Protein landmarks for diversity assessment in wheat genotypes

Grain proteins from 20 Indian wheat genotypes were evaluated for diversity assessment based seed storage protein profiling on sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). Genetic diversity was evaluated using Nei’s index, Shannon index and Unweighted pair group method with arithmetic mean (UPGMA) cluster analysis by constructing dendrogram of fractions of proteins, which were used for the calculation of similarity coefficients between these varieties. Diversity analysis attributes exhibited the importance of seed storage as a marker system. The similarity ranged from 32.14% to as high as 100% between genotypes. Adoption of this technology would be useful to plant protection regulatory systems, especially for plant variety identification and registration of new plant varieties, breeding programs and protection purposes.Keywords: Sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), genetic diversity, population diversity index, coefficient of similarity.African Journal of Biotechnology Vol. 12(29), pp. 4640-464

A Sensitive Branched DNA HIV-1 Signal Amplification Viral Load Assay with Single Day Turnaround

Branched DNA (bDNA) is a signal amplification technology used in clinical and research laboratories to quantitatively detect nucleic acids. An overnight incubation is a significant drawback of highly sensitive bDNA assays. The VERSANT® HIV-1 RNA 3.0 Assay (bDNA) (“Versant Assay”) currently used in clinical laboratories was modified to allow shorter target incubation, enabling the viral load assay to be run in a single day. To dramatically reduce the target incubation from 16–18 h to 2.5 h, composition of only the “Lysis Diluent” solution was modified. Nucleic acid probes in the assay were unchanged. Performance of the modified assay (assay in development; not commercially available) was evaluated and compared to the Versant Assay. Dilution series replicates (>950 results) were used to demonstrate that analytical sensitivity, linearity, accuracy, and precision for the shorter modified assay are comparable to the Versant Assay. HIV RNA-positive clinical specimens (n = 135) showed no significant difference in quantification between the modified assay and the Versant Assay. Equivalent relative quantification of samples of eight genotypes was demonstrated for the two assays. Elevated levels of several potentially interfering endogenous substances had no effect on quantification or specificity of the modified assay. The modified assay with drastically improved turnaround time demonstrates the viability of signal-amplifying technology, such as bDNA, as an alternative to the PCR-based assays dominating viral load monitoring in clinical laboratories. Highly sensitive bDNA assays with a single day turnaround may be ideal for laboratories with especially stringent cost, contamination, or reliability requirements

Preliminary study of the antioxidant properties of flowers and roots of Pyrostegia venusta (Ker Gawl) Miers

<p>Abstract</p> <p>Background</p> <p>Free radical stress leads to tissue injury and can eventually to arthritis, atherosclerosis, diabetes mellitus, neurodegenerative diseases and carcinogenesis. Several studies are ongoing worldwide to find natural antioxidants of plant origin. We assessed the <it>in-vitro </it>antioxidant activities and screened the phytochemical constituents of methanolic extracts of <it>Pyrostegia venusta </it>(Ker Gawl) <it>Miers</it>.</p> <p>Methods</p> <p>We evaluated the antioxidant potential and phytochemical constituents of <it>P. venusta </it>using 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2, 2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and ferric reducing antioxidant power (FRAP) assays. Gas chromatography-mass spectroscopy (GC-MS) studies were also undertaken to assess the phytochemical composition of the flower extracts.</p> <p>Results</p> <p>Phytochemical analyses revealed the presence of terpenoids, alkaloids, tannins, steroids, and saponins. The reducing ability of both extracts was in the range (in μm Fe(II)/g) of 112.49-3046.98 compared with butylated hydroxytoluene (BHT; 63.56 ± 2.62), catechin (972.02 ± 0.72 μm) and quercetin 3208.27 ± 31.29. A significant inhibitory effect of extracts of flowers (IC₅₀= 0.018 ± 0.69 mg/ml) and roots (IC₅₀= 0.026 ± 0.94 mg/ml) on ABTS free radicals was detected. The antioxidant activity of the extracts of flowers (95%) and roots (94%) on DPPH radicals was comparable with that of ascorbic acid (98.9%) and BHT (97.6%). GC-MS study revealed the presence of myoinositol, hexadecanoic acid, linoleic acid, palmitic acid and oleic acid in the flower extracts.</p> <p>Conclusion</p> <p>These data suggest that <it>P. venusta </it>is a natural source of antioxidants. The extracts of flowers and roots of <it>P. venusta </it>contain significant amounts of phytochemicals with antioxidative properties and could serve as inhibitors or scavengers of free radicals. <it>P. venusta </it>could be exploited as a potential source for plant-based pharmaceutical products. These results could form a sound basis for further investigation in the potential discovery of new natural bioactive compounds.</p

Imprinted CDKN1C Is a Tumor Suppressor in Rhabdoid Tumor and Activated by Restoration of SMARCB1 and Histone Deacetylase Inhibitors

SMARCB1 is deleted in rhabdoid tumor, an aggressive paediatric malignancy affecting the kidney and CNS. We hypothesized that the oncogenic pathway in rhabdoid tumors involved epigenetic silencing of key cell cycle regulators as a consequence of altered chromatin-remodelling, attributable to loss of SMARCB1, and that this hypothesis if proven could provide a biological rationale for testing epigenetic therapies in this disease. We used an inducible expression system to show that the imprinted cell cycle inhibitor CDKN1C is a downstream target for SMARCB1 and is transcriptionally activated by increased histone H3 and H4 acetylation at the promoter. We also show that CDKN1C expression induces cell cycle arrest, CDKN1C knockdown with siRNA is associated with increased proliferation, and is able to compete against the anti-proliferative effect of restored SMARCB1 expression. The histone deacetylase inhibitor (HDACi), Romidepsin, specifically restored CDKN1C expression in rhabdoid tumor cells through promoter histone H3 and H4 acetylation, recapitulating the effect of SMARCB1 on CDKNIC allelic expression, and induced cell cycle arrest in G401 and STM91-01 rhabdoid tumor cell lines. CDKN1C expression was also shown to be generally absent in clinical specimens of rhabdoid tumor, however CDKN1A and CDKN1B expression persisted. Our observations suggest that maintenance of CDKN1C expression plays a critical role in preventing rhabdoid tumor growth. Significantly, we report for the first time, parallels between the molecular pathways of SMARCB1 restoration and Romidepsin treatment, and demonstrate a biological basis for the further exploration of histone deacetylase inhibitors as relevant therapeutic reagents in the treatment of rhabdoid tumor

Removal of Tannic Acid From Aqueous Solution by Cloud Point Extraction and Investigation of Surfactant Regeneration by Microemulsion Extraction

The aim of this work is the extraction of tannic acid (TA) with two commercial nonionic surfactants, separately: Lutensol ON 30 and Triton X-114 (TX-114).The experimental cloud point extraction results are expressed by four responses to surfactant concentration and temperature variations: extent of TA extraction (E), remaining solute (X s,w) and surfactant (X t,w) concentrations in dilute phase and volume fraction of coacervate (Φc) at equilibrium. An empirical smoothing method was used and the results are represented on three dimensional plots. In optimal conditions, the extraction extent of TA reaches 95 and 87 % using TX-114 and Lutensol ON 30, respectively. Sodium sulfate, cetyltrimethylammonium bromide (CTAB) addition and pH effect are also studied. Finally, the possibility of recycling of the surfactant is proved

Screw in Lisfranc Ligament

Category: Trauma Introduction/Purpose: The Lisfranc ligament is an interosseous ligament connecting the medial cuneiform with the second metatarsal. Current treatment of displaced Lisfranc injury is rigid fixation using a transarticular screw. The purpose of this study was to observe the amount of ligamentous disruption with the placement of a transarticular screw from the second metatarsal to the medial cuneiform. Methods: This cadaveric study included a total of 15 preserved cadavers and 23 feet. Blunt dissection down to bone, with removal of soft tissue, was performed on each foot for visualization of the Lisfranc joint. A guide-wire was inserted in a dorsolateral to plantar medial direction from the base of the second metatarsal into the medial cuneiform. Then over the guide- wire a 40 mm, 4.0 partially threaded, cannulated screw was inserted. The Lisfranc ligament was then carefully identified with more dissection. The screw was then removed. Separation of the second metatarsal from medial cuneiform was performed by transecting the dorsal, Lisfranc (interosseous), and plantar ligaments. Digital photographs of the Lisfranc ligament, medial cuneiform and second metatarsal articular surfaces were recorded and measurements were taken. Results: Of the 23 feet, the screw came in contact with the Lifranc ligament in 20 feet. The screw fully penetrated the ligament in 7 feet, partially disrupted it in 4 feet, and had <1 mm of contact in 9 feet. In 3 feet, there was no contact with the ligament with an average distance of 1.5 mm. Conclusion: Our results reveal the amount of disruption a transarticular screw, placed in a dorsolateral to plantar medial direction, will have on the Lisfranc ligament. Although the screw came into contact with the ligament in 20 out of 23 feet, only 13 feet had partial disruption or minimal contact and 3 feet had no contact at all. This is clinically relevant because ligamentous damage due to insertion and/or presence of screw in anatomic location of the ligament may interfere with its healing

Association of Diabetes Mellitus and Immediate Postoperative Complications after Total Ankle Replacement: A Large Database Analysis

Category: Ankle Arthritis; Ankle Introduction/Purpose: Total Ankle Replacement (TAR) has rapidly grown in popularity over the last fifteen years as a treatment for ankle arthritis. With this growth, it is imperative to understand risk factors that may predispose patients to complications following the procedure. This information can be valuable in generating treatment plans and facilitating conversation regarding risks and benefits of those plans. Current literature on TAR-related outcomes is limited in timeframe and scope and does not investigate the association of diabetes with common post operative complications. The objective of this study was to perform a retrospective database analysis to quantify the risk of 90-day complications in patients with diabetes undergoing TAR when compared to patients without diabetes undergoing TAR. Methods: The TriNetX database was utilized for this study. Patients who underwent TAR were identified and stratified by those with a history of diabetes mellitus and those without a history of diabetes mellitus of any type. The cohorts were 1:1 propensity matched based on demographic information and medical history. The 90-day complication rates for readmission, inpatient service use, opioid use, anesthesia use, periprosthetic fracture, deep vein thrombosis, and wound dehiscence were calculated using chi-square analysis. Results: n=5,984 TAR patients were identified. 52% were male and 48% female with mean age of 73±10 years. There was a n=1,331 patients in each the patients with diabetes and without diabetes groups after cohort analysis and propensity matching. TAR patients with diabetes had a significantly higher 90-day odds ratio of inpatient service use (OR: 1.675, 95% CI 1.3082-2.144; p< 0.001). There was no significant difference between groups in opioid use (OR: 1.13, 95% CI 0.941-1.357; p=0.1908), anesthesia use (OR: 1.335, 95% CI 0.906-1.965; p=0.1424), and wound dehiscence (OR: 1.313, 95% CI 0.829-2.08; p=0.2445); Table 1. There was insufficient data to determine the role of diabetes on critical care services, periprosthetic fracture, and DVT within 90 days of TAR. Conclusion: TAR is a developing operation for ankle arthritis and little data exists on diabetes related perioperative complications. The results of this study show that although diabetes is significantly associated with a higher rate of inpatient service use, it may not have as large a role in opioid use, anesthesia use, and wound dehiscence in the immediate postoperative period. More information is still needed to address long term outcomes and the role diabetes plays in other postoperative complications