Biomedical subjects
Publications and source records attributed to S Pal.
Tragia involucrata has been widely used in the traditional medicinal system for the treatment of a variety of diseases. The effect of methanolic extract of T. involucrata was studied in different experimental animal models and it was revealed that the extract possesses significant analgesic and anti-inflammatory activity.
A neural network model is proposed for computation of the convex-hull of a finite planar set. The model is self-organizing in that it adapts itself to the hull-vertices of the convex-hull in an orderly fashion without any supervision. The proposed network consists of three layers of processors. The bottom layer computes the activation functions, the outputs of which are passed onto the middle layer. The middle layer is used for winner selection. These information are passed onto the topmost layer as well as fed back to the bottom layer. The network in the topmost layer self-organizes by labeling the hull-processors in an orderly fashion so that the final convex-hull is obtained from the topmost layer. Time complexities of the proposed model are analyzed and are compared with existing models of similar nature.
ExoS is a type III cytotoxin of Pseudomonas aeruginosa, which modulates two eukaryotic signalling pathways. The N-terminus (residues 1-234) is a GTPase activating protein (GAP) for RhoGTPases, while the C-terminus (residues 232-453) encodes an ADP-ribosyltransferase. Utilizing a series of N-terminal deletion peptides of ExoS and an epitope-tagged full-length ExoS, two independent domains have been identified within the N-terminus of ExoS that are involved in intracellular localization and expression of GAP activity. N-terminal peptides of ExoS localized to the perinuclear region of CHO cells, and a membrane localization domain was localized between residues 36 and 78 of ExoS. The capacity to elicit CHO cell rounding and express GAP activity resided within residues 90-234 of ExoS, which showed that membrane localization was not required to elicit actin reorganization. ExoS was present in CHO cells as a full-length form, which fractionated with membranes, and as an N-terminally processed fragment, which localized to the cytosol. Thus, ExoS localizes in eukaryotic cells to the perinuclear region and is processed to a soluble fragment, which possesses both the GAP and ADP-ribosyltransferase activities.
Calcium and calcium-dependent systems have been long implicated in the induction of epilepsy. We have previously observed that intracellular calcium ([Ca2+]i) levels remain elevated in cells undergoing epileptogenesis in the hippocampal neuronal culture (HNC) model. In this study, we employed the hippocampal neuronal culture (HNC) model of in vitro 'epilepsy' which produces spontaneous recurrent epileptiform discharges (SREDs) for the life of the neurons in culture to investigate alterations in [Ca2+]i homeostatic mechanisms that may be associated with the 'epileptic' phenotype. [Ca2+]i imaging fluorescence microscopy was performed on control and 'epileptic' neurons with two different fluorescent dyes ranging from high to low affinities for [Ca2+]i. We measured baseline [Ca2+]i levels and the ability to restore resting [Ca2+]i levels after a brief 2-min exposure to the excitatory amino acid glutamate in control neurons and neurons with SREDs. Neurons manifesting SREDs had statistically significantly higher baseline [Ca2+]i levels that persisted for the life of the culture. In addition, the 'epileptic' phenotype was associated with an inability to rapidly restore [Ca2+]i levels to baseline following a glutamate induced [Ca2+]i load. The use of the low affinity dye Fura-FF demonstrated that the difference in restoring baseline [Ca2+]i levels was not due to saturation of the high affinity dye Indo-1, which was utilized for evaluating the [Ca2+]i kinetics at lower [Ca2+]i levels. Peak [Ca2+]i levels in response to glutamate were the same in both 'epileptic' and control neurons. While [Ca2+]i levels recovered in approximately 30 min in control cells, it took more than 90 min to reach baseline levels in cells manifesting SREDs. Alterations of [Ca2+]i homeostatic mechanisms observed with the 'epileptic' phenotype were shown to be independent of the presence of continuous SREDs and persisted for the life of the neurons in culture. Epileptogenesis was shown not to affect the degree or duration of glutamate induced neuronal depolarization in comparing control and 'epileptic' neurons. The results indicate that epileptogenesis in this in vitro model produced long-lasting alterations in [Ca2+]i regulation that may underlie the 'epileptic' phenotype and contribute to the persistent neuroplasticity changes associated with epilepsy.
Sialic acids typically present as terminal sugars of oligo-saccharides are reported to be modified by O-acetylation at the C-9 position on lymphoblasts of childhood acute lymphoblastic leukemia (ALL) patients (Sinha et al., 1999a, Leukaemia, 13, 119-125). We now report high titers of IgG antibodies directed against O-acetylated derivatives of sialic acids (O-AcSA) in serum of ALL patients. These antibodies were purified using bovine submaxillary mucin (BSM) and the IgG distribution was confined to IgG(1)and IgG(2)subclasses; their binding was totally abolished with de-O-acetylation confirming their specificity towards O-AcSA determinants. Flow cytometry demonstrated binding of these antibody fractions to peripheral blood mononuclear cells (PBMC) of both T- and B-ALL patients having increased cell surface 9-O-AcSA determinants. Western blotting of membranes derived from PBMC of ALL patients confirmed binding of the antibody to O-acetylated sialoglycoconjugates corresponding to 144, 135, 120, 90, and 36 kDa whereas binding to PBMC from normal individuals corresponded to 144 and 36 kDa. Specificity of the antibody fraction towards 9-O-AcSA was substantiated by hemagglutination and hemagglutination-inhibition assays. The antibody purified from ALL serum selectively mediates complement dependent cytolysis of lymphoblasts expressing O-AcSAs and thereby possibly confers passive protection. The enhanced anti O-AcSA antibody levels allowed for development of a serodiagnostic assay (BSM-ELISA) specific for ALL. Minimal crossreactivity was observed with other hematological disorders like acute myeloid leukemia (n = 16), chronic myeloid leukemia (n = 6), chronic lymphocytic leukemia (n = 7) and non-Hodgkin's lymphoma (n = 3) as well as normal healthy individuals (n = 28). The BSM-ELISA therefore provides a simple, noninvasive alternative diagnostic approach for ALL and merits clinical consideration.
Elicited macrophages from 129sv mice with a functional deletion of the natural-resistance-associated macrophage protein 1 gene (Nramp1) were shown to be as susceptible as wild-type mice to infection with the Chlamydia trachomatis mouse pneumonitis and L3 serovars and to Chlamydia pneumoniae. Furthermore, the two groups of mice were shown to be similarly susceptible to an intranasal infection with these microorganisms. In conclusion, the Nramp1 gene does not appear to play a major role in the regulation of the susceptibility of mice to a chlamydial infection.
The LDL receptor plays a pivotal role in the clearance of pro-atherogenic lipoproteins, and LDL receptor deficiency may be the underlying cause of several primary and secondary dyslipidaemic conditions. Intervention strategies are often targeted to increase hepatic LDL receptor expression. It is difficult to quantitate hepatic LDL receptor activity and to monitor changes post-therapy. In order to avoid liver biopsy, human skin fibroblasts or circulating mononuclear cells have often been used as surrogate markers for the hepatic receptor. Fibroblasts, and particularly mononuclear cells, are relatively easy to isolate and can be stored for extensive lengths of time without significant loss of LDL receptor expression. Leucocytes or fibroblasts are normally probed with isotopically or gold-labelled LDL. However, the specific activity of the LDL conjugate is usually too low to enable accurate quantitation of differences, or changes, in LDL receptor expression. In this study, we describe an enhanced colloidal gold-labelling procedure for the detection of LDL receptor binding activity. The binding of colloidal gold-labelled chylomicron remnants to human hepatocytes (HepG2 cells) was compared with that of gold-conjugated LDL. Labelled remnants bound specifically to a cell surface protein with a molecular weight of approximately 130 kDa. Binding was blocked in the presence of unlabelled remnants, LDL, or antiserum specific to the LDL receptor. The binding of gold-labelled remnants was substantially greater than that of gold-labelled LDL. Compared with gold-labelled LDL, we found a much clearer demarcation of remnant binding with hepatocytes incubated in the presence or absence of sterols. Our observations suggest that, because of the greater affinity of the LDL receptor for lipoproteins containing apolipoprotein E, changes in LDL receptor expression might be more readily identified using gold-labelled remnants. We conclude that gold-conjugated chylomicron remnants might provide a useful means of detecting subtle changes in LDL receptor expression.
Replacement of single tooth using a threaded titanium screw root coated with hydroxyapatite (HA) for faster bone apposition to implant site is common. Uncoated pure titanium is also used for osseointegration. Usually bone fixation of implant takes four to six months in either case. Quite often, a good number of teeth in a mandible or maxilla are replaced. Usually it is often said that bones appear to be well designed from the point of view of structural engineering. The "maximum-minimum law" claimed by Roux is a rational concept that states that bone provides maximum strength with a minimum of construction material. According to this proposition, stress distribution in bone will be almost uniform under a set of loading conditions. This was found to be true in the case of normal human mandibular bone as examined by some Japanese scientists. The present authors are interested in examining the stress distribution during multiple single tooth replacements using 3D-FEM technique to ascertain how the stress pattern changes with such implantation of three screws in a row in the human mandible, as we are in the process of clinical trial of hydroxyapatite coated and uncoated titanium implant. This will be of considerable interest to the dental surgeons, who prompted us to address this problem. Our results depicted the mean values of various stress in cortical and cancellous bone while applying the maximum masticatory load of 50 N in each teeth. The generated stress level is within the safe range of stress for bones. However, such screws cannot be applied to osteoporotic or other diseased mandible where bone strength may be quite low.
To design a temporal mandibular joint (TMJ), the designer should pay attention to the range of movement in the joint, the strength of the joint, and the size of the implant should conform, so that it does not hamper facial configuration. As a number of designs are available, in this study we have considered one of the most common and widely used implants for analysis. The main objective of this study is to examine the stress-strain behavior at the implant and what is happening at the implant bone interface. We have also examined whether implant material can be replaced by UHMWPE (ultra high molecular weight polyethylene) instead of titanium or Co-Cr-Mo alloy. Whether the change of positions of the screw used for fixation has any effect or not, we have modeled it considering actual shape and size, then divided it into number finite elements by using a FEM package. An appropriate surgical construct was modeled and loaded and studied for different parameters. We have shown that the metallic prostheses are good from a stress-strain point of view and UHMWPE cannot be used as such.
Forty-two Rhizobium strains obtained from different culture collections were evaluated quantitatively for poly(3-hydroxy-butyric acid) [PHB] production in shake flask culture. The majority of the strains produced the maximum amount of PHB during the late exponential or stationary phase of growth. Synthesis and accumulation of PHB in different species of Rhizobium were found to vary between 1-38% of their dry biomass. Growth and PHB production by the Rhizobium strain TAL-640 were greatly influenced by the C-source and D-mannitol was fundamental to both processes. The identity and purity of PHB isolated from TAL-640 have also been confirmed by UV-, IR- and 1H-NMR spectroscopic analyses.
Calcium ions and calcium-dependent systems have been implicated in the pathophysiology of status epilepticus (SE). However, the dynamics of intracellular calcium ([Ca2+]i) levels during SE has not yet been studied. We have employed the hippocampal neuronal culture (HNC) model of in vitro SE that produces continuous epileptiform discharges to study spatial and dynamic changes in [Ca2+]i levels utilizing confocal laser scanning microscopy and the calcium binding dye, indo-1. During SE, the average [Ca2+]i levels increased from control levels of 150-200 nM to levels of 450-600 nM. This increased [Ca2+]i was maintained for the duration of SE. Following SE, [Ca2+]i levels gradually returned to basal values. The duration of SE was shown to affect the ability of the neuron to restore resting [Ca2+]i levels. Both N-methyl-D-aspartate (NMDA) receptor-gated and voltage-gated Ca2+ channels (VGCCs) contributed to the increased calcium entry during SE. Moreover, this elevation in [Ca2+]i occurred in both the nucleus and cytosol. These results provide the first dynamic measurement of [Ca2+]i during prolonged electrographic seizure discharges in an in vitro SE model and suggest that prolonged epileptiform discharges give rise to abnormal sustained increases in [Ca2+]i levels that may play a role in the neuronal cell damage and long-term plasticity changes associated with SE.
To compare the ability of elementary bodies (EB) and reticulate bodies (RB) of the Chlamydia trachomatis mouse pneumonitis (MoPn) biovar to induce a protective immune response, two groups of BALB/c mice were inoculated and boosted twice, with UV-inactivated EB or RB in Freund's adjuvant. Two weeks after the last immunization mice were challenged with C. trachomatis in the ovarian bursa. Vaginal cultures collected for 6 weeks after the intrabursal challenge showed that mice inoculated with EB were significantly protected, while mice inoculated with RB were not. Six weeks after the genital challenge mice were mated. Mice immunized with EB showed significant protection as demonstrated by the number of animals which were fertile and the number of embryos present in the uterine horns. In contrast, no significant protection against infertility was observed in the mice immunized with RB.
We have shown that the domain V of bacterial 23 S rRNA could fold denatured proteins to their active state. This segment of 23 S rRNA could further be split into two parts. One part containing mainly the central loop of domain V could bind denatured human carbonic anhydrase I stably. This association could be reversed by adding the other part of domain V. The released enzyme was directed in such a way by the central loop of domain V that it could now fold by itself to active form. This agrees with our earlier observation that proteins fold within the cell posttranslationally, a process that is completed after release of the newly synthesized polypeptide from the ribosome (Chattopadhyay, S., Pal, S., Chandra, S., Sarkar, D., and DasGupta, C. (1999) Biochim. Biophys. Acta 1429, 293-298).
The presence of three major virulence genes toxR, tcpA and ctxA as well as expression of several putative virulence factors were compared in 12 Vibrio cholerae O139 and non-O1,non-O139 strains of clinical and environmental origin. All the strains possessed the gene encoding the regulatory protein TOXR. None of the non-O1, non-O139 strains as well as one of the O139 environmental strains carried the genes for ctxA and tcpA. Statistically significant differences in hemagglutinin and hemolysin production were observed amongst the strains depending on the source of their isolation. Expression of extracellular enzymes such as protease, elastase, neuraminidase, phospholipase A and phospholipase C, however, did not vary significantly from the groups of strains isolated from different sources.
Chlamydia infections are epidemiologically linked to human heart disease. A peptide from the murine heart muscle-specific alpha myosin heavy chain that has sequence homology to the 60-kilodalton cysteine-rich outer membrane proteins of Chlamydia pneumoniae, C. psittaci, and C. trachomatis was shown to induce autoimmune inflammatory heart disease in mice. Injection of the homologous Chlamydia peptides into mice also induced perivascular inflammation, fibrotic changes, and blood vessel occlusion in the heart, as well as triggering T and B cell reactivity to the homologous endogenous heart muscle-specific peptide. Chlamydia DNA functioned as an adjuvant in the triggering of peptide-induced inflammatory heart disease. Infection with C. trachomatis led to the production of autoantibodies to heart muscle-specific epitopes. Thus, Chlamydia-mediated heart disease is induced by antigenic mimicry of a heart muscle-specific protein.
A DNA plasmid encoding the gene of the major outer membrane protein (MOMP) of the Chlamydia trachomatis mouse pneumonitis (MoPn) serovar and three plasmids containing the variable domains (VD) of the MOMP were constructed. Female mice were inoculated with the plasmids and 60 days later were challenged in the genital tract with C. trachomatis. Six weeks after challenge female mice were caged with male mice and the course of the mating followed. Mice immunized with the MOMP plasmids mounted weak humoral and cell mediated immune responses. However, following the genital challenge no significant differences in vaginal shedding were observed between the groups immunized with the MOMP and control plasmids. In addition, the fertility rates were similar in the experimental and negative control groups. In conclusion, vaccination with DNA plasmids encoding the MOMP elicited a modest immune response but did not protect against infection or disease.
Post-translational control of Escherichia coli ribosome on newly synthesised polypeptide leading to its active conformation (protein folding) has been shown in the case of the enzyme beta-galactosidase. As expected, antibiotics chloramphenicol and lincomycin, which bind to 23S rRNA/50S subunit and kasugamycin and streptomycin which interact with the 30S subunit instantaneously inhibited protein synthesis when they were added to the growing cells. The increase in beta-galactosidase activity, though stopped immediately after the addition of chloramphenicol and lincomycin, went on considerably in the presence of streptomycin and kasugamycin even after the stoppage of protein synthesis.
We have shown previously that the pattern of expression of the transcription factor CREB (cyclic AMP-response element binding protein) in developing oligodendrocytes (OLGs) suggests a role during a period that precedes the peak of myelination in rat brain. We have now investigated the signaling pathways that could be responsible for activating CREB by phosphorylation at different stages along OLG maturation. CREB phosphorylation was studied in short-term cultures of immature OLG precursor cells and young OLGs isolated from 4- and 11-day-old rat cerebrum, respectively. The results indicated that at both developmental stages, CREB phosphorylation could be stimulated by either increased concentrations of cyclic AMP and cyclic AMP-dependent protein kinase activation or increased Ca2+ levels and a protein kinase C activity. The results also showed that CREB phosphorylation in immature OLG precursor cells could be up-regulated by treatment with histamine, carbachol, glutamate, and ATP (neuroligands known to increase Ca2+ levels in these cells), by signaling cascade(s) that involve a protein kinase C activity, as well as the mitogen-activated protein kinase pathway. In contrast, in cells isolated from 11-day-old rats, at a developmental stage that immediately precedes the beginning of the active period of myelin synthesis, CREB phosphorylation was only stimulated by treatment with the beta-adrenergic agonist isoproterenol in a process that appears to be mediated by a cyclic AMP/cyclic AMP-dependent protein kinase-dependent pathway. These results support the idea that CREB could be a mediator of neuronal signals that, coupled to specific signal transduction cascades, may play different regulatory roles at specific stages along OLG differentiation.