31 research outputs found

    Isolating silkworm genomic DNA without liquid nitrogen suitable for marker studies

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    Genomic DNA was isolated from posterior silk gland of silkworms, Antheraea assama. Absolute alcohol was used as tissue fixing solution instead of grinding in liquid nitrogen, which yielded high molecular weight DNA (>40 kb). Samples yielded similar amount of DNA when fixed in absolute alcohol (400 μmg/g of silk gland tissue) and ground in liquid nitrogen (456 μmg/g of silk gland tissue). RAPD profile of the isolated DNA revealed high degree of polymorphism. The silkworms were analysed using 50 random primers among which 36 polymorphic primers gave 309 amplicons. The average amplicons per primer found to be 8.58 and 94.82% amplicons were polymorphic. Cluster analysis based on Jaccard’s similarity coefficients resulted in the formation of two main clusters with S9 on one cluster and the remaining strains on the other cluster. Jaccard’s similarity coefficients ranged from 0.122 to 0.863 indicating a high level of genetic diversity within muga silkworm collection. Isolated DNA was also suitable for cloning and restriction enzyme digestion. This method does not require liquid nitrogen for fixation, grinding or storage at -80°C, making it advantageous over other common protocols.Key words: Genomic, silkworms, muga, molecular breeding

    Alpha 1 antitrypsin gene: A case-control study in chronic obstructive pulmonary disease

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    Alpha-1 antitrypsin (AAT) deficiency is an inherited disorder that causes low levels of, or no AAT in the blood. The most common illness in adults with AAT deficiency is lung disease during the third and fourth decades of life. Most commonly, it is associated with chronic obstructive pulmonary disease (COPD). Mutations in the protease inhibitor (PI) gene, located on chromosome 14, are associated with this genetic disorder. The Z protein is due to a single amino acid substitution of 342 glutamine lysine. Although cigarette smoking is the main environmental risk factor, only about 15% of smokers develop clinically significant disease suggesting other influences on disease expression. The study included hospital based age and sex matched 100 cases of COPD and 100 controls without COPD recruited from Christian Medical Centre, Jorhat, Assam. These cases were recruited from February 2009 to December 2009. Subjects were included in the COPD group on the basis of lung function test. DNA extraction was done by DNA extraction kit and amplification for AAT gene was done by site directed mutagenesis polymerase chain reaction (PCR) method as described by Tazellar et al. (1992). We found that smoking was the prior cause of COPD. A1AT deficiency is not prevalent in our population subset but certain other genes could be the attributable factor for COPD.Key words: Alpha 1-antitrypsin, chronic obstructive pulmonary disorder (COPD)

    Gene Expression Changes in the Prefrontal Cortex, Anterior Cingulate Cortex and Nucleus Accumbens of Mood Disorders Subjects That Committed Suicide

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    Suicidal behaviors are frequent in mood disorders patients but only a subset of them ever complete suicide. Understanding predisposing factors for suicidal behaviors in high risk populations is of major importance for the prevention and treatment of suicidal behaviors. The objective of this project was to investigate gene expression changes associated with suicide in brains of mood disorder patients by microarrays (Affymetrix HG-U133 Plus2.0) in the dorsolateral prefrontal cortex (DLPFC: 6 Non-suicides, 15 suicides), the anterior cingulate cortex (ACC: 6NS, 9S) and the nucleus accumbens (NAcc: 8NS, 13S). ANCOVA was used to control for age, gender, pH and RNA degradation, with P≤0.01 and fold change±1.25 as criteria for significance. Pathway analysis revealed serotonergic signaling alterations in the DLPFC and glucocorticoid signaling alterations in the ACC and NAcc. The gene with the lowest p-value in the DLPFC was the 5-HT2A gene, previously associated both with suicide and mood disorders. In the ACC 6 metallothionein genes were down-regulated in suicide (MT1E, MT1F, MT1G, MT1H, MT1X, MT2A) and three were down-regulated in the NAcc (MT1F, MT1G, MT1H). Differential expression of selected genes was confirmed by qPCR, we confirmed the 5-HT2A alterations and the global down-regulation of members of the metallothionein subfamilies MT 1 and 2 in suicide completers. MTs 1 and 2 are neuro-protective following stress and glucocorticoid stimulations, suggesting that in suicide victims neuroprotective response to stress and cortisol may be diminished. Our results thus suggest that suicide-specific expression changes in mood disorders involve both glucocorticoids regulated metallothioneins and serotonergic signaling in different regions of the brain

    A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

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    Meeting abstrac

    <span style="font-size:11.0pt;font-family: "Times New Roman";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN-GB">Influence of root-knot nematode infestation on antioxidant enzymes, chlorophyll content and growth in <i style="mso-bidi-font-style:normal">Pogostemon cablin</i> (Blanco) Benth.</span>

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    254-261<span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"times="" roman";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">Plants adapt themselves to overcome adverse environmental conditions, and this involves a plethora of concurrent cellular activities. Physiological experiments or metabolic profiling can quantify this response. Among several diseases of Pogostemon cablin (Blanco) Benth. (Patchouli), root-knot nematode infection caused by <i style="mso-bidi-font-style: normal">Meloidogyne incognita (Kofoid and White) Chitwood causes severe damage to the plant and hence, the oil production. In the present study, we identified M. incognita morphologically and at molecular level using sequenced characterized amplified region marker (SCAR). M. incognita was artificially inoculated at different levels of second stage juveniles (J2) to examine the effect on Patchouli plant growth parameters. Peroxidase and polyphenol oxidase enzyme activity and changes in the total phenol and chlorophyll contents in M. incognita was also evaluated in response to infection. The results have demonstrated that nematode infestation leads to increased peroxidase activities in the leaves of the patchouli plants and thereby, increase in phenolic content as a means of defence against nematode infestation. Chlorophyll content was also found decreased but no changes in polyphenol oxidase enzyme activity.</span

    Consumer Reactions to Rival Failure: Examining Glory Out of Reflected Failure

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    This chapter focuses on the Glory Out of Reflected Failure (GORFing) phenomenon and its relationships regarding team identification, rival perceptions, and favorite team behavior intentions. A sample of 555 sport fans provides responses regarding their team identification, the perceptions of rival teams, their likelihood to experience GORFing, and behavioral intentions toward the favorite team when their rival loses to a third, neutral team. Structural model results showed that rival perceptions are associated with the likelihood of experiencing GORFing, which in turn was associated with behavioral intentions following a rival team’s loss to a comparable team, and mediated the relationship between rival perceptions and behavioral intentions. Contributions and implications for researchers and practitioners are discussed, and avenues for future study are introduced
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