Biomedical subjects

O Singh

Publications and source records attributed to O Singh.

At least 19 recordsLinked to original sources

PubMed2006

Mother's iron status, breastmilk iron and lactoferrin--are they related?

OBJECTIVE: Exclusive breastfeeding is recommended till 6 months age. Factors regulating the breastmilk iron and lactoferrin levels are incompletely known. Considering high prevalence of nutritional anemia in lactating mothers, we studied the iron status of lactating mothers, their breastmilk iron and lactoferrin levels to determine any relationship between them. DESIGN: Prospective study with 6 months follow-up. SETTING: Tertiary care referral hospital. SUBJECTS: Hundred nonanemic and 100 anemic mothers with their babies recruited at birth. Fifty-two nonanemic and 50 anemic mothers and their babies completed the 6-month follow-up. INTERVENTIONS: Hemoglobin (Hb), total iron binding capacity (TIBC), percent transferrin saturation (%TS), serum iron (SI) and serum ferritin measured on day 1 and 6 months postpartum. Breastmilk iron and lactoferrin measured on day 1, 14 weeks and 6 months after delivery. RESULTS: Breastmilk iron decreased progressively from day 1 to 14 weeks and at 6 months in both groups, but no significant difference was noted between nonanemic and anemic mothers (P>0.05). Significant decline in breastmilk lactoferrin concentration from day 1 to 14 weeks in nonanemic and anemic mothers (P<0.001) noted. Hemoglobin, TIBC, %TS, SI and serum ferritin of both groups had no correlation with breastmilk iron and lactoferrin concentration on day 1, 14 weeks and 6 months after delivery. CONCLUSIONS: Breastmilk iron and lactoferrin concentration had no relationship with the mother's Hb and iron status.

Adult↗

PubMed2006

New compounds from Commiphora myrrha (Nees) Engl.

Phytochemical investigation of Commiphora myrrha (Nees) Engl. has afforded six new compounds identified as cadina-3-en-15-ol (myrracadinol A) (1), 7, 8-seco-2, 5-dihydroxy-12-acetoxycalam-8-ene (myrracalamene A) (2), 7, 8-seco-2, 3, 5-hydroxy-12-acetoxycalame-8-ene (myrracalamene B) (3), 7, 8-seco-cadin-3, 8-dien-2beta, 12-diol (myrracadinol B) (4), 7, 8-seco-12-hydroxycalam-8-ene (myrracalamene C) (6), 7, 8-seco-cadin-3,7(12)-dien-5alpha,10alpha-diol (myrracadinol C) (7) along with a known compound tria-cont-1-ene (5). Their structures were elucidated on the basis of spectral and chemical analyses.

Acetylation↗

PubMed2001

Enhanced immunogenicity of a contraceptive vaccine using diverse synthetic carriers with permissible adjuvant.

A vaccine directed against human chorionic gonadotropin (hCG) has previously undergone clinical testing demonstrating the feasibility of the approach in preventing pregnancy in women. Some individuals, however, did not respond adequately despite employing highly immunogenic bacterial toxoids as carriers. We investigated the potential of three promiscuous pathogen-derived Th peptides as carriers, employing alum as the adjuvant. While conjugation with each peptide improved the antibody response against hCG in mice of different haplotypes, immunisation with a combination of these peptide-conjugates generated anti-hCG responses higher than those achieved with the individual peptides or tetanus toxoid (TT). Antibodies were of high affinity and capable of neutralising the bioactivity of hCG but were devoid of anti-peptide reactivity. These results have implication for the design of hCG vaccine with improved immunogenicity for diverse population.

Adjuvants, Immunologic↗

PubMed2001

Absence of corpus luteum rescue by chorionic gonadotropin in women immunized with a contraceptive vaccine.

OBJECTIVE: To determine whether antibodies induced by a betahCG vaccine in women are competent to neutralize the luteotropic action of hCG. DESIGN: Prospective clinical study. SETTING: Hospitals and a laboratory at an academic center. PATIENT(S): Six immunized and three control women of reproductive age, participating in a clinical testing of betahCG contraceptive vaccine. INTERVENTION(S): Increasing doses of hCG simulating early pregnancy. MAIN OUTCOME MEASURE(S): Anti-hCG antibody titers, serum progesterone and urinary hCG levels, and onset of menses. RESULT(S): Administration of hCG to mimic early pregnancy sustained serum progesterone concentrations and extended the luteal phase in control women. In contrast, serum progesterone levels declined and the luteal phase was not extended if prevailing antibody titers were > or =40 ng/mL in women who had been immunized with a betahCG based vaccine. No booster effect was seen in anti-hCG titers after hCG challenge. CONCLUSION(S): Antibodies elicited by a betahCG vaccine inactivate hCG and prevent the hormone from rescuing corpus luteum, resulting in progesterone fall and normal menses. Lack of booster in the antibody response confirms the reversibility of the approach.

Adult↗

PubMed2001

High expression of human chorionic gonadotrophin beta-subunit using a synthetic vaccinia virus promoter.

We have constructed a recombinant vaccinia virus to express the beta-subunit of human chorionic gonadotrophin (betahCG), a secretory glycoprotein that is used as an antigen for a contraceptive vaccine. The cDNA encoding the subunit was cloned under the control of a synthetic promoter that could be recognised by a vaccinia virus RNA polymerase to direct transcription. The peak expression level of betahCG directed by a late synthetic promoter (Psyn) was 11.5 microg/ml, a level that was at least sixfold higher than that directed by the p7.5 early/late promoter. The expressed protein was correctly processed post-translationally such that it attained a conformation with correctly folded discontinuous epitope(s) similar to that seen in native betahCG.

Blotting, Southern↗

PubMed2000

Humoral hyporesponsiveness to a conjugate contraceptive vaccine and its bypass by diverse carriers using permissible adjuvant.

A contraceptive vaccine directed against human chorionic gonadotropin (hCG) has previously undergone clinical testing that demonstrated the feasibility of the approach in preventing pregnancy in women. Some immunized volunteers however, did not respond with an adequate anti-hCG antibody response despite employing highly immunogenic bacterial toxoids as carriers. Since there is some evidence that T cell responses to a complex protein typically focus on a few immunodominant epitopes, we investigated the responsiveness to hCG in mice of different haplotypes using the protein carrier diphtheria toxoid (DT). Our data showed a differential carrier effect of DT. With the aim of making a more potent immunogen employing promiscuous pathogen-derived Th peptides as carriers, peptide:antigen stoichiometric ratios were optimized. When tested individually using alum as the adjuvant, three such peptide conjugates improved the anti-hCG response, though not consistently to levels higher than the DT conjugate. Immunization with a combination of these synthetic epitopes generated anti-hCG responses higher than those achieved with DT or with the individual peptides. Antibodies were of high affinity and capable of neutralizing the bioactivity of hCG, but were devoid of anti-peptide reactivity. These results support our view that differential hyporesponsiveness in a diverse population may arise from inadequate carrier effect and that it can be overcome by use of pathogen-derived broadly reactive non-B Th epitopes employing only alum, a permissible adjuvant.

Adjuvants, Immunologic↗

PubMed1999

Prior immunity to a carrier enhances antibody responses to hCG in recipients of an hCG-carrier conjugate vaccine.

Pre-sensitization with carrier often leads to epitopic suppression of subsequent anti-hapten antibody responses. To ascertain whether epitopic suppression occurs in humans, we examined the effect of pre-existing anti-carrier immunity on antibody responses to hCG in volunteers of a clinical trial of an hCG-based conjugate birth-control vaccine. When we studied the correlation between pre-vaccination anti-carrier immunity and post-vaccination anti-hCG responses, we found that prior immunity to the carriers did not lead to epitopic suppression of anti-hCG responses. On the contrary, it was found that prior immunity to TT, one of the two carriers used in this vaccine, led to enhancement of anti-hCG responses. Our data indicates that prior immunity to the carriers may not be detrimental to the performance of conjugate vaccines, and may actually be beneficial in some cases.

Adult↗

PubMed1998

Dihydropyrancarboxamides related to zanamivir: a new series of inhibitors of influenza virus sialidases. 2. Crystallographic and molecular modeling study of complexes of 4-amino-4H-pyran-6-carboxamides and sialidase from influenza virus types A and B.

The first paper in this series (see previous article) described structure-activity studies of carboxamide analogues of zanamivir binding to influenza virus sialidase types A and B and showed that inhibitory activity of these compounds was much greater against influenza A enzyme. To understand the large differences in affinities, a number of protein-ligand complexes have been investigated using crystallography and molecular dynamics. The crystallographic studies show that the binding of ligands containing tertiary amide groups is accompanied by the formation of an intramolecular planar salt bridge between two amino acid residues in the active site of the enzyme. It is proposed that the unexpected strong binding of these inhibitors is a result of the burial of hydrophobic surface area and salt-bridge formation in an environment of low dielectric. In sialidase from type A virus, binding of the carboxamide moeity and salt-bridge formation have only a minor effect on the positions of the surrounding residues, whereas in type B enzyme, significant distortion of the protein is observed. The results suggest that the decreased affinity in enzyme from influenza B is directly correlated with the small changes that occur in the amino acid residue interactions accompanying ligand binding. Molecular dynamics calculations have shown that the tendency for salt-bridge formation is greater in influenza A sialidase than influenza B sialidase and that this tendency is a useful descriptor for the prediction of inhibitor potency.

Acetamides↗

PubMed1998

Regain of fertility and normality of progeny born during below protective threshold antibody titers in women immunized with the HSD-hCG vaccine.

PROBLEM: Phase II clinical trials with the heterospecies dimer of beta hCG and alpha-subunit of ovine luteinizing hormone (HSD)-human chorionic gonadotrophin (hCG) vaccine showed that pregnancy was prevented at and above 50 ng/ml titers, whereas conceptions occurred below 35 ng/ml of hCG bioneutralization capacity. The effect of below-protective threshold anti-hCG antibodies on the progression of pregnancy and the normality of progeny was studied. METHOD OF STUDY: Four women enrolled by informed consent in the Phase II trials. The women studied were immunized with the HCG vaccine and did not receive booster injections in consideration of their desire to have another child. They were examined clinically at least once every month until delivery. The babies were followed up from 2 to 3.5 years, and their anthropometric indices were compared with elder siblings. RESULTS: The pregnancies progressed to 35 to 38 weeks with the usual obstetric problems, and the children developed normally. CONCLUSIONS: The anti-fertility effect of the HSD-hCG vaccine is reversible and low titers of antibodies below the protective threshold have no apparent side effects on the progression of pregnancy and on the early development of the progeny.

Adult↗

PubMed1998

Stabilization of dichloromethane-induced protein denaturation during microencapsulation.

This paper describes the denaturation of protein drugs by dichloromethane (DCM) during the primary emulsification step of the microencapsulation process using biodegradable polymer matrix for controlled-release application. It was found that interaction of proteins such as tetanus toxoid (TT), diphtheria toxoid (DT), ovine growth hormone (oGH), and human chorionic gonadotropin-based antifertility vaccine (beta-hCG-TT) with DCM during primary emulsification stages of particle formulation led to the precipitation of the proteins at the aqueous organic interface with concomitant reduction in their immunoreactivity. On the other hand, the B subunit of E. coli enterotoxin (LTB) was found to be comparatively stable toward the denaturing action of DCM. Attempts were made to overcome the DCM-induced denaturation by incorporation of stabilizers during the primary emulsification step of the particle formulation. Of the many additives tested to overcome the DCM-induced denaturation of proteins, serum albumins and polyvinyl alcohol (PVA) showed promising results in terms of retention of the immunoreactivity of the protein. TT stabilized by the incorporation of serum albumin during the primary emulsification step not only showed immunoreactivity in vitro, but also invoked antibody titers in rats comparable to those obtained for the native protein molecules. Incorporation of 2.5% of serum albumins in the internal aqueous phase not only protected the protein from the degradative action of DCM but also led to stabilized primary emulsion, which is necessary for uniform entrapment of protein drugs in the polymer matrix.

Animals↗

PubMed1997

Helix-loop-helix motif in GnRH associated peptide is critical for negative regulation of prolactin secretion.

The GnRH associated prolactin inhibiting factor (GAP) reveals the signature sequence associated with the helix-loop-helix structural motif. A number of different peptide fragments of GAP were designed, synthesized and analysed by circular dichroism and by an in vivo assay for prolactin secretion inhibiting activity. Peptides corresponding to the two individual alpha-helices and a 44-residue peptide comprising the entire helix-loop-helix motif show significant helical propensity in circular dichroism spectra. However, a peptide corresponding to the loop sequence shows no helical propensity. Albeit, the peptide corresponding to helix-loop-helix motif was found to inhibit prolactin secretion and augment circulating levels of gonadotropins in the in vivo assay; other shorter peptides did not show such activity. The activity profile of the 44-residue peptide was biphasic and very similar to that of the recombinant GAP. Thus, the prolactin inhibiting activity of this factor is defined by its helix-loop-helix motif as in the case of the transcription factors of developmental genes. The structural features of a homology-based model of GAP in complex with E47, a ubiquitous HLH-type developmental gene regulator, are consistent with the structural requirements of the negative regulation of transcription by helix-loop-helix proteins.

Amino Acid Sequence↗

PubMed1997

Recombinant adenovirus synthesizing cell surface-anchored beta hCG induces bioneutralizing antibodies in rats.

A recombinant adenovirus (re-Ad) has been constructed that synthesizes a cell surface-anchored form of the beta-subunit of human chorionic gonadotropin (beta hCG). This was achieved by in-frame fusion of beta hCG cDNA at its C terminus with the gene sequences coding for the vesicular stomatitis virus glycoprotein (VSVg) transmembrane domain. The fusion protein gene was placed under the control of human cytomegalovirus (hCMV) immediate early promoter and this expression cassette was inserted into the E1 region of Ad type 5 by homologous recombination. In vitro experiments using re-Ad-infected 293 cells showed that beta hCG fusion protein was made as early as 6 h post infection and the protein was anchored to the cell membrane as seen by immunofluorescence staining. The re-Ad induced bioneutralizing antibodies (Ab) to hCG when inoculated in rats through intraperitoneal or intramuscular routes. The Ab were made in a dose-dependent manner. However, orally delivered re-Ad failed to generate any significant immune response.

Adenoviridae↗

PubMed1997

Biphasic testosterone delivery profile observed with two different transdermal formulations.

PURPOSE: Our long-term goal is to develop formulations for pulsatile testosterone (T) delivery. T has been reported earlier to show biphasic pharmacokinetics in humans by Mazer et al, as well as biphasic permeation across excised rat skin by our group. We examined two kinds of formulations to evaluate their delivery profiles and to assess whether differences in the formulation approach affect pharmacokinetics in animal models. METHODS: One formulation consisted of T and a polymer blend dissolved in isopropanol; administered by dispensing the solution on the skin to cast a film in situ. The other was an adhesive-dispersion patch. In vitro release from the patch was evaluated using a flow-through cell interfaced with an HPLC pump and UV detector. Single dose pharmacokinetics were evaluated in castrated Wistar rats and bonnet monkeys immunized against gonadotropin-releasing hormone to deplete endogenous T. RESULTS: Two maximas were observed in the T release profile from the patch and in serum concentration versus time profiles in both animal models on application of either formulation. The relative magnitudes of the two maximas and the time interval separating them were different in the case of each formulation. CONCLUSIONS: Both formulations result in biphasic pharmacokinetics of T in the animal models studied. Discrete maximas presumably correlate with "burst" and "sustained" phases of drug release.

Animals↗

PubMed1995

Biodegradable delivery system for a birth control vaccine: immunogenicity studies in rats and monkeys.

PURPOSE: The purpose of this study was to develop a single administration delivery system for a model birth control vaccine, in order to reduce the need for multiple injections and enhance immunogenicity. METHODS: The immunogen-loaded microspheres were prepared by solvent evaporation method and characterized for loading levels, size distribution and in vitro release kinetics. The microspheres were immunized intramuscularly in wistar rats and bonnet monkeys, and the antibody response was compared to that obtained with the same total dose of the immunogen on alum given at a monthly interval. RESULTS: Results indicated that a single injection of the immunogen entrapped in the microspheres generated a response comparable to that obtained by the same immunogen on alum injected at a monthly interval. The antibodies generated by the microspheres in the monkeys also had a good bioneutralization capacity indicating immunogen integrity during the microencapsulation process. CONCLUSIONS: Biodegradable microspheres served as an effective delivery system for a model immunogen used in this study to reduce the need for frequent immunizations and enhance immunogenicity.

Animals↗

PubMed1995

Development of contraceptive vaccines for humans using antigens derived from gametes (spermatozoa and zona pellucida) and hormones (human chorionic gonadotrophin): current status.

Contraceptive research has entered a new phase of development with the advent of hybridoma and DNA recombinant technologies. During the past 5 years, significant advances have been made in this area and now it seems that realistic prospects exist for the development of contraceptive vaccines for use in humans and animals (veterinary, wild and domestic), applicable to both the female and male sexes. Contraceptive vaccines will be valuable supplements to the presently available methods of family planning, and, due to high specificity, the occurrence of limited side-effects if any, low cost and infrequent administration, contraceptive vaccines may have greater acceptability than the currently available methods. Mammalian reproduction starts with the unison of gametes contributed by the male and female partners. Both spermatozoon and oocyte have antigens on the cell surface that are unique, tissue-specific, immunogenic and accessible to antibodies, and binding of the antibodies to these antigens can cause inhibition of gamete function, resulting in a failure of fertilization. Fertilization is followed by embryogenesis, with the early embryo producing several proteins, some of which, e.g. human chorionic gonadotrophin (HCG), have a vital role in the establishment and maintenance of early pregnancy. Again, these proteins are accessible to antibodies, and their immunoneutralization can cause anti-fertility effects with loss of early embryo. Thus, the antigens derived from proteins on spermatozoa, oocyte and early embryo, especially HCG, constitute interesting molecules for the development of contraceptive vaccines. The aim of the present article is to review the current status of development of contraceptive vaccines based on antigens derived from sperm cell, oocyte zona pellucida and HCG, and to discuss their relative merits and future development.

Acrosome↗

PubMed1995

Targeting vaccinia virus-expressed secretory beta subunit of human chorionic gonadotropin to the cell surface induces antibodies.

We carried out experiments designed to study the effect of a protein's localization on its immunogenicity. A novel cell-surface protein was generated from a small, glycosylated secretory protein. The DNA sequence encoding the entire precursor of the human chorionic gonadotropin beta (beta hCG) subunit was fused in the correct reading frame to the DNA sequence encoding the transmembrane and cytoplasmic domains of vesicular stomatitis virus glycoprotein. This chimeric gene was introduced into the vaccinia virus genome to generate a recombinant virus. The recombinant virus, when used to infect animal cells, expressed a 135-amino-acid beta hCG subunit anchored in cellular membranes by the 48 carboxy-terminal amino acids of vesicular stomatitis virus glycoprotein. The immunogenicity of this recombinant virus with respect to its ability to generate anti-hCG antibodies was compared with that of a second recombinant vaccinia virus expressing the native secretory form of beta hCG. All animals immunized with the vaccinia virus expressing beta hCG on the cell surface elicited high titers of anti-hCG antibodies. Even after a single immunization with the recombinant vaccinia virus, the anti-hCG antibody titers persisted for a long period of time (more than 6 months). None of the animals immunized with vaccinia virus expressing the native secretory form of beta hCG showed any hCG-specific antibody response.

Animals↗

PubMed1995

Safety of intrauterine administration of purified neem seed oil (Praneem Vilci) in women & effect of its co-administration with the heterospecies dimer birth control vaccine on antibody response to human chorionic gonadotropin.

Praneem Vilci (PV), purified neem oil was reported to exercise a reversible antifertility effect after a single intrauterine instillation in rodents and primates without any adverse effects. After toxicology, drug regulatory and ethical clearances, a phase I clinical trial was conducted on PV. Eighteen healthy tubectomised women were enrolled to evaluate the safety of a single intrauterine instillation of PV and to determine the effect of its co-administration on anti-hCG response to the heterospecies dimer (HSD) hCG vaccine. Eight women received PV alone and ten women were given the HSD-hCG vaccine in addition. Base-line and post-treatment haematological and biochemical profiles were determined as also the mid-luteal serum progesterone. Endometrial biopsies were examined to assess ovulatory status and the effect of intrauterine treatment with PV on the endometrium. Anti-hCG antibody titres were estimated in women who were concurrently immunized with the HSD vaccine. No untoward reaction was observed in any woman. Menstrual pattern and ovulatory status remained unaltered. Endometrial biopsy after PV instillation in one woman showed non-specific endometritis but she remained asymptomatic. Mild eosinophilia was seen in two women and this reverted to normal on its own. All women receiving PV and the HSD vaccine generated antibodies against hCG. Our data show that intrauterine administration of PV is safe and does not prevent the antibody response to HSD-hCG vaccine.

Adult↗

PubMed1994

A vaccine that prevents pregnancy in women.

We report here results of clinical trials on a birth control vaccine, consisting of a heterospecies dimer of the beta subunit of human chorionic gonadotropin (hCG) associated noncovalently with the alpha subunit of ovine luteinizing hormone and conjugated to tetanus and diphtheria toxoids as carriers, that induces antibodies of high avidity (K(a) approximately 10(10) M-1) against hCG. Fertile women exposed to conception over 1224 cycles recorded only one pregnancy at antibody titers of > 50 ng/ml (hCG bioneutralization capacity). The antibody response declines with time; fertility was regained when titers fell to < 35 ng/ml. This study presents evidence of the feasibility of a vaccine for control of human fertility.

Adult↗