A study of median nerve entrapment neuropathy at wrist in uremic patients.

Carpal tunnel syndrome (CTS) is the most common entrapment neuropathy seen in uremic patients. The study was undertaken to estimate the frequency of CTS in uremic patients and to identify the most sensitive electrodiagnostic test. Study was conducted on 80 subjects of age 30-60 years. End-stage kidney disease patients were recruited for the clinical evaluation, motor nerve conduction studies (NCS), sensory NCS, F wave study and median-versus-ulnar comparison studies (palm-to-wrist mixed comparison study, digit 4 sensory latencies study and lumbrical-interossei comparison study). Among three different diagnostic modalities, frequency of CTS was found to be 17.5% with clinical evaluation, 15% with routine NCS studies and 25% with median-versus-ulnar comparison studies. Among the median-versus-ulnar comparison studies, lumbrical-interossei comparison study was found to be most sensitive (90%). The comparative tests for CTS are more sensitive compared to routine NCS and clinical examination. Among the comparative tests, lumbrical-interossei comparison study is the most sensitive. Early diagnosis of CTS may help patients of uremia to seek proper treatment at an appropriate time.

Exploration of 2, 4-diaminopyrimidine and 2, 4-diamino-s-triazine derivatives as potential antifilarial agents.

In view of the mandate from the World Health Organization (WHO) for developing novel drug candidates against human lymphatic filariasis, dihydrofolate reductase (DHFR) inhibitors are explored as potential antifilarial agents. The in vitro biological evaluation of an in-house library of 12 diverse antifolate compounds with 2,4-diaminopyrimidine and 2,4-diamino-s-triazine structural features against Brugia malayi is reported. To confirm the DHFR inhibitory potential of these compounds, reversal studies using folic acid and folinic acid were undertaken. Inhibition of DHFR can induce apoptosis; in this light, preliminary evidence of apoptosis by test compounds was detected using ethidium bromide-acridine orange staining and the poly(adenosine diphosphate-ribose) polymerase (PARP) inhibition assay. Among the evaluated compounds, 3 showed significant activity against both microfilariae and adult worms. The effects of 2 of these compounds were mostly reversed by folic acid, validating DHFR inhibitory activity. Partial reversal of the effect of 2 compounds by folinic acid and non-reversal of the effect of the third compound both by folic and folinic acids are discussed. This study opens new avenues for the discovery of lead molecules by exploiting the folate pathway against one of the major neglected tropical diseases, filariasis.

Possible implication of oxidative stress in anti filarial effect of certain traditionally used medicinal plants in vitro against Brugia malayi microfilariae.

INTRODUCTION: Tropical disease research scheme of World Health Organization has duly recognized traditional medicine as alternative for antifilarial drug development. Polyphenolic compounds present in traditionally used herbal medicines are natural antioxidants; however, paradoxically they may exert pro-oxidant effect. Popular drug diethyl carbamazine citrate harnesses the innate inflammatory response and the consequent oxidative assault to combat invading microbes. METHODS: With this perspective, extracts of Vitex negundo L. (roots), Butea monosperma L. (leaves), Aegle marmelos Corr. (leaves), and Ricinus communis L. (leaves) were selected to explore the possible role of oxidative rationale in the antifilarial effect in vitro. RESULTS: Apart from the last, other three plant extracts were reported to have polyphenolic compounds. Dose-dependent increase was found in the levels of lipid peroxidation and protein carbonylation for all the three plant extracts except Ricinus communis L. (leaves). Such increase in oxidative parameters also showed some degree of plant-specific predilection in terms of relatively higher level of particular oxidative parameter. High degree of correlation was observed between the antifilarial effect and the levels of corresponding oxidative stress parameters for these three plants. However, extracts of Ricinus communis L. (leaves) which is relatively deficient in polyphenolic ingredients recorded maximum 30% loss of motility and also did not show any significant difference in various stress parameters from corresponding control levels. CONCLUSION: These results reveal that targeted oxidative stress might be crucial in the pharmacodynamics.

Clinical study of the Immunoglobululin Enhancing effect of "Bala compound" on Infants.

Kaumarbhritya a branch of Asthanga Ayurveda deals with neonatal, infant and child health care. Multicentric studies conducted in various developed and developing countries have indicated that Infant Mortality Rate (I.M.R.) is very high in developing countries, and infection has been observed as the major cause. Immune system in neonates is not yet fully functional. Bala compound having the ingredients ofAtibala (Abutilon indicum Linn), Amalaki (Emblica officinalis Linn), Vidanga (Emblica ribes burn), Guduchi (Tinospora cordifolia Welld Miers), Pippali (Piperlongum linn), Yashtimadhu (Glycyrrhiza glabra Linn), Shankhapuspi (Convolvulus pluricaulis Chois ), Vacha (Acorus calamus Linn), Musta (Cyperus rotundus Linn) and Ativisha (Aconitum heterophyllum wall) are Medhya as well as Rasayana drugs mention in Ayurvedic classics. 'Bala compound" was tried in infants in the form of oral drops for a period of six months and result was assessed for serum immuoglobulins IgG, IgM, IgA for three months of interval of two follow ups (i.e., third and six month of infant). There is significant increase of immunoglobulins observed after six months administration of 'Bala compoumd"

Impact of field vaccination with a Theileria annulata schizont cell culture vaccine on the epidemiology of tropical theileriosis.

Tropical theileriosis, caused by Theileria annulata, is an important tick-borne disease of cattle. A cell culture attenuated vaccine has been developed in our laboratory by long-term in vitro propagation of the schizont stage of the parasite. A longitudinal study was conducted at selected farms housing indigenous, cross-bred and exotic animals to investigate the effect of vaccination on the epidemiology of the disease. A total of 120 animals in 4 age groups were vaccinated with the vaccine before the onset of disease season. An equal number of age-matched animals were kept as controls at the same sites. Animals were monitored for 14 months at monthly intervals. The 97.5% vaccinated animals showed a rise in antibody titres 1 month post-vaccination, as determined by single dilution ELISA. The 78.3% of non-vaccinated animals became sero-positive over the period of observation. Mean antibody titres were significantly higher in vaccinated than non-vaccinated animals. Cross-bred animals showed higher antibody titres followed by exotic and indigenous animals in both the vaccinated and non-vaccinated groups. However, the antibody titres in animals of different ages were similar. The 36.7% vaccinated and 64.2% non-vaccinated animals became carriers (<0.5% piroplasms in erythrocytes) during the observation period. Clinical cases of theileriosis were recorded only in the non-vaccinated group suggesting that vaccinated animals were sufficiently immune to withstand field tick challenge for at least 14 months.

Effect of vaccination in the field with the Theileria annulata (Hisar) cell culture vaccine on young calves born during the winter season.

The responses were monitored of young crossbred calves vaccinated against tropical theileriosis during the winter against a field tick challenge in the disease season. Thirty-eight calves below 2 months of age, born after the end of the disease season, were selected at an organized farm. Twenty-five animals were vaccinated with Theileria annulata (Hisar) cell culture vaccine (developed at CCS HAU Hisar laboratory) after the end of the disease season and 13 calves were kept as non-vaccinated controls. These calves were observed for their susceptibility to theileriosis in the new disease season. There was an increase in antibody titre in 18 of the 25 vaccinated animals one month after vaccination. The antibody titre then declined gradually, but remained higher than those of the non-vaccinated animals at month 0. No fever or other clinical signs of tropical theileriosis were observed in any of the vaccinated animals. Nine out of 25 (36%) vaccinated calves showed occasional piroplasms (<0.5%) in blood smears. All the vaccinated animals withstood the field tick challenge. On the other hand, 9 of the 13 (69%) unvaccinated calves exhibited occasional piroplasms, and included three clinical cases of tropical theileriosis. These observations suggest that young crossbred calves vaccinated with the T. annulata (Hisar) cell culture vaccine at the end of the disease season were relatively resistant during the next disease season.

Single dilution ELISAs using soluble piroplasm, cellular schizont and soluble schizont antigens for the detection of antibodies against Theileria annulata.

Single dilution ELISAs were standardised for the determination of antibody titres against Theileria annulata using three antigens namely soluble piroplasm, cellular schizont or soluble schizont antigens. Antibody titres of 20 cattle serum samples of known identity were determined by multi-dilution ELISA using the three antigens. The ratio of the optical density (OD) of known positive and known negative sera at different serum dilutions were calculated and termed as positive/negative (P/N) ratios. Coefficients of correlation (r) were calculated between the P/N ratios at different dilutions of known sera and their log10 antibody titres by multi-dilution ELISA. The value of "r" was the highest at the dilution of 1:400. From the log10 antibody titres of known sera and their P/N ratios at the dilution of 1:400, regression equations (Y = a + bX, where Y = predicted log10 titre, X = the P/N ratio at 1:400 dilution) were calculated separately for the three antigens. Thus, the equations Y = 1.63 + 1.35X for soluble piroplasm, Y = 2.67 + 0.547X for cellular schizont and Y = 1.817 + 0.663X for soluble schizont antigens were derived. Test sera were diluted to 1:400 and their OD were read in duplicate wells and converted to P/N ratios. The antibody titres were predicted from the P/N ratios using the above mentioned regression equations. Twenty randomly selected sera tested by single and multidilution ELISAs showed non-significant differences (P < 0.01) between antibody titres. Antibody titres of 90 unknown field sera of cattle were determined by single dilution ELISA. The piroplasm antigen detected higher antibody titres followed by cellular schizont and soluble schizont antigens. The study revealed that a single dilution ELISA could be successfully used for field epidemiological studies of tropical theileriosis.

Determination of duration of immunity of calves vaccinated with the Theileria annulata schizont cell culture vaccine.

Bovine tropical theileriosis caused by Theileria annulata is a serious haemoprotozoan disease of cattle affecting exotic cattle, their crossbreeds and young indigenous calves. Cell culture vaccines have been developed and used effectively in various countries for the control of this disease. However, the duration of immunity provided by these vaccines is poorly understood. The present experiments were planned to study the duration of immunity in animals after vaccination with the T. annulata (Hisar) schizont cell culture vaccine. Two groups of calves were vaccinated and challenged after a period of 3 and 6 months, respectively. There was no fever in any of the vaccinated calves after challenge. However, the vaccinated animals exhibited mild to moderate enlargement of lymph nodes and parasitological reactions. The parasitological reactions were very mild in calves challenged after 3 months and moderate in calves challenged after 6 months. There was a mild but significant decrease in the haematological values of calves after challenge. A significant rise in the anti-theilerial antibody titres was observed in all calves after vaccination, which increased further, by many folds after challenge. On the other hand, all the challenge control calves showed symptoms of acute theileriosis and died. The observations suggested that the T. annulata (Hisar) schizont cell culture vaccine provided immunity in vaccinated animals for at least 6 months in the absence of field tick challenge. However, there was some decline in immunity after 6 months, if the animals are not exposed to ticks during this period.

Comparison of cellular schizont, soluble schizont and soluble piroplasm antigens in ELISA for detecting antibodies against Theileria annulata.

The efficacy and suitability of cellular schizont, soluble schizont and soluble piroplasm antigens was compared for detecting antibodies against Theileria annulata. Fifty bovine sera of known identity were evaluated in ELISA using the above mentioned antigens. Antibody titres of 1:100 to 1:51,200 were detected while using soluble piroplasm and cellular schizont antigen in ELISA. The titres ranged between 1:100 to 1:25,600 with the soluble schizont antigen. Soluble piroplasm antigen exhibited the highest antibody titres followed by cellular schizont and soluble schizont antigens. Cellular schizont antigen proved to be better than soluble schizont antigen for detecting anti-schizontal antibodies. Antibody titres obtained by the three antigens exhibited a good linear correlation amongst each other. The study showed that soluble piroplasm and cellular schizont antigens can be used successfully for detecting antibodies against piroplasm and schizont stages of T. annulata, respectively in bovine sera.

Monoclonal antibodies against conformationally dependent epitopes on porcine reproductive and respiratory syndrome virus.

Monoclonal antibodies (MAbs) against porcine reproductive and respiratory syndrome virus (PRRSV) were prepared and characterized. Four MAbs were developed from the mice immunized with the recombinant GP4 protein expressed in insect cells, and six MAbs were derived from the immunization with recombinant GP5 protein. All of the MAbs showed strong perinuclear fluorescence in PRRSV VR2385 infected cells by immunofluorescence staining. Among the MAbs to GP5 protein, one showed strong reactivity in ELISA and recognized a 26 kDa band of PRRSV in a western blot assay, while another showed neutralizing activity against the VR2385 isolate. Out of the four MAbs to GP4 protein, one showed mild reactivity in ELISA with detergent extracted antigen, but had no reactivity in a western-blot assay. The failure of MAb binding to detergent extracted antigen in ELISA or in western-blot analysis indicated that the MAbs were against conformationally dependent epitopes. Reactivity patterns of the MAbs with PRRSV field isolates tested by fixed-cell ELISA showed that there are antigenic variations in PRRSV GP4 and GP5 proteins. Development of these MAbs will benefit further studies on PRRSV structural proteins as well as in understanding their roles in PRRSV pathogenesis.

Successful long-term in vitro cultivation of Theileria annulata schizonts in media supplemented with homologous and heterologous sera.

The efficacy of medium RPMI-1640 supplemented with either foetal bovine, normal bovine, goat or sheep sera was compared for prolonged in vitro propagation of Theileria annulata (Hisar) schizonts. Medium RPMI-1640 supplemented with 20% foetal bovine serum (standard growth medium) resulted in optimum growth of T. annulata (Hisar) schizonts in vitro. Comparable viability and non-viability counts were observed in growth media supplemented with normal bovine or goat sera. However, viability counts in medium supplemented with sheep serum were significantly lower than that of the standard medium. Mitotic indices of cultures of T. annulata (Hisar) schizonts were directly related to the extent of cell growth and were lower in various growth media supplemented with normal bovine, goat or sheep sera than in that of the standard medium. The results suggested that normal bovine and goat sera could be successfully used in place of foetal bovine serum in the growth medium for long-term in vitro propagation of T. annulata schizonts. The study will help in reducing the cost of large-scale in vitro propagation of T. annulata aimed at mass production of the cell culture vaccine.

Susceptibility to tropical theileriosis of calves born to dams immunized with Theileria annulata (Hisar) cell culture vaccine.

The susceptibility/immune status to tropical theileriosis of calves born of immunized dams was evaluated. Six cows were vaccinated with the Theileria annulata cell culture vaccine in the eighth month of pregnancy. Sera from the immunized dams exhibited very high post-vaccination antibody titres as determined by the indirect fluorescent antibody (IFA) test. The calves born to these dams did not show antibodies against T. annulata at the time of birth (IFA titres of < 1:20). The new-born calves were fed colostrum from their mothers and were challenged with T. annulata-infected ground tick supernate at 5-7 days of age. All the calves developed fever (from day 5-6 onwards) and parasitological reactions (from day 8-9 onwards) after challenge. There was a significant decrease in the haemoglobin and packed cell volume of the calves after challenge. All the calves showed signs of acute theileriosis by day 9-10 after challenge and had to be treated with buparvaquone in order to save their lives. The study indicated that detectable levels of anti-theilerial antibodies were not transferred from immune dams to their offspring. All the calves born to immunized dams were fully susceptible to theileriosis and thus themselves needed vaccination.

In vitro propagation of Theileria annulata infected schizonts in different media supplemented with heterologous sera.

Efficacy of medium RPMI-1640 (supplied by Gibco USA, Centron and Hi-media) supplemented with horse, donkey, sheep and goat sera was evaluated for in vitro propagation of Theileria annulata (Hisar) infected bovine mononuclear cells. The results were compared with the growth rate in RPMI-1640 supplemented with foetal bovine serum (Gibco). RPMI-1640 (Gibco) proved to be the best medium for in vitro cultivation of the parasite infected cells. Foetal bovine serum could be easily, safely and reliably substituted with goat and sheep sera in the growth medium. Horse and donkey sera also gave comparable growth of T. annulata infected cells in vitro. Successful use of heterologous sera greatly helped in reducing the cost of in vitro cultivation of T. annulata schizonts. These findings have important implications on mass production of an attenuated cell culture vaccine for the control of bovine tropical theileriosis.

Responses in animals vaccinated with the Theileria annulata (Hisar) cell culture vaccine.

Bovine tropical theileriosis caused by Theileria annulata is an economically important disease of cattle in India. The disease has assumed paramount importance with the intensification of cross-breeding programmes aimed at enhancing milk production in the country. To control this disease, a cell culture vaccine was developed in this department by continuous passaging of T. annulata (Hisar) schizonts in vitro. Current work in this department has concentrated on the epidemiology of theileriosis: development of the cell culture vaccine for very young calves and pregnant cows; evaluation of serological responses using immunofluorescent antibody (IFA) tests and Enzyme-linked immunosorbent antibody assays (ELISA); studies on the duration of immunity stimulated by the cell culture vaccine; the immune/susceptible status of calves born to vaccinated dams. Results have shown the following. Clinical cases of theileriosis were mainly observed in young calves below two months of age followed by adults in exotic and cross-bred animals. Amongst indigenous animals, only young calves below two months of age suffered from clinical disease. Clinical cases of theileriosis mainly occurred between the months of April to October. The T. annulata schizont cell culture vaccine developed in the department was extensively used in the susceptible calves and pregnant/lactating cows in the field. Sufficiently high antibody titres were detected by both schizont as well as piroplasm antigen using both ELISA and IFAT. The results indicated that the vaccine was safe, potent and effective for all breeds and age groups of cattle under field conditions. ELISA was standardised for T. annulata using three antigens, viz.: soluble piroplasm, soluble schizont and cellular schizont antigens. Comparison of results with IFAT showed that ELISA is more sensitive, objective, reliable and specific as well as less cumbersome than IFAT. Piroplasm, cellular schizont and soluble schizont antigens were found to be suitable for the detection of antitheilerial antibodies as per their order in ELISA. Studies on the duration of immunity stimulated by the T. annulata schizont cell culture vaccine indicated that immunity started waning after six months. Calves born of dams immunised against T. annulata with the cell culture vaccine were found to be fully susceptible to theileriosis soon after birth. This indicated that there was no passive transfer of immunity from dams to their offspring through colostrum.

An immunisation trial with in vitro produced Babesia bigemina exoantigens.

Bovine babesiosis, caused by Babesia bigemina, is an important tick-transmitted haemoprotozoan disease in the tropics. This study evaluated the immunoprotective efficacy of in vitro produced B. bigemina exoantigens in bovine calves. The calves inoculated with B. bigemina exoantigens did not show any clinical, parasitological or hypersensitivity reactions after inoculation. They withstood challenge without showing any clinical symptoms except a transient thermal reaction. In contrast, two out of four control calves exhibited clinical symptoms of babesiosis and one died. On challenge, there was a significant reduction in the haematological values of both groups. However, this was more pronounced in the control animals. Challenge resulted into a normocytic hypochromic anaemia. The vaccinated animals revealed a significant rise in antibody titres after vaccination as well as after challenge as detected by a single dilution ELISA. The rise in antibody titres of control animals was only moderate. Inoculation of B. bigemina exoantigens induced a protective immune response in the vaccinated animals which could protect them from infected blood challenge.

Efficacy and safety of unguided percutaneous fine needle aspiration cytology in peripheral lung lesions.

The efficacy and safety of unguided percutaneous fine needle aspiration cytology in peripheral lung lesions were studied in 42 patients. The diagnostic yield of unguided aspiration was 83.3%. The yield in malignant lesion was 85.7% while in benign lesion it was 78.6%. There was no false positive report. Complications seen were asymptomatic pneumothorax and blood tinged sputum in a few cases following aspiration. The procedure was well accepted by the patients. Unguided FNAC is a simple, quick, economical, safe and reliable procedure for the diagnosis of peripheral lung lesions.

Non-aqueous cerimetric determination of dithiocarbamates and its application to the determination of amines.

A non-aqueous titrimetric method for determination of dithiocarbamates with cerium(IV), is described. The compounds are titrated at room temperature, with visual and potentiometric end-point detection. In visual titrations, the reagent serves as self-indicator and turns the solution yellow at the end-point. Methyl Red can also be used as the indicator. The method has been applied to the determination of amines after their quantitative conversion into dithiocarbamates by reaction with carbon disulphide. The proposed method is accurate to +/- 0.8% with a relative standard deviation of +/- 0.7%.

Mortality differentials among immigrant populations in Canada.

PIP: Mortality among different groups of migrants to Canada is compared with that of native-born Canadians and with trends in the migrants' countries of origin, using data for the period 1981-1985. Results show that life expectancy at birth among migrants in Canada is generally higher than that of the populations at place of origin and destination. (SUMMARY IN FRE AND SPA)^ieng