POTENTIAL PROTECTIVE EFFECT OF APOCYNIN IN ETHYLENE GLYCOL-INDUCED HEPATIC DAMAGE BY ATTENUATION OF MITOCHONDRIAL OXIDATIVE STRESS

ABSTRACTObjective: The present study was carried out to investigate the protective role of apocynin (APO), an nicotinamide adenine dinucleotide phosphateoxidase inhibitor, against ethylene glycol (EG)-induced hepatotoxicity in rats.Methods: Male Sprague-Dawley rats were divided into three groups with six animals in each group. Control group; EG group, in which hyperoxaluriawas induced by 0.4% EG in drinking water for 9 days; and EG+APO group, 0.4% EG in drinking water for 9 days along with APO at a dosage of200 mg/kg body weight/day, intraperitoneal. All the experimental animals were sacrificed on day 10. Serum and the liver homogenates were analyzedfor various biochemical parameters. Mitochondria from liver were isolated by differential centrifugation and were diagnosed for vital biochemicalparameters.Results: Hyperoxaluric animals have shown significantly increased levels of serum glutamic oxaloacetic transaminase, serum glutamic pyruvictransaminase, alkaline phosphatase, and lactate dehydrogenase, thus suggesting liver dysfunction. Declined activities of respiratory chain enzymesshowed mitochondrial dysfunction in EG treated rats. In addition, mitochondrial oxidative stress was evident by decreased levels of superoxidedismutase, reduced glutathione, and an increased lipid peroxidation (LPO). APO (200 mg/kg/day), significantly decreased EG-induced oxidativestress by reducing LPO and restoring antioxidant enzymes activities in liver tissue. Also, reduction in the impairment of liver mitochondria functioningwas detected in APO treated rats. Histological analysis depicted that APO treatment decreased liver epithelial damage, increased Kupffer cells, andrestored normal hepatocyte morphology.Conclusion: The results demonstrated the potential beneficial effects of APO in reducing EG-induced liver damage that might be through attenuationof mitochondrial oxidative stress.Keywords: Ethylene glycol, Liver, Oxidative stress, Mitochondrion, Apocynin, Antioxidant

THE ROLE OF NATURAL ANTIOXIDANTS AS POTENTIAL THERAPEUTIC AGENT IN NEPHROLITHIASIS

Renal injury and inflammation caused by ROS play a major role in stone formation. Under the hyperoxaluric condition, crystal deposition results in angiotensin II (Ang II) activation. NADPH Oxidase is stimulated by activated Ang II, leading to ROS production, which can damage renal cells. Oxidative stress also results in mitochondrial dysfunction and release of pro-apoptotic factors from depolarized mitochondria that result in apoptosis that leads to renal injury. Crystal retention in the kidney requires tubular epithelial injury accompanied by luminal expression of HA, OPN, and CD44. The expression of these molecules turns a non–crystal-binding epithelium into a crystal-binding one, thereby setting the stage for crystal retention. Recently many antioxidants have been studied that prevent hyperoxaluria mediated nephrolithiasis. Antioxidant treatment significantly reduces CaOx crystal deposition in kidneys. Naturally occurring antioxidants such as Vitamin E, Apocynin, Phycocyanin, Fucoidin, Gallotannins, Rottlerin, Lupeol, Curcumin, etc. have shown significant effect in combating renal injury which is an early event in nephrolithiasis. These findings point towards a great potential for the therapeutic application of antioxidants and free radical scavengers to reduce stone occurrence particularly under hyperoxaluric conditions. This review article attempts to compile various naturally occurring antioxidants used in treatment of nephrolithiasis. Keywords: Calcium oxalate, Oxidative stress, Hyperoxaluria, Reactive oxygen species, antioxidant

Differential responsiveness of MET inhibition in non-small-cell lung cancer with altered CBL.

Casitas B-lineage lymphoma (CBL) is an E3 ubiquitin ligase and a molecule of adaptor that we have shown is important for non-small-cell lung cancer (NSCLC). We investigated if MET is a target of CBL and if enhanced in CBL-altered NSCLC. We showed that CBL wildtype cells have lower MET expression than CBL mutant cells. Ubiquitination of MET was also decreased in CBL mutant cells compared to wildtype cells. Mutant cells were also more sensitive to MET inhibitor SU11274 than wild-type cells. sh-RNA-mediated knockdown of CBL enhanced cell motility and colony formation in NSCLC cells, and these activities were inhibited by SU11274. Assessment of the phospho-kinome showed decreased phosphorylation of pathways involving MET, paxillin, EPHA2, and VEGFR. When CBL was knocked down in the mutant cell line H1975 (erlotinib-resistant), it became sensitive to MET inhibition. Our findings suggest that CBL status is a potential positive indicator for MET-targeted therapeutics in NSCLC

Myeloid suppressor cell depletion augments antitumor activity in lung cancer.

BackgroundMyeloid derived suppressor cells (MDSC) are important regulators of immune responses. We evaluated the mechanistic role of MDSC depletion on antigen presenting cell (APC), NK, T cell activities and therapeutic vaccination responses in murine models of lung cancer.Principal findingsIndividual antibody mediated depletion of MDSC (anti-Gr1 or anti-Ly6G) enhanced the antitumor activity against lung cancer. In comparison to controls, MDSC depletion enhanced the APC activity and increased the frequency and activity of the NK and T cell effectors in the tumor. Compared to controls, the anti-Gr1 or anti-Ly6G treatment led to increased: (i) CD8 T cells, (ii) NK cells, (iii) CD8 T or NK intracytoplasmic expression of IFNγ, perforin and granzyme (iv) CD3 T cells expressing the activation marker CD107a and CXCR3, (v) reduced CD8 T cell IL-10 production in the tumors (vi) reduced tumor angiogenic (VEGF, CXCL2, CXCL5, and Angiopoietin1&2) but enhanced anti-angiogenic (CXCL9 and CXCL10) expression and (vii) reduced tumor staining of endothelial marker Meca 32. Immunocytochemistry of tumor sections showed reduced Gr1 expressing cells with increased CD3 T cell infiltrates in the anti-Gr1 or anti-Ly6G groups. MDSC depletion led to a marked inhibition in tumor growth, enhanced tumor cell apoptosis and reduced migration of the tumors from the primary site to the lung compared to controls. Therapeutic vaccination responses were enhanced in vivo following MDSC depletion with 50% of treated mice completely eradicating established tumors. Treated mice that rejected their primary tumors acquired immunological memory against a secondary tumor challenge. The remaining 50% of mice in this group had 20 fold reductions in tumor burden compared to controls.SignificanceOur data demonstrate that targeting MDSC can improve antitumor immune responses suggesting a broad applicability of combined immune based approaches against cancer. This multifaceted approach may prove useful against tumors where MDSC play a role in tumor immune evasion

Regional variation in pig farmer awareness and actions regarding Japanese encephalitis in Nepal : implications for public health education

The objective was to explore regional variations in farmer awareness and actions towards Japanese Encephalitis (JE) in Nepal; the association of awareness and actions with farm and farmer variables; and the implications for public health education and extension services. Social factors such as literacy, gender, and cultural practices were associated with farmer attitudes, knowledge and practices for JE control. The low uptake of vaccine and lack of infrastructure or financial capacity to house pigs adequately suggest that farmer personal protection should be a priority for education. JE is a mosquito-borne zoonotic disease with pigs as the amplifying hosts

Novel CCL21-Vault Nanocapsule Intratumoral Delivery Inhibits Lung Cancer Growth

Based on our preclinical findings, we are assessing the efficacy of intratumoral injection of dendritic cells (DC) transduced with an adenoviral vector expressing the secondary lymphoid chemokine (CCL21) gene (Ad-CCL21-DC) in a phase I trial in advanced non-small cell lung cancer (NSCLC). While this approach shows immune enhancement, the preparation of autologous DC for CCL21 genetic modification is cumbersome, expensive and time consuming. We are evaluating a non-DC based approach which utilizes vault nanoparticles for intratumoral CCL21 delivery to mediate antitumor activity in lung cancer.Here we describe that vault nanocapsule platform for CCL21 delivery elicits antitumor activity with inhibition of lung cancer growth. Vault nanocapsule packaged CCL21 (CCL21-vaults) demonstrated functional activity in chemotactic and antigen presenting activity assays. Recombinant vaults impacted chemotactic migration of T cells and this effect was predominantly CCL21 dependent as CCL21 neutralization abrogated the CCL21 mediated enhancement in chemotaxis. Intratumoral administration of CCL21-vaults in mice bearing lung cancer enhanced leukocytic infiltrates (CXCR3(+)T, CCR7(+)T, IFNγ(+)T lymphocytes, DEC205(+) DC), inhibited lung cancer tumor growth and reduced the frequencies of immune suppressive cells [myeloid derived suppressor cells (MDSC), T regulatory cells (Treg), IL-10 T cells]. CCL21-vaults induced systemic antitumor responses by augmenting splenic T cell lytic activity against parental tumor cells.This study demonstrates that the vault nanocapsule can efficiently deliver CCL21 to sustain antitumor activity and inhibit lung cancer growth. The vault nanocapsule can serve as an "off the shelf" approach to deliver antitumor cytokines to treat a broad range of malignancies

Knowledge Synthesis and Dissemination of Ecohealth Research Projects : 8th International Conference on Urban Health (ICUH) in Nairobi, Kenya, October 19-23, 2009

International Development Research Centre (IDRC) supported research study on Echinococcosis/Hydatidosis which was carried out between 1992 and 2001 in Kathmandu Metropolitan City Nepal. The research work was carried out in two distinct project stages in wards 19 and 20 of KMC one from 1992-1996(applying a traditional epidemiology approach) and a second was Urban Ecosystem Health Project (UEHP) Approach from 1998-2006. Overall, there were three main step of activities in the development of the UEHP in Nepal: First is an exploratory/analytical systemic steps focusing on linkages between social, ecological, and health variables (1998-2001); The second a community action step employing a variety of systemic, narrative, and participatory-action research tools (2003-2006); and the third phase of urban ecosystem health project started from 2007 to 2009 which is now running. Community participation or participatory action research is a key element of ecosystem health programmes. Participatory Action Research approaches have three main goals. The first step is to describe what is there? What are the physical and socio-economic possibilities of this person or place? If we use the analogy of a person. The Second step, for ecosystems as for individuals, health is not just a physical state, but what we might call a spiritual state. It has been said that in any part of the world no solutions will be sustainable in the long run unless they are rooted in the communities where the problems occur, drawing on the people in these communities and their many skills, resources, and important knowledge, and those communities feel empowered and supported by higher levels of government. If such approaches can be worked out between local communities in Indian Subcontinent, non-government organizations, businesses, regional institutions of government and university, and with outside input only as necessary, then truly sustainable solutions will be found, and Kathmandu

Experience with Livaditis circular myotomy in management of long gap TEF

Background: Management of long gap oesophageal atresia with tracheoesophageal fistula (OA TEF) is challenging. Various intra-operative and pre-operative manures have been described to tackle this challenge. We reviewed our experiences with livaditis circular myotomy. The aim of this study was to evaluate long-term outcomes in cases of long gap OA TEF managed primarily with livaditis circular myotomy. Materials and Methods: This is a cross-sectional study including cases of long gap oesophagus managed by livaditis circular myotomy between January 1998 and October 2012. Their case records were evaluated for operative and post-operative data. The anthropometric data of these cases were collected. All these cases were subjected to barium swallow and manometry. Those cases with other associated neurological anomalies, multiple congenital anomalies, parents refusing consent for the study, less than 6 months of follow up or incomplete data were excluded from the study. Results: Out of the total of 109 patients of OA TEF managed, long gaps OA TEF were 37. Out of the 37 cases, 13 were managed by primary repair with livaditis circular myotomy. Of these 13 cases, 11 formed the study group. Mean age at evaluation was 36 ± 9 months. Mean age at primary surgery was 3 ± 2.5 days of life. Minor leak in the immediate post-operative period was present in 2/11 cases. Manometry was done in all the cases and revealed motility disorder in the form of un-coordinated contraction in 4/11 cases. Remaining 7/11 cases were normal. Conclusion: Livaditis circular myotomy is a viable option in the management of long gap OA TEF with good comparable long-term results