Comparison of in vitro and in vivo fertilizing potential of buffalo bull semen frozen in egg yolk-, soya bean lecithin- and liposome-based extenders.

The objective of this study was to compare different extenders for post-thaw in vitro sperm function and in vivo fertility of buffalo semen. Accordingly, sperm of 30 ejaculates extended in egg yolk (TRIS with 20% egg yolk; EY), two soya lecithin-based (SL-1; AndroMed® and SL-2; Bioxcell® ) and a liposome-based extender (LS; OptiXcell® ) were tested. The post-thaw semen was evaluated for computer-assisted sperm analysis (CASA), sperm viability, membrane and acrosome integrity, DNA integrity and acrosome reaction and first service pregnancy rate (FSPR) in a fixed-time artificial insemination programme. Total motility and VCL were the only CASA-based parameters that exhibited significantly higher (p < .05) percentage in LS among these extenders. Post-thaw percentage of acrosome integrity (55.9 ± 1.4, 58.1 ± 2.0, 55.8 ± 2.0, 56.6 ± 2.3) and DNA integrity (68.8 ± 2.0, 69.2 ± 2.3, 71.3 ± 2.1, 69.1 ± 2.1) did not differ (p > .05) in EY, SL-1, SL-2 and LS extender, respectively. However, a variable response in terms of efficacy of different extenders for sperm viability and plasma membrane integrity was observed. Assessment of inducibility of acrosome reaction showed significant differences between extenders (51.9 ± 2.1, 44.3 ± 2.4, 46.1 ± 2.3 and 58.1 ± 3.1%, respectively, for EY, SL-1, SL-2 and LS). Furthermore, field trials revealed significantly higher (p < .05) FSPR of LS-extended semen as compared to that for EY, SL-1 and SL-2 extender (46.3%, 41.2%, 31.2% and 29.7%, respectively). It is concluded that the liposome-based extender is more effective than egg yolk- and soya lecithin-based extenders and may be used for cryopreservation of buffalo semen in the future.

Cytomorphological studies in some members of tribe Paniceae (Poaceae) from district Kangra of Himachal Pradesh (Western Himalayas).

The present paper deals with cytological studies on the population basis of 21 species belonging to 9 genera of tribe Paniceae of family Poaceae from cytologically unexplored area of Western Himalayas i.e. district Kangra of Himachal Pradesh for the assessment of genetic diversity of grass flora. On world-wide basis, the chromosome counts have been made for the first time for three species such as Brachiaria remota (n = 16), Digitaria granularis (n = 36) and Isachne albens (n = 5). Similarly, on India basis, altogether new records are made for two species such as Echinochloa cruspavonis (n = 27) and Paspalum distichum (2n = 50). A comparison of the different euploid cytotypes studied at present for Digitaria adscendens, D. setigera and Oplismenus compositus revealed significant variations in their morphology, depicting increase in some of the characters of polyploid cytotypes. The course of meiosis has been observed to be normal in all the studied populations with high pollen fertility except for two species such as Paspalum dilatatum and P. distichum marked with abnormal meiosis and reduced pollen fertility.

Effect of supplementation of vitamin E, copper and zinc on the in vitro phagocytic activity and lymphocyte proliferation index of peripartum Sahiwal (Bos indicus) cows.

To study the effect of vitamin E (VE), copper (Cu) and zinc (Zn) supplementation on the in vitro phagocytic activity (PA) and lymphocyte proliferation response (LPR) of blood neutrophils and lymphocytes, thirty Sahiwal pregnant cows (six in each group) in their late gestation at 30 days before the expected date of calving were selected from the NDRI experimental herd and supplemented with various micronutrients from 30 days before calving to 45 days after calving. Cows were supplemented individually with VE (1000 IU/cow/day), Cu (20 ppm/cow/day) and Zn (80 ppm/cow/day) and also with a combination of VE, Cu and Zn to study cumulative effect of all micronutrients. One group without any supplementation acted as a control. Blood neutrophils and lymphocytes were isolated and studied for their PA and LPR. Supplementation of micronutrients like VE, Cu, Zn and a combination of all these nutrients significantly (p < 0.01) increased the PA of experimental cows as compared to control (unsupplemented) cows during the pre-partum period. During post-partum, all the micronutrients (VE, Cu, Zn and their combination) showed a significant (p < 0.01) increase in the PA of experimental cows as compared to control cows. Of all the groups, significant (p < 0.01) and maximum PA was observed in the combination group followed by Zn-supplemented group during both the pre- and post-partum period. A significant (p < 0.01) increase in LPR of B lymphocytes was observed in combination-supplemented group during the pre-partum period and during both the pre- and post-partum period in the Cu-supplemented group.

Effect of vitamin E supplementation on mRNA expression of superoxide dismutase and interleukin-2 in arsenic exposed goat leukocytes.

The aim of this study was to quantify the expression level of genes involved in antioxidant defenses during inorganic arsenic (iAs) exposure in the blood of goats and to evaluate the regulative activity on these genes of antioxidant vitamin E in the diet. Twenty-four crossbred lactating goats (Alpine × Beetal) were distributed randomly into four equal groups (Control, T(1), T(2) and T(3)) of six in each, on the basis of average body weight (36.10 ± 0.11 kg) and milk yield (1.61 ± 0.004 kg/day). The animals in T(1), T(2) and T(3) were given 50 mg/kg dry matter arsenic daily, while in T(2) and T(3), vitamin E @100 IU and 150 IU/kg dry matter, respectively, was also supplemented additionally for the period of 12 months. Blood was sampled at 0 day then at 3 months interval and analyzed for the expression level of superoxide dismutase (Cu/Zn SOD) and interleukin-2 (IL-2) using real-time PCR technique. Initially there was no difference (p > 0.05) in relative expression of the two genes. But, at 3 months, relative expression of Cu/Zn SOD increased (p < 0.05) in T(1) groups then, at 6 and 9 months expression was decreased (p < 0.05) in all the iAs treated groups whereas at 12 months, vitamin E supplementation increased (p < 0.05) the expression which is comparable to control groups. IL-2 mRNA expression was decreased (p < 0.05) at 6 months in all iAs treated groups, at 9 months there was decline trend but not significantly different whereas at 12 months decline trend was less (p < 0.05) in vitamin E supplemented groups. The result suggests that vitamin E may have a controlling effect on oxidative stress through modulation of SOD and IL-2 expression.

Effect of vitamin E supplementation on arsenic induced oxidative stress in goats.

The present study was designed to assess whether supplementation of different levels of vitamin E to long-term arsenic exposed goats affords protection against the oxidative stress caused by the metalloid. Twenty-four crossbred lactating goats were distributed randomly into four groups (control, T(1), T(2) and T(3)) of six in each. The animals in T(1), T(2) and T(3) were given 50 mg/kg DM arsenic daily, while in T(2) and T(3), vitamin E @100 IU and 150 IU/kg DM, respectively, was also supplemented additionally for the period of 12 months. Compared to control, significant (p < 0.05) decline in SOD (45 %), CAT activities of erythrocytes (63 %), plasma total Ig (22 %) and total antioxidant activity (24 %) was observed in only arsenic treated groups and vitamin E supplementation in both doses produced partial mitigation effect against SOD (23 %, 20 %) and CAT (39 %, 48 %) while complete mitigation against total Ig (16 %, 7 %) and antioxidant activity (10 %, 8 %) was found. Average lymphocyte stimulation index at the end of experiment was (p < 0.05) lower in arsenic exposed groups (1.003 ± 0.01) and significant (p < 0.05) recovery was observed in response of vitamin E supplementation at higher doses (1.138 ± 0.03). So, vitamin E is helpful in reducing the burden of arsenic induced oxidative stress and activities of antioxidant enzymes in goats.

Activation of natural killer-like YT-INDY cells by oligodeoxynucleotides and binding by homologous pattern recognition proteins.

The present study was designed to examine the binding and signalling effects of single base and CpG dinucleotide phosphodiester (Po) oligodeoxynucleotides (ODN) on the human natural killer (NK)-like cell line (YT-INDY). Single base Po ODN composed of 20-mers of guanosine (dG20), adenosine (dA20), cytosine (dC20) or thymidine (dT20) as well as 'conventional' Po CpG ODN were examined for their ability to bind and activate YT-INDY cells. Binding by dG20 and CpG ODN to YT-INDY cells was saturable and specific. dG20 binding was competitively inhibited by homologous dG20 and heterologous CpG ODN but not by dC20 and dA20. Two different YT-INDY membrane proteins (18 and 29 kDa) were identified by ligand (Southwestern) blotting with biotinylated dG20 and CpG. The specificity of the ODN-binding protein(s) was further confirmed by ODN depletion experiments using a teleost recombinant protein orthologue [nonspecific cytotoxic cells (NCC) cationic antimicrobial protein-1 (ncamp-1)] known to bind CpG and dG20. Cell proliferation and activation studies showed that dG20 and CpG treatment of YT-INDY cells induced cellular DNA synthesis (i.e. G1 to S-phase conversion). This signalling function was accompanied in dG20-treated cells by proliferation 10 h posttreatment. Both dG20 and CpG ODN binding induced a calcium flux in YT-INDY cells within seconds of treatment. These experiments demonstrated that Po single base dG20 and CpG ODN bind to a (potential) new class of cell-surface proteins that mediate the activation of YT-INDY cells.

Synergistic antimicrobial activity of tea & antibiotics.

Tea leaves are known for its antibacterial activity against many microorganisms. In this study we attempted to describe the synergistic antimicrobial activity of tea and antibiotics against enteropathogens. Antimicrobial activity of boiled water tea extract and organic solvent extract were studied against Salmonella typhimurium 1402/84, S. typhi, S. typhi Ty2a, Shigella dysenteriae, Yersinia enterocolitica C770, and Escherichia coli (EPEC P2 1265) determining minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and death rate kinetics at MBC of tea extract in presence of subinhibitory concentration of antibiotic. Both green tea or black tea extracts effectively inhibited the growth of S. typhimurium 1402/84, S. typhi, S. typhi Ty2a, S. dysenteriae, Y. enterocolitica C770, and E.coli (EPEC P2 1265). However, the growth inhibitory concentration of tea extract was lower for green tea as compared to black tea extract. Antimicrobial activity of green tea tea methanol: water extract tea was better as compared to boiled water tea extract of green tea. Based on death rate kinetics results, S.typhi Ty2a appeared to be highly sensitive and Y. enterocolitica C770 the most resistant. Chloramphenicol and tea extract in combination inhibited the growth of S.dysenteriae at 2.5 microg/ml chloramphenicol (MIC 5 microg/ml) and 5.094 mg/ml black tea extract (MIC 9.089 mg/ml). Tea extract showed synergistic activity with chloramphenicol and other antibiotics like gentamycin, methicillin and nalidixic acid against test strains.

Neuronal conduction studies of the median nerve in non-impaired humans: a comparison of accepted techniques.

BACKGROUND AND PURPOSE: There are two commonly accepted techniques used for distal electro-stimulation placement when performing median motor nerve conduction studies. The purpose of this study was to compare latency using two commonly accepted sites of distal stimulation of the median nerve when performing motor nerve conduction studies on non-impaired adult humans. PARTICIPANTS: The sample consisted of 36 non-impaired participants (15 female, 21 male) aged 20 to 40 years. METHODS: Participants were randomly assigned to two groups and tested bilaterally for the median motor nerve. For distal stimulation of the median motor nerve, in the first group, 8 cm was measured from the center of the muscle diagonally to arrive at a point between the flexor carpi radialis and plamaris longus tendons. In the second group, 3.5 cm was measured from the distal wrist crease proximally along the median nerve for the distal stimulation of the median motor nerve. Distal latency of both techniques was obtained. Surface skin temperature of the palm was recorded throughout the procedures. RESULTS: No significant differences were found between the 8 cm and 3.5 cm techniques at p < or = 0. 05 level. COMMENT: Even though no differences were found between the two techniques, the 3.5-cm technique is recommended because of its consistency as an anatomical landmark reducing the potential for measurement error.

Evaluation of the pesticidal potential of the congress grass, Parthenium hysterophorus Linn. on the mustard aphid, Lipaphis erysimi (Kalt.).

Petroleum ether extracts of leaves, stem and inflorescence of Parthenium hysterophorus Linn. at 500, 1000, 2000 and 5000 ppm concentrations were tested in the laboratory for their toxic effects on the mean life span and progeny production of adults of the mustard aphid, Lipaphis erysimi (Kalt.). The investigations revealed a significant decrease in life span and progeny production with treatment. Among the three plant parts tested for their efficacy, the leaf extract showed the most significant effect in causing a dose dependent decline in both the life span and progeny production.

Inhibition of peroxynitrite dependent DNA base modification and tyrosine nitration by the extra virgin olive oil-derived antioxidant hydroxytyrosol.

Hydroxytyrosol is one of the o-diphenolic compounds in extra virgin olive oil and has been suggested to be a potent antioxidant. The superoxide radical (O2*-) and nitric oxide (NO*) can react very rapidly to form peroxynitrite (ONOO ), a reactive tissue damaging species thought to be involved in the pathology of several chronic diseases. Hydroxytyrosol was highly protective against the peroxynitrite-dependent nitration of tyrosine and DNA damage by peroxynitrite in vitro. Given that extra virgin olive oil is consumed daily by many humans, hydroxytyrosol derived from this diet could conceivably provide a defense against damage by oxidants in vivo. The biological activity of hydroxytyrosol in vivo will depend on its intake, uptake and access to cellular compartments.

Maternal mortality -- aetiological factors: analytic study from a teaching hospital of Punjab.

PIP: A review of maternal deaths at Rajendra Hospital, Punjab, from January 1978 to December 1991 yielded important data for the planning of maternal health services in this area of India, During the 14 year study period, there were 33,160 births and 339 deaths, for a maternal mortality rate of 1002/100,000 live births. Women who had received no prenatal care accounted for 47.4% of deliveries but 92.8% of maternal deaths. In addition, a disproportionate number of deaths involved rural women (74.6%) and poor women (76.4%). 57.8% of maternal deaths involved women 21-30 years of age; 37.1% occurred among primigravidas. Direct obstetrical causes were considered the etiologic factor in 83.1% of these deaths. Primary among these causes were sepsis (37.1%), obstetric hemorrhage (26.2%), hypertensive disorders of pregnancy (21.4%), and obstructed labor (15.3%). 30.6% of deaths occurred during pregnancy, 50.3% during labor, and 19.1% in the postpartum period. Indirect obstetrical causes, notably severe anemia and anesthesia complications, were implicated in 15.3% of the maternal deaths. Critical analysis of the maternal deaths in this series suggested that 89.6% were totally preventable, 9.6% were probably preventable, and only 0.8% were not avoidable. Factors that would reduce the high rate of maternal mortality in this region include more widespread use of prenatal care, training of traditional birth attendants in asepsis, referral of high-risk pregnancies, and improved transportation in rural areas.^ieng

meso-Tetra(hydroxyphenyl)porphyrins, a new class of potent tumour photosensitisers with favourable selectivity.

We compared para-, meta- and ortho-isomers of meso-tetra(hydroxyphenyl)porphyrin (p-, m- and o-THPP) and the potassium salt of the para compound (K-p-THPP) with haematoporphyrin derivative (HpD) and Photofrin II in their ability to sensitise tumours, skin and brain to light. HpD and Photofrin II induced modest tumour photosensitisation at the cost of substantial skin and brain sensitisation. At doses low enough to keep sensitisation of these normal tissues within acceptable limits, tumour sensitisation was sufficient to give necrosis only approximately 2 mm deep after exposure to 10 J cm-2 light. In contrast, doses of p-THPP, K-p-THPP and m-THPP that produced skin and brain sensitivity within acceptable limits sensitised tumours enough to give 4-9 mm necrosis after 10 J cm-2 light. m-THPP was, on a molar basis, about 25-30 times as potent as HpD and Photofrin II in sensitising tumours. o-THPP was also a potent tumour photosensitiser, but induced a prohibitive degree of skin photosensitivity even at low doses. It is unlikely that these differences in relative selectivity are due to differences in such photophysical parameters as optimum activating wavelength (which would affect tissue penetration by light), or light absorption, and physicochemical factors that determine tissue localisation may be involved. The high tumour sensitising potency and favourable tissue selectivity of m-THPP, p-THPP and K-p-THPP make them promising candidates for clinical tumour phototherapy.