2009 Vienna - European Society of Human Genetics

Therapy for genetic disorders 0
baseline data of endothelial function, assessed by measurement of
changes induced by reactive hyperemia in pulsatile volume at fingertips,
through a non invasive pletismographic method (EndoPAT Itamar
Israel) and redox state as determined by blood thiol concentrations.
Preliminary results show that CADASIL patients have a lower endothelium-dependent
vasodilatation (PAT index) than healthy controls
with/without conventional cardiovascular risk factors (RF). Patient
categorization according to their RF profile shows an interaction between
CADASIL and the coexisting RFs: PAT index is lower in patients
with ≥1 RF [n=19, 1.68 (1.44-1.92)] vs those with no RF [n=28, 2.01
(1.77-2.60)] (P≤0.03). Thiol profile showed significantly increased cysteinylglycine
(37±7 vs 32±9 μmol/l, P=0.02) and decreased glutathione
(5.5±2 vs 7.7±3 μmol/l, P=0.02) concentrations in CADASIL patients
vs healthy controls.
These findings underscore the importance of specific assessment of
ED and redox state and accurate management of RF in CADASIL patients.
P14.04
The benefit of an intensive rehabilitation program in patients
with cerebral palsy
E. Sirbu1 , D. Stoicanescu2 , R. Mihaescu2 , V. Belengeanu2 ;
1 2 West University of Timişoara, Timisoara, Romania, University of Medicine &
Pharmacy, Timisoara, Romania.
Background: Cerebral palsy is defined as a group of permanent disorders
of the development of movement and posture, causing activity
limitation, attributed to non-progressive disturbances that occurred in
the developing brain. Cerebral palsy may be due to many causal factors
that act during prenatal, perinatal or postnatal life, most cases
seeming to be of early prenatal origin.
Objective: The aim of the present study was to demonstrate that application
of an intensive program of physical therapy considerably improves
the global motor function.
Material and methods: We studied a lot of 20 children with cerebral
palsy (12 with pyramidal tetraparesis and 8 with pyramidal hemiparesis),
mean of age 4.4 years, beneficiaries of a physical kinetic program,
specific for each type. The lot was divided in two groups: control group
(10 subjects) and experiment group (10 subjects). Subjects included in
control group followed a rehabilitation program for 6 months, 3 times
a week, whereas subjects included in experiment group followed an
intensive rehabilitation program - 2 times a day, for 6 months. The
global gross motor function was evaluated for each child before and
after therapeutic intervention (GMFM 88).
Results: The gross motor function of children from both groups improved
significantly after this intervention. Children from the experiment
group performed better and showed significantly greater improvement
than those from the control group.
Conclusions: Application of an intensive rehabilitation program determines
increasing of motor independence in children with infantile
cerebral palsy and allows creating of a favorable climate for their integration
in society.
P14.05
Enzyme-Free and High-throughput cloning means for production
of recombinant protein and gene therapy
H. Sadeghi 1 ;
1 jacobs-university, Bremen, Germany, 2 *, Tehran, Islamic Republic of Iran.
DNA cloning is one of the most utilized techniques in molecular biology
research. Using the traditional DNA cloning methods, which include
usage of restriction endonucleases and DNA ligase, have many limitations
including unavailability of unique restriction sites in both insert
and vector. LIC (Ligase-Independent Cloning) methods have been
developed to overcome such problems, they use different enzymes
to generate long enough “sticky ends” that can hold insert and vector
together. Reparative mechanism of cell can join the nicks in both
strands and final construct can be formed.
Here we describe a simple method for the cloning of PCR products
without the need for restriction enzyme and any post enzymatic treatment.
PCR products containing phosphorothiate bonds are used to
create complementary single stranded overhangs on both insert and
vector by a post-PCR iodine treatment. These single stranded overhangs
are designed to allow directional cloning without using any enzyme
(Enzyme-Free cloning). After hybridization and transformation
step construct can be isolated from cells. Our procedure is faster than
the existing cloning strategies and also is highly efficient, directional
and reliable. Besides that since two different cohesive ends are used,
the orientation of the insert in relation to the vector can be controlled.
Flexibility in choice of vector and insert makes this method exceptionally
suitable for high-throughput cloning. This characterization is particularly
useful for ensuring that a DNA fragment is directionally cloned
into the correct reading frame for protein expression or for the creation
of fusion proteins.
P14.06
Improvement of life quality in patients with cystic fibrosis
through kinetotherapy and physical exercises
B. Almajan Guta 1,2 , V. Almajan-Guta 3 , E. Sirbu 4,5 ;
1 University “Politehnica” Timisoara, Timisoara, Romania, 2 National Cystic Fibrosis
Centre, Timisoara, Romania, 3 “Speranta” Special Care Centre, Timisoara,
Romania, 4 West University of Timisoara, Physical Education and Sport Faculty,
Timisoara, Romania, 5 Clinic Municipal Hospital, Timisoara, Romania.
Background: The physical treatment is one of the most important aspects
of the management of Cystic Fibrosis. The problems of the lungs
remain the major difficulty which has to be dealt with on a day-to-day
basis.
The aim of study was to prove the efficiency of several physiotherapy
techniques and to adapt them according to the age and adherence of
patients.
Methods: 71 patients aged between 2 month and 18 years were followed
up over the last 4 years, 2004-2008 in the National Cystic Fibrosis
Centre and divided in 3 groups according to age and compliance:
group I: 15 infants age between 2 months-3 years old, group II: 14
children between 3-6 years old, group III: 42 children between 6-18
years old. We have assessed: clinical status, nutritional condition, Xray,
bacteriological exam, MEF 25-75% , FEV 1, PEF.
Results: The result at the first group has indicated an improvement in
all parameters. At the second group the maximum compliance was in
physical exercises 86%, and the maximum efficiency 72% was in the
combined techniques. The statistical briefing of data in the third group
shows the fact that there are significant statistical difference (p< 0,05),
before and after treatment in all ventilator index.
Conclusions: The chosen technique proved to be very efficient, in improving
of respiratory symptoms and ventilator parameters. It is extremely
important to make sure that the treatment advised for each
individual patient is that which is most suitable to them for their age,
cultural background and disease severity.
P14.07
Effects of early rehabilitation in a plurimalformative syndrome
due to deletions of chromosomes 13 and 18-case report
C. Avram1 , D. Stoicanescu2 , M. Cevei1 ;
1 2 Faculty of Medicine & Pharmacy, Oradea, Romania, University of Medicine &
Pharmacy, Timisoara, Romania.
Deletion of the long arm of chromosome 18 is a chromosomal disorder
with a phenotype that may vary considerably in range and severity,
depending on the type of deletion and location of the breakpoints.
Children have characteristic features including short stature; mental
retardation; hypotonia, malformations of the hands and feet; craniofacial
abnormalities and numerous neurologic deficiencies with a high
incidence of dysmyelination. Deletion of long arm of chromosome 13
is characterized by malformations of the craniofacial region, skeletal
abnormalities, other physical abnormalities and intellectual disability.
In this paper we report the case of a female infant with multiple congenital
abnormalities, craniofacial dysmorphism, heteroataxia, severe
mental retardation and severe hypotonia, who was found to have deletions
of the long arm of chromosomes 13 and 18. We included her in
a rehabilitation program from the age of eleven months. Rehabilitation
programs aimed improving hypotonia as well as stimulating the development
of motor skills. We observed the child for a period of one
year, periodic monitoring of muscle tone and performance along with
the neurological status showing significant motor and mental improvement.
Conclusions: Rehabilitation treatment is effective and must be
an early intervention.