Bilateral otogenic temporal lobe and post-aural abscesses.
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Adult↗
Biomedical subjects
S K Gupta
Publications and source records attributed to S K Gupta.
Symptoms experienced by patients with carpal tunnel syndrome.
BACKGROUND: Patients with carpal tunnel syndrome (CTS) sometimes report sensory symptoms outside the median nerve distribution. This study was designed to provide a more detailed assessment of these symptoms. METHODS: Patients with clinical suspicion of upper limb neuromuscular lesions were divided into those with electrodiagnostic (EDX) evidence of CTS, and those without. CTS patients with superimposed nerve abnormalities were excluded. Motor and sensory symptoms were assessed in the exclusive CTS patients. RESULTS: Over 50% of patients with exclusive CTS reported tingling or numbness over the whole hand, ulnar or radial nerve distributions. Some patients reported symptoms proximal to the wrist. Sensory signs did not extend beyond the median nerve distribution. Numbness and nocturnal pain were predictive of positive EDX evidence of CTS. CONCLUSIONS: Sensory symptoms outside the distribution of the median nerve are common in CTS. For enhanced sensitivity in diagnosis it is useful to be aware of these "atypical" symptoms. Reports of numbness and nocturnal pain are strong indicators of CTS.
Adult↗
B cell responses to a peptide epitope. II: Multiple levels of selection during maturation of primary responses.
This report analyses murine primary humoral recognition of a linear domain (MEP 17-31) within a 100 amino acid polypeptide, MEP-1. An analysis of the early primary IgM response revealed that MEP 17-31 presented at least two distinct domains for pre-immune B cell recognition represented by MEP-1 residues 19-23 and 26-28. However, subsequent maturation into an IgG response saw an exclusive selection for the anti-MEP 19-23 component with loss of all alternate specificities. The IgM response to MEP 19-23 was oligoclonal and composed of diverse paratope phenotypes as evidenced by varied heavy chains of immunoglobulin V-D-J combinations and CDR3 sequences. In contrast to the oligoclonality of IgM mAb, the mature IgG response to MEP 19-23 appeared to derive predominantly from a single progenitor. It therefore appears that maturation of primary humoral responses to polypeptide antigens involves two distinct levels of selection. While there is selection for a restricted subset of the initially induced antibody fine-specificities, progression of the response also entails a reduction in clonal heterogeneity of B cells responding to the dominant epitope.
Amino Acid Sequence↗
Preincubation storage of turkey eggs: impact on rate of early embryonic development.
1. The effect of cooling and storage on preincubation development of turkey embryos was examined. 2. Fresh laid eggs were stored at 15 degrees C for 3, 7, or 14 d (group 1) or held initially at room temperature (21 degrees C) for 6 to 9 h and subsequently stored at 15 degrees C for 3, 7, or 14 d (group 2). 3. Most embryos in eggs from both groups progressed developmentally to Stage-8, just prior to onset of hypoblast formation. There were no significant differences in embryo development either within or between groups. 4. It is concluded that under the conditions of this study, egg cooling and storage augment development of the embryo, albeit only slightly.
Animal Husbandry↗
Production of monoclonal antibodies against Nosema bombycis and their utility for detection of pebrine infection in Bombyx mori L.
Latex agglutination assay based on monoclonal antibodies (MCAs) described in this communication may be useful for detection of Pebrine infection in silkworm. Four murine MCAs were produced against Nosema bombycis spore. In ELISA all 4 MCAs (IgM isotype) reacted with alkali treated Nosema spores and to variable extent with acetone precipitated surface protein. However, MA-310 and MA-542 showed a low degree of cross reactivity with BmNPV. In contrast, MA-503 and MA-515 were devoid of reactivity with BmNPV, B. thuringiensis, S. marcescens, Azotobactor, Rhizobium and normal hemolymph protein in ELISA. Latex beads sensitized with a combination of MA-503 and MA-515 (50 micrograms each per ml of 0.4% latex beads) could detect 1 x 10(5) Nosema spores per test. Sensitization of the latex beads with the cocktail of these two MCAs through protein-A bridge further led to a 10-fold increase in the sensitivity (1 x 10(4) spores/test) of the assay. No agglutination was observed in presence of BmNPV, Rhizobium, Azotobactor, E. coli, B. thuringiensis, S. marcescens and normal hemolymph protein indicating the specificity of the test. The results obtained by latex agglutination assay on hemolymph samples of infected as well as normal larvae collected from field, II instar larvae infected in the laboratory and from infected mother moth revealed 100% correlation with results by microscopic examination.
Animals↗
Chronic extradural haematoma with delayed expansion.
A 35-year-old man became symptomatic 26 months after a head injury. CT showed an occipital-suboccipital chronic extradural haematoma (EDH). A surrounding membrane, with neovascularisation, was evident, both, on CT and at surgery. This unusually long interval between trauma and onset of symptoms suggests that delayed expansion of a chronic EDH should be kept in mind if planning conservative management. An EDH can be considered 'cured' only after disappearance of the membrane and haematoma on CT.
Adult↗
Effect of temperature and cardiopulmonary bypass on the pharmacokinetics of remifentanil.
Sixteen patients undergoing coronary revascularization requiring cardiopulmonary bypass received remifentanil 2 micrograms kg-1 or 5 micrograms kg-1 by infusion over 1 min after sternotomy but before commencing cardiopulmonary bypass, during hypothermic cardiopulmonary bypass and during cardiopulmonary bypass after rewarming. Hypothermic cardiopulmonary bypass reduced the clearance of remifentanil by an average of 20%, and this was attributed to the effect of temperature on blood and tissue esterase activity. Reductions in arterial pressure occurred with administration of both doses during normothermia only.
Adolescent↗
Immunoreactivity and in-vitro effect on human sperm-egg binding of antibodies against peptides corresponding to bonnet monkey zona pellucida-3 glycoprotein.
The following synthetic peptides were made as immunogens for development of a zona-based contraceptive vaccine: P1, KQPFWLLQGGASRAETSVQPVLVE [amino acids (aa) 23-45 with an additional K at the N-terminus]; P2, FSEEKLVFSLRLMEENC (aa 164-179 with an additional C at the C-terminus and T170 replaced by V); and P3, CSFSKSSNSWFPVEGPADICQCC (aa 300-322). The aa are numbered on the basis of bonnet monkey ZP3 precursor protein. Antibodies against an additional peptide P4, KGDCGTPSHSRRQPHVVSQWSRSA (aa 324-347), significantly inhibits human sperm-oocyte binding. In addition, antibodies against cocktail of peptide-diphtheria toxoid conjugates also significantly inhibit the binding of spermatozoa to zona pellucida in a hemizona assay. These results will further help in the design of an immunocontraceptive vaccine based upon synthetic peptides corresponding to ZP3.
Amino Acid Sequence↗
Comparative development of the turkey and chicken embryo from cleavage through hypoblast formation.
The development of the turkey and chicken embryo from the first cleavage division through hypoblast formation is described. The early development of the chicken embryo has been categorized into 14 stages. A similar staging sequence for the turkey was not proposed until 1993, when we described the early development of the turkey embryo, which was divided into 11 stages. Comparatively, differences in the temporal and spatial development of the turkey and chicken blastoderm were evident. Of significance is the observation that at oviposition the turkey is in Stage VII and characterized by the first signs of area pellucida formation. In contrast, the chicken embryo at oviposition is in Stage X and area pellucida formation is completed. Similarly, the hypoblast, which is already apparent in the Stage X chicken embryo, does not appear in the turkey embryo until the egg is incubated. Furthermore, the anterior-posterior (head-tail) axis in the early embryo is achieved prior to oviposition in the chicken but after the onset of incubation in the turkey. It is apparent that the turkey embryo is less mature than the chicken embryo at oviposition. Whether this distinction is related to differences between the hatchability of turkey and chicken eggs is not yet known.
Animals↗
Sequence of complementary deoxyribonucleic acid encoding bonnet monkey (Macaca radiata) zona pellucida glycoprotein-ZP1 and its high-level expression in Escherichia coli.
Zona pellucida (ZP) glycoproteins have been proposed as candidate antigens for immunocontraception. Studies on this potential use can be facilitated by the availability of recombinant proteins. A cDNA lambda gt11 library was constructed using poly(A)+ mRNA isolated from bonnet monkey (Macaca radiata) ovaries and was screened for bonnet monkey ZP1 using a 404-basepair (bp) human ZP1 fragment (nucleotides 818-1221) as probe. Bonnet monkey ZP1 cDNA comprises 1617 nucleotides and encodes a polypeptide of 539 amino acid residues that share 92.0% identity with human ZP1. The major difference between bonnet monkey ZP1 and human ZP1 is the deletion of a 28-amino acid domain (amino acid residues 100-127 corresponding to human ZP1). An internal fragment (1317 bp) of bonnet monkey ZP1, excluding the N-terminus signal sequence and the C-terminus transmembrane-like domain, was amplified by polymerase chain reaction. The amplified Sac I and Kpn I restricted fragment was cloned in a frame downstream of the T5 promoter under the lac operator control for expression in the pQE-30 vector. Recombinant ZP1 (r-ZP1) was expressed as a polyhistidine fusion protein in Escherichia coli strains SG13009[pREP4] and ompT and Ion protease-deficient BL21 (plysS). SDS-PAGE analysis and immunoblotting with a murine monoclonal antibody, MA-410 (raised against porcine ZP3alpha--a homologue of bonnet monkey ZP1--and cross-reactive with bonnet monkey zona pellucida), revealed major bands of 51 and 40 kDa besides truncated fragments. Optimum expression of r-ZP1 was observed at 0.5 mM isopropyl beta-D-thiogalactopyranoside (IPTG). Immunization of male rabbits with r-ZP1 purified on nickel-nitrilotriacetic acid (NTA) resin under denaturing conditions and of female rabbits with r-ZP1 conjugated with diphtheria toxoid-generated antibodies reactive with r-ZP1 in ELISA. Moreover, immune sera, when tested by indirect immunofluorescence on bonnet monkey ovarian sections, showed positive fluorescence with zona pellucida. The information on the sequence of bonnet monkey ZP1 and the availability of the recombinant protein will help toward better understanding and evaluation of the contraceptive potential of homologous immunization in a nonhuman primate model.
Amino Acid Sequence↗
Remifentanil and pulmonary extraction during and after cardiac anesthesia.
We measured the apparent blood clearance and pulmonary extraction ratio of remifentanil in 10 adult patients undergoing elective myocardial revascularization for the first time with hypothermic cardiopulmonary bypass (CPB). Patients received continuous infusions of remifentanil 1.0, 1.5 or 2.0 microg x kg(-1) x min(-1). After surgery, remifentanil was infused at 1.0 microg x kg(-1) x min(-1) in all patients. Remifentanil concentrations were measured in pulmonary and radial artery blood by gas chromatography with high resolution mass spectrometry before and after CPB and 165 min (60 SD) after surgery. Cardiac output was measured by thermodilution at the time of blood sampling. The mean pulmonary extraction ratio of remifentanil was 5.7% (13.1% SD), which was not significantly different from zero. However, pulmonary extraction ratio was related inversely to the pulmonary artery hydrogen ion concentration and directly to the percent of nonionized form of the base in the pulmonary artery. Remifentanil concentrations in pulmonary and radial artery blood were related directly to infusion rate, but not to duration of infusion. There was no evidence of accumulation or sequestration. Mean apparent blood remifentanil clearance was 2.03 L/min (0.35 SD) and, in contrast to remifentanil pulmonary extraction ratio, was related directly to cardiac index and oxygen delivery. Increased tissue perfusion increased blood remifentanil clearance. We found predictable blood remifentanil levels with no evidence of accumulation or pulmonary extraction.
Adult↗
The HSD-hCG vaccine prevents pregnancy in women: feasibility study of a reversible safe contraceptive vaccine.
PROBLEM: To develop a vaccine for reversible control of fertility in women. MATERIALS AND PROTOCOLS: Purified beta subunit of hCG annealed to purified alpha subunit of ovine LH linked chemically to tetanus toxoid (TT) and diphtheria (DT); vaccine employed at 300 micrograms gonadotropin equivalent per injection adsorbed on alhydrogel with 1 mg SPLPS added in the first injection; Phase I safety trials in 47 women with elective tubal ligation; Phase II efficacy studies in 148 proven fertile women (2 children), sexually active, desirous of family planning using IUD; IUD removed when anti-hCG titres exceed 50 ng/ml hCG bioneutralization capacity; boosters given to maintain above threshold antibody levels; post coital tests conducted in 8 volunteers; sera of protected women analysed for immuno-determinants recognized by competitive enzyme immunoassays employing a panel of monoclonal antibodies and by direct binding to synthetic peptides; recombinant vaccines expressing beta hCG as a secreted product or as a fused protein anchored on membrane. RESULTS: Immunization was well tolerated with no significant changes in endocrine, metabolic and hematological indices. Normal ovulatory cycles were maintained as indicated by menstrual regulation. The vaccine was highly effective in preventing pregnancy (1 pregnancy in 1224 cycles ) at and above antibody titres of 50 ng/ml. Antibodies declined in course of time in absence of boosters, with conceptions occurring below 35 ng/ml titres indicating regain of fertility. Ability of antibodies to prevent pregnancy was confirmed by post coital tests. High avidity (10(10) M-1) and other characteristics of antibodies generated by the vaccine are described and compared with those induced by two other hCG vaccines having undergone Phase I trials. The antibody response of the HSD vaccine in humans is characterized predominantly to an epitope recognized by the monoclonals 206 and P3W80. The antibodies had low or no reactivity with the carboxy terminal peptide and 38-57 region peptide. Live recombinant vaccines expressing beta hCG as a membrane anchored peptide generated antibody response to hCG in all animals following a single injection. CONCLUSIONS: Reversible fertility control is feasible with the HSD-hCG vaccine without impairment of ovulation or disturbance of menstrual regularity. Suggestions have been made for further optimization of the vaccine, which include replacement of TT and DT by a panel of T non B determinants communicating with the entire MHC spectrum and development of recombinant vaccine expressing beta hCG along with membrane anchored carrier.
Animals↗
Identification of epitopes of monoclonal antibodies to porcine zona pellucida 3 beta glycoprotein, a homologue of the mouse/human sperm receptor.
PROBLEM: Immunization with zona pellucida (ZP) glycoproteins leads to a block in fertility with a variable degree of ovarian dysfunction. To avoid autoimmune oophoritis, synthetic peptides corresponding to B cell epitope(s) and devoid of oophoritogenic T cell epitopes as immunogens have been proposed. The main objective of the present study is to define the epitopes recognized by monoclonal antibodies (mAbs) generated against porcine ZP3 beta, a homologue of the designated primary sperm receptor in mice and humans. METHODS: A multipin synthetic peptides approach has been used to map the epitopes recognized by mAbs. Dodecapeptides with an overlap of 6 amino acids corresponding to a precursor pZP3 beta-deduced amino acid sequence (excluding the signal sequence) were synthesized on polypropylene pins and were tested for their reactivity with mAbs by enzyme-linked immunoadsorbent assay (ELISA). The ability of synthetic peptides corresponding to the identified epitopes to inhibit the binding of mAbs to pZP3 beta in a competitive inhibition ELISA was investigated to confirm the above findings. RESULTS: Reactivity of the mAbs with the pin-bound peptides in ELISA-identified epitopes for mAb-451 to EEKLVF (166-171) and mAb-462/470 to FKAPRP (250-255) amino acid residues. mAb-30 recognized QPVWQDEGQRLR (23-34) and VICRCC (316-321) amino acid residues. Competitive inhibition with synthetic peptides encompassing the motifs corresponding to 23-34 and 316-321 for binding of mAb-30 to pZP3 beta revealed the epitopic domain to be 23-34 amino acids. Synthesis of overlapping octapeptides further identified WQDE as the minimum motif for binding of mAb-30, and the replacement of one amino acid at a time with glycine revealed tryptophan as the critical residue. CONCLUSIONS: Collectively, these results describe peptide epitopes that will help in the design of an immunocontraceptive vaccine based on synthetic peptides corresponding to pZP3 beta or its homologues in other species.
Amino Acid Sequence↗
Pseudomonas aeruginosa lipopolysaccharide binds galectin-3 and other human corneal epithelial proteins.
The aim of this study was to test whether galectin-3 is present in human corneal epithelium and whether lipopolysaccharide (LPS) purified from Pseudomonas aeruginosa ATCC 19660 binds to this animal lectin and/or to another human corneal epithelial protein(s) (HCEP) and to confirm which component of LPS (inner or outer core or lipid A) is important in bacterial binding by using the eye in organ culture. LPS isolated and purified from P. aeruginosa ATCC 19660 and a commercial LPS (serotype 10) differed in polyacrylamide gel analysis but bound similarly to blotted HCEP. Binding was determined to be a receptor-ligand type of interaction by the solid-phase assay, because it was both specific and saturable. Several LPS binding proteins in HCEP were identified by an overlay method. Western blotting with antibody against galectin-3 revealed the presence of this protein in both freshly isolated and cultured transformed human corneal epithelium. Binding inhibition assays showed that antibody specific for the outer core region of LPS and an anti-galectin antibody significantly inhibited bacterial binding in vitro. These data provide further evidence that LPS is an important adhesin of P. aeruginosa, that it binds to protein receptor molecules in HCEP, that one of the LPS binding proteins is galectin-3, and that the outer core portion of the molecule appears to be critical for LPS binding to the eye.
Antigens, Differentiation↗
Microdissection of stained archival tissue.
In many tissues the preinvasive stage of neoplastic progression can be identified histologically as dysplasia or in situ disease. There is much interest in defining the molecular events associated with the early stages of neoplasia. Retrieval of histologically recognisable preinvasive neoplastic tissue uncontaminated by inflammatory or stromal cells is important for genetic studies using polymerase chain reaction (PCR) assay. A novel method for microdissection is described in which 10 microns sections are dewaxed, stained with haematoxylin and eosin, dried, covered with Sellotape, and the tissue cut out using a scalpel blade under direct visual control. The method is quick, eliminates problems of operator tremor, preserves the architecture of the micro-dissected tissue (for photographic documentation) and requires no special equipment. The presence of Sellotape and adhesive in the reaction mixture has no detrimental effect on the ability to extract DNA or to perform PCR.
Breast Neoplasms↗
Curbing trade of human organs--status of current policy regarding registration of hospitals.
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Certification↗
Clinical dysautonomia in diabetes mellitus.
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Autonomic Nervous System Diseases↗
Prevalence and risk factors of cataract in India: a review.
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Blindness↗