Adaptation strategies of yak to seasonally driven environmental temperatures in its natural habitat.

The gradual increase of ambient temperature (TA) at high altitude can cause heat stress as an effect of climate change and may shift the traditional habitat of yak to further higher altitude. Therefore, an attempt has been made in this study to evaluate the thermo-adaptability of yaks to different seasons at high altitude. The adaptive capabilities of yaks were assessed based on different heat tolerance tests in relation to changes in rectal temperature (RT; °F), respiration rate (RR; breaths/min), pulse rate (PR; beats/min), and plasma heat shock protein (HSP) profile. The experiment was conducted in 24 yaks, divided into three groups based on age as calf (n = 8), adult (n = 8), and lactating cow (n = 8). Thermal adaptability was determined by temperature humidity index (THI), dairy search index (DSI), and Benezra's thermal comfort index (BTCI) along with HSP70 profile. The THI was higher (P < 0.01) in summer than winter which increased from lowest (40.87) to highest (61.03) in summer by 20 points, where yaks were under heat load beyond THI 52. The RT (100.09 ± 0.18 °F), RR (21.76 ± 0.18), and PR (59.78 ± 0.32) increased by 23-35%, and this was correlated to the higher values of DSI exceeding 1 in calves (1.35 ± 0.03), lactating cows (1.29 ± 0.04), and adults (1.23 ± 0.32) during summer in comparison to winter (0.98 ± 0.02). The BTCI also showed values greater (P < 0.01) than 2 in calves (3.47 ± 0.27), lactating cows (3.23 ± 0.28), and adults (2.98 ± 0.29) which reflected 49-75% increase in rectal temperature and respiration rate during summer. Further, heat stress was substantiated by threefold higher (P < 0.01) level of plasma HSP70 in calves (189.61 ± 3.90 pg/ml) followed by lactating cows (168.62 ± 3.03 pg/ml) and adults (155.33 ± 2.30 pg/ml) against the winter average of 87.92 ± 3.19 pg/ml. Present results revealed that yaks were experiencing heat stress in summer at an altitude of 3000 m above sea level and calves were more prone to heat stress followed by lactating cows and adults.

Changes in expression of monocarboxylate transporters, heat shock proteins and meat quality of Large White Yorkshire and Ghungroo pigs during hot summer period.

OBJECTIVE: Present study explores the effect of hot summer period on the glycolytic rate of early post-mortem meat quality of Ghungroo and Large White Yorkshire (LWY) pig and comparative adaptability to high temperature between above breeds by shifting the expression of stress related genes like mono-carboxylate transporters (MCTs) and heat shock proteins (HSPs). METHODS: Healthy pigs of two different breeds, viz., LYW and Ghungroo (20 from each) were maintained during hot summer period (May to June) with a mean temperature of about 38°C. The pigs were slaughtered and meat samples from the longissimus dorsi (LD) muscles were analyzed for pH, glycogen and lactate content and mRNA expression. Following 24 h of chilling, LD muscle was also taken from the carcasses to evaluate protein solubility and different meat quality measurements. RESULTS: LWY exhibited significantly (p<0.01) higher plasma cortisol and lactate dehydrogenase concentration than Ghungroo indicating their higher sensitivity to high temperature. LD muscle from LWY pigs revealed lower initial and ultimate pH values and higher drip loss compared to Ghungroo, indicating a faster rate of pH fall. LD muscle of Ghungroo had significantly lower lactate content at 45 min postmortem indicating normal postmortem glycolysis and much slower glycolytic rate at early postmortem. LD muscle of LWY showed rapid postmortem glycolysis, higher drip loss and higher degrees of protein denaturation. Ghungroo exhibited slightly better water holding capacity, lower cooking loss and higher protein solubility. All HSPs (HSP27, HSP70, and HSP90) and MCTs (MCT1, MCT2, and MCT4) in the LD muscle of pigs inclined to increase more in Ghungroo than LWY when exposed to high temperature. CONCLUSION: Effect of high temperature on the variation of HSPs and MCTs may play a crucial role in thermal tolerance and adaptation to different climatic conditions, pH regulation, muscle acidification, drip loss, protein denaturation and also in postmortem meat quality development.

Clinical evaluation of purified Shilajit on testosterone levels in healthy volunteers.

Purified Shilajit, an Ayurvedic rasayana, was evaluated in healthy volunteers of age between 45 and 55 years for its effect on male androgenic hormone viz. testosterone in a randomised, double-blind, placebo-controlled clinical study at a dose of 250 mg twice a day. Treatment with Shilajit for consecutive 90 days revealed that it has significantly (P < 0.05) increased total testosterone, free testosterone and dehydroepiandrosterone (DHEAS) compared with placebo. Gonadotropic hormones (LH and FSH) levels were well maintained.

Hypokalemic periodic paralysis, facial dysmorphism and ventricular arrhythmia (clinical triad of Andersen-Tawil syndrome).

Andersen-Tawil Syndrome (ATS) is a rare potassium channel disorder, characterized by episodic weakness, ventricular arrhythmias and dysmorphic features (short stature, scoliosis, clinodactyly, hypertelorism, small or prominent low set ears, micrognathia and broad forehead). We report a case of hypokalemic periodic paralysis with dysmorphic facial features and ventricular arrhythmia resembling Andersen-Tawil syndrome.

Clinical evaluation of spermatogenic activity of processed Shilajit in oligospermia.

The safety and spermatogenic activity of processed Shilajit (PS) were evaluated in oligospermic patients. Initially, 60 infertile male patients were assessed and those having total sperm counts below 20 million ml(-1) semen were considered oligospermic and enrolled in the study (n = 35). PS capsule (100 mg) was administered twice daily after major meals for 90 days. Total semenogram and serum testosterone, luteinising hormone and follicle-stimulating hormone were estimated before and at the end of the treatment. Malondialdehyde (MDA), a marker for oxidative stress, content of semen and biochemical parameters for safety were also evaluated. Twenty-eight patients who completed the treatment showed significant (P < 0.001) improvement in spermia (+37.6%), total sperm count (+61.4%), motility (12.4-17.4% after different time intervals), normal sperm count (+18.9%) with concomitant decrease in pus and epithelial cell count compared with baseline value. Significant decrease of semen MDA content (-18.7%) was observed. Moreover, serum testosterone (+23.5%; P < 0.001) and FSH (+9.4%; P < 0.05) levels significantly increased. HPLC chromatogram revealed inclusion of PS constituents in semen. Unaltered hepatic and renal profiles of patients indicated that PS was safe at the given dose. The present findings provide further evidence of the spermatogenic nature of Shilajit, as attributed in Ayurvedic medicine, particularly when administered as PS.

Serological evidence of antibodies against Chlamydophila abortus in free-ranging yak (Poephagus grunniens) in Arunachal Pradesh, India.

Serum samples were randomly collected from 172 free-ranging yak (Poephagus grunniens, presently Bos grunniens) from six different yak tracts of Arunachal Pradesh, India, and subjected to enzyme-linked immunosorbent assay to detect the presence of specific antibodies against Chlamydophila abortus. The overall prevalence of this disease in yak was 35%. The prevalence of Cp. abortus-specific antibodies was significantly higher in yak cows (41%) than among bulls (25%). The highest prevalence (39%: 95% confidence interval [Cl] = 27, 55) was found in yak between one and three years of age, while the lowest prevalence (20%: 95% CI = 10, 41) was reported in yak below one year of age.

A serological survey of antibodies against bovine herpesvirus-1 in yak (Poephagus grunniens) in Arunachal Pradesh in India.

Serum samples were collected from 254 yak (Poephagus grunniens, presently Bos grunniens) in different yak tracts of India. These samples were then screened by virus neutralisation test (VNT) and avidin-biotin enzyme-linked immunosorbent assay (AB-ELISA) to study the seroprevalence of antibodies against bovine herpesvirus type 1 (BHV-1). The overall seroprevalence in yak was found to be 41% (105) by VNT and AB-ELISA. The sex of the animal, whether it was on a farm or free-ranging and the location of the different yak tracts did not seem to have any effect on seroprevalence. However, seroprevalence was found to increase with the age of the animals, being highest in yak older than three years of age (49%). Yak generally share feeding, watering and grazing areas with other domestic and wild animals and this common ecological niche is thought to be a possible avenue of infection. This is the first time that the seroprevalence of antibodies against BHV-1 has been studied in yak in India.

Transcriptional commitment of mitochondrial RNA polymerase from Saccharomyces cerevisiae.

The transcriptional commitment of mitochondrial RNA (mtRNA) polymerase and the conditions required for the formation of a stable ternary complex have been determined by in vitro transcription study. Four different transcription complexes were made in vitro by incubating purified mtRNA polymerase, cloned synthetic mitochondrial promoters and selective ribonucleotides. The responses of these complexes to heparin, an inhibitor of unbound mtRNA polymerase, have been examined to determine their involvement in transcription. This study leads to the following observations. (1) Under normal reaction conditions, 40 nM-heparin completely inhibited mitochondrial transcription. (2) A preinitiation mitochondrial DNA-RNA polymerase complex (complex 0) showed partial resistance to heparin (approximately 25% resistant to 40 nm-heparin) when heparin and ribonucleoside triphosphates (rNTPs) were added together to the preformed complex. This complex was rapidly inactivated when preincubated with heparin before the addition of rNTPs. (3) The early initiation (complexes 2 and 4) containing DNA template, RNA polymerase and a short RNA product showed more resistance (approx. 40 to 50%) to 40 nM-heparin but destabilized upon further incubation with heparin before addition of the rest of the rNTPs. (4) After generation of ten or more phosphodiester bonds (complex 11), the early transcription complex is converted into a stable initiation complex, leading to the polymerase consignment to elongation. On the basis of stability and heparin sensitivity, three initial steps of mitochondrial transcription have been defined: polymerase-promoter interaction, initiation, and the transition from initiation to elongation. The formation of preinitiation complex is the rate-limiting step t 1/2 approx. 50 s), whereas the initiation and elongation reactions are very fast processes (t 1/2 greater than 5 s) in mitochondrial transcription.

Position-specific inhibition of yeast mitochondrial transcription by a poly(T) sequence.

The 3' flanking nucleotide(s) of the octanucleotide promoter sequence regulates transcriptional efficiency of some mitochondrial genes in Saccharomyces cerevisiae. To understand this regulation the in vitro transcriptional activity of various synthetic mitochondrial promoters carrying different 3' flanking sequences was examined. The results presented here demonstrate that consecutive thymidine residues, but no other polynucleotides or secondary structure, in the promoter-proximal non-transcribed DNA strand inhibited mitochondrial transcription. The location and the number of T residues in the cluster as well as the concentration of UTP in the transcription reaction are the important factors determining this transcriptional inhibition. For example, a pair of thymidine nucleotides at positions +2 and +3 is sufficient for inactivation of mitochondrial transcription, whereas more than three consecutive thymidine nucleotides beyond these positions are required for inhibition of mitochondrial transcription. However, a cluster of six to 12 thymidine residues beyond position +11, a point where mtRNA polymerase has been shown to form a stable transcription complex, did not interfere with mitochondrial transcription. Interestingly, at low UTP concentration the mtRNA polymerase generates a large quantity of aborted initiation products on a template carrying promoter-proximal poly(T) sequence probably due to the inability of the polymerase to clear this promoter. On the other hand at high UTP concentration the same mtRNA polymerase on the same mitochondrial promoter produces a higher level of productive initiation complex. These observations suggest that the mechanism of poly(T) inhibition of mitochondrial transcription is a UTP-limited transcriptional attenuation at the promoter site, which might occur under specific physiological conditions (i.e. glucose repression-derepression, switching of aerobic-anaerobic conditions).

Expression of the mitochondrial RNase P RNA subunit-encoding gene from a variant promoter sequence in Saccharomyces cerevisiae.

Ribonuclease P (RNase P) is a common tRNA processing enzyme that removes the 5' leader sequence of precursor tRNAs. This activity is identified in yeast mitochondria as a separate enzyme from the nuclear RNase P. Like other RNase P enzymes, the mitochondrial (mt) RNase P is also a ribonucleoprotein composed of both RNA and protein subunits. The RNA subunit is encoded by a mt gene and the protein subunit is supplied by a nuclear gene. Earlier studies described one active promoter (FP1) located 5' to the mt tRNA(fMet)-RNase P RNA-tRNA(Pro) gene cluster, so that the mitochondrially encoded RNA subunit was thought to be co-transcribed with two of its substrate tRNAs. However, the results of in vitro transcription and primer extension experiments presented here demonstrate that the mt RNase P RNA subunit-encoding gene (RPM1) is transcribed from a new promoter (SP)which is located between the tRNA(fMet) and RPM1 genes. The sequence [5'-TATAAGAA(+1)] of the new promoter varies from the conserved promoter sequence [5'-TATAAGTA(+1)], but is one of the sequences that is active in the in vitro transcription assay to determine the consensus promoter sequence [5'-T A T/a A A/g/c G T/a/c N(+1)]. This result demonstrates that a naturally occurring variant promoter is used by RPM1. Identification of the novel SP promoter suggests that the synthesis of the mt RNase P RNA subunit might be uncoupled from the expression of upstream tRNA(fMet) gene, and that RPM1 might be independently transcribed in Saccharomyces cerevisiae.

Usage of non-canonical promoter sequence by the yeast mitochondrial RNA polymerase.

Prior work has demonstrated that a conserved nonanucleotide [5'-TATAAGTAA(+2)] promoter sequence is used by the mitochondrial [mt]1 RNA polymerase in Saccharomyces cerevisiae. However, the highly AT-rich yeast mt genome carries many other promoter-like sequences, but only a fraction of them are involved in gene-specific transcription. To examine the sequence variability of this nonanucleotide promoter motif, single or multiple nt substitutions were introduced into the canonical promoter sequence. The transcriptional activity of these altered promoter sequences was examined under the in-vitro reaction conditions. The results presented here determined that several variant promoter sequences (i. e. TAAAAGTAA, TATAAGAAA, TATAAGTAG, TATAAGAAG, TATAAGAGA, TATAAGGGA, TATAAGTGG, TAAAAGTAG) were efficiently used by the mtRNA polymerase. However, a single (i.e. AATAAGTAA, TTTAAGTAA, TATTAGTAA, TATAACTAA, TATAAGGAA, TATAAGTAT) or multiple (TATAGGAAA, TAAAAGGAA, TATAGGGAA, TAAAGGAAA, TAAAGGGAA) nt substitution(s) in other locations drastically reduced mt promoter function. Interestingly, some of these poorly or partially active promoter variants (i.e. TATAAGGAA, TATAAGTAT, TATAAGTCA) became fully functional in the presence of sequence-specific dinucleotide primer. Since dinucleotide primer bypasses the first phosphodiester bond formation in transcription, it is suggested that the -1T-->G, +1A-->C and +2A-->T mutations affect mt transcription at the level of initiation rather than polymerase binding.

Characterization of the mitochondrial DNA polymerase from Saccharomyces cerevisiae.

The mitochondrial DNA (mtDNA) polymerase was isolated from a protease-deficient yeast strain (PY2), and purified about 3000 fold by a column chromatography on phosphocellulose, heparin-agarose, and single-stranded DNA cellulose. The purified polymerase was characterized with respect to optimal nucleotide concentrations, template-primer specificity and sensitivity to some inhibitors. These results were compared with the nuclear DNA polymerase I activity. Both polymerases showed similar requirement of deoxynucleotide concentrations (Km < 1 microM), and highest activity with poly(dA-dT) template. However, the mtDNA polymerase was more sensitive to ddTTP, EtBr and Mn2+ inhibition in comparison to the nuclear DNA polymerase I. The mtDNA polymerase did not need ATP as an energy source for in vitro DNA synthesis. This mtDNA polymerase preparation also showed 3'-->5' exonuclease activity.

Properties of mitochondrial DNA polymerase in mitochondrial DNA synthesis in yeast.

Mitochondrial DNA polymerase from Saccharomyces cerevisiae, purified 3500 fold, was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis into three polypeptides. The major 150 kDa polypeptide was probably the catalytic subunit of the mitochondrial (mt) DNA polymerase and the other two polypeptides could be either proteolytic cleavage products of the polymerase, other subunits of the enzyme or protein contaminants. The mtDNA polymerase preferred an A+T-rich DNA template and did not require any RNA primer for DNA synthesis, at least under in vitro reaction conditions. It showed higher processivity on a double-stranded linear DNA template than on a single-stranded circular DNA template, and was capable of synthesizing at least about 1200 nucleotide primer-extended products without any major pause on a double-stranded DNA template.

Purification of acid phosphatase I from germinating seeds of Vigna sinensis.

Acid phosphatase I (AP-I) is the major isoform of Vigna acid phosphatase. It is constitutively expressed in seed cotyledons during germination. AP-I was separated from other isoforms and purified to homogeneity by three simple purification steps; (NH4)2SO4 precipitation, and phosphocellulose and DEAE-cellulose column chromatography. The activity of AP-I was not affected by 1 mM Mg2+, Mn2+, Ca2+, Co2+ or Pb2+, but severely inhibited by 1 mM Cu2+, Fe3+, Hg2+, Mo6+ or Zn2+. AP-I has both phosphatase and pyrophosphatase activities, and is highly stable even at 50 degrees.

Chemical and pharmacological evaluation of different ayurvedic preparations of iron.

Ayurvedic preparations of metallic iron commonly categorised as different 'putas' of 'Louha Bhasma' was chemically analysed and pharmacologically investigated in iron deficiency anemia. Atomic absorption spectral (AAS) study of different putas of Louha Bhasma revealed the presence of various proportions of important metals along with varied concentration of iron in it. The effect of a representative puta viz. 50 puta of Louha Bhasma in the management of agar gel diet and phlebotomy induced iron deficiency anemia in animal model was found to be statistically highly significant (P < 0.001) in comparison to the control and standard drug Fefol treated groups.

Immunoclinical correlation in diabetes mellitus. A preliminary report on the lectin-agglutination test.

Twenty cases with insulin dependent diabetes mellitus (IDDM) were studied and compared with a control group with non insulin dependent diabetes mellitus (NIDDM) and another group of nondiabetic healthy persons. Lymphocytes of each group were tested for agglutination with three sets of lectins: concavalin A (ConA), soyabean agglutinin (SBA) and peanut agglutinin (PNA). SBA test, being highly positive in IDDM and persistently negative in NIDDM, is the most significant of the three tests for differentiating between the two types of diabetes mellitus.

Study of foetal antigen in haematological malignancy.

Forty patients with different varieties of leukaemia and lymphoma were studied before and after therapy. Red cells and lymphocytes from each patient were tested for foetal antigen by lectin-agglutination test. The antigen was detectable on red cells in all untreated cases, the highest titre being found in chronic myeloid leukaemia. The titre showed significant reduction after treatment in all cases. We conclude that foetal antigen on red cells is a useful diagnostic aid in haematological malignancy and is a good indicator of the outcome of therapy.

Study of serum myoglobin and serum electrolytes in acute uncomplicated myocardial infarction.

Forty patients with uncomplicated acute myocardial infarction were studied within 6-18 hours after the infarction. Serum myoglobin was elevated in all the cases and was markedly high in cases studied 18 hours after the acute infarction, though the level did not show any relation with the severity of the attack. Myoglobin level showed no correlation with SGOT level, which did not rise appreciably within 6 hours. Serum sodium and potassium levels did not show any change, even in the most severe cases. Serum myoglobin estimation is thus a good diagnostic test in the early hours of acute myocardial infarction.

Abdominal aortic aneurysm with Leriche syndrome.

An interesting and unusual case of abdominal aortic aneurysm with Leriche syndrome that presented with multiple vasculopathies and peptic ulcer is being reported.

A randomised multicentre study to compare the safety and efficacy of albendazole and metronidazole in the treatment of giardiasis in children.

A randomised control multicentre study to compare the safety and efficacy of albendazole and metronidazole in the treatment of giardiasis in children is reported. One hundred and fifty children of either sex (age range: 2-10 years) were randomised to receive either a single dose of 400 mg of albendazole suspension, or 22.5 mg/kg/day of metronidazole in 3 divided doses for 5 consecutive days. At the end of therapy, majority of children in both treatment groups were symptom free. Two days after completion of therapy, 97% of children in both treatment groups were giardia free in the stools. Side effects were noted in 3 children in the albendazole group, and in 20 children in the metronidazole group. We conclude that albendazole suspension is as effective as metronidazole in the treatment of giardial infection in children. It is safe and has fewer side effects as compared to metronidazole.