Levels of immunoglobulins and complement C3 in protein-energy malnutrition.
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Case-Control Studies↗
Biomedical subjects
B K Das
Publications and source records attributed to B K Das.
Class I histiocytosis: response to combination of etoposide and prednisolone.
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Child, Preschool↗
Screening of sputum: an experience in a tertiary care hospital.
In order to reduce the work load on the clinical laboratory, it has been recommended that sputum samples, before accepting for culture, should be looked for the presence of polymorphs and squamous epithelial cells. (An appropriate sample should have more than 25 polymorphs per low power field and less than 10 epithelial cells per low power field; others are labelled as inappropriate). We examined this criteria for it's suitability on 1043 samples received at the clinical bacteriology laboratory at the All India Institute of Medical Sciences (AIIMS) for a period of one year (September, 1996 to August, 1997). Four hundred samples were found appropriate while 643 were inappropriate as per recommended criteria. Amongst the 400 appropriate samples, 215 were culture positive and 185 grew normal flora. However, we found that out of 643 inappropriate samples, 195 were culture positive (p < 0.01, statistically significant). The data was further divided into hospitalized and OPD cases. The distribution of culture positive cases in the appropriate samples was again found to be highly significant. It is apparent from our results that an attempt to process an inappropriate sample does not provide useful information.
Bacteriological Techniques↗
Neem (Azadirachta indica) extract as an antibacterial agent against fish pathogenic bacteria.
Aquaneem, an emulsified product prepared from the neem (A. indica) kernel was tested against four pathogenic bacteria of fish (i.e. Aeromonas hydrophila, Pseudomonas fluorescens, Escherichia coli and Myxobacteria spp.) to test its efficacy as an antibacterial agent. Growth inhibitory property of the product at 10, 15 and 20 ppm has been noticed and recorded. The percentage reduction of bacterial cell population was noted to be maximum on 9th day at 20 ppm concentration (i.e. 70.14%, 74.15% and 61.75% for A. hydrophila, P. fluorescens and E. coli respectively) with the only exception of myxobacteria which showed maximum reduction percentage (63.90%) on 15th day. Among all the bacteria tested A. hydrophila, P. fluorescens and Myxobacteria spp. exhibited maximum sensitivity to Aquaneem in terms of percentage reduction of bacterial cell population in comparison to E. coli.
Aeromonas hydrophila↗
Thermodynamic and kinetic characterization of calf lens gammaF-crystallin.
Gamma-crystallin is reported to be conformationally stable because of its internal structural symmetry, and gammaF (gammaIVa) is the most stable among the various gamma-crystallin gene products. However, there is no detailed report on its thermodynamic and kinetic stability. In the present study, detailed unfolding of gammaF-crystallin was investigated by equilibrium and kinetics methods with fluorescence and far-UV CD spectroscopic measurements. The GdnHCl-induced unfolding curves probed by Trp emission maximum and intensity showed a sharp single-step transition. Upon widening the unfolding transition with the use of urea in 1.5 M GdnHCl, a more proper fit for thermodynamic analysis was obtained. GammaF-Crystallin underwent a straightforward two-state process (N <==> U) without showing any measurable amount of intermediate. The conformational stability, as measured by deltaG(D)H2O (approximately 9 kcal/mol), indicates that gammaF-crystallin is a very stable protein. The high activation energy deltaG++H2O (approximately 24 kcal/mol), calculated from unfolding kinetics monitored by far-UV CD at 218 nm, also indicates that the native and unfolded states are separated by a high activation energy barrier.
Animals↗
Concentration of total serum IgE in parasitized children and the effects of the antiparasitic therapy on IgE levels.
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Animals↗
Blood sugar and serum insulin response in protein-energy malnutrition.
Blood sugar and serum insulin levels in the fasting state and following an oral glucose load in children with protein-energy malnutrition (PEM) were studied. Twenty-nine children with PEM (15 marasmus, 7 kwashiorkor, and 7 marasmic kwashiorkor) and eight healthy children aged between 6 and 60 months were the subjects of the study. Fasting samples were collected after a 6 h fast. Post-glucose samples were collected after an oral glucose load of 1.75 g/kg. Serum insulin was estimated by radioimmunoassay and glucose by the glucose oxidase method. In malnourished children, the mean fasting blood glucose levels were significantly lower. Two hours following an oral glucose load, only marasmus and marasmic kwashiorkor patients showed significantly higher blood glucose levels. Similarly, the fasting serum insulin levels were significantly lower in malnourished children. Two hours after the oral glucose load, serum insulin levels increased significantly in malnourished children but fell well short of the control values. The insulin:glucose ratio was consistently low in all cases but was more marked in PEM patients, both in the basal state as well as oral glucose loading
Blood Glucose↗
Parturient fundal height as a predictor of low birth weight.
The usefulness of parturient fundal height as a predictor of low birth weight (LBW) was studied in 151 singleton pregnancies. The symphysis to fundus (S-F) distance was measured in early labour and was plotted against the birth weight of the newborns. A significant positive correlation was observed between the two parameters (r = +0.740). For the prediction of LBW, the critical limit of the fundal height was 31 cm, meaning that a S-F distance of more than 31 cm is reasonably safe while lesser values predict a higher chance of a LBW infant. From these observations, the use of a colour-coded tape by peripheral health workers and traditional birth attendants is suggested: red for S-F < 28 cm, yellow for S-F = 28-31 cm, and green for S-F > 31 cm. Mothers who have a fundal height in the 'red zone' are at risk of delivering LBW infants.
Adult↗
Blood glucose and serum insulin response in protein-energy malnutrition following nutritional rehabilitation.
Fifteen children with protein-energy malnutrition and eight healthy children between the ages of 6 months and 60 months were studied for blood glucose and serum insulin levels in the fasting state and 120 minutes following oral glucose load. The measurements were repeated after normalization of the body weight following 6 weeks of nutritional rehabilitation. The fasting blood glucose increased significantly in the post-treatment period and became comparable to the control value. The abnormal blood glucose response to oral glucose load also normalized. The serum insulin level rose significantly (p < 0.001) in the post-treatment period but failed to normalize. The response to oral glucose load was similar. The observed lower insulin response, despite normoglycemia, after 6 weeks of nutritional rehabilitation indicates persistence of hormonal imbalance which may need a longer duration of rehabilitation for full recovery.
Blood Glucose↗
Lack of maternal antibodies to P serotypes may predispose neonates to infections with unusual rotavirus strains.
Rotavirus (RV) strains infecting newborns often have unique neutralization antigens (P serotypes) on their outer capsids that are distinct from those found on RV strains that cause diarrhea in older children. We examined the hypothesis that unusual RV strains preferentially infect newborns because the newborns lack maternal neutralizing antibodies to these strains. To test this hypothesis, sera and saliva samples collected from neonates infected with 116E-like (P[11]G9) strains in the maternity ward of the All India Institute of Medical Sciences (AIIMS) hospital in New Delhi were tested for neutralizing antibodies against common RV strains and those infecting newborns and these titers were compared with those of newborns who did not become infected (controls). The infected neonates had significantly lower levels of cord blood neutralizing antibodies to 116E than the controls, suggesting that immunity to neonatal RV infection is acquired transplacentally through maternal antibodies. Further, this study confirmed the immunogenicity of the AIIMS neonatal strain 116E, a vaccine candidate, in its ability to evoke a potent RV-specific immunoglobulin A and neutralizing antibody response in serum and saliva among the infected babies. Our findings have important implications for the development of an effective RV vaccine. In India, where G9 strains are common in the community, the use of 116E as a vaccine, together with the rhesus tetravalent vaccine, may provide a broader protection against all the circulating RV serotypes, including serotype G9, which is not represented in the current rhesus RV tetravalent vaccine (G1-G4).
Antibodies, Viral↗
Fluorescence and immunochemical studies of advanced glycation-related lens pigments.
PURPOSE: To establish whether advanced glycation is the major mechanism for yellowing of lens proteins. METHODS: Synchronous fluorescence (SF) and immunochemical assays were used to study glycation in vitro and in vivo. In the in vitro study, advanced glycation end products (AGEs) were prepared and used as antigens to induce antibodies to AGEs. The in vitro AGEs and classified nuclear cataracts were analyzed by SF and immunochemical assays. RESULTS: In vitro AGEs generated from various glycating agents and carrier proteins displayed strong SF above 350 nm; the spectra were well resolved with major bands at 380 nm and 420 nm. Samples from human lenses manifested a band at 395 nm in addition to the two bands shown by in vitro AGEs. SF intensity is greater for the water-insoluble (WI) than water-soluble (WS) fraction, but both increased with increasing nuclear color. The immunoreactivity data also showed that the WI fraction contained more AGEs than the WS fraction and that the amount of AGEs increased with increasing nuclear color. CONCLUSIONS: Fluorescence and immunoassays indicated that pigmented AGEs contributed to yellowing of the crystalline lens nucleus.
Animals↗
Detection and characterization of alpha-crystallin intermediate with maximal chaperone-like activity.
Lens alpha-crystallin has been reported to act like a chaperone molecule, with the chaperone-like activity enhanced by partial unfolding. The nature of the partial unfolding, however, is not fully understood. In this project, the unfolding and refolding process of alpha-crystallin was studied with guanidine hydrochloride (GdnHCl). Trp fluorescence (tertiary structure) and far-ultraviolet circular dichroism (UVCD) (secondary structure) demonstrated the presence of an intermediate in the unfolding pathway. ANS (1-anilino-8-naphthalenesulfonate) fluorescence clearly indicated a two-step transition in the unfolding-refolding process and showed that maximum hydrophobicity of the alpha-crystallin occurred at 0.8-1.0 M GdnHCl. This alpha-crystallin intermediate appears to be in a molten globule state; conformational study by near- and far-UVCD measurements indicated that alpha-crystallin intermediate exhibited tertiary structure which was significantly altered from that of the native protein, but had nearly the same secondary structure. Quaternary structure (size of aggregate) of the intermediate also remained unchanged from that of the native protein, as shown by FPLC size exclusion chromatography. The maximal hydrophobicity of the alpha-crystallin intermediate in the unfolding-refolding pathway was accompanied by maximal protection of betaH-crystallin from aggregation. However, an adverse effect of partial unfolding is that the alpha-crystallin intermediate aggregates at high concentrations. Together, these results clearly demonstrated the biological significance of the alpha-crystallin intermediate: it is a more effective chaperone than native alpha-crystallin.
Animals↗
Conformational and functional differences between recombinant human lens alphaA- and alphaB-crystallin.
Human and other mammalian lens proteins are composed of three major crystallins: alpha-, beta-, and gamma-crystallin. alpha-Crystallin plays a prominent role in the supramolecular assembly required to maintain lens transparency. With age, the crystallins, especially alpha-crystallin, undergo posttranslational modifications that may disrupt the supramolecular assembly, and the lens becomes susceptible to other stresses resulting in cataract formation. Because these modifications occur even at a relatively young age, it is difficult to obtain pure, unmodified crystallins for in vitro experiments. alpha-Crystallin is composed of two subunits, alphaA and alphaB. Before the application of recombinant DNA technology, these two alpha-crystallin subunits were separated from calf lens in the denatured state and reconstituted by the removal of the denaturant, but they were not refolded properly. In the present studies, we applied the recombinant DNA technology to prepare native, unmodified alphaA- and alphaB-crystallins for conformational and functional studies. The expressed proteins from Escherichia coli are in the native state and can be studied directly. First, alphaA and alphaB cDNAs were isolated from a human lens epithelial cell cDNA library. The cDNAs were cloned into a pAED4 expression vector and then expressed in E. coli strain BL21(DE3). Pure recombinant alphaA- and alphaB-crystallins were obtained after purification by gel filtration and DEAE liquid chromatography. They were subjected to conformational studies involving various spectroscopic measurements and an assessment of chaperone-like activity. alphaA- and alphaB-crystallins have not only different secondary structure, but also tertiary structure. 1-Anilino-8-naphthalene sulfonate fluorescence indicates that alphaB-crystallin is more hydrophobic than alphaA-crystallin. The chaperone-like activity, as measured by the ability to protect insulin aggregation, is about 4 times greater for alphaB- than for alphaA-crystallin. The resulting data provide a base line for further studies of human lens alpha-crystallin.
Blotting, Western↗
Sequence analysis demonstrates that VP6, NSP1 and NSP4 genes of Indian neonatal rotavirus strain 116E are of human origin.
We have sequenced the genes encoding the inner capsid protein VP6 and the nonstructural proteins NSP1 and NSP4 of the Indian neonatal serotype P8[11]G9 human/bovine reassortant candidate vaccine rotavirus strain 116E. These three genes share a high degree of sequence and deduced amino acid homology with human prototype strain Wa. Our results confirm and extend those of previous RNA-RNA hybridization studies which suggested that these genes are of human origin, and will facilitate examination of the host immune response to 116E induced by natural infection and vaccination.
Amino Acid Sequence↗
Heat-induced conformational change and increased chaperone activity of lens alpha-crystallin.
PURPOSE: Alpha-crystallin is the major structural protein of the eye lens known to have chaperone-like activity. Our objective is to elucidate the nature of the thermal transition that alpha-crystallin undergoes at 60 degrees C and the effect of this transition on the chaperone activity. METHODS: FPLC size exclusion chromatography, far- and near-ultraviolet circular dichroism, and tryptophan (Trp) and 1-anilino-8-naphthalenesulfonate (ANS) fluorescence were used to study conformational change. Turbidity of dithiothreitol (DTT)-reduced insulin was used to study chaperone activity. RESULTS: The thermal transition was identified as a conformational change in mainly tertiary (partial unfolding) and quaternary high-molecular-weight (HMW) aggregation structures, along with a loss of 10 percentage points of secondary structure (beta-sheet). Initial partial perturbation in tertiary structure increased chaperone activity, but the increase was less in the HMW aggregate. Similar results were observed in in vivo-formed HMW alpha-crystallin. CONCLUSIONS: The conformational change and HMW aggregation of alpha-crystallin observed at 60 degrees C, as well as in vivo-formed HMW aggregates, increased chaperone activity.
Anilino Naphthalenesulfonates↗
Immunoglobulins and circulating immune complexes in nephrotic syndrome.
Serum immunoglobulin (IgG, IgA and IgM) levels were measured in 40 children with idiopathic nephrotic syndrome and 20 healthy controls. Significantly decreased IgG (P < 0.001), normal IgA and increased IgM (P < 0.05), averaging 92 per cent, 99 per cent and 104 per cent of normal mean, respectively, were observed in active nephrotic syndrome (ANS). The mean immunoglobulin levels did not differ significantly among different subgroups of ANS cases. In remission, a significant increase in IgG (P < 0.01), a decrease in IgA (P < 0.05) and in IgM (P < 0.01) levels were found in comparison to their corresponding ANS values. Their concentrations were significantly abnormal also when compared with controls. The circulating immune complexes (CIC), detected by polyethylene glycol (PEG) per cent index, demonstrated that overall mean serum level was significantly higher (P < 0.01, 168 per cent of normal) in ANS patients. Among different subgroups, the mean level was significantly elevated (P < 0.001, 205 per cent of normal) only in relapsing nephrotic syndrome cases. Thus, the abnormal levels of immunoglobulins were found both in active as well as in remission stage of the disease, while elevated levels of CIC were seen only in patients with relapse.
Antigen-Antibody Complex↗
Sequence analysis of NSP4 gene of human rotavirus allows classification into two main genetic groups.
The rotavirus nonstructural glycoprotein NSP4 may represent the first identified viral enterotoxin. We have sequenced reverse transcription-polymerase chain reaction (RT-PCR)-generated fragments of 16 NSP4 genes of human rotavirus (HRV) strains from six different countries, representing seven different G and P type combinations. Based on the amount of sequence divergence between these and 11 previously sequenced NSP4 genes of human and animal rotaviruses, three distinct genetic groups could be recognized. Most strains within a group were closely related to each other at the nucleotide (nt) and amino acid (aa) levels (usually <10% divergence) but more distantly related (maximum 30.0% nt divergence and 24.7% aa divergence) to members of the other groups. Intergroup variation occurred in two highly variable regions of NSP4 (aa 16-34 and aa 131-148). The NSP4 "toxic peptide" (aa 114-135) exhibited aa variation at its carboxy terminus both within and between genetic groups. The largest group (genetic group II) contained HRV strains of subgroup II specificity (including genotypes P[8]G1, P[8]G3, P[6]G3, and P[8]G5 and serotype P8[11]G9), and the smaller group (genetic group I) contained HRV strains of subgroup I specificity (genotype P[4]G2). The NSP4 sequence of the rhesus rotavirus vaccine strain was distinct from all other strains and formed the third group (genetic group III). The NSP4 genes of animal rotaviruses UK, NCDV, and SA11 (genetic group I) and YM (genetic group II) and two possible human-animal rotavirus reassortant strains, Brazilian P[8]G5 and Indian P[11]G9 (genetic group II), could also be classified into one of these groups, suggesting a close evolutionary relationship between human and animal NSP4 genes. These results will facilitate studies of the host immune response to NSP4, which may be relevant to future HRV vaccine design.
Amino Acid Sequence↗
Detection of rice tungro bacilliform virus in field and glasshouse samples from India using the polymerase chain reaction.
Rice tungro bacilliform virus (RTBV) together with rice tungro spherical virus (RTSV) is the causal agent for the rice tungro disease. A rapid technique was developed to detect RTBV DNA in the crude extract of freshly collected leaf samples by polymerase chain reaction (PCR). This technique can detect the viral DNA in 1000-fold diluted leaf extract. Detection has been possible in samples stored upto 5 days after the collection. This technique may have wide application for the field diagnosis of RTBV infection.
Badnavirus↗