Cloning and characterization of HindIII fragments of bovine herpesvirus-1 DNA from an Indian isolate.
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Biomedical subjects
Publications and source records attributed to A Rai.
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A 74-year-old male claudicant had undergone operative repair of his bilateral varicose veins 22 years earlier. His records confirmed that on the right side he had only had a saphenofemoral disconnection. Duplex scanning and angiography of his right leg demonstrated an occluded superficial femoral artery (SFA) and a network of channels between the proximal SFA stump and the long saphenous vein (LSV). Distal SFA reconstitution occurred through two distal arteriovenous fistulas. We believe that this is the first case of a spontaneous femorodistal bypass graft secondary to neovascularisation recorded in the literature.
Isatin (2,3-dioxoindole), one of the components of tribulin, which has been postulated to function as an endogenous marker of stress and anxiety, was shown to induce a dose-related attenuation of learning acquisition in an active avoidance test and inhibition of learning retention, or memory, in a step-down passive avoidance paradigm and transfer latency in an elevated plus-maze, in rats. Earlier studies have indicated that isatin functions as a 5-hydroxytryptamine (5-HT)3 receptor agonist in its anxiogenic activity in rats and is an antagonist at mammalian atrial natriuretic peptide (ANP) receptors. Since 5-HT3 receptor antagonists and centrally administered ANP have been shown to facilitate learning and memory, the observed memory dysfunction induced by isatin can be attributed to its receptor activity at 5-HT3 and ANP receptors. The investigation also indicates that anxiogenic agents are likely to disrupt memory functions.
A number of factors contribute to the pathogenesis of osteoarthritis. Genetic factors are predominant in some cases, including the syndrome of hereditary arthro-ophthalmopathy. The clinical features of the syndrome and the concept of genetic mutations in matrix components are discussed.
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Benign soft tissue swelling affecting the interphalangeal joints can sometimes be misdiagnosed as an arthropathy. Garrods fatty pads (knuckle pad syndrome) is a relatively common and harmless condition characterized by discrete fibromatous skin pads over the dorsum of the interphalangeal joints. We describe a case which clinically resembles the rare but benign condition of pachydermodactyly with some pathological features of the distinct syndrome of pachydermoperiostosis. The possibility of an overlap syndrome is discussed.
Over a 2-month period in an accident and emergency department, the clinical benefits and cost-effectiveness of radiology of the knee after trauma was audited. In all, 178 patients had radiographs as part of their initial assessment, the majority of which were subsequently reported by the radiologist as entirely normal (90.4 per cent of films after a twisting injury and 80.6 per cent after a direct blow). Early and appropriate clinical review by a senior doctor is proposed as a viable alternative to perhaps unneeded radiographs where the physical signs of injury are minimal. The clinical and financial benefits of such a policy are discussed.
The actions of peptides (helospectin I, helodermin, exendin-3, exendin-4) that have been isolated from the venoms of Helodermatidae lizards were examined using dispersed chief cells from guinea pig stomach. These actions were compared with those of mammalian glucagon-like peptides, particularly truncated glucagon-like peptide 1 (TGLP-1), a peptide that shares 53% homology with exendin-4. The Helodermatidae venom peptides and TGLP-1 caused a two- to threefold increase in chief cell adenosine 3',5'-cyclic monophosphate and pepsinogen secretion. Exendin-3 and exendin-4 were 100 times more potent than helospectin I and helodermin and 10 times more potent than TGLP-1. Helospectin I and helodermin, but not exendin-4 or TGLP-1, inhibited the binding of 125I-labeled vasoactive intestinal peptide (VIP) and 125I-secretin to dispersed chief cells. The actions of exendin-3, exendin-4, and TGLP-1, but not those of helospectin I, helodermin, VIP, or secretin, were progressively inhibited by increasing concentrations of an exendin-receptor antagonist, exendin-(9-39)-NH2. These data indicate that in gastric chief cells, whereas the actions of helospectin I and helodermin are mediated by interaction with high-affinity secretin (low-affinity VIP) receptors, the actions of exendin-3, exendin-4, and TGLP-1 are mediated by interaction with exendin receptors.
An Indian isolate of BHV-1 was analyzed using several restriction endonucleases. The restriction pattern of BHV-1 DNA with HindIII, BamHI and EcoRI was found to be similar to other reported restriction endonuclease patterns for respiratory isolates of BHV-1. Thus, this isolate could be grouped under BHV-1.1 (respiratory form of BHV-1). Few restriction endonucleases yielded distinct restriction profile of BHV-1 DNA. The total molecular size of BHV-1 DNA for the isolate studied was also estimated to be within 136 kb to 138 kb.
The DNA from an Indian isolate of BHV-1 was cleaved with the restriction enzyme PstI and cloned into pUC9 plasmid. Two recombinant plasmids were then analyzed using restriction endonucleases PstI, EcoRI, BamHI, SmaI, SalI, HindIII, BglI, HaeIII, XhoI, RsaI and StuI. On the basis of digested fragments of recombinant plasmids, two plasmids could be mapped for restriction enzymes SalI, StuI, SmaI and XhoI using single and double digestion.
Bovine herpesvirus 1 DNA has been isolated by SDS lysis of the virus purified from potassium tartrate (10-50%) density gradient centrifugation. The quality and quantity of viral DNA was checked by UV spectrophotometry and ethidium bromide stained agarose gel electrophoresis. The 0.4 kb Hin dIII'O' fragment of BHV-1 DNA was selectively cloned into Hin dIII cut pUC9 plasmid DNA (2.665 kb). Recombinants were screened by white/blue colonies as well as Hin dIII restriction enzyme analysis. On restriction endonuclease analysis of recombinant plasmid DNA (p-BH-0) with several restriction enzymes, viz., Sau 3A, Hin fI, Rsa I, Sal I, Dra I, Bgl I, Bgl II, Sma I, Hpa I, Stu I, Mlu I, Xho I, Kpn I, Hae III, Eco RI, Bam HI, Pst I, Pal I, revealed insert viral DNA having sites for Hin fI, Hae III, Rsa I, Sma I, only. Further, the partial restriction map of the recombinant plasmid DNA was constructed using above enzymes.
OBJECTIVE: To determine whether published cost-effectiveness and cost-benefit analyses have adhered to basic analytic principles. DESIGN: Structured methodologic review of published articles. STUDY SAMPLE: Seventy-seven articles published either from 1978 to 1980 or from 1985 to 1987 in general medical, general surgical, and medical subspecialty journals. MAIN OUTCOME MEASUREMENTS: Articles were reviewed to assess the use and reporting of six fundamental principles of analysis. These principles were derived by reviewing widely cited textbooks and articles describing the methods for performing economic analyses and by selecting the methods universally recommended. MAIN RESULTS: Overall performance was only fair. Three articles adhered to all six principles, and the median number of principles to which articles adhered was three. Among the problems noted were failure to make underlying assumptions explicit and, therefore, verifiable, and failure to test assumptions with sensitivity analyses. No improvement in performance was observed between 1978 and 1987. Articles in general medical journals, however, were more likely to use analytic methods appropriately than articles in the general surgical or medical subspecialty literature. CONCLUSIONS: Greater attention should be devoted to ensuring the appropriate use of analytic methods for economic analyses, and readers should make note of the methods used when interpreting the results of economic analyses.
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Previous work has shown that renal metabolism of 25-dihydroxyvitamin D3 (25(OH)D3) to the active metabolite, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) is stimulated by prostaglandin E2 and inhibited by acetylsalicylate (aspirin). As prostaglandins are primary inflammatory mediators and synovial fluid macrophages are known to synthesise 1,25(OH)2D3 in vitro, the effects of prostaglandin E1, prostaglandin E2, and aspirin on the metabolism of 25(OH)D3 by cells cultured from synovial fluid of patients with inflammatory arthritis were investigated. Most cultures contained non-proliferating macrophages which formed 1,25(OH)2D3; however, two of 13 cultures contained colonies of rapidly proliferating fibroblast-like cells which formed 24,25(OH)2D3 (24,25(OH)2D3). Prostaglandin E1 and prostaglandin E2 (0.01-10 mumol/l) induced marked inhibition of 1,25(OH)2D3 synthesis (up to 94%) in a dose dependent manner after preincubations of 24 hours but not over straightforward six hour incubations. Exposure of macrophages to aspirin (1 mumol/l-1 mmol/l) for 24 hours did not affect 1,25(OH)2D3 synthesis unless the cells had been pretreated with lipopolysaccharides, in which instance 1 mM aspirin increased 1,25(OH)2D3 synthesis. Lipopolysaccharide is a macrophage activating factor which stimulates macrophages to form 1,25(OH)2D3, and it also induces prostaglandin synthesis which would be inhibited by aspirin. Taken together these results suggest that prostaglandin E1 and prostaglandin E2 synthesised by macrophages may act in an autocrine manner to attenuate the ability of macrophage activating factors, such as lipopolysaccharide, to stimulate 1,25(OH)2D3 synthesis. Prostaglandins synthesised by other inflammatory cells may also inhibit 1,25(OH)2D3 synthesis in a paracrine manner. In contrast, prostaglandin E2 and aspirin had limited effects on fibroblast 24,25(OH)2D3 synthesis. This study shows that the effects of prostaglandin E1, prostaglandin E2, and aspirin in macrophages contrast with those previously reported for the renal 25(OH)D3-1alpha-hydroxylase, where prostaglandin E2 stimulated and aspirin inhibited enzyme activity. These results further emphasise that synthesis of 1,25(OH)2D3 in non-renal sites is independently regulated, which is consistent with it having an immunological role at a local level rather than playing a part in systemic calcium homeostasis.
Choice of an ideal, cost-effective and rapid diagnostic test for HIV infection is of immense value in developing countries like India where resources are limited. A number of commercial HIV antibody testing kits are now available with varying sensitivities and specificities. Six different commercial HIV kits namely, Wellcozyme, Flow HIV-TEKG, Abbott HIV EIA, Abbott VIA, Dip-stick EIA and Abbott env/core recombinant EIA were evaluated. Du-Pont Western blot (W.B.) kit was used as gold standard to compare the results. Of the 376 sera from various high-risk individuals screened, Wellcozyme kit yielded 100 per cent concordant results with W.B. Abbott VIA and Abbott env/core also yielded results in confirmation with W.B., excepting the fact that both detected one extra sample positive, which was negative in W.B. Abbott EIA yielded 4 false positive results. Dip-stick kit yielded the maximum number of false positives. The study indicated that 3 kits, namely Wellcozyme, Abbott VIA and Abbott EIA could be used to achieve optimum and acceptable results.
A cross-sectional sample of 2987 children in the age group of 1 to 10 years were selected from urban slums of Delhi for measurement of weight and height. Males and females were 53 and 47% respectively. Mean weight and height were calculated for both the sexes. Comparison of percentiles of weight and height with NCHS and ICMR standards showed that the 75th percentiles weight and height of the present study were comparable with 80 and 90% of 50th percentile of NCHS respectively. Hence, upper 25% of the sample arranged in ascending order of magnitude were used for the construction of reference standards of weight and height. Comparison of percentiles drawn from the top 25% of the sample demonstrated that 50th percentile of the present study corresponded to 80% of 50th percentile of the NCHS for weight and 90% of 50th percentile of NCHS of the height for both boys and girls. Also the 50th percentile of ICMR for weight and height for both the sexes. Standards constructed from such sample would be better suited for two important uses: (i) monitoring of Nutrition Programme, and (ii) to detect a child with abnormal growth at the earliest.
The incidence of mange in dairy buffalo in India has increased significantly in recent years. The authors record an outbreak of mange affecting a dairy herd stocking about 30,000 buffalo and 1000 cows. The mange mites were either Sarcoptes scabiei or Psoroptes ovis, or a mixed infestation of both. The morbidity rate was 5-30% varying from group to group, with 100% in a severely affected group. Signs noticed were progressive dermatitis, alopecia, keratinization, skin thickened and wrinkled, intense itching and marked loss of condition often ending in death. Great losses of young animals from mange and gastrointestinal nematodes are very common in dairy herds in India. In view of their economic importance, the activity of ivermectin against naturally occurring mange and parasitic infections of adult buffalo and buffalo calves was determined. Ivermectin was administered by subcutaneous injection (IVOMEC 1% w/v - MSD AGVET) at a dose of 200 mcg kg-1 body weight. The efficacy was ascertained from the disappearance of mites from skin scrapings and the reduction in numbers of worm eggs in the faeces. The results of the treatment were dramatic: the mites disappeared within 2 weeks of the drug being administered in the majority of animals, with marked improvement in skin lesions. Four buffalo which had their entire body surface affected with mange needed a second dose on Day 28 for complete recovery. The effect on the nematodes was equally spectacular, with infections of Neoascaris vitulorum, Trichostrongylidae, Oesophagostomum spp. and Bunostomum being eliminated within 1 week of treatment.
Piglets treated with cyclophosphamide (50 mg/kg body wt, iv) before infection/vaccination with swinepox virus showed a strong suppression of humoral immune response. Cell mediated immune response was also affected to some extent and the cyclophosphamide treated piglets did not resist the challenge on 21 day post infection/vaccination and developed mild lesions.