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Biomedical subjects

V Singh

Publications and source records attributed to V Singh.

At least 271 records · Page 15Linked to original sources

Repeated dermal toxicity of technical HCH and methyl parathion (50EC) to female rats (Rattus norvigicus).

Repeated dermal application of hexachlorocyclohexane (HCH; 100 mg/kg/day) or methyl parathion (2 mg/kg/day) individually or in combination for 7, 15 and 30 days produced pathomorphological changes in skin, liver, kidney and brain of female rats along with significant enzymatic alterations in the activity of transaminase, alkaline phosphatase lactic dehydrogenase and acetylcholinesterase. The two insecticides in combination though produced severe toxicity on day 30 than at other periods, the changes were not suggestive of any additive or potentiation effect at the test doses.

Administration, Cutaneous↗

Comparison of four different ovarian stimulation protocols in an in vitro fertilisation & embryo transfer programme.

Four different ovarian stimulation protocols were evaluated in an in vitro fertilisation and embryo transfer programme in 208 women (228 treatment cycles). In the rigid protocol (RP), 100 mg of clomiphene citrate (CC) was given from day 3 to day 7 of the menstrual cycle and 300 IU of human menopausal gonadotropin (hMG) was given from day 5 of the menstrual cycle. In the individualised protocol (IP) the same drugs and doses were used as in RP, but the day of initiation of CC depended on the length of the individual's menstrual cycle and hMG was administered from the last day of CC. In the programmed protocol (PP), ovarian function was suppressed with oral contraceptive pills (ethinyl estradiol 30 micrograms and norethisterone 1 mg) started on day 5 of the menstrual cycle for 45 to 70 days. Considering the last day of pill intake as day 0, CC was given for 5 days from day 5 and hMG (300 IU) from day 7. In the alternate day protocol (ADP), 100 mg of CC was administered from day 2 to day 6 and hMG (300 IU) was given on alternate days from day 2 to day 8 or day 10 of the cycle. In all the women, hCG (5000 IU) was administered when the diameter of at least 2 follicles was greater than or equal to 16 mm and estradiol levels were 300 pg/ml/dominant follicle. Patients not showing such a response were not treated further. The cardinal events of IVF-ET such as number of good responders, incidence of oocytes harvested, fertilised and embryos transferred per cycle were compared and it was concluded that the pregnancy rates were highest in women treated by the PP.

Adult↗

Effect of thiolation of amino groups of gelonin on its protein-biosynthesis inhibitor activity.

Gelonin was purified from the dry seeds of Gelonium multiflorum by ammonium sulfate fractionation followed by cation-exchange and gel-filtration chromatography in order to minimize extraction of non-proteineous material. Gelonin was characterized for its purity, homogeneity and molecular weight determination by RP-HPLC and SDS-PAGE analysis respectively. The amino groups of pure gelonin were thiolated by a hererobifunctional cross-linking agent, SPDP which is used in the design of cytotoxic hybrid molecules. Therefore, an attempt has been made to study the effect of thiolation on the ribosome inactivating property of gelonin. Thiolation of one amino group resulted in the loss of about 90% protein synthesis inhibition activity. Further modification of 2-3 amino groups further hampered the bioactivity (greater than 95-99.5%) of gelonin, suggesting that a 1:1 molar ratio of carrier-toxin conjugate would be highly active against the target cells.

Amino Acids↗

In-vitro selective killing of gonadal cells by a hormonotoxin composed of ovine luteinizing hormone linked by a disulfide bond to the ribosome-inactivating protein, gelonin.

With the aim to synthesize a bioeffective hormonotoxin for selective targeting to specific cells in the gonads, a single chain RIP was conjugated to oLH with the use of SPDP which generated oLH-S-S-gelonin hormonotoxin. Extensive physico-chemical, immunochemical and biochemical characterization reveal that 1:1 oLH-gelonin was linked through the alpha-subunit of the oLH. The hormonotoxin retained substantial receptor binding and steroidogenic activity in the leydig tumor cells. The competitive displacement analysis indicate that the binding occurs via the hormone. The presently described hormonotoxin exhibited higher receptor binding and cytotoxicity to the target cells than that of others reported so far.

Animals↗

Effect on immunoreactivity of gonadotropin releasing hormone (GnRH) in the GnRH-BSA conjugates of variable stoichiometry.

In order to render hypothalamic 'self' decapeptide GnRH immunogenic, the GnRH and its azotized derivative were covalently coupled to the carrier protein, BSA. The azotization at histidine and or tyrosine of GnRH was carried out and characterized extensively. Three different molar ratios of GnRH/azo-GnRH with BSA were prepared and characterized by physico-chemical techniques. The conjugates purified by gel-filtration chromatography on G-100 and on gel permeation column using HPLC were subjected to molar ratio determination by amino acid analysis. The immunoreactivity of GnRH/azo-GnRH towards monoclonal and polyclonal antibodies was drastically hampered after conjugation. It is therefore suggested that in the GnRH-carrier conjugate, GnRH concentration should be determined carefully by using specific radioimmunoassay.

Amino Acid Sequence↗

Hormonotoxins: abrogation of ribosome inactivating property of gelonin in the disulfide linked ovine luteinizing hormone-gelonin conjugates.

In order to synthesize a bioeffective hormonotoxin for selective targeting to specific cells in the gonads, gelonin, a single chain ribosome-inactivating protein obtained from an Indian plant called Gelonium multiflorum was covalently linked to ovine luteinizing hormone (oLH) by a disulfide bond. Ovine LH-S-S-gelonin conjugates of different molar ratios were subjected to determine the ribosome-inactivating property in a cell-free translation assay using rabbit reticulocyte lysate system. A single amino group modification with N-succinimidyl-3-(2-pyridyldithio)propionate resulted in a loss of 90% protein synthesis inhibition activity. Upon conjugation of gelonin to oLH, the activity was further inhibited ranging from 2.5-6.4%. A 1:1 to 1:1.5 molar ratio (oLH-S-S-gelonin) conjugates showed 2.5-4.6% activity while 1:2.8 to 1:2.2 molar ratio exhibited 5.5-6.4% inhibition ability.

Animals↗

Hormonotoxins: synthesis, characterization and bioefficacy of some defined disulfide linked conjugates of ovine LH with a ribosome inactivating protein, gelonin.

In order to synthesise a bioeffective hormonotoxin for selective targeting to specific cells in the gonads, gelonin, a single chain RIP obtained from an Indian plant, Gelonium multiflorum of Euphorbeaceae family was covalently linked to oLH with the use of N-succinimidyl-3-(2-pyridyldithio)propionate, generating a linkage containing a disulfide bond and a amide bond. The hormonotoxins were separated according to their molecular weight (indirectly according to oLH:gelonin molar ratio) and a complete biochemical analysis was performed. The linkage occurred through the epsilon-NH2 group of alpha oLH as judged from RP-HPLC analysis. The conjugates were devoid of ingredients as determined by SDS-PAGE and RP-HPLC analysis. The hormonotoxins retained substantial receptor binding, steroidogenic activity and immunoreactivity of oLH and gelonin to their antibodies. Hormonotoxins bind to the Leydig tumour cells via oLH part leaving gelonin free as judged by competitive displacement analysis. The hormonotoxin was internalized to the sufficient degree to effectively inhibit protein synthesis. The cytotoxicity of 1:1 molar ratio conjugate was relatively higher than that of others. The cytotoxicity of presently described more defined hormonotoxins exhibited higher receptor binding and cytotoxicity than the hormonotoxins reported earlier [Singh, et al., J Biol Chem, 264 (1989) 3089].

Animals↗

Craniofacial suture stenosis: morphologic effects.

Craniofacial anomalies, such as Apert's and Crouzon's syndromes, are presumed to be related to premature growth arrest of cranial base growth sites. However, premature growth arrest at cranial vault sutures in animals appears to play a causative role in the development of cranial deformities characteristic of single-suture, or simple, craniosynostosis in humans. To study the possible causative role of cranial vault and other (interface) suture stenoses on the development of craniofacial deformity, a vault suture and an interface suture between the cranial vault and facial skeleton were simultaneously immobilized. Thirty-one New Zealand White rabbits at 9 days of age underwent implantation of dental amalgam growth markers adjacent to cranial vault and facial sutures. In the experimental group (n = 15), methylcyanoacrylate adhesive was applied over the coronal (vault) and frontonasal (interface suture between vault and facial skeleton) sutures to immobilize them. The remaining 16 animals served as sham-treated controls. All animals underwent serial radiographic cephalometry to document growth effects in the cranial vault, cranial base, and facial skeleton. Application of adhesive resulted in statistically significant (p less than 0.05) reduction in growth at the coronal and frontonasal sutures. This was accompanied by an overall significant reduction in neurocranial vault length during the first 30 days of development.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Liquid membrane phenomenon in the actions of digitalis.

The liquid membrane phenomenon in the actions of digitalis glycosides (digitoxin, digoxin and ouabain) has been studied. Formation of liquid membranes, in series with a supporting membrane, by digitalis alone and by digitalis in association with lecithin and cholesterol has been demonstrated. The results obtained on the transport of relevant permeants, viz. sodium, potassium and calcium ions and dopamine, adrenaline, noradrenaline and serotonin, in the presence of the liquid membrane generated by digitalis in association with lecithin and cholesterol indicate that the liquid membrane barrier to transport may have a relevance with the biological actions of digitalis.

Cholesterol↗

Effect of yoga breathing exercises (pranayama) on airway reactivity in subjects with asthma.

The effects of two pranayama yoga breathing exercises on airway reactivity, airway calibre, symptom scores, and medication use in patients with mild asthma were assessed in a randomised, double-blind, placebo-controlled, crossover trial. After baseline assessment over 1 week, 18 patients with mild asthma practised slow deep breathing for 15 min twice a day for two consecutive 2-week periods. During the active period, subjects were asked to breathe through a Pink City lung (PCL) exerciser--a device which imposes slowing of breathing and a 1:2 inspiration:expiration duration ratio equivalent to pranayama breathing methods; during the control period, subjects breathed through a matched placebo device. Mean forced expiratory volume in 1 s (FEV1), peak expiratory flow rate, symptom score, and inhaler use over the last 3 days of each treatment period were assessed in comparison with the baseline assessment period; all improved more with the PCL exerciser than with the placebo device, but the differences were not significant. There was a statistically significant increase in the dose of histamine needed to provoke a 20% reduction in FEV1 (PD20) during pranayama breathing but not with the placebo device. The usefulness of controlled ventilation exercises in the control of asthma should be further investigated.

Adult↗

Risk factors related to biological behaviour of precancerous lesions of the uterine cervix.

In a study of factors related to cervical carcinogenesis, a cohort of 1,107 cervical dysplasia along with 1,077 controls matched for age and parity were followed up prospectively. During the follow up 75 dysplasia cases progressed to carcinoma in situ. The overall rate of progression of dysplasia to malignancy was observed to be 15.7% at the end of 108 months of follow-up. The analysis of progression rates in relation to various factors revealed significantly higher progression rates for initially higher grade of dysplastic lesions, and early age at consummation of marriage (ACM). The other factors, such as religion, literacy status of the patient, number of pregnancies, presence of cervical erosion, history of fetal loss and positivity to HSV-II antibodies, did not reveal statistical significance. The case-control comparison for detection of HPV 16/18 by in situ hybridisation revealed the presence of HPV 16/18 sequences in 67.3% of the dysplasia subjects progressed to carcinoma in situ while 27.3% of precancerous cases regressed to normalcy. The difference was found to be statistically significant (P less than 0.001).

Adult↗

Relative merits of various rapid biopsy urease tests for diagnosis of Helicobacter pylori (Campylobacter pylori).

Three rapid urease tests, i.e., liquid urea broth containing phenol red as indicator, liquid urea broth containing bromothymol blue as indicator and CLO gel were compared in 109 patients of dyspepsia for the diagnosis of Campylobacter pylori (Helicobacter pylori) infection. Mean time taken for positive reaction in liquid broth with phenol was 3 minutes (range 0.6 to 5.3 minutes) with bromothymol blue was 3.5 minutes (range 0.4 to 5.5 minutes) while with CLO gel it was 101 minutes (range 11-261 minutes). There was no difference in results of liquid urea broth containing phenol red and bromothymol blue. The difference in timing of urea broth containing phenol red and bromothymol blue was statistically significant as compared to CLO gel (p less than 0.05). Rapid urease tests employing liquid urea broth are quick, simple and reliable for the diagnosis of Helicobacer pylori infection.

Adolescent↗

Serodiagnosis of gut tuberculosis.

The efficacy of enzyme-linked immunosorbent assay (ELISA) in the diagnosis of gut tuberculosis was evaluated. The study included 95 patients with gut tuberculosis, 40 with pulmonary tuberculosis, 25 with non-tubercular intestinal diseases and 40 healthy controls. Mantoux response and antibody titre against PPD using ELISA were studied. The ELISA sensitivity in gut tuberculosis was 80% and in pulmonary tuberculosis 90%. It was also positive in 12% of patients with non-tubercular intestinal diseases and 5% of healthy controls. The sensitivity of Mantoux test in gut (77%) and pulmonary (84%) tuberculosis was comparable to that of ELISA. However, Mantoux test lacked diagnostic significance due to high positivity in non-tubercular disease patients (55%) and healthy controls (56%), whereas ELISA was highly specific (93%) for the diagnosis of gut tuberculosis.

Antibodies, Bacterial↗