Gene transfer technology : a tool for studying gene function and role in corneal pathogenesis [abstract]

Transforming growth factor β (TGFb) is associated with many corneal pathologies, diseases and dystrophies. The function of TGFb in adult corneas cannot be studied using conventional transgenic approach because TGFb1 and TGFb2 deficient transgenic animals suffer multiple inflammatory diseases, severe developmental defects, and death by 3-4 weeks of age. This study tested the hypothesis that selective tissue-targeted gene transfer approaches will permit examination of TGFb gene function in the adult cornea without altering TGFb expression in vital organs

Role of neostigmine and polyvalent antivenom in Indian common krait (Bungarus caeruleus) bite

SummaryBungarus caeruleus (Indian common krait) bite during monsoons is common in Northwest India. This study was undertaken to find the effectiveness of neostigmine and polyvalent antivenom in improving neuromuscular paralysis following bite. All the consecutive patients admitted between June 2007 and December 2008 with common krait bite, identified either from brought snake or circumstantial evidence were studied. Ten vials of polyvalent antivenom and three doses of 2.5mg neostigmine at 30min intervals after administration of 0.6mg of atropine were administered I.V. and patients were assessed for any improvement in neuroparalysis. Seventy-two patients were admitted during the study period. All the patients except two came from rural areas and were brought between June and September. Sixty-two patients were bitten during the day while clearing bricks, cutting grass or walking. The mean time interval between bite and arrival to hospital was 4.5h. None of the patients showed any improvement following treatment and all patients developed respiratory paralysis, requiring assisted ventilation. Seventy survived and two died. Neostigmine is ineffective in reversing or improving neuroparalytic features in patients with B. caeruleus bite even at higher dose than normally recommended

PRODUCTION OF INDUSTRIALLY IMPORTANT ENZYMES BY THERMOBACILLI ISOLATED FROM HOT SPRINGS OF INDIA

Enzymes from thermophilic bacteria have received great attention for their potential applications in various industrial sectors. The present study deals with the production of five thermozymes (amylase, lipase, xylanase, protease and cellulase) from 10 thermophilic bacterial species, originally isolated from two hot springs namely Soldhar and Ringigad in Uttarakhand Himalaya, India. The bacterial isolate GBPI_25 produced maximum amylase (1217.86 U/ml) at 45 °C and 5 pH, GBPI 3 produced maximum lipase (22.59 U/ml) at 65 °C and 9 pH, GBPI_25 produced maximum xylanase (98.07 U/ml) at45 °C and 9 pH, GBPI_35 produced maximum protease (16.66 U/ml) at 55 °C and 9 pH, and GBPI 4 produced maximum cellulose (108.68 U/ml) at 45 °C and 5 pH. Crude enzyme preparations showed thermal and pH activities at broad temperature and pH range between 10-100 °C and 3-11 pH, respectively, with different temperature and pH optima. Amylase, xylanase and cellulase showed maximum activity at 50 °C while lipase and protease showed higher activity at 40 and 60 °C, respectively. Enzyme activity at wide temperature range-cellulase and protease from 10-100 °C, amylase and xylanasefrom10-90 °C, and lipase activity from 10-80 °C were the remarkable records from this study. Similarly, pH range for amylase and lipase activity was recorded from 4-11, for xylanase from 3-9, and for protease and cellulase from 3-10. All the thermozymes showed maximum stability at 40 °C and pH 5 except cellulase that showed higher stability at40 °C and neutral pH

Circadian rhythm disruption and Alzheimer’s disease: The dynamics of a vicious cycle

All mammalian cells exhibit circadian rhythm in cellular metabolism and energetics. Autonomous cellular clocks are modulated by various pathways that are essential for robust time keeping. In addition to the canonical transcriptional translational feedback loop, several new pathways of circadian timekeeping - non-transcriptional oscillations, post-translational modifications, epigenetics and cellular signaling in the circadian clock - have been identified. The physiology of circadian rhythm is expansive, and its link to the neurodegeneration is multifactorial. Circadian rhythm disruption is prevelant in contamporary society where light-noise, shift-work, and transmeridian travel are commonplace, and is also reported from the early stages of Alzheimer's disease (AD). Circadian alignment by bright light therapy in conjunction with chronobiotics is beneficial for treating sundowning syndrome and other cognitive symptoms in advanced AD patients. We performed a comprehensive analysis of the clinical and translational reports to review the physiology of the circadian clock, delineate its dysfunction in AD, and unravel the dynamics of the vicious cycle between two pathologies. The review delineates the role of putative targets like clock proteins PER, CLOCK, BMAL1, ROR, and clock-controlled proteins like AVP, SIRT1, FOXO, and PK2 towards future approaches for management of AD. Furthermore, the role of circadian rhythm disruption in aging is delineated

Outcomes of treatment of unresectable esophageal carcinoma treated with chemoradiotherapy and oral metronomic chemotherapy: An experience from a rural cancer center

Introduction: Esophageal carcinoma is the eight most common cancer in the world. The management of locally advancedcarcinoma esophagus is mainly palliative with chemoradiotherapy. The outcome data of such a modality along with oralmetronomic chemotherapy after treatment completion are sparse. Here, we present the outcomes of treatment of locally advancedunresectable esophageal cancer after palliative chemoradiotherapy and oral metronomic therapy from a rural setting in India.Methods: Retrospective analysis of all patients of locally advanced unresectable nonmetastatic esophageal carcinoma treatedwith short course of induction chemotherapy followed by radiotherapy/chemoradiotherapy and oral metronomic chemotherapywas performed. The primary aim was estimation of progression free-survival (PFS) and overall survival (OS). Results: A total of45 patients were analyzed. Mean age was 55 years (30-85 years). A total of 32 patients had tumors in upper and middle esophagus,with the most common histology being squamous cell carcinoma (N-41). The estimated 2 year PFS is 47.2% and the estimated2 years OS is 57.8%. Conclusion: Combined modality therapy with adjuvant oral metronomic therapy shows promising results inthe management and should be the basis of further trials

RF Based Remote Control for Electrical Appliances

This work presented here is to control electrical appliances through RF based remote system. From anywhere without any line of sight, RF based wireless remote control system can change the state of the electrical appliances either in off state or in on state. The controlling circuit is built around RF transmitter and RF Receiver modules which are operating at certain frequency along with a encoder and a decoder with few passive components. The four different channels at the encoder IC are used as input switches and the four channels at the decoder output are connected to the electrical devices through a relay. Here the transmission technique is amplitude shift keying (ASK) and the circuit is powered with 9 Volt. The main objective of this work is to control electrical appliances without line of sight requirement using the RF technology. It has many applications like we can control any independent electrical appliance such as T.V, room light, fan just from a remote. Operating them manually is a tedious job and become hectic sometimes. If one can control devices like fan, TV, lights and music system with a remote from a distance place just by pressing the button, life will become simpler. This will make our life more comfortable and easier

N/P ratio in the PEI2-GNP-DNA complex affects transgene delivery in the human cornea in vitro

Recently, we discovered that polyethylenimine-conjugated gold nanoparticles (PEI2- GNP) could be used as gene therapy vector for the cornea. It was hypothesized that DNA concentration, incubation timing and PEI monomer amount in transfection solution affect gene transfer efficiency and toxicity. The aims of this study were to test whether molar ratio of PEI2 nitrogen (N) and phosphate (P) of DNA in PEI2-GNP transfection solution regulates transgene delivery in human corneal fibroblasts in vitro, and examine PEI2-GNP toxicity, uptake and clearance for the cornea in vivo."National Eye Institute, NIH, Bethesda for RO1EY017294 (RRM), Diversity (RRM) and Veteran Health Affairs Merit (RRM) grants and Unrestricted grant from Research to Prevent Blindness, New York

Probing Magnetoelastic Coupling and Structural Changes in Magnetoelectric Gallium Ferrite

Temperature dependent X-ray diffraction and Raman spectroscopic studies were carried out on the flux grown single crystals of gallium ferrite with Ga:Fe ratio of 0.9:1.1. Site occupancy calculations from the Rietveld refinement of the X-ray data led to the estimated magnetic moment of ~0.60 \muB /f.u. which was in good agreement with the experimental data. Combination of these two measurements indicates that there is no structural phase transition in the material between 18 K to 700 K. A detailed line shape analysis of the Raman mode at ~375 cm^-1 revealed a discontinuity in the peak position data indicating the presence of spin-phonon coupling in gallium ferrite. A correlation of the peak frequency with the magnetization data led to two distinct regions across a temperature ~180 K with appreciable change in the spin-phonon coupling strength from ~ 0.9 cm^-1 (T < 180 K) to 0.12 cm-1 (180 K < T < Tc). This abrupt change in the coupling strength at ~180 K strongly suggests an altered spin dynamics across this temperature.Comment: 16 pages, 4 figure

Toward Silicon-Matched Singlet Fission: Energy-Level Modifications Through Steric Twisting of Organic Semiconductors

Singlet fission (SF) is a potential avenue for augmenting the performance of silicon photovoltaics, but the scarcity of SF materials energy-matched to silicon represents a barrier to the commercial realization of this technology. In this work, a molecular engineering approach is described to increase the energy of the S1 and T1 energy levels of diketopyrrolopyrrole derivatives such that the energy-level requirements for exothermic SF and energy-transfer to silicon are met. Time-resolved photoluminescence studies show that the silicon-matched materials are SF active in the solid state, forming a correlated triplet pair 1(TT) – a crucial intermediate in the SF process – as observed through Herzberg-Teller emission from 1(TT) at both 77 K and room temperature. Transient electron paramagnetic resonance studies show that the correlated triplet pair does not readily separate into the unbound triplets, which is a requirement for energy harvesting by silicon. The fact that the triplet pair do not separate into free triplets is attributed to the intermolecular crystal packing within the thin films. Nevertheless, these results demonstrate a promising route for energy-tuning silicon-matched SF materials

Efficacious and Safe Tissue-Selective Controlled Gene Therapy Approaches for the Cornea

Untargeted and uncontrolled gene delivery is a major cause of gene therapy failure. This study aimed to define efficient and safe tissue-selective targeted gene therapy approaches for delivering genes into keratocytes of the cornea in vivo using a normal or diseased rabbit model. New Zealand White rabbits, adeno-associated virus serotype 5 (AAV5), and a minimally invasive hair-dryer based vector-delivery technique were used. Fifty microliters of AAV5 titer (6.5×1012 vg/ml) expressing green fluorescent protein gene (GFP) was topically applied onto normal or diseased (fibrotic or neovascularized) rabbit corneas for 2-minutes with a custom vector-delivery technique. Corneal fibrosis and neovascularization in rabbit eyes were induced with photorefractive keratectomy using excimer laser and VEGF (630 ng) using micropocket assay, respectively. Slit-lamp biomicroscopy and immunocytochemistry were used to confirm fibrosis and neovascularization in rabbit corneas. The levels, location and duration of delivered-GFP gene expression in the rabbit stroma were measured with immunocytochemistry and/or western blotting. Slot-blot measured delivered-GFP gene copy number. Confocal microscopy performed in whole-mounts of cornea and thick corneal sections determined geometric and spatial localization of delivered-GFP in three-dimensional arrangement. AAV5 toxicity and safety were evaluated with clinical eye exam, stereomicroscopy, slit-lamp biomicroscopy, and H&E staining. A single 2-minute AAV5 topical application via custom delivery-technique efficiently and selectively transduced keratocytes in the anterior stroma of normal and diseased rabbit corneas as evident from immunocytochemistry and confocal microscopy. Transgene expression was first detected at day 3, peaked at day 7, and was maintained up to 16 weeks (longest tested time point). Clinical and slit-lamp eye examination in live rabbits and H&E staining did not reveal any significant changes between AAV5-treated and untreated control corneas. These findings suggest that defined gene therapy approaches are safe for delivering genes into keratocytes in vivo and has potential for treating corneal disorders in human patients