Genotypes of the human papillomavirus: Relevance to Indian field trials of the vaccine.

Highly effective HPV vaccines are likely to become available for use in India shortly. The availability and validation of the vaccines to prevent oncogenic HPV infection associated lesions from progressing to cancer has clearly offered a cost effective long-term strategy to reduce the cervical cancer burden, particularly for developing countries where effective screening programmes are not available. The enthusiasm for these new vaccines duly justified, are we in reality targeting the actual delinquent by prompting these vaccines in India? The answer would be no as we may not be essentially implementing a rational fool-proof vaccine program which will aid in preventing HPV-related diseases, for the simple reason that there is a lack of understanding about the underlying HPV types responsible for cervical cancer in India. Field trials involving large populations form a major part of continued analysis in understanding any disease and India is still short of such a trial as far as cervical cancer is concerned. Conducting such studies, might also act as prevention programs that will save lives and improve public health in a substantive manner.

Effect of Bacopa monniera (Linn.) on lipid peroxidation and lipofuscinogenesis in prostate gland of D-galactose induced aging mice, Mus musculus.

The levels of malondialdehyde and lipofuscin pigments were increased in the prostate of D-galactose (0.5 ml/day, sc, for 20 days) induced aging mice. After B. monniera (40 mg/kg body weight for 20 days) ethanol leaf extract administration levels of both the parameters were reduced significantly. The results suggest that B. monniera prevents formation of malondialdehyde and lipofuscin pigments which are the indicators of aging.

Expression of matrix metalloproteinases (MMP-1, MMP-2 and MMP-9) and their inhibitors (TIMP-1 and TIMP-2) in oral submucous fibrosis.

Immunohistochemical staining of formalin fixed, paraffin embedded tissue sections of OSF for MMPs-1,2,9 and their tissue inhibitors TIMP-1and 2 was performed using monospecific antibodies coupled with gelatin zymography (MMP-2 and 9) for measuring enzymatic activity quantitatively and for distinguishing the active from the inactive variants of enzymes. The present study, contrary to earlier reports, recorded statistically significant increase in the levels of stromal expression of MMP-1, MMP-2 and MMP-9 and TIMP-1 and TIMP-2 using monospecific antibodies reacting against tissue antigens.The simultaneous increase in reactivity of MMPs and TIMPs poise difficulty in interpretingthe results of this study. The possible reasons for this result, against the backdrop of existing knowledge, were attempted in this study.

Radiolabelled monoclonal antibodies against human cardiac myosin.

Myocardial infarction (MI) can be monitored using several protein markers including human cardiac myosin (HCM). Monoclonal antibodies were raised against HCM by hybridoma technique. Antimyosin antibody producing clones were identified by ELISA and monoclonality was established by limiting dilution. The antibodies were purified, isotyped and their cross reactions with myosin from other species were estimated. All the clones showed negligible cross reaction with rabbit myosin, but reacted with bovine skeletal myosin to different extents (40-100%). The most avid antibody Mab 4G4 which also strongly reacted with rat cardiac myosin, was labelled with 125I using different oxidising agents such as iodogen, chloramine-T and lactoperoxidase. More than 95% pure radiolabelled antibody could be obtained by gel filtration. The immunoreactivity was retained. Mab 4G4 was also labelled with 99mTc using stannous tartrate as the reducing agent. Radiolabelling yield was approximately 60%, the purity was >95%. Both the radiolabelled preparations were tested for biodistribution in rats--both normal and those with induced MI. Approximately 0.7 % of the injected activity/g was found in the infarcted region and the accumulation of activity in the infarcted heart was 1.5 times that in the normal heart. A very high percentage of activity (80%) accumulated in the thyroid. With further optimisation of labelling and use of F(ab')2 fragments, better delineation of the infarct sites may become possible.

Apoptosis: future directions in cancer therapy.

Cancer as a multifactorial disease results in gain of immortality due to defective apoptosis. The primary mode of cell death by apoptosis induced by various modes of treatment often fail in vivo. The in vitro environment is less complex while the in vivo environment is influenced by various external regulatory signals besides the existence of multiple, parallel and independent apoptotic pathways. Further, specific preference for an apoptotic pathway in a certain cell type would significantly alter the apoptotic responses. Identification of defects in preferred pathways and choosing alternative and potentially inducible pathways would help in deciding on apoptosis-based treatment protocols. Mechanisms involved in the execution of apoptosis may also not be unique to apoptotic pathways since similar events, possibly with strict control, do occur during mitosis. Further evaluation may yield new dimensions to apoptosis and apoptosis-based therapy.

Use of Traditional Eye Medicines by corneal ulcer patients presenting to a hospital in South India.

PURPOSE: To investigate the nature and frequency of use of Traditional Eye Medicine (TEM) for corneal ulcer in patients from predominantly rural background. METHODS: We documented the the use of TEM by corneal ulcer patients presenting to a tertiary eye-care centre in South India during two months of 1996. RESULTS: Of 283 patients enrolled in the study, 135 (47.7%) of the patients used TEM. There was no difference with regard to age and sex distribution of patients using TEM and those who did. Patients with history of trauma were more likely to use TEM. Common forms of TEM used were human breast milk 61(45.2%), leafy matter 40(29.6%), castor oil 16 (11.9%), and hen's blood 8 (5.9%). CONCLUSION: Though the awareness of intraocular lens implantation for cataract surgery is very high in this segment of the population, it is still tragic that an awareness of primary eye care following trauma has not been created. Health education is mandatory to prevent this avoidable cause of blindness.

Characteristics of a 60K gelatinase involved in rat mammary gland involution.

In order to study the role of matrix degrading enzymes in modulating cell matrix interaction, an understanding of the characteristics and regulation of their activity is useful. A number of matrix degrading metalloproteinases are involved in modulating the cell-ECM interactions during the involutory phase of mammary gland resulting in its remodelling. Zymographic studies showed that three types of gelatinases (60K, 68K and 130K) occur during the different phases of involution. The 60K gelatinase which appeared on the fifth day of involution has been purified by affinity chromatography over gelatin sepharose. Zymographic and radiolabelled substrate digestion studies at different pH and in presence of different cations showed that the activated form of this gelatinase is a Ca2+ dependent neutral matrix metalloproteinase capable of cleaving collagen I and collagen IV. Immunocytochemical studies showed that the enzyme is localised at pericellular/extracellular sites.

Comparison of clinical outcome of fixed-dose subcutaneous low molecular weight heparin (tinzaparin) with conventional heparin in unstable angina: a pilot study.

Forty patients who were hospitalized for unstable angina were randomized to receive treatment with either regular heparin (Group I) in conventional dose as continuous infusion for 5 days or fixed-dose low molecular weight heparin (LMWH) (Group II), 3500 units subcutaneous twice daily for a period of 5 days. Both the groups were evenly matched with regard to age, sex presence of risk factors and adjunctive drug therapy. The clinical endpoints at the end of 5 day therapy were: recurrence of angina, occurrence of myocardial infarction and need for urgent revascularization. In Group I, 6 out of 20 patients had recurrence of angina, of whom 3 required urgent coronary angiography and revascularization. In Group II, 4 out of 20 patients had recurrence of angina, of whom one patient required urgent angiography and angioplasty. There were no bleeding complications in either of the groups. The recurrent anginal episodes in the conventional heparin group correlated with low aPTT values at the time of angina. Thus, this pilot study suggests that LMWH is equally effective in the treatment of unstable angina, the advantage of LMWH being the ease of administration and no need for monitoring aPTT levels during therapy.

Microplate assay of elevated esterase activity in individual pyrethroid-resistant mosquitoes.

Involvement of esterase-mediated hydrolysis as a mechanism of pyrethroidresistance in three species of mosquitoes, viz., Aedes aegypti, Culex quinquefasciatus and Anopheles Stephensi was investigated by microplate assay of β-esterases in individual larva and adult female and male mosquitoes. Assuming an absorbance value of 0·4 and above at 555 nm as the threshold level of elevated esterase activity which confers resistance, frequency distributions of such individual test mosquitoes were constructed in resistant and susceptible populations. The results indicate the involvement of ester hydrolysis of Pyrethroids as a predominant mechanism of pyrethroid-resistance in the larvae of Culex quinquefasciatus but not in Aedes aegypti. However, a marginal role of esterases is indicated in the larvae of Anopheles stephensi.

Intracranial haematoma resulting from lightning stroke.

Intra-cerebral haemorrhage due to lightning stroke is extremely rare. We report a 45 year old woman who developed intracranial haemorrhage due to a direct lightning stroke. This was proved by CT scan. The haematoma was evacuated surgically, resulting in full neurological recovery of the patient.

Involvement of mono-oxygenases as a major mechanism of deltamethrin-resistance in larvae of three species of mosquitoes.

Role of mono-oxygenases as a mechanism of resistance to the synthetic pyrethroid, deltamethrin in the larvae of Culex quinquefasciatus Say, Aedes aegypti L. and Anopheles stephensi Liston developed by laboratory selections with deltamethrin, DDT or deltamethrin and the synergist, piperonyl butoxide (PBO) in the ratio of 1:5, was investigated. There was a significant correlation with mono-oxygenase activity and larval LC50 to deltamethrin in various strains of all the three species. In addition, the activity of glucose-6-phosphate dehydrogenase (G6PD), the main NADPH generating enzyme for mono-oxygenases, also showed enhanced activity in deltamethrin and DDT-selected strains. The present data, therefore, clearly suggest that deltamethrin resistance in the larvae of Cx. quinquefasciatus, Ae. aegypti and An. stephensi is mainly due to the detoxification of deltamethrin by microsomal mono-oxygenases. High activity of G6PD observed in DDT-selected strains seems to be related to its role as a rate-limiting enzyme in GSH-dependent dehydrochlorination of DDT.

Effect of ayurvedic medicines on beta-glucuronidase activity of Brunner's glands during recovery from cysteamine induced duodenal ulcers in rats.

Biochemical and histochemical studies revealed decreased beta-glucuronidase activity in the Brunner's glands of duodenal ulcerated rats. The enzyme activity showed gradual increase during recovery. Rats treated with a mixture of Ayurvedic medicines (Glycyrrhiza glabra, Terminalia chebula, Piper longum and Shanka Bhasma) recovered faster with concomitant increase in beta-glucuronidase activity in the Brunner's glands. It can be concluded that Ayurvedic medicines used do not act as antacid but improve the secretory status of Brunner's glands involved in the protection against duodenal ulcer.