In vitro and in vivo approaches for the functional characterization of the scaffold protein, GRASP

Studies using the pluripotent embryonic carcinoma cell line, P19, as a retinoic acid (RA)-responsive model system have been instrumental towards our understanding of the RA-dependent signaling pathways in development and homeostasis. Grp1-associated scaffold protein (GRASP; also known as Tamalin) was first identified by our group, as a gene robustly induced by RA treatment in P19 cells. GRASP was reported to influence the cell-surface expression of interacting membrane receptors in vitro albeit by an unknown mechanism. Furthermore, the in vivo role of GRASP is poorly understood. Mice with a germline deletion of Grasp (GRASP-/-) do not exhibit any gross morphological, behavioral or sexual deficits and exhibit mild, altered sensitivities to morphine and cocaine administration. The goals of our studies herein were to investigate the mechanistic details of GRASP for the role in trafficking and cell-surface, localization of membrane receptors in vitro and the role of GRASP in vivo. Previously, our group had identified a strong interaction of GRASP with Grp1, a guanine nucleotide exchange factor for the small G protein, Arf6. In this study we show GRASP localizes in intracellular recycling endosomal compartment(s), recruits Grp1 to these structures, and facilitates activation of Arf6. Activation of Arf6 regulates key aspects of vesicular trafficking, actin reorganization and cellular migration. Furthermore, we show that GRASP preferentially regulates intracellular membrane trafficking by the Arf6-dependent/clathrin-independent pathway. We have shown that GRASP is expressed in the adult murine skin. Our studies demonstrate a robust induction of GRASP transcripts in murine dermis and epidermis following exposure to ultraviolet (UV) rays. We also report the generation of novel mice with germline deletion of Grasp (GRASP-/-), that exhibit altered proliferative and apoptotic responses to UV exposure. Ongoing investigations suggest that the observed phenotypes of GRASP-/- mice after UV exposure maybe due to the dysregulation of the subcellular localization of the tumor suppressor protein, p53. Taken together, our in vitro and in vivo approaches have provided new insight in the role of GRASP as scaffold protein that regulates intracellular trafficking pathways

Social Determinants Predicting the Community Pharmacists' Workforce Preparedness for, and Response to, the Public Health Emergencies

Objectives:To identify the predicting factors that contribute to preparedness for public health emergencies among community pharmacists in India.Methods:Multistage cluster sampling was done. The geographic breakdown was done based on villages and areas and used as clusters. A simple random method was done in the first stage to select the villages as clusters. From each selected village, a simple random method was done in the second stage to select the areas. From each selected area, all the community pharmacies were selected. The survey questionnaire had 3 sections with 43 items: (A) demographic information, (B) preparedness, (C) response toward infectious diseases. The participants chose “Yes/No”, in sections B and C. A score of 1 was given for “Yes”, and a score of zero was given for “No”.Results:Multiple correlation analyses were conducted between participants’ preparedness and response (PR) scores and independent variables. The independent variables such as “More than one Pharmacist working in a pharmacy”, “Pharmacists who are trained more than once on disaster management”, and encountered more than 1 patient with the infectious disease were positively and significantly correlated with the dependent variable (PR scores).Conclusions:Community pharmacists were aware of the issues they may face in their community concerning public health emergencies. They believed that the medications available in their pharmacy are sufficient to face any emergency. They could identify the clinical manifestations of public health emergency conditions and provide counselling to the customers toward them. Community pharmacists who were trained more than once in disaster management were the strongest predicting factor.<br/

Synthetical experiments in the chromone group part XXI. synthesis of gentisin, the colouring matter of gentian root

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A new synthesis of 4-hydroxycoumarins

4-Hydroxynaphthocoumarin has been synthesised by the internal condensation ofO-carbethoxy-2-acetyl-1-naphthol, thus suggesting a new general method for the synthesis of 4-hydroxycoumarins. The behaviour of 4-hydroxynaphthocoumarin has been studied; its derivatives including the corresponding dicoumarin have been prepared. By the action of potassium carbonate or sodamide on the carbethoxy ester of 2-acetyl-1-naphthol, a second product was obtained, the constitution of which is under examination

High-Risk Histopathological Features of Retinoblastoma following Primary Enucleation: A Global Study of 1426 Patients from 5 Continents

Purpose: To evaluate high-risk histopathological features (HRHF) following primary enucleation of eyes with retinoblastoma (RB) and assess the patient outcomes across continents // Methods: Retrospective study of 1426 primarily enucleated RB eyes from five continents // Results: Of all, 923 (65%) were from Asia (AS), 27 (2%) from Australia (AUS), 120 (8%) from Europe (EUR), 162 (11%) from North America (NA), and 194 (14%) from South America (SA). Based on the continent (AS vs. AUS vs. EUR vs. NA vs. SA), the histopathology features included massive choroidal invasion (31% vs. 7% vs. 13% vs. 19% vs. 27%, p=0.001), post-laminar optic nerve invasion (27% vs. 0% vs. 16% vs. 21% vs. 19%, p=0.0006), scleral infiltration (5% vs. 0% vs. 4% vs. 2% vs. 7%, p=0.13), and microscopic extrascleral infiltration (4% vs. 0% vs. <1% vs. <1% vs. 4%, p=0.68). Adjuvant chemotherapy with/without orbital radiotherapy was given in 761 (53%) patients. Based on Kaplan-Meier estimates in different continents (AS vs. AUS vs. EUR vs. NA vs. SA), the 6-year risk of orbital tumor recurrence was 5% vs. 2% vs. 0% vs. 0% vs. 12% (p<0.001), systemic metastasis was reported in 8% vs. 5% vs. 2% vs. 0% vs. 13% (p=0.001), and death in 10% vs. 3% vs. 2% vs. 0% vs. 11% (p<0.001) patients. // Conclusion: There is a wide variation in the infiltrative histopathology features of RB across continents, resulting in variable outcomes. SA and AS had a higher risk of orbital tumor recurrence, systemic metastasis, and death compared to AUS, EUR, and NA

Dry Sliding-Friction and Wear Behavior of Hot-Extruded Al6061/Si3N4/Cf Hybrid Metal Matrix Composite.

The effects of reinforcement addition and hot extrusion on the microstructures, micro hardness, friction, and wear behavior of aluminium (Al) hybrid composite were investigated. Al6061 dispersed with electroless nickel-coated Si3N4 (6wt.%) and copper-coated carbon fiber (Cf) (1wt.%) hybrid composites was developed through stir casting followed by hot extrusion. Optical micro structural studies confirmed that the size of reinforcements decreased, and their orientations were in the extrusion direction. The decrease in the grain size (29%) of hybrid composites was larger than that in the grain size of matrix alloys under hot-extruded conditions. The synthesized hot-extruded Al6061 hybrid composite exhibited a lower coefficient of friction (51%) and high wear resistance (39%) compared with the hotextruded Al6061base alloy