Virus Purification Framework And Enhancement In Aqueous Two-Phase System

Viral infections regularly pose detrimental health risks to humans. Preventing viral infections through global immunization requires the production of large doses of vaccines. The increasing demand for vaccines, especially during pandemics such as COVID-19, has challenged current manufacturing strategy to develop advanced unit operations with high throughput capability. Over the decade, the upstream processing responsible for synthesizing viral products in cell cultures has shown significant success in yielding high titers of viruses and virus-like particles. The progress in the upstream stage has now shifted the bottleneck to the downstream processing (DSP). Overlooked for decades, the DSP responsible for viral product purification from the cell culture contaminants requires a makeover with the development of new purification strategies and an upgrade in the traditional unit operations. The current DSP train employing chromatography and filtration methods have been suboptimal in efficiently processing comparatively complex and fragile viral particles. Thus, the lack of platform technology for viral vaccine and biotherapeutic DSP has led to a search for alternative and innovative methods that have not only high-throughput capabilities but also have potential for continuous operation. In the pool of potential technologies, aqueous two-phase system (ATPS) has shown to be a promising candidate with the numerous advantages over conventional methods. However, an unambiguous and complex biomolecule partitioning mechanism has required a large experimental setup for optimizing virus purification. This work focused on a framework utilizing a phase diagram of a rationalized polyethylene glycol-citrate system to optimize virus purification. The partitioning behavior of two non-enveloped viruses, porcine parvovirus (PPV) and human rhinovirus-14 (HRV), were studied in various system compositions. A tie-line length framework was utilized to define the systems and relate the partitioning behavior of viruses with different surface physicochemical characteristics

Modified SPLICE and its Extension to Non-Stereo Data for Noise Robust Speech Recognition

In this paper, a modification to the training process of the popular SPLICE algorithm has been proposed for noise robust speech recognition. The modification is based on feature correlations, and enables this stereo-based algorithm to improve the performance in all noise conditions, especially in unseen cases. Further, the modified framework is extended to work for non-stereo datasets where clean and noisy training utterances, but not stereo counterparts, are required. Finally, an MLLR-based computationally efficient run-time noise adaptation method in SPLICE framework has been proposed. The modified SPLICE shows 8.6% absolute improvement over SPLICE in Test C of Aurora-2 database, and 2.93% overall. Non-stereo method shows 10.37% and 6.93% absolute improvements over Aurora-2 and Aurora-4 baseline models respectively. Run-time adaptation shows 9.89% absolute improvement in modified framework as compared to SPLICE for Test C, and 4.96% overall w.r.t. standard MLLR adaptation on HMMs.Comment: Submitted to Automatic Speech Recognition and Understanding (ASRU) 2013 Worksho

Various sacral indices: role in study of sexual dimorphism

Background: Skeleton is an excellent material in living and non-living population for genetic, anthropological and forensic investigations. Sound anatomical knowledge of human osteology in the evaluation of death may help to shed light on legal issues such as sex and age determination, parentage and racial identity. It has widely recognized that skeletal characters vary among different racial groups and even within the same racial groups. Sacrum is one of the commonest part of skeleton used to opine the sex. The study documents effort to determine sex by using different measurements of the sacrum and calculating the different indices based on these measurements. The most useful aspect of such study is to evaluate the role of different indices of sacrum in sex determination.Methods: Study was carried out on 141 adult, dry sacra in Department of Anatomy, BVDU Medical College, Pune. 141 sacra were classified into male and female bones by observing the parameters like - length of sacrum, width of sacrum, transverse diameter of body of S1, anteroposterior diameter of S1, length of ala of sacrum.Results: The indices like sacral index, curvature index, corpobasal index and alar Index were found to be statistically significant with p value 0.05 and was statistically not significant.Conclusions: The most useful aspect of such studies was to determine appropriate indices of sexual dimorphism for sacral bone. The study concluded that no single index can identify sex of sacrum with 100% accuracy. So multiple indices should be used for determination of sex by sacrum with 100% accuracy.

Association Mapping for Improvement of Quantitative Traits in Plant Breeding Populations

DNA-based molecular markers have been extensively utilized for mapping of genes and quantitative trait loci (QTL) of interest based on linkage analysis in mapping populations. This is in contrast to human genetics that use of linkage disequilibrium (LD)-based mapping for fine mapping of QTLs using single nucleotide polymorphisms. LD based association mapping (AM) has promise to be used in plants. Possible use of such approach may be for fine mapping of genes / QTLs, identifying favorable alleles for marker aided selection and cross validation of results from linkage mapping for precise location of genes / QTLs of interest. In the present review, we discuss different mapping populations, approaches, prospects and limitations of using association mapping in plant breeding populations. This is expected to create awareness in plant breeders in use of AM in crop improvement activities.Key words: Association mapping; plant breeding; DNA marker; quantitative trait lociDOI: http://dx.doi.org/10.3126/njb.v2i1.5686  Nepal Journal of Biotechnology Jan.2012, Vol.2(1): 72-8

Are counselling psychology training courses developing multicultural competencies in their trainees? Comparing cultural competency in health care and counselling psychology trainees

A Portfolio submitted to the University of Wolverhampton for the Practitioner Doctorate in Counselling Psychology.The need for professionals to develop multicultural awareness, knowledge and skills has been recognised in literature and mental health practitioner programmes since the early 1970s. Despite the professional growth of the Doctorate in Counselling Psychology (DCoP), recent research still suggests that cultural competency skills must be further emphasised and developed in training courses. Following an integrative philosophical model, this study adopts a mixed methods approach including interviews and cross-sectional surveys to explore how cultural competency skills are facilitated in DCoP training or acquired by professionals compared to biomedical trainees and professionals. Interviews were conducted with the DCoP course directors, while the Lie scale (Eysenck, 1976) measuring social desirability and the Healthcare Provider Cultural Competence Instrument (HPCCI) (Schwarz, Witte, Sellers, Luzadis, Weiner, Domingo-Snyder, 2015) measuring level of cultural competency skills, were administered to the DCoP and biomedical groups. I hypothesised that the DCoP trainees will score significantly higher on cultural competency compared to trainees from other training programmes and this proved not significant. The results have implications for further developments of cultural competencies in counselling psychology and biomedical training courses

IN VITRO ANTHELMINTIC ACTIVITY OF MAYTENUS EMARGINATA STEM BARK ON INDIAN ADULT EARTHWORM

Objective: The development of anthelmintic activity and the high cost of synthetic anthelmintic drugs it leads to the evaluation of medicinal plants as an alternative source of anthelmintics. In the current study, in vitro experiments were conducted to determine the possible anthelmintic effects of crude methanolic, aqueous, and hydroalcoholic extracts of the stem bark of Maytenus emarginata on eggs and adult Pheretima posthuma and Ascaridia galli. Methods: Three concentrations (10, 25, and 50 mg/ml) of methanolic, aqueous, and hydroalcoholic extracts of plant M. emarginata were studied in activity which involved the determination of the time of paralysis (called as vermifuge) and time of death (called as vermicidal) of the worms. Piperazine citrate in same concentration as those of extract was included as standard reference and normal saline water with 1% carboxymethyl cellulose as control. Results: The methanolic, aqueous, and hydroalcoholic extracts exhibited significant anthelmintic activity at a concentration of 50 mg/ml. Peak activity was exhibited by the methanolic extract at a concentration of 50 mg/ml. Conclusion: The overall findings of the present study have shown that M. emarginata contain possible anthelmintic compounds and further evaluation of these plants should be carried out. The traditional use of anthelmintic activity of this plant is genuine

The pathophysiological significance of Factor Seven (FVII) activating protease (FSAP) in Ischemic stroke

Factor Seven Activating Protease (FSAP) is a circulating protease with a role in coagulation, fibrinolysis and inflammation. The FSAP-Marburg I polymorphism, with low proteolytic activity, is associated with an increased risk of stroke and late complications of carotid stenosis in humans. We recently showed that FSAP antigen and activity levels are elevated in patients with Ischemic Stroke. In order to find a mechanistic explanation for this we have investigated the effect of FSAP on primary mouse brain microvascular endothelial cells, astrocytes and neurons. We demonstrate that FSAP can regulate endothelial permeability in an in-vitro model of the blood brain barrier (BBB) by stabilizing junctional localization of ZO-1 and by preventing the opening of the endothelial tight junctions after oxygen glucose deprivation (OGD)/reoxygenation. Furthermore, FSAP can cross the BBB and exert a protective effect on cortical astrocytes and neurons exposed to conditions mimicking stroke by activating the PI3K-Akt pathway. This protective effect was mediated by PAR-1 dependent activation of Akt pathway and dependent on its proteolytic activity. In a mouse in-situ thromboembolic stroke model, we observed that infarction volumes at 24 hours were significantly larger in FSAP-/- mice and neurological score worse. However, no difference was observed in the either cerebral reperfusion or cerebral vasculature. Increased FSAP antigen and activity levels were detected in WT mice after stroke in accordance with the clinical findings. In the absence of endogenous FSAP, there was a tendency towards a pro-inflammatory phenotype post stroke. Also, the lack of FSAP after stroke resulted in decreased Akt phosphorylation and Bcl-2 levels while increased levels of p53 were observed. Thus, from the data presented in this thesis, we propose that FSAP is a novel neuroprotective agent of the neurovascular unit and loss of FSAP activity could suggest the possible mechanism which leads to less protection in Marburg I polymorphism patients.Die Faktor VII-aktivierende Protease (FSAP) ist eine zirkulierende Protease mit einer Rolle in der Koagulation, der Fibrinolyse und in Entzündungsprozessen. Der FSAP-Marburg-I-Polymorphismus, ohne proteolytisch Aktivität ist mit einem erhöhten Risiko für Hirn-Infarkte und späteren Komplikationen wie Karotisstenosen assoziiert. Wir zeigen, dass FSAP-Antigen und -Aktivität in Patienten mit ischämischem Schlaganfall erhöht sind. Um eine mechanistische Erklärung dafür zu finden, haben wir die Wirkung von FSAP auf primäre mikrovaskuläre Hirn-Endothelzellen, Astrozyten und Neuronen untersucht. In dieser Studie zeigen wir, dass FSAP die endotheliale Permeabilität in einem in-vitro-Modell der Blut-Hirn-Schranke (BBB) durch Stabilisierung der junktionalen Lokalisierung von ZO- 1 und durch die Öffnung der endothelialen tight-junctions nach Sauerstoff-Glucose-Deprivation (OGD)/ Reoxygenierung verhindern regulieren bzw kann. Darüber hinaus kann FSAP die BBB überqueren und somit eine schützende Wirkung auf kortikale Astrozyten und Neuronen ausüben, die einem Schlaganfall nach Aktivierung des PI3K-Akt-Signalweges ausgesetzt sind. Dieser schützende Effekt wird durch PAR-1-abhängige Aktivierung des Akt-Signalweges vermittelt. In einem in-situ thromboembolischen Schlaganfall-Modell der Maus beobachteten wir, dass das Infarktvolumen nach 24 Stunden deutlich größer und das neurologische Ergebnis in FSAP-/- Mäusen deutlich schlechter war. Es wurde kein Unterschied in der zerebralen Reperfusion oder in zerebralen Gefäßen beobachtet. In WT-Mäusen wurde mehr FSAP-Antigen und eine erhöhte FSAP-Aktivität festgestellt. In Abwesenheit von endogenem FSAP gab es eine Tendenz hin zu einem pro-inflammatorischen Phänotyp nach einem Schlaganfall. Auch das Fehlen von FSAP nach einem Schlaganfall führte zu einer verringerten Akt- Phosphorylierung und veringerten Bcl-2-Werten während erhöhte p53-Werte beobachtet wurden. So gibes eine neue Rolle für FSAP als neuroprotektives Agens in der neurovaskulären Einheit und der Verlust der FSAPAktivität könnte eine mögliche Ursache des geringeren Schutzes in Marburg-IPolymorphismus Patienten sein

THE ROLE OF SDF-1[alpha] AS A VASCULOGENIC CHEMOKINE AND ENDOTHELIUM-ASSOCIATED CELL ADHESION MOLECULE FOR THE RECRUITMENT OF BONE MARROW-DERIVED PROGENITOR CELLS TO DEVELOPING TUMORS

Tumor vascularization is an important control point in cancer progression and its inhibition is a promising approach to cancer therapy. Stromal-derived factor-1 alpha (SDF-1 alpha or CXCL12) is a chemoattractant for lymphocytes, hematopoietic progenitor cells, and vascular endothelial cells. SDF-1 alpha binds to the CXCR4 receptor on cell surfaces and to heparan sulfate in cell membranes and the extracellular matrix. We hypothesized that SDF-1 alpha is an important mediator of tumor vasculogenesis, recruiting bone marrow-derived endothelial progenitor cells to tumors. We evaluated the role of SDF-1 alpha in the adhesion of bone marrow-derived lineage-negative cells to the murine Lewis lung adenocarcinoma (LLCaB subclone), both in vitro and in vivo. LLCaB lung and liver metastases expressed significantly elevated levels of SDF-1 alpha compared to normal tissue from naive controls and primary tumors. SDF-1 alpha expression inversely correlated with tumor size, implicating involvement in early tumor development. SDF-1 alpha was detected on ~30% of blood vessels in primary and metastatic tumors by double immunocytochemistry staining and on endothelial cells within primary cultures of dissociated LLCaB tumors (DTC). Lineage-negative BMC enriched for Sca1+cKit+ (LSK) cells, including endothelial progenitors and CXCR4+ cells, were used for subsequent adhesion studies. LSK cells preferentially adhered in vitro to DTC, compared to other target cell types. Pre-incubation with anti-CXCR4 antibody or the CXCR4 antagonist AMD3100 decreased adhesion of LSK cells by 50%. Biolocalization of adoptively-transferred EGFP-Lineage-negative BMC in tumor-bearing wild-type mice was evaluated by qPCR for the egfp-transgene. A significantly greater number of Lineage-negative BMC localized to metastases 2-3 days after intravenous injection than to adjacent healthy tissue, normal organs, or primary tumors. Control CXCR4-negative EGFP-dermal fibroblasts showed significantly less tumor localization. One-week post-injection, Lineage-negative BMC were still detected in every metastasis, suggesting that BMC were retained and/or had replicated. These data support the hypothesis that the biolocalization of circulating Lineage-negative BMC to tumors is SDF-1 alpha-mediated. We have also put forth the novel proposal that SDF-1 alpha promotes early tumor vasculogenesis as a tumor endothelium-associated adhesion molecule for circulating CXCR4+ Lineage-negative BMC. These findings may aid our understanding of the mechanism of tumor vascularization and the design of tumor-targeted therapy.  Ph.D

Kvercetin potpomaže akaricidnu aktivnost ivermektina u slučaju svinja prirodno oboljelih od sarkoptoze

Sarcoptic mange triggers continuous oxidative onslaughts, resulting in severe oxidative stress in pigs and, to date, no antioxidant has been evaluated for the treatment of naturally infested pigs. This randomized clinical trial aimed to assess the ameliorative potential of the antioxidant quercetin (QR) when integrated with ivermectin (IVM) in the treatment of sarcoptic mange in pigs. The control group (T0 , n=10) consisted of healthy subjects. The first treatment group (T1 , n=10) consisted of infested pigs receiving the standard treatment (subcutaneous IVM only) while the second treatment group (T2 , n=10) consisted of infested pigs receiving integrated treatment (subcutaneous IVM plus oral QR). On day 0, the circulating malondialdehyde (MDA) was significantly higher and superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), total antioxidant capacity (TAC) and antioxidative minerals (zinc, copper, iron) were lower in all infested pigs compared to the healthy subjects. On day 14 post-treatment, maximum recovery was observed in the MDA, SOD, GSH, CAT, TAC, zinc, copper and iron in group T2 and the results returned to normal earlier in group T2 than in T1 . Likewise, more significant improvements in parasitological cure rate, scratching index and skin score were recorded after treatment in group T2 than group T1 . These results suggest the greater effectiveness of IVM plus QR than IVM alone against sarcoptic mange, and quercetin may be recommended as an ancillary therapy with IVM to negate severe oxidative stress, improve post-therapy convalescence and produce a speedy recovery in pigs.Sarkoptoza u svinja pokreće poremećaje koji rezultiraju teškim oksidacijskim stresom za koji još uvijek nije otkriven antioksidans kojim bi se prirodno infestirane svinje tretirale. Cilj je ovog randomiziranog kliničkog istraživanja bio procijeniti antioksidacijski potencijal kvercetina (QR) u kombinaciji s ivermektinom (IVM) u liječenju sarkoptoze u svinja. U kontrolnoj su skupini (T0 , n=10) bile zdrave jedinke. U prvoj su pokusnoj skupini (T1 , n = 10) infestirane svinje dobile standardnu terapiju (samo IVM primijenjen supkutano), dok su infestirane svinje u drugoj pokusnoj skupini (T2 , n = 10) primile integriranu terapiju (supkutano IVM i oralno QR). Nulti dan cirkulacijski je malondialdehid (MDA) bio znakovito veći, dok su superoksidna dismutaza (SOD), reducirani glutation (GSH), katalaza (CAT), ukupan anitoksidacijski kapacitet (TAC) i antioksidacijski minerali (cink, bakar i željezo) bili smanjeni u infestiranih svinja u usporedbi sa zdravim jedinkama. Četrnaesti dan poslije liječenja uočen je maksimalan oporavak u pogledu pokazatelja MDA, SOD, GSH, CAT, TAC, cinka, bakra i željeza u skupini T2 te njihov raniji povratak na uobičajene vrijednosti u skupini T2 u odnosu na skupinu T1. Osim toga, u skupini T2 u odnosu na skupinu T1 zapaženo je znakovito poboljšanje u stopi izliječenosti parazitoze, indeksu grebenja i bodovanju promjena na koži. Ovi rezultati upućuju na veću učinkovitost IVM-a u kombinaciji s QR-om nego IVM-a upotrijebljenog kao samostalna terapija sarkoptoze u svinja. Zaključuje se da bi kvercetin mogao biti dodatna terapija uz IVM kako bi se poništili teški učinci oksidacijskog stresa, poboljšala poslijeterapijska rekonvalescencija i ubrzao oporavak svinja

Availability and use of magnesium sulphate at health care facilities in two selected districts of North Karnataka, India

Background: Pre-eclampsia and eclampsia are major causes of maternal morbidity and mortality. Magnesium sulphate is accepted as the anticonvulsant of choice in these conditions and is present on the WHO essential medicines list and the Indian National List of Essential Medicines, 2015. Despite this, magnesium sulphate is not widely used in India for pre-eclampsia and eclampsia. In addition to other factors, lack of availability may be a reason for sub-optimal usage. This study was undertaken to assess the availability and use of magnesium sulphate at public and private health care facilities in two districts of North Karnataka, India. Methods: A facility assessment survey was undertaken as part of the Community Level Interventions for Pre-eclampsia (CLIP) Feasibility Study which was undertaken prior to the CLIP Trials (NCT01911494). This study was undertaken in 12 areas of Belagavi and Bagalkote districts of North Karnataka, India and included a survey of 88 facilities. Data were collected in all facilities by interviewing the health care providers and analysed using Excel.Results: Of the 88 facilities, 28 were public, and 60 were private. In the public facilities, magnesium sulphate was available in six out of 10 Primary Health Centres (60%), in all eight taluka (sub-district) hospitals (100%), five of eight community health centres (63%) and both district hospitals (100%). Fifty-five of 60 private facilities (92%) reported availability of magnesium sulphate. Stock outs were reported in six facilities in the preceding six months – five public and one private. Twenty-five percent weight/volume and 50% weight/volume concentration formulations were available variably across the public and private facilities. Sixty-eight facilities (77%) used the drug for severe pre-eclampsia and 12 facilities (13.6%) did not use the drug even for eclampsia. Varied dosing schedules were reported from facility to facility.Conclusions: Poor availability of magnesium sulphate was identified in many facilities, and stock outs in some. Individual differences in usage were identified. Ensuring a reliable supply of magnesium sulphate, standard formulations and recommendations of dosage schedules and training may help improve use; and decrease morbidity and mortality due to pre-eclampsia/ eclampsia