Genetic Characterization of Aeromonas hydrophila using Protein Profiling and RAPD PCR

Genetic relationship among Aeromonas hydrophila isolates from fish and traditional brackishwater farms were evaluated through Randomly Amplified Polymorphic DNA (RAPD) analysis as well as Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS PAGE) profile of cellular proteins. PCR amplification of the DNA from the bacterial isolates using five random primers (OPA-01, 03, 04, 05 and 10) produced 46 amplicons, which were scorable as distinct bands on agarose gel electrophoresis. Absolute polymorphism of RAPD profile was evident with a unique pattern for each isolate, indicating its usefulness as an ideal tool for evaluation of genetic heterogeneity within the species, which are not revealed by other methods like morphological and biochemical characterization and cellular protein profile. Cellular protein profile did not reveal significant polymorphism as all the isolates revealed uniform pattern indicating its usefulness as a tool for species level identification. Four protein bands of molecular size viz., 19.5 kDa, 25.6 kDa, 29 kDa and 65.6 kDa were shared by all the isolates in the study

Stock structuring in Asian green mussel Perna viridis population along the Indian coast based on shell morphometrics and RAPD markers

In this study, shell morphometrics and random amplified polymorphic DNA (RAPD) techniques were used to evaluate the genetic variability in 4 wild populations of Perna viridis (H≈2500 km apart) along the Indian coast. The phenotypic stocks of mussels were separated using canonical discriminant function analysis. Scatter plots developed with CDA depicted overlapping clusters, indicative of the morphological uniformity of the species along the Indian coast, while Andaman population was forming a separate cluster. RAPD profiles generated from green mussels using 6 primers amplified a total of 43 different fragments ranging from <300 bp to 2500 bp, of which 30 were polymorphic. High genetic variability and moderate genetic differentiation (0.126) was noted among the populations. Percentage of polymorphism among green mussels ranged from 30.2% (Dona Paula) to 79.1% (Andamans). The observed genetic homogeneity among the green mussels of Indian coast indicated their high level of genetic mixing and it may be attributed to the pelagic larval dispersal along with coastal currents

Effects of antiplatelet therapy after stroke due to intracerebral haemorrhage (RESTART): a randomised, open-label trial

Background Antiplatelet therapy reduces the risk of major vascular events for people with occlusive vascular disease, although it might increase the risk of intracranial haemorrhage. Patients surviving the commonest subtype of intracranial haemorrhage, intracerebral haemorrhage, are at risk of both haemorrhagic and occlusive vascular events, but whether antiplatelet therapy can be used safely is unclear. We aimed to estimate the relative and absolute effects of antiplatelet therapy on recurrent intracerebral haemorrhage and whether this risk might exceed any reduction of occlusive vascular events. Methods The REstart or STop Antithrombotics Randomised Trial (RESTART) was a prospective, randomised, open-label, blinded endpoint, parallel-group trial at 122 hospitals in the UK. We recruited adults (≥18 years) who were taking antithrombotic (antiplatelet or anticoagulant) therapy for the prevention of occlusive vascular disease when they developed intracerebral haemorrhage, discontinued antithrombotic therapy, and survived for 24 h. Computerised randomisation incorporating minimisation allocated participants (1:1) to start or avoid antiplatelet therapy. We followed participants for the primary outcome (recurrent symptomatic intracerebral haemorrhage) for up to 5 years. We analysed data from all randomised participants using Cox proportional hazards regression, adjusted for minimisation covariates. This trial is registered with ISRCTN (number ISRCTN71907627). Findings Between May 22, 2013, and May 31, 2018, 537 participants were recruited a median of 76 days (IQR 29–146) after intracerebral haemorrhage onset: 268 were assigned to start and 269 (one withdrew) to avoid antiplatelet therapy. Participants were followed for a median of 2·0 years (IQR [1·0– 3·0]; completeness 99·3%). 12 (4%) of 268 participants allocated to antiplatelet therapy had recurrence of intracerebral haemorrhage compared with 23 (9%) of 268 participants allocated to avoid antiplatelet therapy (adjusted hazard ratio 0·51 [95% CI 0·25–1·03]; p=0·060). 18 (7%) participants allocated to antiplatelet therapy experienced major haemorrhagic events compared with 25 (9%) participants allocated to avoid antiplatelet therapy (0·71 [0·39–1·30]; p=0·27), and 39 [15%] participants allocated to antiplatelet therapy had major occlusive vascular events compared with 38 [14%] allocated to avoid antiplatelet therapy (1·02 [0·65–1·60]; p=0·92). Interpretation These results exclude all but a very modest increase in the risk of recurrent intracerebral haemorrhage with antiplatelet therapy for patients on antithrombotic therapy for the prevention of occlusive vascular disease when they developed intracerebral haemorrhage. The risk of recurrent intracerebral haemorrhage is probably too small to exceed the established benefits of antiplatelet therapy for secondary prevention

Effects of fluoxetine on functional outcomes after acute stroke (FOCUS): a pragmatic, double-blind, randomised, controlled trial

Background Results of small trials indicate that fluoxetine might improve functional outcomes after stroke. The FOCUS trial aimed to provide a precise estimate of these effects. Methods FOCUS was a pragmatic, multicentre, parallel group, double-blind, randomised, placebo-controlled trial done at 103 hospitals in the UK. Patients were eligible if they were aged 18 years or older, had a clinical stroke diagnosis, were enrolled and randomly assigned between 2 days and 15 days after onset, and had focal neurological deficits. Patients were randomly allocated fluoxetine 20 mg or matching placebo orally once daily for 6 months via a web-based system by use of a minimisation algorithm. The primary outcome was functional status, measured with the modified Rankin Scale (mRS), at 6 months. Patients, carers, health-care staff, and the trial team were masked to treatment allocation. Functional status was assessed at 6 months and 12 months after randomisation. Patients were analysed according to their treatment allocation. This trial is registered with the ISRCTN registry, number ISRCTN83290762. Findings Between Sept 10, 2012, and March 31, 2017, 3127 patients were recruited. 1564 patients were allocated fluoxetine and 1563 allocated placebo. mRS data at 6 months were available for 1553 (99·3%) patients in each treatment group. The distribution across mRS categories at 6 months was similar in the fluoxetine and placebo groups (common odds ratio adjusted for minimisation variables 0·951 [95% CI 0·839–1·079]; p=0·439). Patients allocated fluoxetine were less likely than those allocated placebo to develop new depression by 6 months (210 [13·43%] patients vs 269 [17·21%]; difference 3·78% [95% CI 1·26–6·30]; p=0·0033), but they had more bone fractures (45 [2·88%] vs 23 [1·47%]; difference 1·41% [95% CI 0·38–2·43]; p=0·0070). There were no significant differences in any other event at 6 or 12 months. Interpretation Fluoxetine 20 mg given daily for 6 months after acute stroke does not seem to improve functional outcomes. Although the treatment reduced the occurrence of depression, it increased the frequency of bone fractures. These results do not support the routine use of fluoxetine either for the prevention of post-stroke depression or to promote recovery of function. Funding UK Stroke Association and NIHR Health Technology Assessment Programme

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Not AvailableGenetic relationship among Aeromonas hydrophila isolates from fish and traditional brackishwater farms were evaluated through Randomly Amplified Polymorphic DNA (RAPD) analysis as well as Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS PAGE) profile of cellular proteins. PCR amplification of the DNA from the bacterial isolates using five random primers (OPA-01, 03, 04, 05 and 10) produced 46 amplicons, which were scorable as distinct bands on agarose gel electrophoresis. Absolute polymorphism of RAPD profile was evident with a unique pattern for each isolate, indicating its usefulness as an ideal tool for evaluation of genetic heterogeneity within the species, which are not revealed by other methods like morphological and biochemical characterization and cellular protein profile. Cellular protein profile did not reveal significant polymorphism as all the isolates revealed uniform pattern indicating its usefulness as a tool for species level identification. Four protein bands of molecular size viz., 19.5 kDa, 25.6 kDa, 29 kDa and 65.6 kDa were shared by all the isolates in the study.Not Availabl

Not Available

Not AvailableIn this study, shell morphometrics and random amplified polymorphic DNA (RAPD) techniques were used to evaluate the genetic variability in 4 wild populations of Perna viridis (H≈2500 km apart) along the Indian coast. The phenotypic stocks of mussels were separated using canonical discriminant function analysis. Scatter plots developed with CDA depicted overlapping clusters, indicative of the morphological uniformity of the species along the Indian coast, while Andaman population was forming a separate cluster. RAPD profiles generated from green mussels using 6 primers amplified a total of 43 different fragments ranging from <300 bp to 2500 bp, of which 30 were polymorphic. High genetic variability and moderate genetic differentiation (0.126) was noted among the populations. Percentage of polymorphism among green mussels ranged from 30.2% (Dona Paula) to 79.1% (Andamans). The observed genetic homogeneity among the green mussels of Indian coast indicated their high level of genetic mixing and it may be attributed to the pelagic larval dispersal along with coastal currents.Not Availabl