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

The Relevance of Thrombo-Inflammatory Biomarkers and Their Relationship with Circulating Glycosaminoglycans in End-Stage Renal Disease Patients

En-stage renal disease (ESRD) is a growing public health problem. The atherosclerotic cardiovascular complications are the leading causes of mortality and morbidity in the ESRD. In this study, we sought to quantify the levels of thrombo-inflammatory biomarkers in an ESRD patients in comparison to healthy controls to determine their relevance in thrombo-inflammation and adverse outcomes. The levels of D-Dimer, C-reactive protein (CRP), plasminogen activator inhibitor 1 (PAI-1) antigen, functional PAI-1, thrombin activatable fibrinolysis inhibitor, tissue plasminogen activator, von Willebrand factor, and anti-PF4 IgG and microparticle (MP) activity were quantified by using commercially available ELISA immunoassays for each of the ESRD ( n = 73) and control plasma samples ( n = 10). The levels of endogenous glycosaminoglycans (GAGs) were quantified by utilizing a Heparin Red Probe (Redprobes UG, Germany). The collected data were analyzed to demonstrate the relationship between various parameters. All the tested biomarkers were increased in ESRD patients in comparison to healthy controls ( p < 0.05). These biomarkers have shown significant correlations within each other except for anti-PF4 Ig G and MPs. The CRP levels were significantly higher in patients who had coronary artery disease (CAD) ( p < 0.05), but there was no significant difference in other biomarkers according to the cardiovascular outcomes. In the multivariate analysis, the CRP (odds ratio: 1.19; 95% confidence interval: 1.01–1.41; p : 0.03) value was an independent predictor of CAD. In this study, we demonstrated increased levels of 10 different biomarkers in ESRD patients. The CRP levels can be a good predictor of CAD in ESRD patients

Does Hallux Valgus Correction with Chevron-Akin Osteotomies Reduce the Length of the First Ray

Category: Bunion; Midfoot/Forefoot Introduction/Purpose: Patients who undergo a successful surgical correction of hallux valgus have relieved toe pain and improvements in foot appearance. Prior studies have shown hallux valgus correction using a distal Chevron osteotomy with a concomitant Akin osteotomy has the potential to reduce forefoot width. There are no studies evaluating if similar changes occur in the length of the foot. The objective of this study is to check if similar reduction occurs in the patient's foot length. Methods: The operating room schedule of Dr. Panchbhavi and Dr. Chen were reviewed for Chevron/Akin Osteotomies from 2017-2022. Patient charts were excluded for contaminant operations of the foot. The preoperative and postoperative radiographs were then evaluated by one medical student with oversight from a Board-Certified Foot and Ankle Orthopedic Surgeon. The Hallux Valgus Angle (HVA), Intermetatarsal Angle (IMA), Metatarsal Span (MS), first/second ray of the foot, and difference between first and second ray were all be measured based on predetermined methods. All variables were analyzed utilizing an unpaired t-test. Results: In total 12 Chevron/Akin patients met the criteria. The results indicated that with chevron/akin osteotomy measurements of the foot were affected in HVA, IMA, MS, and length of first ray relative to second ray. The average results for each of these values were a decrease of 9.2 degrees (SD 4.32), 7.93 degrees (SD 4.33), 11.63mm (SD 2.26) respectively. For results of change in difference of first and second ray, average results for those who underwent Chevron/Akin Osteotomy was -2.23mm (SD 2.23, p = 0.0281). Both changes to MS and length were statistically significant (p < 0.05). Conclusion: Our results were consistent with existing evidence, Chevron with contaminant Akin Osteotomy will produce statistically significant changes in width of the foot; however, also a significant decrease in the length of the first ray relative to the second ray. All measurements were done utilizing PACS software and can be reliably used in clinics as a clinical tool to gauge post- operative outcomes alongside providing adequate patient education on the potential changes in foot length and width

Peroneus Brevis Tenodesis: Side-to-Side or Weave?

Background: Inversion ankle injuries are extremely common, sometimes causing injury to the peroneus brevis tendon. If more than 50% of the tendon is injured, it oftentimes requires tenodesis to the adjacent peroneus longus tendon. Both Pulvertaft (PT) and side-to-side (SS) techniques have been used for joining the 2 tendons. The purpose of this study was to compare the strength and stiffness of these 2 techniques. Methods: Five matched pairs of cadaver ankle specimens were randomized to receive either an SS or PT tenodesis of the peroneus brevis to longus tendons. Following the tenodesis, the specimens were tested for failure load, displacement, energy absorbed at failure, and peak load. Stiffness was also calculated. Paired t tests were performed to detect differences between the 2 conditions. Results: There were no statistically significant differences between the SS and PT tenodesis for any of the metrics measured. For stiffness, the techniques were very similar (SS = 10.14 [4.35], PT = 12.85 [1.72]). Conclusion: There is no difference in failure load, displacement, energy absorbed at failure, peak load or stiffness between the PT and SS techniques for peroneal tenodesis. Level of Evidence: Level V, cadaver study