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

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CHAPTER 4.3 CHANGES IN THE TESTES OF M. NATALENSIS INFECTED WITH 43.1 MATERIALS AND METHODS 4.3.1.1 Testosterone B. MALAY1 Testosterone Radioimmunoassay was done using Testosterone Double Antibody kit obtained from Diagnostics Products Corporation, Los Angeles, USA. The basic principle of FUA involves competition between a radioactive and non- radioactive antigen for a fixed number of antibody binding sites in a fixed time. The amount of labeled analyte bound to the antibody (goat anti-rabbit gamma globulin) is inversely proportional to the concentration of the analyte present in the sample. The antiserum is very specific for testosterone with an extremely low cross-reactivity to other steroids. Martomys rats were infected wth B. rnalayi as desnibed earlier. The testes was excised after 0, 30, 60, 90, 120 days post inoculation, washed with phosphate buffered saline. pH 7.4. homogenized and centrifuged at 2000 rpm for 20 minutes. The pellet was discarded and the supernatant was used as sample for RIA. Reagents 1. Testosterone antiserum 2. ('"1) Testosterone 3. Calibrators (conc. 5, 10,20,50, 100 & 200 pgtube) 4. Goat anti-rabbit gamma globulin 5. Polyethylene glycol 6. Borate buffer for testosterone
Tubes were labeled for calibrators, blank, maximum binding tube, total counts and samples (tissue homogenate) in duplicate. Two hundred pl of the buffer was taken into the blank tube and 100 pl ihto the maximum binding tube. Similarly 100 pi of the calibrators and the sample was taken in the calibrator tubes and sample tubes. Then 100 p1 of the [I2' I] testosterone was added into all the tubes, followed by 100 p1 of testosterone antiserum to all the tubes except the blank tubes. The tubes were vortexed and incubated for a minimum of 60 minutes at R.T., which was followed by the addition of 100 pi of goat anti-rabbit gamma globulin to all the tubes. The contents of the tubes were vortexed and incubated for I hr at R.T. then 2.0 rnl of cold 4% PEG-saline solution was added and centrifuged at 2,000g for 20 minutes. The supernatant was decanted, the rim of each tube was blotted dry and counted in a Gamma Counter for 1 minute. Calibration curve was prepared as indicated by the kit procedure. The testosterone levels were expressed in nanornoles/mg protein. 43.1.2 7-GTP 1-GTP was assayed according to the method of Orlowski and Meister (1965) and modified by Rosalki and Rau (1972) as described earlier. 43.13 LDH-X The testes of M.naralemis infected with B.rnalayi were excised on different days of post inoculation (0,30, 60, 90. 120). washed in chilled potassium phosphate buffer (0.1 M, pH 8.3) and homogenised in the same buffer. The homogenate was centrifuged at 10,000g for 30 minutes at 4°C and the supernatant was used for LDH-X isoenzyrne assay. LDH-X isorymes were visualized on agar gel by following the method of Hanis and Hopkinson ( 1978). Reagents for staining 1. 0.05 M Tris-HCl buffer. pH 8.0 : 20 ml 2. Calcium l,actate (pentahydrate) mixture). 3. NAD : 100 mg (8 mM final concentration in reaction
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EFFECT OF INFECTION OF THE FILARIAL
Page 4 and 5:
I wish to express my deep sense of
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1. INTRODUCTlON 2. REVIEW OF LlTERA
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As stated earlier, the host immune
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13 Testicular changes Testosterone
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2. REVIEW OF LITERATURE Filariasis
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2.2 Clinical signs Inflammalation o
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the tegument by the release of thei
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the compound kills the filarial wor
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3. MATERIALS AND METHODS COLONY MAI
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Ragenb 1. Tris-HCI buffer . : 0.1 M
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3. Reduced glutathione : 30 mM To I
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4.1.1.6 Glucose-6-phosphate dehydro
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absorbance of the clear supernatant
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-eats I. Lowry's reagent :Solution
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4.1.2.1 Changes in antioxidant enzy
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Fig.3: Effect of B.malayi on SOD in
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Fig.9: Effect of B.rnalayi on GST i
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Fig.12: Effect of 6.rnalayi on GPx
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Fig. 18: Effect of B.malayi on G6PD
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12- 3 m- 15- lor, Fig.21: Effect of
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Fig.27: Effect of B.malayi on Ascor
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4.1.2.3 Change in the antioxidant e
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Fig.35: Effect of B.malayi on CAT i
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m 20 0) - 15 9 g 10 il! 5 0 Fig.39:
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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: with i n d lymphocytes and decrease
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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