Tuberculosis Case Notification by Registered Private Medical Practitioners in Kolkata: A Mixed-Methods Study

Background: Under?notification of tuberculosis(TB) cases remains a persistent problem that impedes accurate estimation of the disease burden. India’s private health sector contributes to only one?fourth of the total TB notifications. Objectives: The present study was conducted among registered private practitioners in Kolkata to assess their knowledge, attitude and practice on TB notification, to find the socio?demographic and work-related factors associated with it, to identify the barriers faced by them in notifying TB cases and to elicit suggested solutions in overcoming these barriers. Materials and Methods: It was an observational study, cross-sectional in design following explanatory sequential mixed?methods approach conducted among 426 private practitioners in Kolkata Municipal Corporation area over 2 years (July 2019–October 2021). Quantitative data were analyzed using SPSS 25.0 with suitable descriptive and inferential statistics. Qualitative data were analyzed using Atlas.ti 7.1 and data were represented in the form of themes, codes, and verbatims. Results: Out of 426, 295 (69.2%) of the study population had adequate knowledge, 385 (90.4%) had positive attitude and only 115 (27.0%) had satisfactory practice. Lack of awareness, inadequate communication, and breaching patient confidentiality were the main barriers identified. Suggested solutions to improve engagement of private sector were organizing more continuing medical educations, active case finding, and acknowledgement to private practitioners on notification. Conclusion: Private practitioners had adequate knowledge on TB notification, their attitude was very positive but practice was poor. Training and sensitization of private practitioners on notification are recommended with feedback from both ends.

A Cross-Sectional Study on Anxiety Due to COVID-19 and its Predictors among Undergraduate Medical Students in a Tertiary Care Teaching Hospital of Kolkata, West Bengal

Background: Medical education is recognized as stressful globally. COVID‑19 pandemic is an additional source of anxiety to the medical students. Objectives: This study was conducted to assess the prevalence and to identify the factors associated with anxiety due to COVID‑19 among undergraduate medical students in a teaching hospital of Kolkata, West Bengal. Methods: An observational cross‑sectional study was conducted among 363 undergraduate medical students using the stratified random sampling of a medical college from June to July 2021. Data were collected using a predesigned, pretested, and structured online questionnaire, including “Coronavirus Anxiety Scale.” Descriptive statistics were used to estimate the prevalence of anxiety. Pearson’s Chi‑square test was performed to find out the factors associated with anxiety due to COVID‑19. Results: About 25.6% of the medical students were found to have anxiety due to COVID‑19. About 28.9% of them reported COVID‑19 infection in family in recent past and 11.0% had themselves tested positive. Nearly 20% reported loss of family members, relatives, and close friends due to COVID‑19. The factors associated with anxiety due to pandemic were socioeconomic status, social stigma, sleep disturbances, history of COVID‑19 in family, loss of job. and vaccination status of family members missing practical classes and exam‑related anxiety. Conclusion: The study found that one‑fourth of the medical students had anxiety due to COVID‑19. Social stigma due to COVID‑19 and loss of job of parents were the most significant predictors. It is recommended that targeted psychological and clinical interventions need to be taken to alleviate students’ anxiety due to COVID.

Molecular characterisation of Aspergillus flavus isolates from peanut fields in India using AFLP

Aflatoxin contamination of peanut, due to infection by Aspergillus flavus, is a major problem of rain-fed agriculture in India. In the present study, molecular characterisation of 187 Aspergillus flavus isolates, which were sampled from the peanut fields of Gujarat state in India, was performed using AFLP markers. On a pooled cluster analysis, the markers could successfully discriminate among the ‘A’, ‘B’ and ‘G’ group A. flavus isolates. PCoA analysis also showed equivalent results to the cluster analysis. Most of the isolates from one district could be clustered together, which indicated genetic similarity among the isolates. Further, a lot of genetic variability was observed within a district and within a group. The results of AMOVA test revealed that the variance within a population (84%) was more than that between two populations (16%). The isolates, when tested by indirect competitive ELISA, showed about 68.5% of them to be atoxigenic. Composite analysis between the aflatoxin production and AFLP data was found to be ineffective in separating the isolate types by aflatoxigenicity. Certain unique fragments, with respect to individual isolates, were also identified that may be used for development of SCAR marker to aid in rapid and precise identification of isolates.

Study of Microbial Population and Enzyme Activities in Intercropped Peanut Rhizosphere with Different Nutrient Application.

The aim of this study was to investigate the effect of different levels of chemical fertilizers (CF) alone or in combination with farmyard manure (FYM) under lime or no lime amelioration on biological properties of the rhizosphere soil of peanut, grown as intercrop with sabai grass (Eulaliopsis binata (Retz.) C.E. Hubb) in an acid lateritic soil. The effect of five fertilization levels viz., no CF (F0), CF @ 20:40:30 (F1), CF @ 40:80:60 (F2) kg ha-1 NPK, F1 + 2.5 t FYM ha-1 (F3) and F2 + 5 t FYM ha-1 (F4) with (2 t ha-1) and without lime application were studied on the bacterial density, dehydrogenase and phosphatase enzyme activities (i.e. acid and alkaline phosphomonoesterases), nitrogen accumulation in nodules and phosphorus solubilizing power at 25, 50, 75 and 100 days after sowing of peanut for two years. Results showed significant effects of fertilization levels and growth stages of the crop on the microbial activities. Populations of symbiotic nitrogen fixing and phosphorus solubilizing bacteria, soil enzyme activities, nitrogen accumulation in nodules and phosphorus solubilizing power in the FYM+CF treated plots significantly increased compared to sole CF treatments under both lime and no lime application. Lime application improved the activity of dehydrogenase and alkaline phosphomonoesterase enzymes, while decreased acid phosphomonoesterase activity. This study revealed that integrated application of optimum level of inorganic fertilizer, farmyard manure along with lime could improve the biological properties of an acid lateritic soil as well as the growth of peanut under sabai grass-peanut intercropping system.

Yield, Fruit Body Diameter and Cropping Duration of Oyster Mushroom (Pleurotus sajor caju) Grown on Different Grasses and Paddy Straw as Substrates.

The present experiment aimed at finding the suitability of some grasses as cost effective alternative substrates, for cultivation of one species of oyster mushroom viz., Pleurotus sajor caju (Fr.) Singer in eastern India. Relative efficacy three grasses viz., kash grass (Saccharum spontaneum L.), sabai grass (Eulaliopsis binata C.E. Hubb (Retz.)) and lemon grass (Cymbopogon citrates Stapf.) was tested by using each of them either as whole substrate or in combination with the conventional substrate i.e., paddy straw in 3:1, 1:1, and 1:3 ratios. Results revealed that the maximum yield of mushroom was recorded under paddy straw with biological efficiency of 85.9%. However, no significant difference in yield was found when 25% or 50% of the conventional substrate (paddy straw) was replaced by lemon grass and sabai grass. The results indicated that grasses which are available in plenty in the forests and wastelands of lateritic uplands of eastern India can be utilized successfully as promising substrate for the commercial cultivation of Pleurotus sajor caju.

Influence of Soil Ameliorants, Manures and Fertilizers on Bacterial Populations, Enzyme Activities, N Fixation and P Solubilization in Peanut Rhizosphere under Lateritic Soil.

The aim of this study was to investigate relative efficacy of different organic wastes like farmyard manure (FYM) and water hyacinth (WH) and industrial wastes like paper factory sludge (PFS) on balancing with chemical fertilizers (CF) along with soil ameliorants viz., lime (L) or rice husk ash (RHA), another industrial waste, on dry matter production and biological properties of the rhizosphere soil of peanut (Arachis hypogaea), grown as intercrop with sabai grass (Eulaliopsis binata) in acid lateritic soil. Population of symbiotic nitrogen fixing and phosphorus solubilizing bacteria, activity of dehydrogenase and phosphatase enzymes (i.e. acid and alkaline phosphomonoesterases), nitrogen accumulation in nodules and phosphorus solubilizing power of rhizosphere soil were measured after 25, 50, 75 and 100 days after sowing (DAS) of peanut for two years. Results showed significant effects of nutrient sources and growth stages of the crop on the microbial activities. Higher values of all the biological properties and plant growth parameters were recorded significantly under the integrated application of CF and any of the organic or industrial wastes over sole application of CF. Among three organic or industrial wastes WH was superior to others regarding microbial activities at 25 DAS, whereas PFS became superior at 50, 75 and 100 DAS. Application of lime or RHA improved the activity of dehydrogenase and alkaline phosphomonoesterase enzymes, while decreased acid phosphomonoesterase activity. This study revealed that integrated application of organic or industrial wastes, soil ameliorants and inorganic fertilizer, could improve the biological properties of an acid lateritic soil as well as the dry matter production of peanut, intercropped with sabai grass under lateritic soil.

Profile of pediatric dengue cases from a tertiary care hospital in Kolkata.

A study was conducted on the 52 serologically positive cases of dengue, admitted to the Dept. of Paediatrics, R.G. Kar Medical College & Hospital, from an outbreak in Kolkata. The most unusual feature observed in this study was that the rash in some cases was urticarial and intensely pruritic. The shock appeared early in the course of the disease and it was less commonly associated with bleeding (22%). One out of three dengue cases was a severe disease. It was not possible to predict a severe disease from the early symptomatology.

Morphological variation, histology, histochemistry and nutrient contents of Frankia root nodules in Casuarina equisetifolia.

Root nodules were collected from the existing plantation of Casuarina equisetifolia from three different agroclimatic zones of Tamil Nadu, India. Morphological variation, histology, histochemistry and nutrient contents of the nodules were studied. The nodules were found to be simple, dichotomously branched and coralloid. The stele in simple and branched nodules reached up to the tip of the nodules. Structure of Frankia nodules was described using histochemical methods to elucidate the nature of micro-symbiont within the nodules. The nodule consisted of a distinct periderm enclosing the cortex. The cortex showed patches of infected cells interspersed among the uninfected ones. Inside the cortex was a stele bound by an endodermis, which contained high amount of tannin. The micro-symbiont was observed in the nodule tisues in hyphal form and spore packed sporocorp was also seen in the cortex. The histochemical tests revealed the presence of proteins, insoluble polysaccharides and polyphenols.

Effect of angiotensin II on liposome uptake by the rat brain in vivo.

Studies have been performed to assess the possibility of using small unilamellar liposomes as therapeutic carriers to the brain of hypertensive rats. Rats were made temporal hypertensive by the infusion of angiotensin II (AII; 15 micrograms in 1 ml) through their right common carotid artery. Another control group was infused with physiological saline. Free 125iodine-BSA (125I-BSA) and 125I-BSA encapsulated liposomes (average diameter approximately equal to 100 nm) were injected in the tail vein 2 min after the infusion of AII or saline. Plasma radioactivity was monitored at different times up to 15 min when the cerebral uptake of 125I-BSA was determined. While a little variation in plasma clearance pattern of liposomes in hypertensive and control group was noticed, the uptake by cerebral tissues was markedly higher in hypertensive group. Analysis of pharmacokinetic parameters in relation to cerebral uptake indicated AII induced a short term opening of the blood-brain barrier (BBB) resulting in an increased cerebral uptake. Positively charged liposomes was found to be most effective in hypertensive state.

Targeting of piperine intercalated in mannose-coated liposomes in experimental leishmaniasis.

The leishmanicidal property of piperine intercalated in liposomes and in mannose-coated liposomes was tested in experimental visceral leishmaniasis in hamsters. Mannose-coated liposomal piperine eliminated intracellular amastigotes of Leishmania donovani in splenic macrophages much more efficiently than did the liposomal piperine or free piperine. At a dose equivalent to 6 mg/kg body wt every 4th day for a total of 4 doses in 12 days, the mannose-coated liposomal piperine was found to reduce spleen parasite load to the extent of 90% in comparison to that achieved by liposomal piperine (77%) or free piperine (29%). Histological examination of spleen and liver function tests showed that the toxicity of piperine was reduced when mannosylated liposomal piperine was administered.

Cloning and expression of SAT-3 involved in SA-Le(x) biosynthesis: inhibition studies with polyclonal antibody against GST-SAT-3 fusion protein.

The SAT-3 activity (CMP-NeuAc:Gal beta 1-4GlcNAc beta 1-3 Gal beta 1-4Glc-ceramide alpha 2-3 sialytransferase) involved in the biosynthesis of sialy Le(x) has been characterized in human colon carcinoma cells and embryonic chicken brains. Using RT-PCR-based strategy, we have isolated partial cDNA clones of SAT-3 from ECB and Colo-205 mRNAs. Suitable primers from sialylmotif and N-terminal sequence of human placenta SAT-3 (HP-SAT-3) were used. The 800 bp cDNA fragment encoding a region (90%) of alpha 2-3 sialyltransferase (SAT-3) catalytic domain from ECB has been expressed as a glutathione S-transferase (GST) soluble fusion protein (62 kDa) in E. coli and purified over glutathione-agarose affinity matrix. Polyclonal antibody has been produced against affinity-purified catalytic domain of SAT-3 (GST-SAT-3 fusion protein). A concentration-dependent polydonal antibody binding to native SAT-3 has also been demonstrated by measuring the residual SAT-3 activity in the enzyme fractions from Colo-205. The marked inhibition (> 80%) of SAT-3 activity and relatively less inhibition (< 20%) of SAT-4 activity (CMP-NeuAc:GgOse4Cer alpha 2-3sialyl transferase) suggests strongly the existence of two different gene products (SAT-3 and SAT-4) in human colon carcinoma Colo-205 cells and in embryonic chicken brains (ECB).

Ceramide glycanase from rat mammary tissues: inhibition by PPMP(D-/L-) and its probable role in signal transduction.

A ceramide glycanase (CGase activity has been characterized from lactating rat mammary tissue which cleaves the glycosidic bond between sphingosine and the glucose chain of a glycosphingolipid (GSL) thus liberating the intact oligosaccharide chain from a GSL. The majority (65%) of the hydrolase activity was detected in the supernatant fraction when the rat mammary tissue homogenate was centrifuged at 100,000 x g. Attempts to purify the enzyme indicated that the CGase protein is of hydrophobic nature as it binds to hydrophobic columns. The enzyme has been partially purified using hydrophobic columns in tandem. The partially purified protein was found to be immunoreactive to the antibody raised against the purified clam CGase. The immunostained band corresponded to a 64 kDa protein as also found with the clam enzyme. This immuno cross-reactivity indicated probable structural similarities between CGase proteins isolated from widely separated species in the evolutionary tree. The rat CGase was found to have a specific detergent requirement for optimal activity, and the pH optimum was found to be between 5 and 6. The enzyme activity is partially heat stable. It is not a divalent cation requiring enzyme; however, the activity is totally inhibited in the presence of mercury, indicative of a sulfhydryl group in the active site of the enzyme. The rat mammary CGase activity is inhibited in the presence of both D- and L-PPMP (1-phenyl-2-hexadecanoylamino-3-morpholino-1-propanol. HCl), homologs of PDMP (1-phenyl-2-decanoylamino-3-morpholino-1-propanol. HCl), a well-known inhibitor of GlcT-1 (Ceramide: UDP-Glc Glucosyltransferase), an enzyme in the glycolipid synthetic pathway. The inhibition seems to be of a competitive nature and the same type of inhibition is also observed with clam CGase. The CGase activity was found to be highest in lactating tissue compared to the activity found in either pregnant or post-lactating rat mammary tissues. Tissue survey indicated the presence of high levels of CGase in lactating rat liver, uterus, and ovary; moderate activity was detected in kidney and spleen. Both virgin and male rat mammary tissue also indicated a basic level of CGase activity. However, newborn spleen and mammary tissue showed a comparable level of activity to that found in lactating rat tissues. This report is mainly concerned with the characterization of CGase activity from a mammalian source and its importance in cellular processes.

The changing profile of Plasmodium falciparum malaria.

Resurgence of malaria has been noted in the Rohtak district (Haryana, India) after the recent floods. The profile of 66 patients of P. falciparum infection who were admitted to our hospital over one month in October 1995 is reported. While only a minority of cases (< 15%) presented with an uncomplicated course, all others developed one or more complication(s), some of them very rare. The usual manifestations viz, cerebral malaria, black water fever and algid malaria seen in the past were observed in less than half the patients. The remaining presented with unusual complications like haemolytic anaemia (46.2%), severe anaemia (37.9%), thrombocytopaenia (18.2%), pancytopaenia (6%), adult respiratory distress syndrome (4.5%) often not seen in sporadic cases of falciparum malaria which occurred in the past in this district. Similarly all deaths (15.1%) were noted in patients with rarer manifestations and only one patient died of cerebral malaria. This study confirms the occurrence of severe and complicated falciparum malaria in this part of the country.

Glycoside-bearing liposomal delivery systems against macrophage-associated disorders involving Mycobacterium leprae and Mycobacterium tuberculosis.

Asiaticoside, a plant glycoside with rhamnose as end sugar and having microbicidal properties was tested against Mycobacterium leprae and Mycobacterium tuberculosis both in vivo and in vitro. As rhamnose is reported to have no tissue specificity, corchorusin D having glucose as end sugar was used for targeting with an equimolar proportion of asiaticoside in liposomal form for testing the drug value. Results showed that liposomal asiaticoside had better microbicidal property against M. leprae and M. tuberculosis when compared to that of free asiaticoside whereas liposomes containing asiaticoside and corchorusin D were found to be equally or more active in comparison to liposomal asiaticoside alone. It is inferred that appropriate glycosides, if used in liposomal form (incorporated or covalently grafted) have enhanced drug efficacy and such glycoside bearing liposomes as targeted delivery systems could be used for chemotherapeutic control of several other diseases.

Regulation of expression of cell surface neolacto-glycolipids and cloning of embryonic chicken brain GalT-4 (UDP-Gal: GlcNAc-R beta 1-4 galactosyltransferase).

The biosynthesis of GM1 ganglioside (Gal beta 1-3GalNAc beta 1-4 (NeuAc alpha 2-3) Gal beta 1-4Glc-Cer) and nLcOse4Cer is catalyzed by two different beta-galactosyltransferases GalT-3 (UDP-Gal: GM2 beta 1-3GalT) and GalT-4 (UDP-Gal: Lc3 beta 1-4GalT) respectively. Solubilized GalT-3 and GalT-4 have been purified 3,000-fold and 22,000-fold, respectively, from 11-19-day-old embryonic chicken brain. The purified GalT-3 transfers galactose to GM2 very actively (Km 33 microM), whereas acetyl GM2 is not an active substrate (Km 350 microM), GalT-3 and GalT-4 are classified as HYCARS (hydrophobic recognition site) and CARS (carbohydrate recognition sites), respectively. An anion-transport inhibitor DIDS (diisothiocyanato stilbene 4,4'-disulphonate), irreversibly inhibits both GalT-3 and GalT-4 activities by binding to a UDP binding site. Polyclonal antibodies against purified GalT-3 and GalT-4 inhibited these two purified activities and showed no cross reactivity on the western blots. RNA-PCR of 11-day-old embryonic chicken brain mRNA with PCR primers designed from the homologous coding regions of cloned sequences of beta 1-4 GalT of human, bovine, and murine-tissues produced a -600 bp cDNA fragment. Dideoxy-sequences of this fragment reveals it to be 74% similar to the nucleotide sequences of the cloned beta 1-4GalT from human liver. The cloned-600 bp cDNA was used to identify two mRNA transcripts (1.4 and 2.3 kb) from ECB by Northern blot analysis and four genomic DNA EcoRI fragments (18.6, 12.9, 10.5 and 3.7 kb) on a Southern blot analysis.

Biosynthesis and regulation of Le(x) and SA-Le(x) glycolipids in metastatic human colon carcinoma cells.

This report concerns the stepwise biosynthesis in vitro of Sialyl Lewis X, (SA-Le(x)), a carcinoembryonic antigen, in human colon carcinoma KM12 cells exhibiting different metastatic behaviors. The significance of SA-Le(x) has become even more apparent since the detection of its terminal epitope NeuAc(alpha 2-3)Gal beta 1-4(Fuc alpha 1-3)GlcNAc-, as the binding ligand of the selectin family member ELAM-1. The activity level of galactosyltransferase GalT-4 which catalyzes the formation of core nLcOse4Cer (Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc-Cer) is very high in all the metastatic lines tested with highly metastatic lines (KM12-SM) exhibiting the highest activity. The same activity pattern for galactosyltransferase is also observed when tested with iLcOse5Cer (GlcNAc beta 1-3nLcOse4Cer), the precursor for polylactosamine glycolipid. Sialyltransferase SAT-3 which catalyzes the formation of LM1 (NeuAc alpha 2-3nLcOse4Cer), the precursor for SA-Le(x), is also present in all the metastatic cell lines although the activity levels are much lower compared to galactosyltransferase. The fucosyltransferase FucT-3, which catalyzes the formation of R'-Gal-Fuc(alpha 1-3)GlcNAc-R linkage, is active with both nonsialylated substrate, nLcOse4Cer, and sialylated substrate, LM1 (NeuAc alpha 2-3nLcOse4Cer) with the formation of either Le(x) (Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc-Cer) or SA-Le(x) (NeuAc alpha 2-3nLcOse4Cer). However, the sialylated substrate LM1 is preferred to enzymatic activity since it exhibited lower Km (46 microM) than that of nLcOse4Cer (67 microM).(ABSTRACT TRUNCATED AT 250 WORDS)

Lipid peroxidation in hepatic microsomal membranes isolated from mice in health and in experimental leishmaniasis.

Normal hepatic microsomal membranes when exposed in vitro to different free radicals, cause membrane damage by lipid peroxidation which could be monitored by the analysis of malonaldehyde formation and measurement of membrane microviscosity. Lipid peroxidation in vivo, when examined in hepatic microsomal membranes in experimental Leishmaniasis, reveals a direct relationship between membrane microviscosity and the extent of lipid peroxidation. Scavengers of free radicals and peroxides such as superoxide dismutase (SOD) for O2.-, mannitol for (OH.) and catalase for H2O2 in modest amounts were used for preventing the membrane damage caused by lipid peroxidation.

(Dr. P. C. Sen Award on rural health practice). Infant and early childhood mortality in some rural ICDS blocks of west Bengal.

This one year prospective study was carried out to determine the CBR, CDR and different underfive mortality rates in some selected rural ICDS blocks of West Bengal and also to find out the common causes of infant and childhood mortality in these areas. It was revealed that most of the above mentioned indicators in ICDS blocks were considerably lower than that of National figures, but more or less similar to those of rural Bengal as recorded in routine Govt. reports. Compared to the picture of rural West Bengal, both still birth & perinatal mortality rates were found higher in all ICDS blocks. The causes of mortality trends among under fives' were found similar to that of rural India pattern i.e. prematurity, acute respiratory infections and diarrhoea being the leaders. Neonates, who contributed the largest share of infant mortality died mainly due to prematurity. So, observations like high perinatal mortality & still birth rates, huge proportion of neonatal death during infancy, same IMR but low 5MR, and death of neonates due to prematurity etc. claim the necessity of improving maternal component of ICDS services, their nutritional care in particular.