effect of infection of the filarial parasite brugia malayi - Pondicherry ...

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Reagent6 1. DTNB : 0.6 mM in 0.2 M phosphate buffer, pH 8.0. 2. Phosphate buffer : 0.2 M, pH 8.0. 3. TCA : 5% 4. Standard : 10 mg of reduced glutathione in I00 ml water. This contained 10pgm of glutathione in 0. I ml solution. To 0.1 mi of the homogenate, 1.0 ml of water and 2.0 ml of precipitating agent were added, mixed thoroughly and centrifuged after 5 minutes. To an aliquot of the supernatant, 2.0 ml of DTNB reagent was added and made upto a final volume of 3.0 ml with phosphate. buffer. The absorbance was read at 412 nm against a reagent blank The amount of glutathione was exprrssed as pgm of glutathionel mg protein. 4.1.1.8 Total thiol groups (1 %8). Total thlol groups were estimated acconhg to the method of Sedlack and Lindsay The sulphydryl pups in tissues were determined by using the Ellman's reagent. In this method, DTNB is reduced by -SH groups to form 1 mole of 2-nitro, 5-mercaptobenzoic acid per mole of -SH. Reagents 1.0.01 MDTNEI : 99 mg DTNEI dissolved in 25 ml of absolute methanol. 2. Tris-HCl buffer : 0.2 M, pH 8.2 containing 0.02 M EDTA. The sampk was pnpartd by homogenising 100 mg of tissue in 4.0 ml of 0.02 M EDTA. To an aliquot of the tissue homogenate, 1.5 ml of Tris-HCI buffer @H 8.2) and 0.1 ml of DTNB was added, mixed and made upto 10.0 ml with absolute methanol. A reagent blank without the sample and sample blank without DTNEI were prepared in the same manna. The test tubes were stoppered and allowed to stand with occasional hkmg for 15 minutes. The reaction mixture was cenhifuged at 3000g at R.* for 15 minutes. The
absorbance of the clear supernatant was read at 412 nm. Calibration curves were obtained with reduced glutathione as standard. Values were expressed as pgm of glutathiondmg protein. 4.1.1.9 Ascorbic acid Ascorbic acid was estimated by the method of Omaye ef al. (1979). Ascorbic acid is oxidzed by copper to form &hydroascorbic acid and diketogluconic acid. When treated with 2.4, dmirophenyl hydrazie, it reacts to form the derivative bis 2,4, dinitrophenyl hydrame. This compound in strong sulphuric acid undergoes a rearrangement to form a product with an absorption band that is measured at 520 nm. The reaction is run in the presence of thlourea to provide a mildly reducing medium, which helps to prevent the ~nterference from non-ascorbic chromogens. Reagents 1. TCA : 5% 2. DTC reagent : 3 gms DNPH (ditropbenyl hydrazine), 0.4 gms thiourea and 0.05 gms of copper sulphate were dissolved in 10 ml of 9N sulphuric acid and made upto 100 ml with the same solution. An aliquot of the homogate was precipitated with ice-cold TCA, cenhihged for 20 minutes at 3,500g and 1.0 ml of the supernatant was made upto 3.0 ml with 5% TCA and then, treated with 0.2 ml of DTC and incubated for 3 hrs at 37'C. Then 1.5 ml of icecold 65% sulphuric acid was added, mixed and kept at RT. for an additional 30 minutes. Blank contained only 3.0 ml of TCA. Standards in the range of 10-50pgms were treated in the same way. The intensity of the color was measured at 520 nm. Results were expressed as pgm of ascorbic acid! mg protein.
Page 1 and 2: EFFECT OF INFECTION OF THE FILARIAL
Page 4 and 5: I wish to express my deep sense of
Page 6: 1. INTRODUCTlON 2. REVIEW OF LlTERA
Page 9 and 10: As stated earlier, the host immune
Page 11 and 12: 13 Testicular changes Testosterone
Page 14 and 15: 2. REVIEW OF LITERATURE Filariasis
Page 16 and 17: 2.2 Clinical signs Inflammalation o
Page 18 and 19: the tegument by the release of thei
Page 20 and 21: the compound kills the filarial wor
Page 22: 3. MATERIALS AND METHODS COLONY MAI
Page 25 and 26: Ragenb 1. Tris-HCI buffer . : 0.1 M
Page 27 and 28: 3. Reduced glutathione : 30 mM To I
Page 29: 4.1.1.6 Glucose-6-phosphate dehydro
Page 33 and 34: -eats I. Lowry's reagent :Solution
Page 35 and 36: 4.1.2.1 Changes in antioxidant enzy
Page 37 and 38: Fig.3: Effect of B.malayi on SOD in
Page 39 and 40: Fig.9: Effect of B.rnalayi on GST i
Page 41 and 42: Fig.12: Effect of 6.rnalayi on GPx
Page 43 and 44: Fig. 18: Effect of B.malayi on G6PD
Page 45 and 46: 12- 3 m- 15- lor, Fig.21: Effect of
Page 47 and 48: Fig.27: Effect of B.malayi on Ascor
Page 49 and 50: 4.1.2.3 Change in the antioxidant e
Page 51 and 52: Fig.35: Effect of B.malayi on CAT i
Page 53 and 54: m 20 0) - 15 9 g 10 il! 5 0 Fig.39:
Page 55 and 56: 30 25 8 - a 20 2 15 C I 10 I 5 0 35
Page 57 and 58: Fig.45: Effect of B.maiayi on GSH i
Page 59 and 60: Fig.49: Effect of B.malayi on Vit-C
Page 61 and 62: 4.1.3 DISCUSSION 4.1 3.1.1 Toxic ox
Page 63 and 64: 1973). The two major systems of LPO
Page 65 and 66: 1981), and their increase in human
Page 67 and 68: 1 0 Fig.58:Relationship between cha
Page 69 and 70: 1 - 0- ' 1 5 - 6 - 7 - Fig.63:Relat
Page 71 and 72: Catalase had been reported to be re
Page 73 and 74: - Fig.68:Relationship between chang
Page 75 and 76: esults in its reduced activity (Con
Page 77 and 78: g 20- :I-. F1g.70:Relationship betw
Page 79 and 80: 12, - I 0 U) Q 4 2 0 - + - Fig.75:R
Page 81 and 82:
$ Fig.80:Relationship between chang
Page 83 and 84:
Fig.85:Relationship between change
Page 85 and 86:
V) (3 :; - Fig.9O:Relationship betw
Page 87 and 88:
A reduction in the total thiol cont
Page 89 and 90:
2 5- Fig.98:Relationship between ch
Page 91 and 92:
Fig. 103:Relationship between chang
Page 93 and 94:
4.2.1.1 N ay ATPase Reagents Nay AT
Page 95 and 96:
Reagents 1. Acetylcholine bromide :
Page 97 and 98:
To 1.5 ml of carbonate buffer, 1.5
Page 99 and 100:
4.2.1.10 Differential leucocyte cou
Page 101 and 102:
increase in the control animals (Fi
Page 103 and 104:
- Fig.110: Effect of 6. malayi on H
Page 105 and 106:
efflux and thereby alter the membra
Page 107 and 108:
with i n d lymphocytes and decrease
Page 109 and 110:
Tubes were labeled for calibrators,
Page 111 and 112:
Fig.114: Effect of B, malayi on GTP
Page 113 and 114:
433 DISCUSSION Testosterone is a se
Page 115 and 116:
CHAPTER 4.4 HISTOPATHOLOGICAL CHANG
Page 117 and 118:
Platc 2 Plate 3 Plate 4 Plate 5 Pla
Page 119 and 120:
Platc X Plate 9 Plate 10 Plate 11 P
Page 121 and 122:
Plate w: Section of Testes of norma
Page 123 and 124:
4.4.2.1.5 Brain The control animals
Page 126 and 127:
Plate ICt Section of Kidney of infe
Page 128 and 129:
4.4.2.2 Histopathological changes i
Page 130 and 131:
Plate 32
Page 132:
The mf could not be seen in spleen
Page 135 and 136:
ain, it decreased from 0 day itself
Page 137:
inflammatory nature indicative of c
Page 141 and 142:
BIBLIOGRAPHY Ackerman, S.J., Gleich
Page 143 and 144:
Case, T., Leis, B., Witte, M., Way,
Page 145 and 146:
Drabkin, D.L.R. and Austin, J.H. (1
Page 147 and 148:
Fukuota, M., Zhou, Y. and Tanaka.(l
Page 149 and 150:
Jacob, H.S. and Lux, S.E. (1968). D
Page 151 and 152:
Linda, I. and Mayeda, B. (1974). Th
Page 153 and 154:
Murray, H.W. (1981). Susceptibility
Page 155 and 156:
Santiago- Stevenson, D., Oliver-Gon
Page 157 and 158:
Tate, S.S., Thomson, G.A. and Meist
Page 159 and 160:
Zaini, M., Ramachandran, C.P. and E
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effect of infection of the filarial parasite brugia malayi - Pondicherry ...

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