Plant growth stimulation of wheat (Triticum aestivum L.) by inoculation of salinity tolerant Azotobacter strains.

Five salinity tolerant Azotobacter strains i.e., ST3, ST6, ST9, ST17 and ST24 were obtained from saline soils. These Azotobacter strains were used as inoculant for wheat variety WH157 in earthen pots containing saline soil under pot house conditions, using three fertilizer treatment doses i.e., control (no fertilizer, no inoculation), 90 Kg N ha(-1) and 120 Kg N ha(-1). Inoculation with salinity tolerant Azotobacter strains caused significant increase in total nitrogen, biomass and grain yield of wheat. Maximum increase in plant growth parameters were obtained after inoculation with Azotobacter strain ST24 at fertilization dose of 120 kg N ha(-1) and its inoculation resulted in attaining 89.9 cms plant height, 6.1 g seed yield, 12.0 g shoot dry weight and 0.7 % total nitrogen. The survival of Azotobacter strain ST24 in the soil was also highest in all the treatments at 30, 60 and 90 days after sowing (DAS). However, the population of Azotobacter decreased on 90 DAS as compared to counts observed at 60 DAS at all the fertilization treatments.

Prevalence of bronchial asthma in school children of 6-13 years of age in Shimla city.

BACKGROUND: Asthma is a major public health problem worldwide. However, there is limited data on asthma prevalence among school children in Shimla. The present study was undertaken to determine the prevalence of asthma in Shimla city and also to examine the different factors influencing its occurrence. METHODS: This was a questionnaire-based study (International Study of Asthma and Allergies in Childhood [ISAAC] protocol) carried out in four randomly selected schools in the city of Shimla. The questionnaire was pre-tested in 50 school children. The age group included in the study was 6-13 years. The questionnaire was distributed to all the children (n = 1136). A response rate of 89.5% was achieved. The socio-economic status was determined on the basis of monthly income (cumulative square root value method). The ISAAC questionnaire was given standard scores. Various factors influencing the prevalence of asthma, such as socio-economic status, history of asthma and other allergic disorders in the family, type of cooking fuel used, history of active and passive smoking were elucidated from the history. RESULTS: Overall prevalence of asthma in our study was found to be 2.3 percent. Boys had a higher prevalence (3.1%) than girls (1.4%). We found a significant association between asthma prevalence and family history of asthma and other atopic manifestations. There was no significant association of socio-economic factors and exposure to smoking with prevalence of asthma. CONCLUSION: In the present study, the prevalence of bronchial asthma in school children in the age group of 6-13 years is 2.3 percent.

Infectious Disease Markers in Blood Donors.

BACKGROUND: The infectious disease markers for which blood donors are screened include anti-human immunodeficiency virus (HIV), hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (HCV), rapid plasma reagin (RPR) card test for syphilis and malarial parasites. METHODS: A total of 6751 donors were screened over three years to assess the prevalence of infectious disease markers. Screening for anti-HIV I and II, HBsAg and anti-HCV was carried out by enzyme linked immunosorbent assay (ELISA). Syphilis was tested using RPR card test. Malarial parasite was tested by detection of genus specific plasmodium lactate dehydrogenase. RESULT: The overall seropositivity for anti-HIV I and II was nine (0.13%), for HBsAg 67 (0.99%), for anti-HCV 13 (0.19%) and for syphilis 42 (0.62%). No sample showed malarial parasites. There was no significant difference (p>0.05) in the seropositivity of various markers between voluntary and replacement donors. There was a significant decline (p<0.05) in the prevalence of seropositivity for HCV and syphilis, but not for HIV and HBsAg over the three year period of the study. CONCLUSION: The prevalence of infectious disease markers was similar to that reported by other studies. However, no significant difference was seen in the marker positivity in voluntary and replacement donors, which is at variance from other studies.

Characterization of genetic polymorphism of the bovine lymphocyte antigen DRB3.2 locus in Kankrej cattle (Bos indicus).

Bovine lymphocyte antigen DRB 3.2 (BoLA-DRB3.2) gene encodes for the beta chain of the major histocompatibility complex (MHC) class II molecule in cattle, which is a glycoprotein present on the surface of antigen-presenting cells. This locus shows extensive polymorphism in it. The objective of the present study was to genotype the BoLA-DRB3.2 locus in Kankrej cattle (n = 50) by PCR-RFLP. Bovine DNA was isolated from aliquots of whole blood. Primers specific for exon 2 of the bovine lymphocyte antigen (BoLA)-DRB3 gene were used to amplify the region. The 304-bp amplified product of the DRB3 gene was separately digested with restriction endonucleases RsaI, BstYI, and Hae III. Twenty-four BoLA-DRB 3.2 alleles were identified with frequencies ranging from 1 to 22.0%. Twenty-one alleles of the total 24 alleles were similar to those reported earlier; 3 alleles were new and had not been reported previously. The allele BoLA-DRB3.2*34 occurred at the highest frequency of 22% (approx.) in the Kankrej animals studied. Six alleles (BoLA-DRB3.2 *34, *15, *06, *20, *37, and *20) accounted for almost 71% of the total alleles observed to be present in the Kankrej animals. All the new alleles observed were present at frequencies of 1%. The results obtained in the present study demonstrated that the BoLA DRB3.2 locus is highly polymorphic in the Kankrej cattle.

Genetic relationships of five Indian horse breeds using microsatellite markers.

The genetic relationships of five Indian horse breeds, namely Marwari, Spiti, Bhutia, Manipuri and Zanskari were studied using microsatellite markers. The DNA samples of 189 horses of these breeds were amplified by polymerase chain reaction using 25 microsatellite loci. The total number of alleles varied from five to 10 with a mean heterozygosity of 0.58 ± 0.05. Spiti and Zansakari were the most closely related breeds, whereas, Marwari and Manipuri were most distant apart with Nei's DA genetic distance of 0.071 and 0.186, respectively. In a Nei's DA genetic distances based neighbour joining dendrogram of these breeds and a Thoroughbred horse outgroup, the four pony breeds of Spiti, Bhutia, Manipuri and Zanskari clustered together and then with the Marwari breed. All the Indian breeds clustered independently from Thoroughbreds. The genetic relationships of Indian horse breeds to each other correspond to their geographical/environmental distribution.

Climatically driven emissions of hydrocarbons from marine sediments during deglaciation.

Marine hydrocarbon seepage emits oil and gas, including methane ( approximately 30 Tg of CH(4) per year), to the ocean and atmosphere. Sediments from the California margin contain preserved tar, primarily formed through hydrocarbon weathering at the sea surface. We present a record of variation in the abundance of tar in sediments for the past 32,000 years, providing evidence for increases in hydrocarbon emissions before and during Termination IA [16,000 years ago (16 ka) to 14 ka] and again over Termination IB (11-10 ka). Our study provides direct evidence for increased hydrocarbon seepage associated with deglacial warming through tar abundance in marine sediments, independent of previous geochemical proxies. Climate-sensitive gas hydrates may modulate thermogenic hydrocarbon seepage during deglaciation.

Genetic analysis of Ankamali pigs of India using microsatellite markers and their comparison with other domesticated Indian pig types.

Ankamali pigs, the domesticated native pigs of Kerala province of India were genetically characterized using 23 FAO recommended microsatellite markers and were compared with other native Indian pig types and Large White pigs. Twenty-six blood samples were collected from genetically unrelated animals from their breeding tract and DNA was isolated by standard procedure of phenol/chloroform. The genomic DNA was amplified by polymerase chain reaction (PCR) at these 23 microsatellite loci, which were also used earlier to characterize Desi (North Indian) and Gahuri (North-East Indian) pigs, the other two native domesticated pig types of India. The PCR products were resolved by denaturing urea-polyacrylamide gel electrophoresis and alleles were visualized after silver nitrate staining. The data were analysed for allele size range, number of alleles, allelic frequencies, heterozygosity and polymorphism information content for each locus. The allele size range varied between 92-108 bp at locus S0026 and 280-296 bp at locus IGF-1. The total number of alleles varied between five (S0178 and S0386) and 11 (S0355). The mean observed and expected heterozygosities were found to be 0.74 +/- 0.09 and 0.83 +/- 0.03 respectively. In the neighbour-joining dendrogram based on D(A) genetic distances developed after 1000 bootstraps, the Ankamali pigs did not show genetic closeness either with other native Indian pig types or exotic Large White pigs with high bootstrap values indicating genetic distinctness of Ankamali pigs.

Possible role of neonatal infection with the asymptomatic reassortant rotavirus (RV) strain I321 in the decrease in hospital admissions for RV diarrhea, Bangalore, India, 1988-1999.

We sought to determine the proportion of rotavirus (RV) infections among children with severe diarrhea in Bangalore, India, and to determine the role of neonatal infection with the asymptomatic RV strain I321 in protection against subsequent RV diarrhea. At 2 major hospitals, there was a >42% decrease in diarrhea-specific admissions during the study period. At 6 hospitals, asymptomatic infections were found in 25%-50% of neonates, when screening was performed randomly, and in >58% of neonates, when screening was performed daily, with the majority of infections occurring within the first 7 days of life. All the RVs found in asymptomatic neonates were strain I321. A 24-month follow-up of a cohort of 44 children who had been neonatally infected with strain I321 and 28 children who had not (control group) revealed comparable rates of RV detection but a marked decrease in the number of RV diarrhea episodes in the strain I321-infected group (2.3%), compared with the control group (39.3%) (P<.0001). This preliminary study suggests a possible association between neonatal infection with strain I321 and protection against subsequent RV illness.

Symmetrical peripheral gangrene due to viral gastroenteritis.

A case of multifactorial symmetrical peripheral gangrene due to viral gastroenteritis, shock, dopamine infusion, exposure to low temperature and nonlactose fermenters septicaemia is presented for its rarity and devastating consequences.

Lack of neighborhood effects from a transcriptionally active phosphoglycerate kinase-neo cassette located between the murine beta-major and beta-minor globin genes.

For the treatment of beta-globin gene defects, a homologous recombination-mediated gene correction approach would provide advantages over random integration-based gene therapy strategies. However, "neighborhood effects" from retained selectable marker genes in the targeted locus are among the key issues that must be taken into consideration for any attempt to use this strategy for gene correction. An Ala-to-Ile mutation was created in the beta6 position of the mouse beta-major globin gene (beta(6I)) as a step toward the development of a murine model system that could serve as a platform for therapeutic gene correction studies. The marked beta-major gene can be tracked at the level of DNA, RNA, and protein, allowing investigation of the impact of a retained phosphoglycerate kinase (PGK)-neo cassette located between the mutant beta-major and beta-minor globin genes on expression of these 2 neighboring genes. Although the PGK-neo cassette was expressed at high levels in adult erythroid cells, the abundance of the beta(6I) mRNA was indistinguishable from that of the wild-type counterpart in bone marrow cells. Similarly, the output from the beta-minor globin gene was also normal. Therefore, in this specific location, the retained, transcriptionally active PGK-neo cassette does not disrupt the regulated expression of the adult beta-globin genes. (Blood. 2001;98:65-73)

Establishment of phosphate-solubilizing strains of Azotobacter chroococcum in the rhizosphere and their effect on wheat cultivars under green house conditions.

A pot experiment was conducted in the green house to investigate the establishment of phosphate solubilizing strains of Azotobacter chroococcum, including soil isolates and their mutants, in the rhizosphere and their effect on growth parameters and root biomass of three genetically divergent wheat cultivars (Triticum aestivum L.). Five fertilizer treatments were performed: Control, 90 kg N ha(-1), 90 kg N + 60 kg P2O5 ha(-1), 120 kg N ha(-1) and 120 kg N + 60 kg P2O5 ha(-1). Phosphate solubilizing and phytohormone producing parent soil isolates and mutant strains of A. chroococcum were isolated and selected by an enrichment method. In vitro phosphate solubilization and growth hormone production by mutant strains was increased compared with soil isolates. Seed inoculation of wheat varieties with P solubilizing and phytohormone producing A. chroococcum showed better response compared with controls. Mutant strains of A. chroococcum showed higher increase in grain (12.6%) and straw (11.4%) yield over control and their survival (12-14%) in the rhizosphere as compared to their parent soil isolate (P4). Mutant strain M37 performed better in all three varieties in terms of increase in grain yield (14.0%) and root biomass (11.4%) over control.

Effect of epidermal growth factor on steroidogenesis by caprine granulosa cells in culture: interaction with FSH.

Besides gonadotrophins various peptide growth factors have been implicated in the ovarian folliculogenesis. In this study, the effect of epidermal growth factor (EGF) (0, 0.1, 1.0, 10ng/ml culture medium) on steroidogenesis by caprine granulosa cells at various stages of maturation was investigated using serum free culture medium. Caprine granulosa cells were obtained from ovarian follicles and classified into three classes: small (<3mm), medium (3-6mm) and large (>6mm in diameter). EGF (10ng/m culture medium) alone reduced estradiol secretion in granulosa cell from small, medium and large follicles by 62, 48 and 29%, respectively, as compared with control. This inhibition was 50, 36 and 21%, respectively, when EGF (10ng/ml culture medium) was applied in combination with FSH (100ng/ml culture medium). EGF alone stimulated the secretion of progesterone in granulosa cells from all the three categories of follicles only at highest dose tested (10ng/ml culture medium). FSH acted synergistically with EGF in stimulating progesterone secretion by cultured granulosa cells. EGF in combination with FSH (100ng/ml culture medium) significantly (P<0.05) stimulated progesterone secretion by cultured granulosa cells from all three categories of follicles even at the lowest dose (0.1ng/ml culture medium) tested. In conclusion, EGF significantly influences the steroidogenesis by caprine granulosa cells in vitro and may play important role in the follicular growth and maturation.

Evaluation of the genetic variability of 13 microsatellite markers in native Indian pigs.

We analysed polymorphism of 13 microsatellites in two Indian domesticated pig types (North Indian and Northeast Indian). Heterozygosity, polymorphism information content, and probability of identity of two random individuals were calculated for all microsatellites in both types. The number of alleles observed at a locus varied between five and 12. The evaluated microsatellites exhibited a very high heterozygosity and polymorphism information content. The probability of identity of two random individuals from different populations taking into account all the 13 microsatellites was as low as 3.51 x 10(-19). On the basis of these results, we propose that these microsatellite markers may be used with reliability for studying the genetic diversity and for identification of individuals in Indian pig types.

Induction of tissue factor procoagulant activity in myelomonocytic cells inoculated by the agent of human granulocytic ehrlichiosis.

Human granulocytic ehrlichiosis (HGE) is a recently recognized rickettsial tick-borne febrile illness that may occasionally be complicated by coagulopathy. The agent of HGE (aHGE) is an obligate intracellular pathogen, which replicates in endosomes within neutrophils and their precursors. We hypothesized that aHGE might cause DIC via induction of monocyte tissue factor procoagulant activity (TF PCA). Peripheral blood mononuclear cells (PBMNC) and HL-60 cells were used to model the effect of aHGE infection on monocytes/macrophages. Mononuclear cells inoculated with aHGE in vitro demonstrated approximately a 12-15-fold increase in TF PCA, with peak activity occurring at 8-12 h. HL-60 cells inoculated with aHGE also manifested a 4-6 fold induction of TF PCA, with maximal activity occurring at about 8 h. By comparison, E. Coli lipopolysaccharide (LPS) also induced an increase in TF PCA of an equivalent magnitude, and with a similar time course. Induction of TF did not require inoculation of HL-60 cells with live organism, since heat-inactivated aHGE still stimulated TF PCA expression in the target cells. Furthermore, filtered supernatants from heat-inactivated organisms induced TF PCA suggesting that the effect is due to a soluble mediator produced by the organism. Although aHGE is a gram negative organism, the soluble mediator did not appear to be classic endotoxin in that the supernatants tested negative for endotoxin by the Limulus Amoebocyte assay, and polymixin had no inhibitory effect on aHGE supernatants. We conclude that aHGE induces cells of the myelo-monocytic lineage to synthesize TF, which may contribute to the clinical coagulopathy that can be observed in this condition. An atypical soluble mediator or cellular component of the organism appears to be critically important in TF induction by aHGE.

Hepatic pyridine nucleotides content in rat--a better indicator for determining available niacin values of food.

After 10 days depletion period with niacin free diet, weanling rats were repleted with either reference diets containing niacin at four levels (2, 4, 8 and 12 mg/kg diet) or test diets containing test material at two levels as source of niacin. Gains in body weight and hepatic pyridine nucleotides content increased with increase in niacin intake. The correlation coefficient for hepatic pyridine nucleotides content and niacin intake was 0.98, whereas for weight gain and niacin intake was 0.89. Dried skim milk and pearl millet were taken as test materials. Dried skim milk with most of its niacin in free form showed 98% niacin equivalent to be bioavailable whereas in pearl millet niacin was in bound form and bioavailability equivalent was only 48 per cent.

Effect of insulin-like growth factor-1 on steroidogenesis in cultured carp ovarian follicles: interactions with estradiol.

The biological action of insulin like growth factor-1 (IGF-1) on follicular steroidogenesis during follicular development in common carp was examined. Studies were carried out by culturing small (1-2 mm diam.) and large (> 2 mm diam.) follicles. IGF-1 (0.3-100 ng/ml) had no effect on progesterone accumulation or aromatase activity during 48 hr culture of small follicles. Progesterone accumulation by large follicles was also unaffected by IGF-1 over the same period, although aromatase activity was stimulated in a dose dependent manner (8-fold increase over basal levels with a maximum stimulatory dose of 30 ng IGF-1/ml). In contrast, small and large follicles responded to IGF-1 in terms of both progesterone accumulation and aromatase activity after longer periods of culture (4 days for progesterone and 6 days for aromatase). Concurrent treatment of small follicles with estradiol (10(-7) M) enhanced the action of IGF-1 on both indices of steroidogenesis and advanced the time at which IGF-1 stimulated activity was first detectable. The effect of estradiol on follicular IGF-1 responsiveness were independent of cell number. In summary, these results demonstrate varied actions of IGF-1 carp ovarian follicular steroidogenesis in vitro. The results indicate that carp follicles acquire responsiveness to IGF-1 in terms of aromatase activity during follicular development in vivo and that estradiol can induce the response in vitro. The results also suggest that estrogen and progesterone biosynthesis by cultured carp ovarian follicles is differentially regulated by IGF-1. Together, these results provide new insights into the biological actions of IGF-1 in fish ovary.

Radiopharmaceutical uptake as a marker of sternal blood supply following internal mammary artery harvesting.

To evaluate the impact of internal mammary artery harvesting on sternal blood supply after open heart surgery, a conventional bone scan was performed 7 days after operation in 30 patients. After administration of 370 MBq of technetium 99 m-medronic acid complex, imaging was carried out at the level of the sternum and including the humerus as a reference. A quantitative analysis of uptake (sternum/humerus uptake index) was performed and compared in three different groups of patients: group A, ten patients who had only vein grafts or valve surgery; group B, ten patients with single internal mammary artery harvesting; and group C, ten patients with bilateral internal mammary artery harvesting. These results were compared with 24 non-surgical subjects as a control (group D). Although intervention had a significant influence in raising the uptake index of the surgical groups (A = 3.34; B = 3.09 and C = 3.48) when compared with normal subjects (D = 2.45) (P < 0.01), there was not a statistically significant difference among the three surgical groups (P > 0.05). It was concluded that the vascular supply of the sternum is not entirely dependent upon the internal mammary arteries and that mobilization of both vessels does not cause per se additional serious impact to the bone vascularization after midline sternotomy, at least beyond day 7 after operation.

Early results of direct repair of aortic interruption by the lateral approach.

Eleven consecutive patients were operated on for interruption of the aortic arch by direct aortoplasty through a lateral approach. Median age was 5 days and median weight, 3.52 kg. All received prostaglandin E. Four of the 11 required preoperative ventilatory and inotropic support. Diagnosis was by echocardiography with one confirmed by angiography. There were five type B and six type A interruptions. Aortic continuity was achieved through a left thoracotomy. The aorta and great vessels were mobilized, and after clamps were placed above and below the interruption, ductal tissue was excised and direct anastomosis performed. There was one death from sepsis. One patient died of pulmonary hypertension early after the second operation for correction of other anomalies, and 1 died of hypoplastic lungs after 6 months of ventilation. In 10 of the 11 patients there was no postoperative gradient. Palliative staged approach for correction of this malformation is advocated. The need for bypass and profound hypothermia is removed. This approach restores aortic continuity without sacrifice of the major vessels, use of ductal tissue, or use of prosthetic grafts that will necessitate reoperation.