Protection of goats against capripox using a subunit vaccine.
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Biomedical subjects
Publications and source records attributed to P Chand.
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The synthesis and evaluation as inhibitors of purine nucleoside phosphorylase of six 9-deazahypoxanthines (2a-f) are reported. In contrast to reports in the literature of other hypoxanthine-guanine analog pairs, these inhibitors (2a-f) are equipotent with the corresponding 9-deazaguanines.
The gene coding for the capripoxvirus structural protein P32 was cloned, expressed in Escherichia coli as a fusion protein with glutathione-S-transferase, and purified on glutathione Sepharose. An indirect enzyme linked immunosorbent assay (ELISA) using this antigen was developed to screen bovine sera for antibodies to capripoxvirus. Sequential serum samples from experimentally infected animals tested by ELISA and by virus neutralisation test (VNT) showed that the ELISA was more sensitive and detected antibodies to capripoxvirus earlier post-infection than the VNT.
This paper reports the development and evaluation of serological tests for the differentiation of antibodies in animals infected with capripox and parapox viruses. Agar-gel immunodiffusion tests using sera from sheep with naturally-acquired infections and from sheep experimentally inoculated with orf or capripox viruses showed cross reactions. Virus-specific antibody responses to structural proteins of the viruses were analysed by Western-blot analysis. This analysis readily differentiated the infections as either capripox or contagious pustular dermatitis. The antibody responses to the 32 kDa and 26 kDa proteins of capripoxvirus provided a firm basis for differentiation.
We estimated individual and population Michaelis-Menten pharmacokinetic parameters for phenytoin (DPH) in epileptic patients attending our neurology clinic using the computer programme. OPT. Our results agreed well with literature values but were lower than those we obtained earlier in a smaller number of patients. The Km was independent of age, weight and sex but there was a weak, correlation between Vm and body weight. We conclude that the use of population Vm and Km in normograms could lead to errors in DPH dose estimations as they correlated very poorly with patient characteristics. OPT was easy to use and sufficiently accurate for deriving dose estimates in routine patients. Its use would enable practitioners to generate their patients' own parameters for use in individual dosage adjustments. The estimates can subsequently be updated as more data become available.
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Four reagent formulations (three provided by a manufacturer; one prepared in-house by mixing equal volumes of two commercial reagents) are used for the assay of phencyclidine (PCP) in urine samples. Performance characteristics evaluated included assay precision and sensitivity at and near the assay cutoff concentration. Data resulting from the reagent prepared in-house are better than those using then commercially available formulations, and are comparable with those obtained using the recently available new commercial formulation.
Enzymatic digestion with beta-glucuronidase (EC 3.2.1.31) was used to release intact oxazepam from urine samples containing the d5-analog internal standard. The resulting specimens were extracted with Du Pont PREP Type W cartridge (processed by a PREP Automated Sample Processor), Bond Elut Certify, and J.T. Baker "spe" columns for comparison of the columns' extraction recovery and overall effectiveness. Methyl iodide/tetrahexylammonium hydrogen sulfate and N,O-bis(trimethylsilyl)trifluoroacetamide/trimethylchlorosilane (10 g/L) were used for the methylation and trimethylsilylation studies. We used a Hewlett-Packard HP 5790 mass-selective detector equipped with a 13-m J & W DB-5 column (5% phenyl polysiloxane phase) for gas chromatography/mass spectroscopy (GC/MS) analysis and the Thru-Put Target software package for data processing. After several exploratory experiments, we adopted the Du Pont PREP system methylation procedure because of its effective recovery, the superior stability of the derivatization product, the possibility of incorporating a clean-up step, and the potential for high throughput. The extraction recovery from a set of control samples was 87%. Coefficients of variation obtained for six replicates of GC/MS analysis and for the overall procedure were 1% and 3%, respectively. Excellent linearity was established in the 50-8000 micrograms/L concentration range studied. With the use of 3-mL samples, a 20-microL final reconstitution volume, oxazepam at 50 micrograms/L was easily detected under the adopted operation conditions.
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The effects of water deprivation and intraperitoneal salt loading on urine volume and on various urinary constituents have been examined in two gerbil species of the Rajasthan desert, the Indian desert gerbil (Meriones hurrianae Jerdon) and the Indian gerbil (Tatera indica indica Hardwicke). During summer, hydrated T. indica excreted 0.782 ml urine.100 g-1.d-1 which was about 60.5% higher than the volume of urine excreted by hydrated M. hurrianae (0.487 ml.100 g-1.d-1). During winter, both species excreted around 1.5 ml urine.100 g-1.d-1. The experimental treatments caused reductions in urine volume in M. hurrianae from 40 to 76% during summer and from 35 to 71% in winter. Similar treatments in T. indica caused reductions in urine volume of 50-82% in summer and 5-60% in winter. The mean increase in urine osmolarity following various salt loading treatments in T. indica ranged from 3800 to 5761 mosmol.l-1 and from 4034 to 6255 mosmol.l-1 during summer and winter, respectively. The mean values of urine osmolarity for hydrated T. indica were 2831 and 3189 mosmol.l-1 during summer and winter, respectively. In M. hurrianae salt loading treatments caused increases of urine osmolarity between 3381 and 5646 mosmol.l-1 and between 4032 and 5434 mosmol.l-1, during summer and winter, respectively, over the values recorded for hydrated animals (summer = 3292; winter = 3294 mosmol.l-1). A maximum urine osmolarity of around 7000 mosmol.l-1 was found in both species when subjected to 2% salt-loading treatment. The treatments used in this study increased urinary urea level in both T. indica (3039-4056 mM) and in M. hurrianae (1900-2180 mM) compared to the level in their respective hydrated controls (T. indica = 1628 mM; M. hurrianae = 1372 mM). The results indicate that T. indica may be better adapted to produce more concentrated urine than M. hurrianae.
A total of 384 sheep serum samples collected from two organised sheep farms was tested by dot immunobinding assay (DIA) and indirect enzyme-linked immunosorbent assay (I-ELISA) for the presence of bluetongue virus (BTV) antibodies. The results of both these assays were compared to find a sensitive, specific, rapid, easily performed and economical test for the diagnosis of bluetongue disease. DIA detected BTV antibodies in 210 samples (54.94%) and I-ELISA detected 157 positive samples (40.88%). Competitive ELISA (C-ELISA) was performed to check the discrepancies in I-ELISA and DIA. On the basis of these tests the overall agreement, relative specificity and sensitivity between ELISA and DIA were 75%, 87.6% and 100%, respectively. DIA was found to be a rapid, sensitive, easily performed and economical test as compared to ELISA.
A dot-immunobinding assay (DIA), using as antigen a sonic extract of Brucella abortus dotted on nitrocellulose bound to a plastic strip, was employed for the detection of Brucella antibodies in 666 sheep sera. The results were compared with the complement fixation test (CFT). All the 242 sera belonging to two flocks were found to be negative by DIA. CFT was negative in 239 cases, whereas three samples showed anti-complementary activity. Of the 424 sera from the remaining three flocks, 98 were positive by both tests and six were positive in DIA, but negative in CFT. In addition, 14 of the 19 anti-complementary sera were also positive by DIA.
A dot Enzyme-linked Immunosorbent Assay (dot-ELISA), using whole cell Brucella abortus antigen dotted on the nitrocellulose membrane bound to a plastic strip (dipstick) was employed for the detection of Brucella antibodies in bovine sera. The results were compared with that of serum agglutination (SAT), Rose Bengal plate agglutination (RBPT) and Complement Fixation test (CFT). All the four tests gave negative reaction in 127 sera obtained from a brucellosis free herd. Testing of 549 sera from a chronically infected herd revealed 57 positive and 447 negative animals in all the four assays. Of the remaining 45 sera, 34 were positive in dot-ELISA. Six of these cases were independently detected by dot-ELISA while 28 showed positive reactions in combination with other tests. When serum samples from 158 aborted cases were subjected to dot-ELISA, 79 were found positive. Of these dot-ELISA positive cases, 71 gave positive reaction in SAT, 72 in RBPT and 78 in CFT. B. abortus biotype 3 was isolated from 34 of the 98 aborted fetuses examined.
A dot-enzyme linked immunosorbent assay (dot-ELISA) for the detection of antibodies in bovine brucellosis using soluble antigens extracted from Brucella abortus S-99 is described in which antigen was deposited on a nitrocellulose sheet (0.5 x 0.5 cm) bound to a plastic strip and the unsaturated sites blocked by a solution of spray-dried milk powder. Sera were tested at dilutions of 1:800 and 1:1600, using rabbit anti-bovine-immunoglobulin or protein-A coupled to peroxidase as conjugates and diamino-benzidine as substrate. A positive reaction was clearly indicated by a brown dot on the nitrocellulose sheet. Using antigen-coated, pre-blocked strips, the test could be completed within 45 minutes.
A prospective study on measles was undertaken during 1974 to 1986 in Hiramanpur village, Varanasi having an initial population of 1538 which rose to 2101 in 1986 enabling to register a total of 23,221 (all population) person years of observation and a total of 9191 (0-14 years) person years of observation. The entire population was covered regularly through domicilliary visits by trained para-medical personnel under direct supervision of medical officers. All cases were verified by doctors. A total of 411 measles cases were detected, which gave average annual incidence rates of 17.7 and 44.7 per thousand population and per thousand children (0-14 yrs), respectively. The maximum number of cases occurred in Ist quarter of the year (69.7 per cent). The Kolmogorov-Smirnov Statistical method validated the seasonal character of the disease (Vn = 5.18, p less than 0.01). The maximum measles cases occurred in children 1-4 yrs (57.4 per cent). Males (54.9 per cent) were affected slightly more than females (45.1 per cent). A total of 246 (59.9 per cent) cases developed complications. The most frequent was respiratory infections (42.2 per cent) followed by infection of gastrointestinal system (32.2 per cent). The complication rates were found higher in infants and older children (greater than 5 yrs.) in both the sexes. Three-fourth of the cases did not get any treatment. The overall case-fatality rate was 2.2 per cent. Females had higher rate (3.3 per cent) than males (1.3 per cent) and the highest case fatality rate was observed in infants, particularly in females.
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The relative body water conservation efficiency of two Indian desert gerbil species, Meriones hurrianae (diurnal/crepuscular) and Tatera indica (nocturnal), has been examined under near-natural conditions in different seasons. A mean urine osmolarity of 3180 mosmol/l (maximum 4645 mosmol/l) in M. hurrianae and a mean value of 5128 mosmol/l (maximum 7547 mosmol/l) in T. indica have been recorded during summer. Urine osmolarity and urea levels indicated that while M. hurrianae remain sufficiently hydrated mainly by virtue of their feeding habit, Tatera indica may depend on the relatively higher concentrating capacity of their kidneys.