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ABSTRACT: A polyherbal vaginal pessary (Praneem) has been formulated that has antimicrobial properties against genital pathogens in addition to spermicidal action. Thus, it has dual potential as a barrier method for contraception and for providing protection against some sexually transmitted infections. The present study reports the findings of a multicentre trial that was conducted to evaluate the safety of this product. Trials were carried out in 23 women in three centres in India: the Postgraduate Institute of Medical Education and Research, Chandigarh; Safdarjang Hospital, New Delhi; and Kamla Nehru Memorial Hospital, Allahabad. Thorough clinical and pelvic examinations were carried out as well as cervical cytology, blood biochemistry and haematology before and after use of the polyherbal pessary intravaginally once daily for 7 consecutive days. No toxicity was observed on clinical examination or by laboratory investigations. Daily intravaginal use of this pessary for 7 days had no adverse effects on cervical cytology or on metabolic and organ functions.
Transactions of the Royal Society of Tropical Medicine and Hygiene 01/2007; 100(12):1164-7. · 2.16 Impact Factor
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Pankaj Sharma,
Rama Mukherjee, G P Talwar,
K G Sarathchandra,
R Walia,
S K Parida,
R M Pandey,
Rajni Rani,
Hemant Kar,
Ashok Mukherjee,
Kiran Katoch,
S K Benara,
Tulsi Singh,
Padam Singh
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ABSTRACT: We report here a large scale, double blind immunoprophylactic trial of a leprosy vaccine based on Mycobacterium w (Mw) in an endemic area of Kanpur Dehat, Uttar Pradesh, India. A population of 420,823 spread over 272 villages was screened where 1226 multibacillary (MB) and 3757 paucibacillary (PB) cases of leprosy were detected. A total of 29,420 household contacts (HHC) of these patients were screened for evidence of active or inactive leprosy. After exclusion of 1622 contacts for any of the different exclusion criteria, a total of 24,060 HHC could be vaccinated for vaccine or placebo under coding (20,194 administered two doses and 3866 received single dose). The vaccine consisted of 1 x 10(9) heat killed bacilli (Mw) in normal saline for the first dose and half of the first dose, i.e. 5 x 10(8) bacilli for the second dose, given 6 months after the first dose. The placebo consisted of 1/8th dose of the normal dose of tetanous toxoid. Both placebo and vaccine were given under double-blind coding, The contacts were followed up during three surveys at 3, 6 and 9 years after the initial vaccination, for detection of post-vaccination cases (PVCs) and observing any side-effects caused as a result of vaccination. The codes were opened on 24th January 2001, after the analysis of the data following completion of the third and final follow-up survey. When only contacts received the vaccine, Mw vaccine showed a protective efficacy (PE) of 68-6% at the end of first, 59% at the end of the second and 39.3% at the end of the third follow-up survey. When both patients and contacts received the vaccine, the protective efficacy observed was 68%, 60% and 28% at the end of the first, second and third surveys, respectively. When patients, and not the contacts, received the vaccine, a PE of 42.9% in the first, 31% in the second and 3% in the third survey was shown. These results suggest that the vaccination of the contacts is more valuable in achieving the objective of immunoprophylaxis than that of patients, and the vaccine effects are noted maximally in children (as compared to adolescents and adults) who constitute the most responsive group The effect of vaccine is sustained for a period of about 7-8 years, following which there is a need to provide a booster vaccination for the sustained protection.
Leprosy review 07/2005; 76(2):127-43. · 1.04 Impact Factor
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ABSTRACT: The feasibility of photothermal treatment and use of salmon gonadotropin releasing hormone analogue (sGnRH-A) for induced spawning of Heteropneustes fossilis outside the spawning season was investigated. Fish were maintained at 30° C and 14 h light (L)/10 h dark (D) for 60 days between the post-spawning season and preparatory season. Fish attained gonadal maturity 4 months earlier compared to the natural spawning season. Mature fish were induced to spawn by D-Lys6 sGnRH-A at a dose of 25 μg kg−1 body weight. The eggs obtained were fertilizable. Injection of D-Lys6 sGnRH-A at different time points during the photodiurnal cycle did not influence the latency period and spawning of H. fossilis.
Journal of Fish Biology 04/2005; 45(6):909 - 915. · 1.68 Impact Factor
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ABSTRACT: Though a number of barrier methods and potent spermicides are available, most of these have nonoxynol-9 (N-9) as the active ingredient which is observed to cause inflammation and genital ulceration on repeated use. The present study was undertaken to develop a safe spermicide with conjoint microbicidal properties.
A polyherbal pessary was formulated with purified ingredients from neem (Azadirachta indica) leaves, Sapindus mukerossi (pericarp of fruit) and Mentha citrata oil. Spermicidal action on human sperm was tested by Sander-Cramer slide test in vitro and by post coital tests in vivo. Contraceptive action was tested in rabbits.
The combination of the three herbal ingredients resulted in the potentiation of the spermicidal action by 8-folds. The post coital tests confirmed the spermicidal properties of the Praneem polyherbal pessary (PPP) in women with high cervical mucous score around mid estrus. It also prevented in most women the migration of sperm into the cervical mucous. In 15 rabbits studied pregnancy was prevented by the intravaginal administration of PPP, whereas 13 of the 15 animals in the control group became pregnant.
The Praneem polyherbal pessary has potent spermicidal action on human sperm in vitro and in vivo. When applied in the vagina before mating, it prevented rabbits from becoming pregnant.
The Indian journal of medical research 04/2001; 113:135-41. · 1.84 Impact Factor
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ABSTRACT: Effects of the native GnRHs and various agonists have been evaluated on the spawning of an Indian catfish, Heteropneustes fossilis. This study tested salmon (s) GnRH agonists and mammalian (m) GnRH agonists where a D-amino acid residue was substituted alone at position 6 or the C-terminal was modified with ethylamide. GnRH agonists with a combination of these structural modifications were also evaluated separately for their effect on the spawning of the catfish. Native sGnRH, [Pro9 NEt]-sGnRH agonist and chicken (c) GnRH-II exhibited similar activity and induced spawning within 14–18 h at a dose of 100 g kg–1 body weight (BW). [D-Lys6]-sGnRH agonist and [D-Lys6 Pro9 NEt]-sGnRH agonist, induced spawning at a dose of 100 g kg–1 BW and 1 g kg–1 BW, respectively. The most notable observation in this study was the ineffectiveness of [D-Ala6]-mGnRH agonist and [Des Gly10 D-Ala6 Pro9 NEt]-mGnRH agonist. The results obtained suggest that substitution at position 6 alone, and in conjunction with an ethylamide-based modification at the C-terminal in the native sGnRH structure, increases the potency of the tested agonists to induce spawning in the catfish. This study also discusses the potential use and incorporation of cGnRH-II for the development of more generic spawning induction therapies.
Aquaculture International 01/2000; 7(6):383-392. · 0.91 Impact Factor
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ABSTRACT: 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.
Vaccine 09/1999; 17(23-24):3116-23. · 3.77 Impact Factor
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ABSTRACT: We identified a 685-nucleotide gene fragment that codes for the transmembrane and cytoplasmic domains of glycoprotein of the LEP strain rabies virus and carried out experiments designed to express a novel fusion protein on the cell surface. The cDNA encoding the membrane anchor sequence was fused in the correct reading frame to the 3' end of the cDNA encoding the beta subunit of human chorionic gonadotropin (beta(h)CG), a secretory glycoprotein that is used as an antigen for a contraceptive vaccine being developed in our laboratory. The fusion gene cassette was placed under the control of a vaccinia virus early promoter and cloned in a host-restricted fowlpox viral vector. The recombinants, when used to infect mammalian cells that do not allow the replication of fowlpox virus, expressed the N-terminal 135 amino acid residues of beta(h)CG anchored in the cell membrane by the 75-amino acid C-terminal sequence derived from rabies virus glycoprotein. This hybrid protein is correctly processed post-translationally and transported efficiently to the plasma membrane of non-permissive cells such that the anchored beta(h)CG molecule retains the correctly folded native antigenic epitope(s).
DNA and Cell Biology 08/1998; 17(7):573-81. · 2.07 Impact Factor
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ABSTRACT: 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.
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.
The pregnancies progressed to 35 to 38 weeks with the usual obstetric problems, and the children developed normally.
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.
American journal of reproductive immunology (New York, N.Y.: 1989) 07/1998; 39(6):395-8. · 3.05 Impact Factor
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ABSTRACT: A novel approach for immunocontraception by intervention of local cell mediated immunity in the reproductive system by using single intrauterine application of neem oil has been described earlier. The reversible block in fertility was reported to last for 107-180 days in female Wistar rats (Upadhyay et al., 1990. Antifertility effects of neem oil by single intrauterine administration: A novel method of contraception. Proceedings Of The Royal Society Of London B 242, 175-180) and 7-11 months in monkeys (Upadhyay et al., 1994. Long term contraceptive effects of intrauterine neem treatment (IUNT) in bonnet monkeys: An alternative to intrauterine contraceptive devices. Contraception 49, 161-167). The present study, describes the identification and characterization of the biologically active fraction from neem seeds (Azadirachta indica A. Juss. Family Meliaceae), responsible for the above activity in adult female Wistar rats. Initial studies with the mechanically extracted oil and solvent extracts of neem seeds have revealed that the antifertility activity was present in constituents of low to intermediate polarity. A hexane extract of neem seeds was reported to be biologically active (Garg et al., 1994. Comparison of extraction procedures on the immunocontraceptive activity of neem seed extracts. Journal of Ethnopharmacology 22, 87-92). Subsequently, hexane extract was sequentially fractionated through the last active fraction using various separation techniques and tested for antifertility activity at each step. Preparative HPLC was used for isolating individual components of the active fraction in quantities, sufficient for characterization. An analytical HPLC method was developed for standardization of the fraction. The active fraction was identified to be a mixture of six components, which comprises of saturated, mono and di-unsaturated free fatty acids and their methyl esters. Dose response study was performed with the last active fractions. The antifertility activity with the active fraction was reversible in nature and it was completely active until 5% concentration. There was no systemic toxic effect following the administration of the active fraction. This study, for the first time, proposes an active fraction from neem seeds, responsible for long term and reversible blocking of fertility after a single intrauterine administration with high efficacy.
Journal of Ethnopharmacology 05/1998; 60(3):235-46. · 3.01 Impact Factor
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Dna and Cell Biology - DNA CELL BIOL. 01/1998; 17(7):573-581.
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ABSTRACT: 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.
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.
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.
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.
Pharmaceutical Research 10/1997; 14(9):1264-8. · 4.09 Impact Factor
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ABSTRACT: 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.
Pharmaceutical Research 08/1997; 14(9):1264-1268. · 4.09 Impact Factor
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G P Talwar,
O M Singh,
S K Gupta,
S E Hasnain,
R Pal,
S S Majumbar,
S Vrati,
A Mukhopadhay,
J Srinivasan,
U Deshmukh,
S Ganga,
A Mandokhot,
A Gupta
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ABSTRACT: 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.
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.
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.
American journal of reproductive immunology (New York, N.Y.: 1989) 03/1997; 37(2):153-60. · 3.05 Impact Factor
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Analytical Biochemistry 11/1996; 241(2):262-4. · 3.00 Impact Factor
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Biotechnology and Bioengineering 05/1996; 50(2):228. · 3.95 Impact Factor
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ABSTRACT: The gonotrophic cycle of female Anopheles was impaired by exposure to volatiles of neem, (Azadirachta indica), reetha, (Sapindus mukorossi), and garlic, (Allium sativum), but not to castor seeds and cotton seed oil. A brief exposure to contact or volatile extracts from neem suppressed rather than inhibited oviposition. Complete inhibition of oviposition was observed by exposure of mosquitoes to neem oil and 1 fraction containing volatile components. Vitellogenesis was impaired irreversably by long-term exposure to neem odor and some extracts. The effect of volatiles on oviposition seems to be regulated by absorption through the cuticle, although passage through the spiracles could not be excluded.
Journal of Medical Entomology 04/1996; 33(2):195-201. · 1.76 Impact Factor
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ABSTRACT: In the course of clinical trials on a birth control vaccine, it was found that some of the immunized women responded poorly to booster immunizations. This vaccine consists of a dimer of the beta chain of human chorionic gonadotropin (beta hCG) and the alpha chain of ovine luteinizing hormone (alpha oLH), linked to tetanus toxoid (TT) as a carrier. Changing this carrier to diphtheria toxoid resulted in reversion to high anti-hCG antibody titers, indicating the extent to which the carrier influences anti-ligand responses in this system. The suppression of anti-hCG responses after booster immunizations was reminiscent of the phenomenon of carrier-induced, epitope-specific regulation. In a mouse model designed to test the effects of preimmunization with TT on anti-hCG responses, we found that a single preimmunization with TT causes reduced anti-hCG antibody responses in two out of four mouse strains, while anti-alpha oLH antibody responses were not affected by the preimmunization with TT. This is particularly interesting considering that beta hCG and alpha oLH were not presented when linked separately to TT. In an effort to devise a strategy to circumvent this carrier-induced, ligand-specific hyporesponsiveness, we investigated the effectiveness of a synthetic T helper epitope from TT as carrier. We show that preimmunization with TT causes a less profound reduction in anti-hCG titers if the preimmunized mice are subsequently injected with alpha oLH-beta hCG conjugated to a synthetic tetanus toxin peptide recognized by TT-induced and peptide-induced T cells.
European Journal of Immunology 01/1996; 25(12):3375-80. · 5.10 Impact Factor
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ABSTRACT: 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.
Infection and Immunity 01/1996; 63(12):4907-11. · 4.16 Impact Factor
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ABSTRACT: Synthesis of the beta-subunit of human chorionic gonado-tropin (betahCG) in Vero cells by the recombinant vaccinia virus has been studied. The yield of betahCG was a function of the multiplicity of infection (MOI), and was highest at 25 MOI. The kinetics of synthesis and initial secretion of betahCG, deduced from the pulse-chase experiments were "zero order." At 30 h postinfection, the relative values of net synthesis and secretion rates were 4.0 AU. mm(2) betahCG/10(6) cells. h and 1.55 AU. mm(2) betahCG/10(6) cells. h, respectively. The time required to secrete 50% of intracellular betahCG was 210 min. Pulse-chase data also showed that 24% of betahCG was degraded intracelluiarly within 10 h, of which 17% was detected in the autoradiogram. Along with 30 kD betahCG, a satellite band of 28 kD was evident among the peptide synthesized in Vero cells. The molecular weight of vaccinia-derived betahCG was 13 kD more that its nonglycosylated form, indicating extensive glycosylation in Vero cells. The mRNA levels in infected Vero cells at different postinfection times were quantified by excess DNA dot-blot hybridization. It appears that the Vero cell possesses some host cell-associated factor(s), which prevented the transcription of early betahCG-mRNA promoted by the early signal of the vaccinia P 7.5 promoter. The half-life of betahCG-mRNA, as determined by follow-up of decay after blocking transcription initiation, was found to be 6.4 h. The synthesized betahCG was immunoreactive as it reacted with monoclonal and polyclonal monospecific antibodies. The subunit was also biologically active, as it combined with native betahCG to form heterodimer betahCG, which competed with (125)I-hCG for radioreceptors and stimulated testosterone synthesis in Leydig cells. (c) 1995 John Wiley & Sons, Inc.
Biotechnology and Bioengineering 10/1995; 48(2):158-68. · 3.95 Impact Factor
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ABSTRACT: 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.
The Indian journal of medical research 09/1995; 102:66-70. · 1.84 Impact Factor
