Pseudomonas Aeruginosa Elastase Induces Restructuring of Actin Cytoskeleton by Phosphorylation of RhoA Proteins

Pseudomonas aeruginosa causes aggressive infection in patients with pre-existing disorders and recurrent pulmonary infections in cystic fibrosis patients. Pathogenesis of P. aeruginosa infections is multifactorial owing to numerous virulence factors. The focus of this thesis research was to investigate whether P. aeruginosa elastase (PE) causes remodeling of the cytoskeleton by increasing the phosphorylation of RhoA GTPase proteins. In addressing our hypothesis, we utilized Small GTPase Immuno-sorbent Activation assays (G-LISA) and Enzyme linked Immuno-sorbent assay (ELISA) to quantitate changes in the total as well as phosphorylated RhoA protein in Calu3 cell lines. Fluorescence microscopy aided in understanding the changes in morphological organization of F-actin. Changes in expression of TJ protein, ZO1, due to PE induced RhoA GTPase activity, was analyzed with SDS PAGE and Western Blot Analysis. Our data from G-LISA and ELISA assays indicate that PE increases the amount of active RhoA protein by 50.8% in comparison to PBS treated control cells. RhoA inhibitor reduced the PE-induced phosphorylation of RhoA proteins by 30.35 % in PE treated cells. Images from fluorescence microscopy revealed that increase in RhoA GTPase activity causes formation of specific morphological protrusions such as stress fibers, lamellipodium and filopodium. Presence of RhoA inhibitor reverses the changes induced by PE. Results from immuno-sorbent assays and fluorescence microscopy indicate that inhibition of EGFR and MAPK significantly reduces the PE-induced GTPase activity of RhoA by 48.07 % and 42.2 % respectively. Data from G-LISA and ELISA assays correlate well with the morphology data obtained by fluorescence microscopy. Taken together, our data indicate that PE, at least in part, activates RhoA kinase via upstream EGFR and downstream MAPK signaling pathways, in vitro. The impact of these PE-induced alterations in the anatomy and physiology of the tight junctional complex, and their impact on the homeostasis of the lungs, demands further investigation

Gendered and Casteist Body: Cast(e)ing and Castigating the Female Body in select Bollywood Films

This study analyzes the lopsided relationship between gender and caste and the intertwining body politics in select Bollywood films. Bandit Queen (1994) and Article 15 (2019) are films that depict marginalized Dalit women—victims of (s)exploitation and twofold oppressions of graded patriarchy. Based upon real incidents, Bandit Queen tells the tale of Phoolan Devi who is gang-raped by the upper caste Thakur Shri Ram and his clans of the village while Article 15 takes recourse to the gruesome Badayun rape case of 2014 and presents the murder and possible rape of two lower caste young girls. In both the films, the marginalized women are imprisoned and ghettoized in the “mutual bracketing” (Guru 112) of caste and gender. Their bodies thus become the ploys of the power dynamics of a caste-ridden society. The body is to be captured, controlled, and incarcerated by both the apparatus of hegemonic masculinity and the hierarchical ladder of the caste system. Dalit women’s bodies are the territories that are to be possessed through the weapons of sexual violence; the gang rape “perpetrated by the conquerors is a metonymic celebration of territorial acquisition” (Spivak 303). Within the framework triad of caste studies, gender studies, and body politics studies, this paper investigates dynamics of power through a detailed analysis of the films and aims to point out whether and how the films make any differentiations from the real incidents. These films produce socially conscious visual landscapes directed at a society that horridly bears spectacular and brutal realities that are often swept under the rug

In vitro Culture of Naïve Human Bone Marrow Mesenchymal Stem Cells: A Stemness Based Approach

Human bone marrow derived mesenchymal stem cells (BM-MSCs) resides in their niches in close proximity to hematopoietic stem cells (HSCs). These naïve MSCs have tremendous potential in regenerative therapeutics, and may also be exploited by cancer and infectious disease agents. Hence, it is important to study the physiological and pathological roles of naïve MSC. However, our knowledge of naïve MSCs is limited by lack of appropriate isolation and in vitro culture methods. Established culture methods use serum rich media, and serial passaging for retrospective isolation of MSCs. These primed MSCs may not reflect the true physiological and pathological roles of naive MSCs (Figure 1). Therefore, there is a strong need for direct isolation and in vitro culture of naïve MSCs to study their stemness (self-renewal and undifferentiated state) and developmental ontogeny. We have taken a niche-based approach on stemness to better maintain naïve MSCs in vitro. In this approach, stemness is broadly divided as niche dependent (extrinsic), niche independent (intrinsic) and niche modulatory (altruistic or competitive). Using this approach, we were able to maintain naïve CD271+/CD133+ BM-MSCs for 2 weeks. Furthermore, this in vitro culture system helped us to identify naïve MSCs as a protective niche site for Mycobacterium tuberculosis, the causative organism of pulmonary tuberculosis. In this review, we discuss the in vitro culture of primed vs. naïve human BM derived MSCs with a special focus on how a stemness based approach could facilitate the study of naïve BM-MSCs

In vitro Culture of Naïve Human Bone Marrow Mesenchymal Stem Cells: A Stemness Based Approach

Einführung Seit etwa zehn Jahren haben die globalen Migrationsbewegungen an Bedeutung gewonnen. Heute, in der Ära der Globalisierung, betreffen diese Migrationsbewegungen praktisch alle Regionen der Welt. In Lateinamerika hat sich die Zahl der Migranten, die in die Vereinigten Staaten, in andere Länder des Kontinents und nach Europa ausgewandert sind, zwischen 1990 und 2000 mehr als verdoppelt. Zu den wichtigsten sozioökonomischen Aspekten der Migrationsbewegungen zählen die Geldmittel und di..

Moments of Gaussian hypergeometric functions over finite fields

We prove explicit formulas for certain first and second moment sums of families of Gaussian hypergeometric functions n+1Fn, n > 1, over finite fields with q elements, where q is an odd prime. This enables us to find an estimate for the value 6F5(1). In addition, we evaluate certain second moments of traces of the family of Clausen elliptic curves in terms of the value 3F2(−1). These formulas also allow us to express the product of certain 2F1 and n+1Fn functions in terms of finite field Appell series which generalizes current formulas for products of 2F1 functions. We finally give closed form expressions for sums of Gaussian hypergeometric functions defined using different multiplicative characters

Posterior mediastinal biphasic synovial sarcoma in a 12 year-old boy: A case report and review of literature

We report a case of biphasic synovial sarcoma of the mediastinum, a very rare tumor, in a 12-year-old boy with left-sided chest pain of 3 years duration at presentation. Chest X-ray showed left-sided opacity with loss of cardiac silhouette and the mediastinum deviated to the opposite side. Computed tomography (CT) of thorax showed left-sided posterior mediastinal mass with left-sided pleural effusion and pleural thickening. CT guided fine needle aspiration cytology (FNAC) from the mass reported it as spindle cell variant of adenocarcinoma. Ultrasonography (USG) of the whole abdomen revealed no abnormality. The mediastinal tumor was resected by left thoracotomy and histopathological report confirmed it to be a biphasic synovial sarcoma with capsule invasion at places

Posterior mediastinal biphasic synovial sarcoma in a 12 year-old boy: A case report and review of literature

We report a case of biphasic synovial sarcoma of the mediastinum, a very rare tumor, in a 12-year-old boy with left-sided chest pain of 3 years duration at presentation. Chest X-ray showed left-sided opacity with loss of cardiac silhouette and the mediastinum deviated to the opposite side. Computed tomography (CT) of thorax showed left-sided posterior mediastinal mass with left-sided pleural effusion and pleural thickening. CT guided fine needle aspiration cytology (FNAC) from the mass reported it as spindle cell variant of adenocarcinoma. Ultrasonography (USG) of the whole abdomen revealed no abnormality. The mediastinal tumor was resected by left thoracotomy and histopathological report confirmed it to be a biphasic synovial sarcoma with capsule invasion at places

An audit of gastrointestinal stromal tumors from a tertiary medical college hospital in Eastern India

Introduction: Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of GI tract. They are recently recognized as a distinct pathological entity although previously they were grouped with sarcomas of smooth muscle origin, i.e., leiomyomas, leiomyoblastomas, or leiomyosarcomas. However, apart from GI tract, GIST can occur in any smooth muscle-like in the urinary bladder. Proper diagnosis by immunohistochemistry stain CD133 and risk stratification by morphological parameters has been the cornerstone of treatment. Materials and Methods: This study was conducted as a retrospective analysis to audit the number of cases presenting as GIST in the tertiary medical colleges of eastern India and find out the patterns of care with the available modalities of therapy. Results: Out of total 15 cases, the median age of presentation was 45 years; the Male: Female (M: F) ratio was 2:3 and persisting dragging prolonged chronic abdominal pain was present in the majority. Intestinal complications were few (20%). All were treated with imatinib mesylate 400 mg once daily. However, two patients progressed for whom the dose of imatinib was escalated and one patient was metastatic at onset who was later switched over to sorafenib even after disease progression with dose escalation with imatinib. The median follow-up was 17.5 months and the median time to response to imatinib was 3.2 months. The 2-year actuarial overall survival was 79.12%, and progression-free survival was 86.67%. Conclusions: The future directions are to determine appropriate duration of imatinib therapy in adjuvant/neoadjuvant and therapeutic setting