Country watch: India.

PIP: At the first nongovernmental organization (NGO) networking meeting on HIV/AIDS in Gujarat State, India (July 1992), the 80 participants concluded that training for grassroots volunteers was urgently needed. The Gujarat AIDS Prevention Unit (GAP-SIRMCE) assumed responsibility for organizing this capacity-building activity. GAP facilitators formulated a training-of-trainers program and outlined a workshop manual. Feedback was obtained from NGOs and experts from training programs so that a manual could be finalized. 200 NGOs were invited to participate; 65 sent delegations for training during the 1-year project. They included NGOs working on health, rural and agricultural development, family welfare, slum development, cooperative movements, and women's groups. The workshops lasted 1-3 days, generally had 10-12 participants (2 trainees per NGO). Topics included basic information on HIV/AIDS, sexuality, and barriers to condom use. There were discussions on negotiating safer sex and identifying fulfilling alternatives to sex acts. Condom demonstrations were also done. GAP then presented important educational messages. Each workshop ended with an evaluation. One of the lessons learned during the project concerned funding of participants. Some NGOs said their participation depended on money received for travel and daily subsistence. It was decided that GAP should not offer such reimbursements because this encourages NGOs to see workshop participation merely as a means of earning money. The Gujarat NGOs in the Indian Network of NGOs on HIV/AIDS have identified 2 major areas for future collaboration. The first was adolescent sex education; more than 98% of NGOs wanted to offer such programs. The second area concerned training in counseling skills in relation to HIV/AIDS and also in family planning, drug addiction, and family and marriage counseling.^ieng

Quantitative topochemistry of rat liver enzymes during postnatal development in relation to activity-rest cycle.

A quantitative histochemical method (Trident) has been adapted to measure the activities of 4 enzymes, succinate dehydrogenase (SD), isocitrate dehydrogenase (ICD), glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6-PGD), within the liver acini of the rat during the postnatal developmental period. Quantitative changes of these enzymes in livers of rats of 25 g and 50 g body weight were studied, with particular emphasis on the activity-rest cycle. The results indicate a time-dependent heterogeneous distribution of enzymes along the acinar zones and the pattern of localization is age-dependent. When the mean enzyme activity from each group in relation to the time of the day are compared, a mirror image of each other could be seen. In general, a high enzyme activity has been observed during the resting phase in 25-g rats and low in 50-g rats. During the developmental period, the mean ICD activity is diminished, whereas G6PD and 6-PGD are augmented, and SD activity remains unchanged.

Topogenesis of glycogen distribution pattern in post-natal rat liver in reference to the activity-rest cycle.

The present study, on immature rats, revealed that in the liver with exclusively diploid cell population (MD), the glycogen content was half of the adult level and the 24 h distribution pattern was reverse, i.e. highest in the evening and lowest in the morning. In these animals, in general, the protein content was high and did not show any circadian rhythm but incidentally showed a time dependent zonal distribution pattern. The low glucose-6-phosphatase activity reciprocates with low glycogen content. The relative number of cells per unit area showed a time dependent distribution pattern. The liver with equal distribution of diploid and tetraploid classes (MD:MT), attained the classical circadian rhythm of glycogen with high morning and low evening oscillation pattern. The TRIDENT measurements manifest the zonal distribution pattern with high values at the perilobular region (PL) and low at the centrolobular region (CL). The relative cell number study along the acinus demonstrated more number of cells around the PL region than that at CL region, indicating a variation in the cell size. Eventually, the protein content showed a circadian rhythm in this group, with high amount in the morning. The zonal distribution pattern always revealed the high content near the perilobular region. This could be due to more number of cells per unit area in this region. The glucose-6-phosphatase showed a circadian rhythm. The typical high glucose-6-phosphatase in the perilobular region could be further subzonated into small groups of high activity surrounded by lighter zones, thus establishing the heterogeneous function of liver parenchymal cells.

Circadian rhythm of blood and urinary copper in presumably healthy subjects of vegetarian food habit.

Circadian rhythm of blood and urinary copper has been studied in presumably healthy subjects of a particular ethnic group in India who are vegetarians. A definite 24-hr variation has been observed for both blood and urinary copper. The peak for blood copper was 1,500 hr and the lowest value was 0600 hr, with values of 0.185 mg/100 ml and 0.106 mg/100 ml respectively. The urinary peak and trough occurred at 0600 and 0300 hr, respectively. Remarkably higher 24-hr copper excretion values were noted (64.49 micrograms/day) with a range of 15-100 micrograms/day. The blood level of copper (0.134 mg/100 ml) remained within the range reported. One subject out of 25 deviated from the group with respect to circadian phasing and amplitude to urinary copper excretion.

Copper and zinc level in biological samples from healthy subjects of vegetarian food habit in reference to community environment.

Many epidemiologists have found a correlation between copper and zinc in the community environment and diseases, such as myocardial and vascular pathologies, and diabetes. The purpose of this study was to investigate the total daily intake of these two metals in cooked food, drinking water and air and their respective levels in blood and urine. A chronobiological methodology has been adopted to establish the reference values of these two metals in biological samples. It has been observed that the daily intake of copper is within the recommended value, whereas its urinary excretion is high. The daily intake of zinc is below the recommended value and its urinary excretion is also high. Both the metals showed a temporal oscillation pattern in blood and urine. A possible chronic zinc deficiency has been anticipated in this particular ethnic group of vegetarian food habit.

Ploidy class-dependent variations during 24 h of glucose-6-phosphate and succinate dehydrogenase activity and single-stranded RNA content in isolated rat hepatocytes.

The time-dependent variations over 24 h of glucose-6-phosphate dehydrogenase (G6PDH) activity, succinate dehydrogenase (SDH) activity and single-stranded RNA (ssRNA) content have been investigated by cytophotometric analysis of cytochemically stained isolated hepatocytes of different ploidy classes from adult male rats. A marked variation of 48% over the day in G6PDH activity of the mononuclear diploid cells was revealed, but no significant variation in the binuclear tetraploid cells. The cells of the inbetween ploidy classes showed an amplitude of variation of 38% (binuclear diploid cells) and 24% (mononuclear tetraploid cells), respectively. All cells showed a maximum activity of the enzyme at the middle of the day and a minimum during the night. The relative enzyme activity per mononuclear diploid cell was significantly higher than the relative activity in the other cells, especially at its maximum. The variation of the SDH activity in hepatocytes isolated from the same rats was similar in all cells, irrespective of their ploidy class. The activity was highest at the end of the activity phase of the animals. The SDH activity per cell was directly proportional to the quantity of genome copies. The ssRNA content of the hepatocytes showed a time-dependent variation with a maximum during the resting phase of the animals and a minimum during their activity phase. The variation was larger in the mononuclear diploid cells than in the cells of other ploidy classes and the ssRNA content was also significantly higher in these cells than in the hepatocytes of other ploidy classes when calculated on the basis of genome copies. It is concluded that the large amplitude of variation over the day and the high relative amount of G6PDH activity and ssRNA content in mononuclear diploid cells is related to the function of these cells as stem cells of the liver parenchyma.

Glucose-6-phosphate dehydrogenase activity in individual rat hepatocytes of different ploidy classes. II. Time-dependent variations during 24 h.

The time-dependent variation of glucose-6-phosphate dehydrogenase (G6PDH) activity during 24 h has been investigated by cytophotometric analysis in cytochemically stained, isolated hepatocytes of different ploidy classes from adult male rats. It was found that the amplitude of the variation in the total amount of parenchyma was 22%, with a maximum in the middle of the resting phase of the animals (13%). This small variation could not be detected subjectively in cryostat sections of liver that had been stained histochemically; nor could a metabolic zonation of the liver parenchyma be observed except the intermediate zone, which stained more strongly for G6PDH activity at 13% and 22%. The quantitative study on the isolated hepatocytes, however, revealed a relatively strong variation of 48% during the day in enzyme activity of the mononuclear diploid cells and no significant variation in the binuclear tetraploid cells. The cells of the in-between ploidy classes showed an amplitude of variation of 38% (binuclear diploid cells) and 24% (mononuclear tetraploid cells). All cells showed a maximum activity at 13% and a minimum activity during the night. It is concluded that this ploidy-dependent variation might play a role in the generation of pentoses for RNA synthesis. The findings indicate that for metabolic studies the heterogeneity of the liver parenchyma should be taken into account with respect to the different ploidy classes, as well as the well-known metabolic zonation of the acinus.

Enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier. Studies on the synthetizing reaction technique, the indigogenic method, the metal salt method, the post-azo-coupling technique, and the tetrazolium salt technique.

A recently developed method for the (quantitative) demonstration of glucose-6-phosphate dehydrogenase activity in individual cells with the use of a polyacrylamide carrier has been extended for other enzyme cytochemical techniques. Isolated hepatocytes have been incorporated in the matrix of a thin transparent polyacrylamide gel prior to incubation in a cytochemical medium. The techniques which have been applied are the synthetizing reaction technique for glycogen phosphorylase, the indigogenic method for nonspecific esterase, the metal salt method for glucose-6-phosphatase, the post-azo-coupling technique for acid phosphatase, and the tetrazolium salt technique for succinate and lactate dehydrogenase activities. In all cases a few major problems which occur in the cytochemistry on single cells seem to be solved. The morphology is very well preserved, the final reaction product seems to be precipitated at the expected site of enzyme activity and the coloured end-product is highly specific for the enzyme activity to be studied, as has been demonstrated well with control experiments. The conclusion is reached, therefore, that this relatively simple device can be used routinely for the optimalization of enzyme cytochemistry of single cells.

A time-dependent distribution pattern of ploidy classes in adult rat liver parenchyma.

The distribution pattern of rat liver parenchymal cells of different ploidy classes has been investigated with isolated liver cell preparations on four different time points of the day. The percentage distribution of different ploidy populations was found to fluctuate along the activity-rest cycle of the animals. The mononuclear tetraploid (MT) cell population showed the strongest time-dependent distribution pattern with a 10% variation around the 24 h mean, the variations in the binuclear diploid (BD) cells being somewhat less obvious, but clearly present. The mononuclear diploid cells and the binuclear tetraploid cells occurred in a virtually constant percentage. An inverse correlationship was noted between MT and BD cells, the increase in the number of MT cells being accompanied by a decrease in the number of BD cells. This interdependency in the presence of the two cell types can be related to their relationship in the process of formation.

Circadian variation of liver esterases.

The circadian rhythm of liver esterase activity has been studied in rats maintained under highly standardized laboratory condition. They were housed in specially constructed climate rooms and subjected to an artificial 12 : 12 light-dark regimen. The total esterase was further differentiated to A esterase (E.C. 1.1.2) by using E 600 (Paraoxan), and a circadian rhythm for this enzyme also has been observed. During the lowest phase of total esterase activity, the higher phase for organophosphate-resistant esterase activity was observed.