Advancing the STMS genomic resources for defining new locations on the intraspecific genetic linkage map of chickpea (Cicer arietinum L.)

<p>Abstract</p> <p>Background</p> <p>Chickpea (<it>Cicer arietinum </it>L.) is an economically important cool season grain legume crop that is valued for its nutritive seeds having high protein content. However, several biotic and abiotic stresses and the low genetic variability in the chickpea genome have continuously hindered the chickpea molecular breeding programs. STMS (Sequence Tagged Microsatellite Sites) markers which are preferred for the construction of saturated linkage maps in several crop species, have also emerged as the most efficient and reliable source for detecting allelic diversity in chickpea. However, the number of STMS markers reported in chickpea is still limited and moreover exhibit low rates of both inter and intraspecific polymorphism, thereby limiting the positions of the SSR markers especially on the intraspecific linkage maps of chickpea. Hence, this study was undertaken with the aim of developing additional STMS markers and utilizing them for advancing the genetic linkage map of chickpea which would have applications in QTL identification, MAS and for <it>de novo </it>assembly of high throughput whole genome sequence data.</p> <p>Results</p> <p>A microsatellite enriched library of chickpea (enriched for <b>(</b>GT/CA)_nand (GA/CT)_nrepeats) was constructed from which 387 putative microsatellite containing clones were identified. From these, 254 STMS primers were designed of which 181 were developed as functional markers. An intraspecific mapping population of chickpea, [ICCV-2 (single podded) × JG-62 (double podded)] and comprising of 126 RILs, was genotyped for mapping. Of the 522 chickpea STMS markers (including the double-podding trait, screened for parental polymorphism, 226 (43.3%) were polymorphic in the parents and were used to genotype the RILs. At a LOD score of 3.5, eight linkage groups defining the position of 138 markers were obtained that spanned 630.9 cM with an average marker density of 4.57 cM. Further, based on the common loci present between the current map and the previously published chickpea intraspecific map, integration of maps was performed which revealed improvement of marker density and saturation of the region in the vicinity of <it>sfl </it>(double-podding) gene thereby bringing about an advancement of the current map.</p> <p>Conclusion</p> <p>An arsenal of 181 new chickpea STMS markers was reported. The developed intraspecific linkage map defined map positions of 138 markers which included 101 new locations.Map integration with a previously published map was carried out which revealed an advanced map with improved density. This study is a major contribution towards providing advanced genomic resources which will facilitate chickpea geneticists and molecular breeders in developing superior genotypes with improved traits.</p

Global Stability For Double-Diffusive Convection In A Couple-Stress Fluid Saturating A Porous Medium

We show that the global non-linear stability threshold for convection in a double-diffusive couple-stress fluid saturating a porous medium is exactly the same as the linear instability boundary. The optimal result is important because it shows that linearized instability theory has captured completely the physics of the onset of convection. It is also found that couple-stress fluid saturating a porous medium is thermally more stable than the ordinary viscous fluid, and the effects of couple-stress parameter (F ) , solute gradient ( S f ) and Brinkman number ( D a ) on the onset of convection is also analyzed

A Hospital Based Prospective Study to Correlate of Microvascular Complications with Different (Fpg, 2hpg & HbA1c) Glycemic Indices Among Type 2 Diabetes Mellitus Patients

Background:Microvascular complications in type 2 diabetes have significant impact on morbidity, mortality and patients’ quality of life. There are few clinical studies in this direction but most of them lack sufficient power and are focused only towards one specific complication. The aim of this study to determine the association of microvascular complications with duration of diabetes and glycemic indices (Fpg, 2hpg &amp;amp; HbA1c) among type 2 diabetes mellitus patients.Materials &amp; Methods: This is a hospital based prospective study done on 100 cases of type 2 diabetes mellitus in upgraded department of medicine, Government Medical College &amp; attached group of Hospitals, Barmer, Rajasthan. Each subject underwent detailed history and complete clinical examination. Diabetes was diagnosed according to American Diabetes Association (ADA) revised criteria. The selected patients were evaluated for presence of micro complications i.e. diabetic retinopathy, diabetic nephropathy and diabetic neuropathy by relevant investigations.Results: Our study showed that The fasting blood glucose level, 2 hr. postprandial glucose level &amp; HbA1c was not significant correlate but &gt;10 years duration of diabetes was higher mean value as compared to less than 5 year duration of diabetes. The spearmen correlation analysis showing association of various risk factors with diabetic neuropathy, diabetic retinopathy &amp; diabetic nephropathy. The fasting blood glucose level, 2 hr. postprandial glucose level &amp; HbA1c was significant correlate (P&lt;0.0001, p&lt;0.0001 &amp; p&lt;0.0001 respectively).Conclusion: We concluded that on applying spearman correlation for microvascular complications, a positive association was observed for age of patients, duration of diabetes, fasting blood sugar, 2 hr. postprandial glucose and HbA1C

Comparison of “Amicus and COBE Spectra” for autologous peripheral blood stem cell harvest: An Indian experience

Background: Adequate collection of peripheral stem cells from the patients depends on the disease condition, efficient mobilization and the equipment used for PBSC harvest. COBE Spectra has been the major platform for collecting these PBSC for more than 2 decades. Now with introduction of PBSC harvest option with Amicus cell separators (which was used primarily for plateletpheresis) in India, it is worth while comparing both the platforms for stem cells collections. Materials and Methods: Our study is a retrospective analysis of autologous PBSC harvest procedures done at our centre. The study included the data of autologous PBSC collections from January 2015 to June 2016. Total 61 patients underwent 85 autologous PBSC harvests for both haematological and non haematological indications. Results: Out 61 patients, 40 patients collected their target number of cell in a single harvest, 18 patient required dual harvests and 3 patients required three consecutive days of harvest. Pre-Apheresis WBC, platelets and CD45/CD34 cell counts were comparable. COBE Spectra collects significantly higher product volume and higher number of platelets in the apheresis product. Whereas WBC counts of the product, total CD45/CD34 cell dose and collection efficiency (CE2) and collection ratio (CR) were comparable on both the platforms. Conclusions: Amicus took more time to harvest the anticipated number of cells in the graft as well as COBE Spectra resulted in higher platelets loss during the process of cell collection. Our analysis is first of its kind from Indian subcontinent and indicates that with gradual phasing out of COBE spectra from the market Amicus offers a comparative platform for PBSC harvest

Outcome of 51 autologous peripheral blood stem cell transplants after uncontrolled-rate freezing (“dump freezing”) using −80°C mechanical freezer

Background and Objective: Controlled-rate freezing is a complicated, expensive, and time-consuming procedure. Therefore, there is a growing interest in uncontrolled-rate freezing (UCF) with −80°C mechanical freezers for cryopreservation of hematopoietic stem cells. This is a retrospective analysis of efficiency of UCF and outcome of autologous peripheral hematopoietic stem cell (PBSC) transplants at our center from December 2011 to June 2016. Materials and Methods: Cryoprotectant solutions used included 5% dimethyl sulfoxide and 5% albumin with 2% hydroxyethyl starch and stored at −80°C mechanical freezer till transplant. Evaluation of cryopreservation was studied by analyzing the variation in cellularity, viability, and CD34+ stem cell dose recovery as well as clinical follow-up with engraftment. Results: A total of 51 patients (23 females and 28 males) underwent autologous PBSC transplantations with a median age of 31 years (range: 3–60 years) for both hematological and nonhematological indications. Mean recovery post by UCF at −80°C mechanical was 92.9% ± 15.5% for nucleated cells, 86.6% ± 15.5% for viability, and 80% ± 21.5% in CD34+ dose. The median day to neutrophil engraftment was 10 (range 5–14 days) and platelets engraftment was 15 (range 8–45 days). The cryopreserved products were stored at −80°C for median 7 days (range 2-41 day) before transplant. Discussion/Conclusion: Our analysis shows that PBSC can be successfully cryopreserved with mechanical uncontrolled rate freezing. This is a cheap and simple method to freeze the stem cells for a short period in resource-constrained setting