A pilot survey on hand washing among some communities of West Bengal.

A cross-sectional observational study was carried out between April to May 2006 by interview method and observation technique with the objective to know the knowledge regarding hand washing in the community and it was done in the slum and nonslum urban areas and also one rural area. The result shows that in urban slum area 98% washed their hands with soap after defecation; Only 36%, 16% and 2% washed their hands with soap before meal, before serving food and before cooking respectively. However, it was observed that 69% used soap and water for hand washing after cleaning the child's faeces. In rural area 71% used soap and water after defecation while 26% used mud or ash. Only 13%, 1%, 1% and 5% used soap and water before meal, before serving food, before cooking and after cleaning the child's faeces. 82.35% of respondents in non slum area and 89% of respondents in rural area considered that diarrhoea and dysentery could be prevented by hand washing while they did not give importance to hand washing in prevention of diarrhoea over other methods like cleanliness, boiling and purification of water. ARI was much higher (25.72%) in rural area followed by slum area (13.77%) and non-slum area (3.87%). Out of 30 observations among 302 interview made on hand-washing only first step i.e. palm washing (transient rubbing the palm with soap) was followed by all the participants observed. Time taken for such hand-washing was only around five seconds (ideal 15-30 seconds) in urban slum and rural areas while in non slum area it varied between 7-10 seconds on an average. No one followed any other steps of hand-washing, recommended by IFH.

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).

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)