective nor safe. Therefore the responsibility lies
in the hands of physicians to control the prescribing
of opioids even though when patients demand to
increase the doses [17].
3.5. Cancer Therapy vs. Pain Therapy
Cancer therapy and pain therapy go hand in hand.
The two important factors to be considered are
cancer’s treatability and cancer’s “non pain”
pathophysiology (pathophysiology that does not
cause pain). Non-pain pathophysiology can give
tough time by precluding oral administration of
drugs, narrowing a patient’s therapeutic index for
analgesics, rendering psychologic pain therapies
ineffective, and limiting the execution of invasive

292 Wasam Liaqat Tarar et al
pain-relieving procedures. Both cancer therapy and
therapy can hinder pain therapy by exacerbating
pain or implicating other adverse effects. On the
other hand it can be of help to pain therapy by
reducing the proliferation of cancer, lending hand
as co-analgesic, and by providing route for IV
administration of drugs according to patients needs.
Likewise pain therapy can improve cancer therapy
by the performance status of patient and surgically
performed interventions can help improve organ
function [18].
3.6. Use of Multiple Opioids
No single opioid is completely safe. As drugs are
only tools, it depends on our expertise how we use
them to get the desired therapeutic outcomes. Dose
escalation with many co-administered opioids leads
to adequate pain relief along with many unwanted
side effects in cancer patients with pain. Drug
switching from one opioid to another is found to be
helpful with no considerable correlation among the
genetics and opioid response [19].
3.7. Morphine vs. Oxycodone
When intravenous dosing of two drugs i.e., morphine
and oxycodone is compared, it is found that equal
analgesic effect is achieved from both drugs. Their
bioavailability is pretty much similar, only a little
higher for oxycodone hydrochloride. The patients
IV dose for oxycodone hydrochloride to produce
same amount of analgesia as morphine is 30%
higher. Nausea and hallucinations are the drawbacks
of morphine use. Otherwise both morphine and
oxycodone are similar in their actions and there is
[20].
3.8. Spinal Administration of Opioids
The spinal administration of opioids may provide
analgesia for longer duration to patients suffering
from bilateral or midline lower abdominal or pelvic
cancer pain. However, there are certain reasons
which make the use of spinally administered
analgesics obscure. Cross-tolerance to orally
and parenterally administered narcotics is of
to spinal narcotics has also markedly limited their
usefulness. Opioids extensively distribute in the
CSF and plasma when administered through the
epidural or intrathecal route. The drug reaching to
brain stem sites may account for many of the toxic
and therapeutic effects of spinal opioids [21].
3.9. Use of Benzodiazepines
Benzodiazepines can be used to indirectly manage
cancer pain and have been found effective. It is
related to their psychotropic effects including
reduction of anxiety and depression in many
instances. Benzodiazepines can serve the purpose
for treating chronic pain, acute muscle spasm and
associated anxiety, and neuropathic pain. Alprazolam
and clonazepam have been found to be drugs of
choice. They should not be always considered as
with potential harmful effects sometimes such as
physical and psychological dependence, cognitive
impairment, worsening depression, overdose, and
many other side effects [22].
4. BARRIERS TO CANCER PAIN
MANAGEMENT
The various obstacles including behaviours which
prevent successful management of pain are referred
fear of addiction and tolerance of analgesics, poor
assessment of pain, communication barrier among
healthcare professionals and patients and some
religious and cultural norms which all cumulatively
lead to poor pain control. [23] [24] Reluctance of
patients to report pain and utilization of analgesics
stand at the top of the list. The patients are concerned
about the addiction causing factor of analgesics
and their side effects. There is also a wrong notion
in patient’s minds that pain is part and parcel of
cancer and ‘good patients do not whine about pain’.
It should be eradicated from patient’s minds. A
educated and patients belonging to lower income
group have these sorts of concerns. Higher pain
intensity and under treatment of pain leads to more
concerns. [25] Other barriers included in effective
pain management are poor assessment of pain,
physicians are reluctant to prescribe opioids and
perceived excessive regulation. The barriers and

Pain Management in Cancer Patients 293
limitation in pain related knowledge and practice
norms within the oncology settings have not been
duly addressed for past few decades, though cancer
pain has been highlighted [14].
5. QUALITY IMPROVEMENT PROGRAMS
Quality improvement programs have been designed
and implemented for the treatment of chronic and
QI programs are; unrelieved cancer pain grabs
providing information regarding analgesics when
orders require so, gaining patient’s trust that they
will be provided analgesic care hence they should
report pain, implementing effective policies for
the use of new era analgesic technologies, and
coordinating and monitoring implementation
of aforesaid measures. Though QI approach is
helpful in improving patient’s satisfaction and also
helps identifying many underrated obstacles to
optimal pain management, yet it is not an answer
and solution to all undertreated pain problems.
QI programs are multidimensional in nature and
can provide a cornerstone that includes physician
and patient education, minimizing errors in drug
use process by designing informational tools, and
overall improved process of assessing and treating
cancer pain [26].
6. KNOWLEDGE AND BEHAVIOURS OF
HEALTHCARE PROVIDERS
The knowledge and behaviours of healthcare
providers towards pain management in cancer plays
a key role in effective management and it varies
widely. Nurses possess better pain assessment
expertise than do doctors or pharmacists. As nurses
are on the bedside of patients and they are more
likely to observe changes in behaviour pattern of
patients. Doctors have better knowledge of clinical
pharmacotherapy. As pharmacists are experts on
drugs therefore they have most knowledge about
opioids pharmacology and kinetics. As these
professionals nurses, physicians and pharmacists
possess expertise in many areas they also lack
help us identify the need for collaborative clinical
teams instead of individual efforts to be involved in
managing cancer pain [27, 28].
7. CONCLUSIONS
Drug therapy is the key to management of pain
in cancer patients. Despite the availability of
guidelines for pain management, many cancer
patients still experience considerable pain and
patients is often inadequate and suboptimal.
This can be accredited to many reasons; pain is
not given priority in overall healthcare, lack of
education, inappropriate pain assessment and the
appropriateness of opioid therapy i.e., restricted
use of strong opioids among health care providers,
patients, and patients’ families. There is a right away
need of effective methods of training oncologists
and holding them accountable for their actions
and ample pain relief. Most oncologists should be
patients with cancer. There should be alliance with
pain experts in relieving pain and implementing
modern analgesic technologies. Improving the
hundreds of patients and their families. It will also
provide a model for better healthcare of the even
larger number of patients who have undertreated
pain due to diseases other than cancer.
8. ACKNOWLEDGMENTS
We express gratitude to our Head of Department, Dr.
Hafeez Ikram, for his kind guidance. We also express
profound appreciation to our honorable teachers for
their thoughtful guidance and sagacious advices. We
are greatly indebted to our parents for their continuous
support, encouragement and supplications at all stages.
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3. van den Beuken-van Everdingen, M.H., J. M. de
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4. Vigano, A., E. Bruera, & M. E. Suarez-Almazor.
Age, pain intensity and opioid dose in patients with

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Opioid escalation in patients with cancer pain:
the effect of age. Journal of Pain and Symptom
Management 32(5): 413-419 (2006).
6. Mercadante, S., F. Roila, O. Berretto, R. Labianca
& S. Casilini. DOMAIN-AIOM study group.
Prevalence and treatment of cancer pain in Italian
oncological wards centres: a cross sectional survey.
Supportive Care in Cancer 16(11):1203-1211
(2008).
7. Allen, R. S., W, E. Haley, B. J. Small & S. C.
McMillan. Pain reports by older hospice cancer
patients and family caregivers: the role of cognitive
functioning. The Gerontologist 42(4):507-514
(2002).
dose and side-effects: a comparison of older
and younger patients with tumor pain. Deutsche
Medizinische Wochenschrift 125(41):1216-1221
(2000).
9. Serlin, R.C., T. R. Mendoza, Y. Nakamura, K. R.
Edwards & C. S. Cleeland. When is cancer pain
its interference with function. Pain 61 (2): 277-284
(1995).
10. Klepstad, P., J. H. Loge, P. C. Borchgrevink, T. R.
Mendoza, C. S. Cleeland & S. Kaasa. The Norwegian
brief pain inventory questionnaire: translation and
validation in cancer pain patients. Journal of Pain
and Symptom Management 24(5):517-525 (2002).
11. Yates, P. M., H. E. Edwards, R. E. Nash, A. M.
Walsh, B. J. Fentiman, H. M. Skerman, et al. Barriers
to effective cancer pain management: a survey of
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of Pain and Symptom Management 23(5):393-405
(2002).
12. Breivik H, Cherny N, Collett B, de Conno F, Filbet
M, Foubert AJ, et al. Cancer-related pain: a Pan-
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patients with cancer pain: A prospective evaluation
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Journal of Pain and Symptom Management 9 (6):
372–382 (1994).
14. Breuer, B., S. B. Fleishman, R. A. Cruciani & R.
K. Portenoy. Medical oncologists’ attitudes and
practice in cancer pain management: a national
survey. Journal of Clinical Oncology 29 (36): 4769-
4775 (2011).
15. Bruera, E., K. Macmillan, J. Hanson & R.
N. MacDonald. The cognitive effects of the
administration of narcotic analgesics in patients
with cancer pain. Pain 39 (1): 13-16 (1989).
16. Ballantyne, J. C. & J. Mao. Opioid therapy for chronic
pain. The New England Journal of Medecine 349:
1943-1953 (2003).
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27-32 (1990).
18. Levy MH. Integration of pain management into
comprehensive cancer care. Cancer 63 (11 Suppl):
2328-2335 (1989).
Portenoy. Individual variability in the response to
Pain 49 (1):
87-91 (1992).
20. Kalso, E. & A. Vainio. Morphine and oxycodone
hydrochloride in the management of cancer pain.
Clin Pharmacol Ther 47 (5): 639-646 (1990).
21. Payne, R. Role of epidural and intrathecal narcotics
and peptides in the management of cancer pain. Med
Clin North Am 71 (2): 313-327 (1987).
22. Reddy, S. & R. B. Patt. The benzodiazepines as
Journal of Pain and Symptom
Management 9 (8): 510-514 (1994).
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and caregivers as a barrier to the pharmacological
control of cancer pain. Clinical Pharmacology and
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Mueller, A. Nolan, D. Pawlik-Plank, et al. Patientrelated
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Quality Improvement guidelines for the treatment of
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1880 (1995).
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Pierce, M. Dolan, L. Roberts, et al. Knowledge and
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Proceedings of the Pakistan Academy of Sciences 49 (4): 295-300 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Research Article
Wavelet Characterization of Turbulence for
Ionospheric Region of the Pakistan Air Space
M. Ayub Khan Yousuf Zai 1 * and Khusro Mian 2
1
Solar-Terrestral and Atmospheric Research Center, Department of Applied Physics,
University Karachi, Karachi, Pakistan
2
Institute of Space and Planetary Astrophysics, University of Karachi,
Karachi, Pakistan
Abstract: It has been observed that solar activity affects the communication system. This effect is due to
the ionospheric turbulence. The turbulence due to the perturbation concentration and parcel velocity were
analyzed in upper and lower layers of the ionosphere by Spectral analysis, Fast Fourier Transformation and
wavelet transformation in one dimension, Haar 5-Levels for approximation and details. Wavelet analysis
is known as the method to study non-stationary data. Wavelet analysis is used to quantify both signal of
presented under changing ionospheric conditions.
Keywords: Ionosphere, turbulence, perturbation, spectral analysis, wavelet, Fast Fourier Transformation
1. INTRODUCTION
Ionosphere is part of magnetosphere, and is created
by the ionization of neutral particles. Ionospheric
perturbations can be enumerated and life time
monitor this important region of our globe because
it is critical for the future progress as it was in the
past. Turbulence means unstable and disorderly
movements and the ultimate result of the current
of air and water whirling against the main current.
It has been observed that Earth Science and Space
Science are linked together in a peculiar way. The
ionosphere lies in the upper atmospheric region of
the earth. The ionosphere has diurnal and seasonal
changes and its density varies with the solar
radiation interaction. The structure of the ionosphere
differs with its altitude and the density of ions and
electrons. At the base of ionosphere, about 80 km
above the earth surface, there are about 10 4 ions
per cubic cm. In comparison, at the same point in
the atmosphere, there are about 2x10 16 non-ionized
or neutral particles per cubic cm,. Thus, at 80 km
————————————————
Received, February 2012; Accepted, December 2012
*Corresponding author: M. Ayub Khan Yousuf Zai; Email: ayubzai@yahoo.com
above the earth, the ratio of neutral particles to
ionosphere were analyzed by spectral analysis
and wavelet analysis. This is a new approach of
exploring, ionospheric turbulence at the Pakistan
air space [1–5].
2. MATERIAL AND METHODS
The study was based on the analyses of ionospheric
turbulence at Pakistan air space such as perturbation
concentration and parcel velocity. These were
analyzed in upper and lower ionospheric layers using
Spectral analysis, Fast Fourier Transformation and
wavelet approach for one dimension Haar 5-Levels
for approximation and details.
3. RESULTS AND DISCUSSION
3.1. Computation of Turbulence Flux
layer at the Pakistan region by using perturbed
electron concentration (N / ) and perturbed parcel

296 M. Ayub Khan Yousuf Zai et al
Fig. 1. Time plot for F 2
layer, ionospheric turbulence.
Fig. 2. Illustration of period-o-gram value and frequency
for ionospheric turbulence F 2
Layer data.
Fig. 3. Illustration of spectral density and frequency for
ionospheric turbulence F 2
-layer data.
Fig. 4. Predicted value vs. observed value obtained
concentration at ionospheric F 2
layer.
velocity (V /
________
is unsteady. The product U / N / represents the
transport of Kinematics Turbulent Flux:
U
N
U
2
N
________ / / / /
/ / E E F 2 F 2
U
N
where
/
U
F 2
is Instantaneous and Local Perturbation Scalar
Velocity content
/
U
E
is Instantaneous and Local Perturbation Scalar
Velocity content
/
N
F 2
is Instantaneous and Local Perturbation
Plasma Concentration content
/
N
E
is Instantaneous and Local Perturbation Plasma
Concentration content [5].
1.2. Spectral Analysis
The ionospheric turbulence clearly appears to
follow a cyclical pattern (Fig. 1), the line spectrum
on periodogram (Fig. 2) constructed to identify the
randomness in the ionospheric turbulence due to
Fig. 3 shows spectral density at ionospheric
turbulence F 2
-layer data. Fig. 4 shows predicted
turbulence and electron concentration at ionospheric
F 2
layer. In fact, it is descried to show seasonality

Wavelet Characterization of Turbulence 297
among the time series event. The purpose of spectral Table 1. Illustration of ionospheric turbulence as
concept is central to spectral analysis technique, the
Fig. 5. Descriptive statistics: (i) Histogram; (ii)
A , , form a period-o-gram [6, 7].
Cumulative histogram of ionospheric turbulence at the
Pakistan air space.
analysis is to present ionosphere that examines
data series in term of their frequency content.
Frequency domain analysis has become much more
——————————————————————
central to decode the cause and effect of upper F P Cos Sin Pgm
atmospheric changes. Fast Fourier Transformation ——————————————————————
(FFT) techniques further aided the application of
frequency domain analysis in upper atmosphere.
0.000
0.002 364
0.003
-0.045
0.000
0.082
0.002
1.607
0.005 182 -0.437 -0.186 41.20
peak frequencies are periodogram peaks. The large
0.008 121 -0.088 -0.092 2.987
0.010 091 -0.071 0.099 2.734
and number of observations after padding is 364 ——————————————————————
and Transformations: Mean = 2.4458 subtracted.
Largest peak frequencies corresponding values
are:
1.4. Wavelet Analysis
We consider ionospheric turbulence data at Pakistan
air space as presented in Fig 5 and descriptive
Value: Frequency
statistics is shown in Table 2 and 3 histogram and
41.21: 0.006
19.28: 0.033
07.64: 0.030
05.19: 0.035
04.01: 0.351
1.3. Fourier Analysis
We have presented ionospheric turbulence in the
form of astrophysical signals as depicted in Fig.
3 and listed their parameters in table.1. These are
Sine, Periodogram, Density, Hamming Weight.
A goal of time series analysis in the frequency
domain is to separate, periodic oscillations from
analysis is one of the famous methods for
identifying periodic components. Fourier series,
f ( t)
f ( t)
[
APCos(
Pt)
BPSin(
Pt)]
P
where f (t)
is the mean value of the function,
A , B P P
function of Sin (
Pt)
and Cos ( Pt)
and the
angular frequency
P
are integer p = 1, 2, 3, …..,
multiples of the total length of the time series. This
P B P

298 M. Ayub Khan Yousuf Zai et al
Decomposition at level 5: s: a5+d5+d4+d3+d2+d1.
S
a 5
d 5
d 4
d 3
d 2
Fig. 7. Reconstructed details analyzed at level 5,
residual part of ionospheric turbulence model at Pakistan
air space.
d 1
50 100 150 200 250 300 350
Fig. 6. Decomposition at level 5,
of ionospheric turbulence at Pakistan air
space.
cumulative histogram of ionospheric turbulence at
Pakistan air space in Fig. 6–8. Wavelet approach is a
recently developed signal processing tool enabling
us on several timescale of the local properties of
complex signals that can present non stationary
variations and also to study how it varies with time
by decomposing time series into frequency space.
Finally, powerful multi resolution with respect to
Haar is carried out. This decomposes actual signal
into different signals to be analyzed into principal
and residual part. By mathematical expression

Wavelet Characterization of Turbulence 299
Residuals
S
A j
D j
j
4
2
0
-2
50 100 150 200 250 300 350
Histogram
Auto correlations
Cumulative histogram
S, A
j
and D
j
are the signal, principal part j level
and residual part j level. Mathematically, the
relation between principal and residual part j level
by mathematical is:
A
j1
A
j
D
j
The wavelet used in the study by Haar of the level 5,
j
the dyadic scale is a 2 for level 5 the resolution
is given by 1/a, or 2 -j . In order to approach the
cyclic study maximum and minimum values, we
have carried out wavelet analysis of ionospheric
turbulence. In Fig. 6 the variation is presented in
the form of different resolutions at level 5 of Haar
wavelet type in the detailed and approximated part
the cyclic variation is also presented at different
level. In the detailed and approximated part at
the lowest resolution several peaks are appeared.
The expression of Haar wavelet in continuous
time series function is represented as follows:
1
{ 1;0 t
2
{ 0; otherwise,
1
2
t
0
f ( t)
{ 1;
t0
t t0
The corresponding wavelet in discrete time series
{ 1; n 1 h { 1;
n 2
h n
{0;otherwise
0
The quantity h 1/ 0
2 is a constant. Fig. 7 shows
wavelet 1-D analysis of ionospheric turbulence at
Pakistan air space. Present data is non-linear there
is local minimum in this period which depicts a
positive skewness of frequency histogram:
FFT - Spectrum
Frequency
Fig. 8. The graph of residual, autocorrelation, FFTspectrum,
histogram, cumulative histogram obtained from
wavelet analysis F 2
layer.for ionospheric turbulence.
s a
5
d5
d
4
d3
d
2
d1
The Fig. 8 manifests the residual, autocorrelation,
FFT-spectrum, histogram, cumulative histogram
of ionospheric turbulence F 2
layer. For an
advance and more recent analysis of ionospheric
turbulence at Pakistan air space, between 1 to 365
components data point, we have constructed model
of ionospheric turbulence. We have used wavelet
transformation which is a common method of
analyzing ionospheric plasma turbulence [8-10].

300 M. Ayub Khan Yousuf Zai et al
Table 2. Descriptive statistics, quantities of ionospheric
turbulence at Pakistan air space.
——————————————————————
Mean 2.5
Median 2.3
Mode 2.2
Max 5.7
Min 0.8
Range 4.9
St. Dev. 0.7
Median AD 0.5
Mean AD 0.6
——————————————————————
Table 3. Descriptive statistics, residual quantities of
ionospheric turbulence at Pakistan air space.
——————————————————————
Mean -0.001
Median -0.051
Mode -0.400
Max 2.306
Min -1.300
Range 3.605
St. Dev. 0.600
Median AD 0.338
Mean AD 0.450
——————————————————————
3. CONCLUSIONS
out the hidden periodicities in different processes
quantifying the ionospheric turbulence that was
analyzed in upper layers of ionosphere by spectral
analysis, FFT and wavelet transformation one
dimension Haar 5-Levels for approximation and
decomposition details at the Pakistan air space. The
more precise results were obtained using tests of
randomness and normality in datasets. These results
might be useful in astrophysical plasma.
4. ACKNOWLEDGEMENTS
The authors are grateful to the technical staff of Space and
Upper Atmosphere Research Commission (SUPARCO)
for providing the Ionospheric data in the form of critical
frequency and other variables recorded by Digital
Ionospheric Sounder DGS-256.
5. REFERENCES
1. Rishbeth, H. Introduction to Ionospheric Physics.
Academic Press, New York, p. 4-31 (1969).
2. May–Britt Kallenrode. Space Physics – An
Introduction `to Plasma and Particles in the
Heliosphere and Magnetospheres, 3 rd ed. Springer-
Verlag, Berlin, p. 301-302, 166-167 (2004).
3. Chen, M. C. Introduction to Plasma Physics. Plenum
Publishing Corporation, New York, p. 243 (1929).
4. Barclay, L. Propagation of Radio Waves. The
Institute of Electrical Engineers, London (2003).
5. Jacobson, M. Z. Fundamentals of Atmospheric
Modeling “IST Cambridge University Press, USA,
p. 60-65 (1999).
6. Pandit, S M. Time Series and System Analysis with
Application. John Wiley & Sons, USA, p. 24, 29,
119 (1983).
The Analysis of Time Series: An
Introduction, 4/e, Chapman & Hall, London, p.105-
35 (1989).
8. Misiti, M., Y. Misiti, G. Oppenheim & Jean-Michel
Poggi Wavelets and Their Applications. ISTE, Ltd.
USA, p.13-23 (2007).
9. Van den Berg, J. C. Wavelets in Physics. Cambridge
University Press, UK, p. 8-15 (2004).
10. Hong, D. Real Analysis with an Introduction to
Wavelets and Applications. Elsevier India, p.123-
213 (2005).

Proceedings of the Pakistan Academy of Sciences 49 (4): 301-307 (2012) Pakistan Academy of Sciences
Copyright © Pakistan Academy of Sciences
ISSN: 0377 - 2969 print / 2306 - 1448 online
Research Article
Hadamard-type Inequalities through -Convexity
Muhammad Iqbal 1* , Muhammad Iqbal Bhatti 1 and Silvestru Sever Dragomir 2
1 Deparment of Mathematics, University of Engineering & Technology, Lahore, Pakistan
2 School of Engineering and Science, Victoria University, Melbourne, Australia
Abstract: Some new inequalities of Hadamard-type for h convex mappings with general point
of line segment are established. Two Lemmas permitting us to establish new inequalities
connected with lower and upper part of the celebrated Hermite-Hadamard inequality, are pointed
out.
Keywords: Hermite-Hadamard inequality, convex, s convex and h convex functions.
1. INTRODUCTION
Let f :[
a,
b]
R
be a convex function. Then
b
a b 1
f ( a)
f ( b)
f f ( x)
dx
(1.1)
2 b a a
2
refer as the Hermite-Hadamard inequality [3, 8, 9].
In recent years, many authors established several
inequalities connected to this famous integral
inequality (1.1). For recent results, refinements,
counterparts, generalizations and new Hermite-
Hadamard type inequalities see the papers [2–7].
Definition 1: [3]Let I be an interval of real
numbers. The function f : I R is said to be
convex if for all x, y I and t [0, 1] one has the
inequality:
f ( tx (1 t)
y)
tf ( x)
(1 t)
f ( y).
If the above inequality is reversed then f is said
to be concave.
Dragomir and Agarwal [4] obtained inequalities
for differentiable convex function which are
connected with the Hermite-Hadamard inequality
(1.1), one of them is pointed out as:
Theorem 1: Let f : I RR
be differentiable
function on I o where a, bI
with a b . If f ' is
convex on [ a , b]
, then
f( a) f( b) 1
f ( x)
dx
2 b a
a
b
a
f '( a) f '( b)
8
Kirmaci [6] gave the following results:
b
(1.2)
Theorem 2: Let f : I RR
be differentiable
function on I o where a, b I with a b . If f '
is convex on [ a , b]
, then
b
1
a
b
f ( x)
dx f
b a
2
a
b
a
f '( a) f '( b)
8
(1.3)
Theorem 3: Let f : I RR
be differentiable
function on I o where a, b I with a b and
p 1 . If
q
f ' is convex on [ a , b]
, then
b
1
a b b a 4
f ( x)
dx f
b a
a 2 16 p 1
1/ p
q
q 1/ q
q q 1/ q
f ( a)
3
f'(
b)
3
f'(
a)
f'(
b)
' (1.4)
where 1 1 1.
p q
————————————————
Received, June 2012; Accepted November 2012
*Corresponding author: Muhammad Iqbal; E-mail: miqbal.bki@gmail.com

302 Muhammad Iqbal et al
Kirmaci et al [7] established the following
new Hadamard-type inequality for concave
functions:
Theorem 4: Let f : I RR
be differentiable
function on I o where a, bI
with a b . If
q
f ' , q 1
is concave on [ a , b]
, then
b
f(
a)
f ( b)
1
b a q 1
f ( x)
dx
2 b a a 4 2q
1
1
1
q
3a
b a 3b
f ' f '
(1.5)
4 4
Alomari et al [2] established the following
Hadamard-type inequality for concave functions:
Theorem 5: Let f : I RR
be differentiable
function on I o where a, bI
with a b . If
q
f ' , q 1
is concave on [ a , b]
, then
1
b
a b
b
f ( x)
dx f
a
a 2
b a q 1
4 2q
1
1
1
q
3a
b a 3b
f ' f '
(1.6)
4 4
Definition 2: A function f : I RR
is said to be
Godunova-Levin function or f belongs to class
Q (I) if f is non-negative and for all x, yI
and
t (0,1), the inequality holds [3]:
1 1
f ( t x (1 t)
y)
f ( x)
f ( y).
t 1 t
Definition 3: A function f : R [0,
]
is said to
be a P function or that f belongs to the class
P (I) , if for all x, yR
and t (0,1)
[3]. we have :
f ( t x (1
t)
y)
f ( x)
f ( y).
Definition 4: [5] Let s be a real number,
s (0, 1]. A function f :[0,
)
[0,
)
is said to
be s function(in the second sense) or f belongs
2
to the class K s , if for all x , y[0,
)
and t[0,1] ,
the following inequality holds:
s
f ( tx (1 t)
y)
t
f ( x)
(1 t)
f ( y).
Varošane [13] defined a large class of nonnegative
functions, the so-called h convex
s
functions. This class contains several well-known
classes of functions such as non-negative convex
functions, s-convex in the second sense,
Godunova-Levin functions and P-functions. The
definition of an h-convex function is stated below:
Definition 5: Let h : JRR
be a non-negative
function. A non-negative function f : I RR
is
said to be h convex function (or f belongs to
class SX ( h,
I)
) if for all x, yI
and t (0,1)
, the
following inequality holds:
f ( t x (1 t)
y)
h(
t)
f ( x)
h(1
t)
f ( y).
If the inequality is reversed then f is said to be
h concave function (or f SV( h,
I)
).
Remark 1: It is noted that if:
h( t)
t , then all the non-negative convex
functions belong to the class SX ( h,
I)
and all
non-negative concave functions belong to the
class SV ( h,
I)
.
1
h( t)
, then SX( h,
I)
Q(
I)
.
t
h ( t)
1, then SX( h,
I)
P(
I)
.
s
2
( h,
I)
K s
h( t)
t , where s (0,1)
, then SX .
Some interesting and important inequalities for
h convex functions can be found in [11,12].
In [12], Sarikaya et. al. established a new
Hadamard-type inequality for h convex
functions.
Theorem 6: Let f SX( h,
I)
, a, bI
with a b
and f L[ a,
b]
, then
1 a
b 1
f
2 h(1/2) 2 b
a
[ f( a) f( b)] h( t)
dt
1
0
b
a
f( x)
dx
In this paper new results of Hadamard-type on the
basis of two established lemmas for h convex
functions will be presented.
2. MAIN RESULTS
Lemma 1: Let f : I RR
be differentiable
o
function on I where a, bI
with a b . If
f ' L[
a,
b]
and , [0,
)
with 0
, then

Hadamard-type Inequalities through h-Convexity 303
f( a) f( b) 1
f( x)
dx
ba
a
ba t t
( t) f ' a b dt
2
0
t
0
t t
f ' a
b
b
dt
Proof: It suffices to note that
I1
( t)
f '
0
=
0
t
a
t
bdt
ba t t
f a b
2
( )
0
1 t
t
f a
bdt
ab
f ( a)
1
= f ( x)
dx.
b a a
and similarly we get
I
2
t f '
0
t
a
b
f ( b)
1
=
b a a
b
t
bdt
f ( x)
dx.
(2.1)
(2.2)
(2.3)
By adding identities (2.2) and (2.3), we get
identity (2.1).
Remark 2: For , Lemma 1 reduces to [1,
Lemma 2.1].
In the following theorems, we shall propose
some new upper bounds for the right-hand side of
Hadamard's inequality for h convex mappings.
Theorem 7: Let f ' be h convex function and
the assumptions of Lemma 1 hold, then
f( a) f( b) 1
ba
f ( xdx )
1
( ba) f '( a)
t
0
b
a
f '( b)
t h(
t)
dt
(2.4)
Proof: By using the property of modulus and from
Lemma 1, we have
f( a) f( b) 1
ba
2 0
b
a
f ( xdx )
ba t t
t f ' a
b
dt
t
t
t f ' a b
dt
(2.5)
0
Since
f ' is h convex
f ( a)
f ( b)
1
b a
2 0
b
a
f ( x)
dx
t
f '( a)
h
b a
t
dt
t
f '( b)
h
t t
t f '( a)
h
f '( b)
h
dt
0
1
( ba) f '( a) t
h( t)
dt
f '( b) th( t)
dt
0
f '( a) t h( t)
dt
0
1
f '( b) t h( t)
dt
Hence the proof is completed.
Remark 3: For , and convex function
inequality (2.4) reduces to inequality (1.2).

304 Muhammad Iqbal et al
q
Theorem 8: Let f ' be h convex function and
the assumptions of Lemma 1 hold, then
f( a) f( b) 1
ba
a
1
1
p
b
a
1/ p
( p 1)
0
b
f( x)
dx
q
q
f '( a) h(1 t) f '( b) h( t)
dt
1
1
1
q
+ p {| f '( a) | h(1 t)
q
| f '( b)| h( t)}
dt
1
q
Proof: By applying Hölder’s inequality and
q
convexity on f ' , we have
1
q
(2.6)
h
Putting the above inequalities in (2.5) we get
(2.6).
Corollary 1: For , and convex function
inequality (2.6) reduces as:
f ( a)
f ( b)
1
2 b a
b
a
f ( x)
dx
3 f '( a) f '( b)
b a
4
4( p 1)
f '( a) 3 f '( b)
4
1
q q q
1/ p
1
q q q
Theorem 8 may be extended to be as follows:
q
Theorem 9: Let f ' be h convex function and
the assumptions of Lemma 1 hold, then
t
0
t t
f ' a
b
dt
f ( a)
f ( b)
1
b a
b
a
f ( x)
dx
1
q
p
p t t
t dt
f ' a b dt
0
0
( )
1
q
q
f '( a) h(1 t)
dt
11/ p
1/ q
0
1/ p
( p 1)
q
Analogously
0
f '( b) h( t)
dt
0
t
t
t f ' a
b
dt
( )
q
f '( a) h(1 t)
dt
11/ p
1/ q
0
1/ p
( p 1)
q
f '( b) h( t)
dt
0
1
q
1
q
1
q
q
b a
t {| f '( a) | h(1 t)
1/ p
2
+
0
q
| f '( b)| h( t)}
1
q
1 q
t {| f '( a) | h(1 t)
(2.7)
q
| f '( b)| h( t)}
Proof. The proof of this theorem is same as proof
2
of Theorem 8 and using facts that 1
and
2
( )
2
( )
reader.
2
1. However, the details are left to the
Now, we give the following Hadamard-type
inequality for h concave mappings.
Theorem 10: Let
q
f ' be h concave function
and the assumptions of Lemma 1 hold, then

Hadamard-type Inequalities through h-Convexity 305
f( a) f( b) 1
ba
f ( xdx )
1/ p
1/ q
b
a 1 1
2
( ) p1 2 h(1/2)
2 2
2 1
f ' a
a
b
2 2( ) 2( )
2 2
2 1
f ' b a
b
2 2( ) 2( )
b
a
(2.8)s
t t
f ' a b 0
2
1
1/ p
( p 1)
2h(1/
2)
1
f '
2
b
q
1/ q
dt
2
2
b a
2( )
2( )
Using these above inequalities in (2.5) we get
(2.8).
Proof: By applying Hölder’s inequality, we have
t t
t
f ' a b
dt
0
1
q
p
p t t
t dt
f ' a b dt
0
0
Since
q
1
q
f ' is h concave on [a, b] , we have
t t
2h(1/
2) f ' a b 0
f 1
'
0
0
t
dt
0
=
f
1
'
2
Therefore
a
t
a
t t
f ' a b 0
2
1
1/ p
( p 1)
2h(1/
2)
1
f '
2
a
Analogously
q
dt
t
bdt
q
t
0
0
dt
2
2
a b
2( )
2( )
q
1/ q
dt
2
2
a b
2( )
2( )
q
Remark 4: For
and concave function, the
inequality (2.8) reduces to inequality (1.5).
Lemma 2: Let assumptions of Lemma 1 be
satisfied, then
1
b a
b
a
b a
( )
0
a
f ( x)
dx f
b
t t
(
t )
f a bdt
0
2
'
t t
( t)
f ' a b
dt
(2.9)
Proof. The proof is consequence of integration by
parts.
In the following theorems, we shall propose
some new upper bounds for the left-hand side of
Hadamard's inequality for h-convex mappings
Theorem 11: Let f ' be h convex function and
the assumptions of Lemma 1 hold, then
1
b a
b
a
a
f ( x)
dx f
b
1
( b a)
f '( a)
t h(
t)
dt (1
t)
h(
t)
dt
0
1
f '(
b)
t h(
t)
dt (1
t)
h(
t)
dt
(2.10)
0
Proof: The proof is similar as proof of Theorem
7.

306 Muhammad Iqbal et al
Remark 5: For and convex function, the
inequality (2.10) reduces to inequality (1.3).
q
Theorem 12: Let f ' be h convex function and
the assumptions of Lemma 1 hold, then
1
b a
b
a
a
f ( x)
dx f
b
f '( a) h(1 t)
dt
1
1
q
b a p
1/ p
q
( p 1)
0 f '( b) h( t)
1
1
q
1
p
f '( a) h(1 t)
dt
q
f '( b) h( t)
1
q
1
q
(2.11)
Proof: The proof is similar as proof of Theorem
8.
Remark 6: For
and convex function, the
inequality (2.11) reduces to inequality (1.4).
q
Theorem 13: Let f ' be h convex function and
the assumptions of Lemma 1 hold, then
1
b a
b
a
a
f ( x)
dx f
b
1
q
2 p
b a
f '( a) h(1 t)
t
dt
1/ p
q
( ) 2
0
f '( b) h( t)
1
q
2 1
p
f '( a) h(1 t)
(1 t)
dt
q
2
f '( b) h( t)
1
q
1
q
(2.12)
Proof: The proof is similar as proof of Theorem
9.
Corollary 2: For , and convex function
inequality (2.12) reduces as:
1 b
a b b
f ( x)
dx f
a
b a
a 2 8
2 f '( a)
q
3
f '( b)
q
1
q
f '( a)
q
2 f '( b)
Theorem 14: Let be h-concave function and
the assumptions of Lemma 1 hold, then
1
ba
b
a
a
b
f( x)
dx f
b
a 1
1/ p
(1 p) 2 h(1 / 2)
2
(
)
2
1
f '
2
a
1
q
2
3
q
1
q
a b
2( )
2( )
2
2
2
1
f'
b a b (2.13)
2
( )
2 2( )
2( )
Proof: The proof is similar as proof of Theorem
10.
Remark 7: For and concave function, the
inequality (2.13) reduces to inequality (1.6).
1
Remark 8: For h( t)
, 1
t
and t s at mid-point,
the inequalities (2.4), (2.6), (2.7) and (2.8) provide
new estimates of right-hand of Hermite-Hadamard
inequality (1.1) for functions of Q (I)
, P (I)
and
2
K s and the inequalities (2.10), (2.11), (2.12) and
(2.13) provide new estimates of left-hand Hermite-
Hadamard inequality (1.1) for functions of Q (I)
,
P (I) and
2
K s respectively.
3. APPLICATIONS TO SPECIAL MEANS
Now, using the results of Section 2, we give some
applications to special means of real numbers
We shall consider the means for arbitrary two real
numbers.
1. Arithmetic mean
a b
Aa,
b ; a,
bR
2
2. Geometric mean
G
a
b
ab
, ; a, bR
3. Logarithmic mean
2

Hadamard-type Inequalities through h-Convexity 307
b a
La,
b ; a, bR
ln b lna
Therefore, by applying the convex mapping
x
f ( x)
e , the following inequalities hold:
Proposition 1: Let a, b I , with a b , we have
b
a
Aab , Lab
,
1/ p
2( p 1)
3a b a 3b
A
,
4 4
L a,
b
q q
1/ q
q q
1/ q
Proof: The proof follows from Corollary 1.
Proposition 2: Let a, bI
, with a b , we have
b
a
La, bGa,
b
4
2a b a 2b
A
,
3 3
L a,
b
q q
1/ q
q q
1/ q
Proof: The proof follows from Corollary 2.
4. ACKNOWLEDGEMENTS
The authors would like to thank the anonymous referees
and Editor-in-Chief for their valuable comments and
suggestions which helped to improve the article. This
study was supported by the Higher Education
Commission of Pakistan and Abdus Salam School of
Mathematical Sciences, Lahore, Pakistan.
5. REFERENCES
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quasi-convex functions with applications to
trapezoidal formula and special means. Computers
and Mathematics with Applications 59: 225-232
(2010).
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inequalities of Hermite-Hadamard Type for s-
convex functions. Acta Mathematica Scientia
31B4: 1643-1652 (2011).
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on Hermite-Hadamard Inequalities and
Applications. RGMIA Monographs, Victoria
University, Melbourne, Australia (2000). Online:
http://www.sta.vu.edu.au/RGMIA/monographs/her
mite_hadamard.html.
4. Dragomir, S. S. & R. P. Agarwal. Two inequalities
for differentiable mappings and applications to
special means of real numbers and trapezoidal
formula. Applied Mathematics Letters 11:91-96
(1998).
5. Hussain, S., M. I. Bhatti & M. Iqbal. Hadamard-
Type inequalities for s-convex functions-I. Punjab
University Journal of Mathematics 41: 51-60
(2009).
6. Kirmaci, U.S. Inequalities for differentiable
mappings and applications to special means of real
numbers and to mid-point formula. Applied
Mathematics and Computation 147: 137-146
(2004).
7. Kirmaci, U.S., M. K. Bakula & M. E. Özdemir &
J. Peari. Hadamard-type inequalities for s-
convex functions. Applied Mathematics and
Computation 193: 26–35 (2007).
8. Niculescue, C. & L. E. Persson. Convex Functions
and Their Application. Canadian Mathematic
Society (2004).
9. Peari, J., F. Proschan & Y. L. Tong. Convex
Functions, Partial Orderings and Statistical
Applications, 187 of Mathematics in Science and
Engineering. Academic Press, Boston,
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10. Pearce, C. E. M. & J. Peari. Inequalities for
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Mathematics Letters 13(2): 51-55 (2000).
11. Sarikaya M. Z., E. Set & M. E. Özdemir. On some
new inequalities of Hadamard type involving h-
convex functions. Acta Math. Univ. Comenianae
LXXIX 2: 265-272 (2010).
12. Sarikaya, M. Z., A. Saglam & H. Yildirim. On
some Hadamard-type inequalities for h-convex
functions. J. Math. Ineq. 2(3): 335-341 (2008).
13. Varošanec, S. On h-convexity. Journal of
Mathematical Analysis and Applications 326: 303-
311 (2007).

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Berlin, Germany, p. 365–410 (2000).
7. Smolen, J.E. & L.A. Boxer. Functions of Europhiles. In: Hematology, 4 th ed. Williams, W.J., E. Butler, &
M.A. Litchman (Eds.), McGraw Hill, New York, USA, p. 103–101 (1991).
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Volume 49, No. 4, December 2012
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Research Articles
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