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Background: Lead (Pb), a heavy metal, and quinolinic acid (QA), a metabolite of the kynurenine pathway of tryptophan metabolism, are known neurotoxicants. Both Pb and QA impair spatial learning and memory. Pb activates astrocytes and microglia, which in turn induce the synthesis of QA. We hypothesized increased QA production in response to Pb exposure as a novel mechanism of Pb-neurotoxicity., Methods: Two experimental paradigms were used. In experiment one, Wistar rat pups were exposed to Pb via their dams' drinking water from postnatal day 1 to 21. Control group was given regular water. In the second protocol, QA (9 mM) or normal saline (as Vehicle Control) was infused into right lateral ventricle of 21-day old rats for 7 days using osmotic pumps. Learning and memory were assessed by Morris water maze test on postnatal day 30 or 45 in both Pb- and QA-exposed rats. QA levels in the Pb exposed rats were measured in blood by ELISA and in the brain by immunohistochemistry on postnatal days 45 and 60. Expression of various molecules involved in learning and memory was analyzed by Western blot. Means of control and experimental groups were compared with two-way repeated measure ANOVA (learning) and t test (all other variables)., Results: Pb exposure increased QA level in the blood (by ~ 58%) and increased (p < 0.05) the number of QA-immunoreactive cells in the cortex, and CA1, CA3 and dentate gyrus regions of the hippocampus, compared to control rats. In separate experiments, QA infusion impaired learning and short-term memory similar to Pb. PSD-95, PP1, and PP2A were decreased (p < 0.05) in the QA-infused rats, whereas tau phosphorylation was increased, compared to vehicle infused rats., Conclusion: Putting together the results of the two experimental paradigms, we propose that increased QA production in response to Pb exposure is a novel mechanism of Pb-induced neurotoxicity.

Dibutyryl cyclic adenosine monophosphate (dBcAMP), a cell-permeable synthetic analog of cAMP, has been shown to induce astrogliosis in culture. However, the exact mechanism underlying how dBcAMP exerts its function in situ is not clear. The objective of this study was to examine the effects of dBcAMP on astrogliosis and survival of neurons in stab wound and kainic acid models of brain injury. Stab wound was done in cerebral cortex of BALB/c male mice. Kainic acid lesion was induced in hippocampus by injecting 1μl kainic acid into the lateral ventricle. Animals in both models of injury were divided into L+dBcAMP and L+PBS groups and treated with dBcAMP or PBS for 3, 5, and 7 days respectively. The brain sections were stained for Cresyl violet and Fluro jade-B to assess the degenerating neurons. Immunostaining for GFAP and Iba-1 was done for assessing the astrogliosis and microglial response respectively. Expression of GFAP and BDNF levels in the tissue were estimated by Western blotting and ELISA respectively. The results showed a gradual increase in the number of both astrocytes and microglia in both injuries with a significant increase in dBcAMP-treated groups. The number of degenerating neurons significantly decreased in dBcAMP treated groups. In addition, it was found that dBcAMP stimulated the expression of GFAP and BDNF in both stab wound and kainic acid injuries. Treatment with BDNF receptor inhibitor AZ-23, showed an increase in the degenerating neurons suggesting the role of BDNF in neuroprotection. This study indicates that dBcAMP protects neurons from degeneration by enhancing the production of BDNF and may be considered for use as therapeutic agent for treatment of brain injuries.

Background Aims: We have previously reported the generation of a current Good Manufacture Practice (cGMP)-compliant induced pluripotent stem cell (iPSC) line for clinical applications. Here we show that multiple cellular products currently being considered for therapy can be generated from a single master cell bank of this or any other clinically compliant iPSC line METHODS: Using a stock at passage 20 prepared from the cGMP-compliant working cell bank (WCB), we tested differentiation into therapeutically relevant cell types of the three germ layers using standardized but generic protocols. Cells that we generated include (i) neural stem cells, dopaminergic neurons and astrocytes; (ii) retinal cells (retinal pigment epithelium and photoreceptors); and (iii) hepatocyte, endothelial and mesenchymal cells. To confirm that these generic protocols can also be used for other iPSC lines, we tested the reproducibility of our methodology with a second clinically compliant line RESULTS: Our results confirmed that well-characterized iPSC lines have broad potency, and, despite allelic variability, the same protocols could be used with minimal modifications with multiple qualified lines. In addition, we introduced a constitutively expressed GFP cassette in Chr13 safe harbor site using a standardized previously described method and observed no significant difference in growth and differentiation between the engineered line and the control line indicating that engineered products can be made using a standardized methodology CONCLUSIONS: We believe that our demonstration that multiple products can be made from the same WCB and that the same protocols can be used with multiple lines offers a path to a cost-effective strategy for developing cellular products from iPSC lines., (Copyright © 2018 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)

Objective: This observational study was a retrospective analysis of prospectively collected Hopkins Lupus Cohort data to compare longterm renal survival in patients with lupus nephritis (LN) who achieved complete (CR), partial (PR), or no remission following standard-of-care LN induction therapy., Methods: Eligible patients with biopsy-proven LN (revised American College of Rheumatology or Systemic Lupus Collaborating Clinics criteria) were identified and categorized into ordinal (CR, PR, or no remission) or binary (response or no response) renal remission categories at 24 months post-diagnosis [modified Aspreva Lupus Management Study (mALMS) and modified Belimumab International Lupus Nephritis Study (mBLISS-LN) criteria]. The primary endpoint was longterm renal survival [without endstage renal disease (ESRD) or death]., Results: In total, 176 patients met the inclusion criteria. At Month 24 postbiopsy, more patients met mALMS remission criteria (CR = 59.1%, PR = 30.1%) than mBLISS-LN criteria (CR = 40.9%, PR = 16.5%). During subsequent followup, 18 patients developed ESRD or died. Kaplan-Meier plots suggested patients with no remission at Month 24 were more likely than those with PR or CR to develop the outcome using either mALMS (p = 0.0038) and mBLISS-LN (p = 0.0097) criteria for remission. Based on Cox regression models adjusted for key confounders, those in CR according to the mBLISS-LN (HR 0.254, 95% CI 0.082-0.787; p = 0.0176) and mALMS criteria (HR 0.228, 95% CI 0.063-0.828; p = 0.0246) were significantly less likely to experience ESRD/mortality than those not in remission., Conclusion: Renal remission status at 24 months following LN diagnosis is a significant predictor of longterm renal survival, and a clinically relevant endpoint.

The discovery of reprogramming and generation of human-induced pluripotent stem cells (iPSCs) has revolutionized the field of regenerative medicine and opened new opportunities in cell replacement therapies. While generation of iPSCs represents a significant breakthrough, the clinical relevance of iPSCs for cell-based therapies requires generation of high-quality specialized cells through robust and reproducible directed differentiation protocols. We have recently reported manufacturing of human iPSC master cell banks (MCB) under current good manufacturing practices (cGMPs). Here, we describe the clinical potential of human iPSCs generated using this cGMP-compliant process by differentiating them into the cells from all three embryonic germ layers including ectoderm, endoderm, and mesoderm. Most importantly, we have shown that our iPSC manufacturing process and cell culture system is not biased toward a specific lineage. Following controlled induction into a specific differentiation lineage, specialized cells with morphological and cellular characteristics of neural stem cells, definitive endoderm, and cardiomyocytes were developed. We believe that these cGMP-compliant iPSCs have the potential to make various clinically relevant products suitable for cell therapy applications.

Lead (Pb) is a neurotoxic heavy metal, but the mechanism of its neurotoxicity is not clearly understood. Expression of metallothioneins (MTs) is induced in response to heavy metal exposure as a protective mechanism against heavy metal toxicity. There are several isoforms of MTs (MT-1 to 4), of which MT-3 is the neuron specific isoform, which also has neurite growth inhibitory effects. Whereas, the induction of MT-1 and 2 in response to Pb has been reported, the effect of Pb on the expression of MT-3 in the brain has not been documented. This study aimed at investigating the effect of Pb exposure on the expression of MT-3 in the cerebrum and hippocampus. Wistar rat pups were exposed to Pb via their dams' drinking water (0.2% lead acetate in deionized water) from postnatal day (PND) 0 to 21 and directly via drinking water until PND30. Expression of MT-3 was measured by Western blot and quantitative RT-PCR. MT-3 localization was done by immunohistochemistry. Divalent metal ions were analysed by atomic absorption spectrophotometry. Levels of Pb in blood and cerebrum were significantly increased, while that of copper (Cu), zinc (Zn) and manganese (Mn) were significantly decreased in the Pb-exposed rats at both PND21 and PND30. MT-3 protein was significantly increased in the cerebrum (by 2.5-fold) and in hippocampus (1.4 to 3.2-fold) in both PND21 and PND30 Pb-exposed rats over controls. MT-3 gene expression also increased in the cerebrum (by 42%), and in the hippocampus (by 65% and 43% in the PND21 and PND30 rats, respectively), in the Pb-exposed rats over controls, but the increase was statistically significant (p < 0.05) only in the PND30 rats. Pb exposure significantly increased (p < 0.05) percentage of MT-3 immunoreactive cells in Cornu Ammonis and dentate gyrus regions in the PND21 rats, and in the Cornu Ammonis 1, dentate gyrus and cortex regions in the PND30 rats. Our data thus provide convincing evidence that exposure to low levels of Pb during preweaning period increases the expression of MT-3 in the brain of rats., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Monoamine oxidase-B (MAOB), a flavin adenine dinucleotide (FAD), is an enzyme which catalyzes the oxidation of amines. MAOB is proposed to play a major role in the pathogenesis of neurodegeneration through the production of reactive oxygen species (ROS) and neurotoxins. The present study was designed to outline the effects of the MAOB inhibitor (MAOB-I) on neuroprotection of spinal neurons, regeneration of sciatic nerve fibers, and recovery of sensory-motor functions in the sciatic nerve crush injury model. Male Wistar rats (4-months-old) were assigned to i) Naïve (N), ii) Sham (S), iii) Sciatic nerve crush and treated with saline (CRUSH + SALINE) and iv) Sciatic nerve crush and treated with MAOB inhibitor (CRUSH + MAOB-I) groups (n = 10/group). In groups iii and iv, the crush injury was produced by crushing the sciatic nerve followed by treatment with saline or MAOB-I (Selegiline ® 2.5 mg/kg) intraperitoneally for 10 days. Behavioral tests were conducted from week 1 to week 6. At the end of the study, sciatic nerve and lumbar spinal cord were examined by immunohistochemistry, light and electron microscopy. MAOB-I treatment showed significant improvement in sensory and motor functions compared to saline treatment (p < 0.05-0.001) in injured nerves. The morphological study showed a significantly increased number of nerve fibers in sciatic nerve distal to the site of injury (p < 0.05), with better myelination pattern in CRUSH + MAOB-I treated group compared to CRUSH + SALINE group. Spinal cord ventral horns showed a significant increase in the number of NeuN-immunoreactive neurons in the MAOB-I treated group compared to Saline treated group (p < 0.01). MAOB-I has a significant potential for protecting the degenerating spinal cord neurons and enhancing the regeneration of injured sciatic nerve fibers following crush injury., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Groundwater depletion has been widely studied in northwest India, but water quality concerns are still poorly constrained. In this study, we explore the hydrochemistry of the top 160m of the aquifer system, through detailed field studies in the Bist-Doab region, considering both anthropogenic and geogenic controls. A detailed comparison is made between sites dominated by urban and agricultural landuse. Salinity, nitrate, chloride and lead concentrations are significantly higher in the shallow (0-50m) groundwater system due to surface anthropogenic contaminant loading from agricultural and urban sources. The widespread occurrence of oxic groundwater within the aquifer system means that denitrification potential is limited and also enhances the mobility of selenium and uranium in groundwater. Geogenic trace elements (e.g. As, Se, F), are generally found at concentrations below WHO guideline drinking water values, however elevated U concentrations (50-70μg/L) are found within the deeper part of the aquifer and shallow urban aquifers associated with higher bicarbonate waters. Higher concentration of Se (10-40μg/L) are found exclusively in the shallow groundwater system where Se is mobilised from soils and transported to depth in the shallow aquifer due to the prevailing oxidising aquifer conditions. New evidence from a range of environmental tracers shows elevated concentrations of anthropogenic contaminants in the deeper part of the aquifer (50-160m deep) and demonstrates vulnerability to vertical migration of contaminants. Continued intensive groundwater abstraction from >100m deep means that water quality risks to the deep aquifer system need to be considered together with water quantity constraints., (Copyright © 2017. Published by Elsevier B.V.)

A 56-year-old man presented with massive right haemothorax 10 days following percutaneous nephrolithotomy (PCNL) for complex, large-bulk, right renal stones. Antiplatelet medication started following coronary stenting 7 months ago was stopped 5 days prior and resumed 2 days following surgery. Stones were cleared through two tracts, one supracostal, with placement of ureteral stent but no nephrostomy. He was discharged the next day with an unremarkable chest X-ray. He developed cough and fever after 1 week. Three days later he presented with acute dyspnoea, blood pressure of 100/60 mm Hg, pulse of 120/min and haemoglobin of 9.0 g/dL. Chest X-ray and CT scan showed a large right haemothorax. Two-litre haemothorax was drained by intercostal drainage with prompt recovery. Haemothorax is a rare complication following PCNL usually after supracostal access. Most occur at or immediately following surgery. Infection and early resumption of antiplatelet medication might have contributed to his presentation with delayed secondary haemorrhage from a pleural injury., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Ltd (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

Personal identification methods may not be efficient when bodies are decomposed, burned, in cases of mass disasters when soft tissue cannot provide reliable information or has been lost. Various methods currently employed in forensic odontology for personal identification include comparing with antemortem dental charts, rugoscopy, denture labeling, DNA analysis from dental pulp, bite mark analysis, etc., Recently, there is growing interest in the study of enamel rod end patterns. These enamel rod end patterns are termed as "Tooth prints" and the study of these prints is known as "Ameloglyphics" (amelo: Enamel, Glyphics: Carvings). This review encompasses about the basis of using enamel rod end patterns, methods of obtaining the patterns and further suggests these tooth prints as an analogy to finger print in personal identification in mass disasters., Competing Interests: There are no conflicts of interest.

Background: Oral health is a vital part of general health and is a valuable asset of every individual. The working population in India usually belong to the lower socioeconomic group., Aim: This study was conducted to assess the oral health status and treatment needs of Gunj marketing yard laborers., Materials and Methods: A descriptive study was conducted among 550 laborers of Gunj marketing yard of Raichur city. A specially designed questionnaire was used to assess the demographic variables and oral hygiene practices. Oral health status was assessed using the WHO assessment form 1997. Simplified oral hygiene index (1964) was used to assess the oral hygiene status., Results: The mean age of the study participants was 35.1 (± 8.02) years and the mean decayed teeth, missing teeth, filled teeth, and decayed, missing, filled teeth was 2.06 (± 1.49), 0.76 (± 2.53), 0.13 (± 0.39), and 2.95 (± 3.02), respectively. The prevalence of dental caries and periodontal disease was 85.7% and 93.5%, respectively. The oral hygiene status was poor in 45.9% of the study participants., Conclusion: This study demonstrates poor oral hygiene and high prevalence of periodontal diseases and dental caries as well as a large proportion of unmet dental needs among these laborers., Competing Interests: There are no conflicts of interest.

Introduction: Breast carcinoma is the most common cancer among women in the urban Indian population. Conventionally, immunohistochemistry (IHC) is done to determine the hormone receptor status of the tumor. Immunocytochemistry (ICC) on fine-needle aspiration cytology (FNAC) was carried out to determine the same hormone receptor status of the tumor., Objective: The study was undertaken to evaluate the diagnostic reliability of performing estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (Her2/neu) receptor status on FNAC by ICC and to compare the results with IHC., Materials and Methods: A 2 years 6 months prospective study conducted in the Department of Pathology, ESIC Medical College and PGIMSR and ESIC Model Hospital, Rajajinagar, Bengaluru, wherein 100 breast carcinoma patients' samples both cytology and histology were collected. IHC and ICC were done by peroxidase antiperoxidase technique. Validations of the receptor status were analyzed using sensitivity, specificity, positive and negative predictive values (PPV and NPV), and kappa statistics for agreements between ICC and IHC., Results: ICC was positive for ER, PR, and Her2/neu in 53, 50, and 22 cases, respectively. For ER, a cytohistologic correlation of 98%, with a sensitivity of 96.3%, specificity of 100%, and PPV and NPV being 100% and 95.7%. For PR, concordance of 97%, with a sensitivity of 94.3%, specificity of 100%, and PPV and NPV being 100% and 94%. Her2/neu had an agreement of 89%, with a sensitivity of 72%, specificity of 95.5%, and PPV and NPV being 85.7% and 90.1%., Conclusion: ICC has been a boon and can be a paramount diagnostic adjunct to the routine investigations.

Lithium is a commonly used drug in psychiatric practice. It is used in the treatment of depression and bipolar disorder. It has a narrow therapeutic index with documented adverse effects even near therapeutic levels. It has myriad of manifestations at toxic levels. The cardiovascular effects range from relatively benign ST-T wave changes to fatal arrhythmias. We describe a case of lithium toxicity which presented as a junctional rhythm and later showed a variety of manifestations such as complete heart block, atrial fibrillation, sinus bradycardia, and finally reverted to sinus rhythm at par with serum lithium levels., Competing Interests: There are no conflicts of interest.

The Indo-Gangetic aquifer is one of the world's most important transboundary water resources, and the most heavily exploited aquifer in the world. To better understand the aquifer system, typologies have been characterized for the aquifer, which integrate existing datasets across the Indo-Gangetic catchment basin at a transboundary scale for the first time, and provide an alternative conceptualization of this aquifer system. Traditionally considered and mapped as a single homogenous aquifer of comparable aquifer properties and groundwater resource at a transboundary scale, the typologies illuminate significant spatial differences in recharge, permeability, storage, and groundwater chemistry across the aquifer system at this transboundary scale. These changes are shown to be systematic, concurrent with large-scale changes in sedimentology of the Pleistocene and Holocene alluvial aquifer, climate, and recent irrigation practices. Seven typologies of the aquifer are presented, each having a distinct set of challenges and opportunities for groundwater development and a different resilience to abstraction and climate change. The seven typologies are: (1) the piedmont margin, (2) the Upper Indus and Upper-Mid Ganges, (3) the Lower Ganges and Mid Brahmaputra, (4) the fluvially influenced deltaic area of the Bengal Basin, (5) the Middle Indus and Upper Ganges, (6) the Lower Indus, and (7) the marine-influenced deltaic areas., (© The Author(s) 2017.)

Asymmetric supercapacitors (ASCs) with aqueous electrolyte medium have recently become the focus of increasing research. For high performance ASCs, selection of cathode materials play a crucial role, and core-shell nanostructures are found to be a good choice. We successfully synthesized, ZnO@MnO 2 core-shell nanofibers (NFs) by modification of high-aspect-ratio-electrospun ZnO NFs hydrothermally with MnO 2 nanoflakes. High conductivity of the ZnO NFs and the exceptionally high pseudocapacitive nature of MnO 2 nanoflakes coating delivered a specific capacitance of 907 Fg -1 at 0.6 Ag -1 for the core-shell NFs. A simple and cost-effective ASC construction was demonstrated with ZnO@MnO 2 NFs as a battery-type cathode material and a commercial-quality activated carbon as a capacitor-type anode material. The fabricated device functioned very well in a voltage window of 0-2.0 V, and a red-LED was illuminated using a single-celled fabricated ASC device. It was found to deliver a maximum energy density of 17 Whkg -1 and a power density of 6.5 kWkg -1 with capacitance retention of 94% and Coulombic efficiency of 100%. The novel architecture of the ZnO@MnO 2 core-shell nanofibrous material implies the importance of using simple design of fiber-based electrode material by mere changes of core and shell counterparts.

Exposure to laser irradiation on seeds brings about the changes in agronomical characteristics of the plants. Solanum melongena L. var. Mattu Gulla, a variety of brinjal is of high economic value due to its unique colour and flavour. The aim of the study was to understand the influence of Helium-Neon (He-Ne) laser irradiation on agronomical characters of Solanum melongena L. var. Mattu Gulla in the field conditions. Various growth characteristics including seed germination percentage, survival rate, plant height, number of branches, and flowers and fruits were estimated during different developmental stages of the brinjal. In addition, the chlorogenic acid content of fruits obtained from the laser irradiated seeds were quantified using reversed-phase high-performance liquid chromatography (RP- HPLC). The plants from the seeds irradiated with different doses (20, 25 and 30J/cm 2 ) of He-Ne laser showed significant enhancement on the growth characteristics when compared to the non-irradiated control groups. He-Ne laser irradiation also improved the yield characteristics of the plants significantly in in vivo conditions in comparison with control group. 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed using methanolic extract of matured fruit of Mattu Gulla on HepG2 and fibroblast cell lines. The IC 50 values of fruit extract from laser irradiated groups were found to be similar to non-irradiated control groups. Chlorogenic acid content was found to be higher in 20J/cm 2 and lower in 30J/cm 2 treated fruit tissue. The current study thus elucidates the role of He-Ne laser as a biostimulator on brinjal var. Mattu Gulla not only in the in vitro conditions but also in the in vivo field conditions., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Introduction: Chronic otitis media is the most common cause of hearing impairment in the developing countries. Hearing loss can have serious effects on day to day life. Ear discharge in chronic otitis media may continue for months to years with increasing hearing impairment and also life threatening infective complications, which is more common in active and also in inactive disease form as well. Myringoplasty is an operative procedure to close the perforation in tympanic membrane., Aim: To compare the success rate of graft uptake in dry and wet ears and to compare the postoperative hearing improvement in dry and wet ear following myringoplasty., Materials and Methods: The comparative study was carried out on total 30 patients with chronic otitis media with central perforation. Of these 15 patients belong to dry ear group and 15 patients with wet ear group. These selected patients on simple random basis were subjected to clinical, audiological, radiological and laboratory investigations and one day before operation, patients were admitted to the hospital and written informed consent was taken in all cases. All patients underwent underlay technique myringoplasty. Postoperatively all patients were evaluated for graft uptake and hearing improvement by pure tone audiometry at 3 rd month follow-up., Results: In our study, the successful graft uptake was seen in 80% in dry ear and 73.3% in wet ear, statistically p-value (χ 2 =1.24) is (p>0.05) which is insignificant. Postoperatively hearing gain was (0-5 dB) in 3 patients (20%) with dry ear and 2 patients (13.5%) with wet ear; (6-10 dB), in 4 patients (26.6%) with dry ear and 6 patients (40%) with wet ear; more than 10 dB in 5 patients (33.3%) with dry ear and 3 patients (20%) with wet ear, statistically p-value is χ 2 =1.24 (p >0.05) which is not significant., Conclusion: In this study the success rate of graft uptake and hearing improvement is found almost equal in dry and wet ear by using underlay technique of myringoplasty, also found statistically insignificant.

We have recently described manufacturing of human induced pluripotent stem cells (iPSC) master cell banks (MCB) generated by a clinically compliant process using cord blood as a starting material (Baghbaderani et al. in Stem Cell Reports, 5(4), 647-659, 2015). In this manuscript, we describe the detailed characterization of the two iPSC clones generated using this process, including whole genome sequencing (WGS), microarray, and comparative genomic hybridization (aCGH) single nucleotide polymorphism (SNP) analysis. We compare their profiles with a proposed calibration material and with a reporter subclone and lines made by a similar process from different donors. We believe that iPSCs are likely to be used to make multiple clinical products. We further believe that the lines used as input material will be used at different sites and, given their immortal status, will be used for many years or even decades. Therefore, it will be important to develop assays to monitor the state of the cells and their drift in culture. We suggest that a detailed characterization of the initial status of the cells, a comparison with some calibration material and the development of reporter sublcones will help determine which set of tests will be most useful in monitoring the cells and establishing criteria for discarding a line.

Dyslipidemia is an established risk factor for cardiovascular disease (CVD). Atherosclerosis begins early in life as suggested by "fatty streaks" observed in coronaries of healthy organ donors aged 20-29 years. Premature occurrence of coronary heart disease (CHD) in Indians, increases the risk for young individuals. Management of Dyslipidemia in the young Indian poses several challenges. In this article we provide in-depth review of prevalence, guidelines' perspective and expert comments on management of Dyslipidemia in the young (20-40 years) Indian.

We report generation of induced pluripotent stem cell (iPSC) lines from ten Parkinson's disease (PD) patients carrying SNCA, PARK2, LRRK2, and GBA mutations, and one age-matched control. After validation of pluripotency, long-term genome stability, and integration-free reprogramming, eight of these lines (one of each SNCA, LRRK2 and GBA, four PARK2 lines, and the control) were differentiated into neural stem cells (NSC) and subsequently to dopaminergic cultures. We did not observe significant differences in the timeline of neural induction and NSC derivation between the patient and control line, nor amongst the patient lines, although we report considerable variability in the efficiency of dopaminergic differentiation among patient lines. We performed whole genome expression analyses of the lines at each stage of differentiation (fibroblast, iPSC, NSC, and dopaminergic culture) in an attempt to identify alterations by large-scale evaluation. While gene expression profiling clearly distinguished cells at different stages of differentiation, no mutation-specific clustering or difference was observed, though consistent changes in patient lines were detected in genes associated mitochondrial biology. We further examined gene expression in a stress model (MPTP-induced dopaminergic neuronal death) using two clones from the SNCA triplication line, and detected changes in genes associated with mitophagy. Our data suggested that even a well-characterized line of a monogenic disease may not be sufficient to determine the cause or mechanism of the disease, and highlights the need to use more focused strategies for large-scale data analysis.

Introduction: Meniere's disease is a disorder of the membranous labyrinth of the inner ear manifesting as vertigo, tinnitus, sensory neural hearing loss and aural fullness of known or unknown origin. Some patients may present after years into typical forms. The endocrine disorders, especially hypothyroidism and the factors like inflammation, trauma, otosclerosis, autoimmunity explains the multifactorial causation for the basis of Meniere's disease. Endolymphatic hydrops is the pathologic term named for the defective regulation of endolymph volume with respect to production and absorption. However, hyper production of endolymph cannot be excluded. The cause of Meniers disease is multifactorial so clinical features of repeated attacks of variable duration cannot be explained satisfactorily based on single causation., Aim: To study the incidence of hypothyroidism in Meniere's disease and to assess the success rate of improvement in subjective symptoms after treatment., Materials and Methods: The study was carried out on total 35 out patients who were clinically diagnosed of Meniere's disease and of these 35 patients, 12 patients were found to have hypothyroidism. All patients with hypothyroidism were treated with oral thyroxin supplements and they were evaluated clinically at 3 weeks, 3 months and 6 months interval for the improvement in subjective symptoms on outpatient basis., Results: In our study, the subjective improvement of all symptoms was seen in all cases of Meniere's disease with hypothyroidism after 12 weeks of treatment., Conclusion: The improvement in clinical symptoms after treatment demonstrates an association between Meniere's disease and hypothyroidism which was found to be significant. Hence, clinicians should consider screening patients with Meniere disease for thyroid dysfunction who are not already taking supplements.

Induced pluripotent stem cells (iPSC) and their differentiated derivatives offer a unique source of human primary cells for toxicity screens. Here, we report on the comparative cytotoxicity of 80 compounds (neurotoxicants, developmental neurotoxicants, and environmental compounds) in iPSC as well as isogenic iPSC-derived neural stem cells (NSC), neurons, and astrocytes. All compounds were tested over a 24-h period at 10 and 100 μM, in duplicate, with cytotoxicity measured using the MTT assay. Of the 80 compounds tested, 50 induced significant cytotoxicity in at least one cell type; per cell type, 32, 38, 46, and 41 induced significant cytotoxicity in iPSC, NSC, neurons, and astrocytes, respectively. Four compounds (valinomycin, 3,3',5,5'-tetrabromobisphenol, deltamethrin, and triphenyl phosphate) were cytotoxic in all four cell types. Retesting these compounds at 1, 10, and 100 μM using the same exposure protocol yielded consistent results as compared with the primary screen. Using rotenone, we extended the testing to seven additional iPSC lines of both genders; no substantial difference in the extent of cytotoxicity was detected among the cell lines. Finally, the cytotoxicity assay was simplified by measuring luciferase activity using lineage-specific luciferase reporter iPSC lines which were generated from the parental iPSC line. This article is part of a Special Issue entitled SI: PSC and the brain., Competing Interests: The authors declare they have no actual or potential competing financial interests., (Copyright © 2015. Published by Elsevier B.V.)

The discovery of induced pluripotent stem cells and the ability to manufacture them using clinically compliant protocols has the potential to revolutionize the field of regenerative medicine. However, realizing this potential requires the development of processes that are reliable, reproducible, and cost-effective and that at the same time do not compromise the safety of the individuals receiving this therapy. In the present report, we discuss how cost reductions can be obtained using our experience with obtaining approval of biologic agents, autologous therapy, and the recent approval of cord blood banks. Significance: For therapy to be widely available, the cost of manufacturing stem cells must be reduced. The steps proposed in the present report, when implemented, have the potential to reduce these costs significantly., (©AlphaMed Press.)

Composite tumors are formed when there is intermingling between two components of separate tumors seen histologically. Cases demonstrating composite tubular adenoma with other types of tumors in the colon are rare. Composite tubular adenomas with nonlymphoid tumors including carcinoids, microcarcinoids, and small cell undifferentiated carcinoma have been reported in the literature. The occurrence of composite lymphoma and tubular adenoma within the colorectal tract is extremely rare. Only three cases have been reported and include one case of mantle cell lymphoma and two cases of diffuse large B-cell lymphoma arising in composite tubular adenomas. We present the first case of composite Epstein-Barr virus-associated B-cell lymphoproliferative disorder and tubular adenoma in a rectal polyp with a benign endoscopic appearance., (© The Author(s) 2015.)

In the mammalian liver, 60 % of the cellular components are hepatocytes while the remainder (35 %) includes biliary epithelium, Kupffer cells, endothelial cells, fat storing cells and connective tissue cells. Although neoplasms of hepatocytes are the most common, a significant number of both benign and malignant primary liver neoplasms arising from other cell types can develop, such as tumors of bile duct epithelium (Table 1). In addition, the liver is one of the most susceptible sites for metastatic tumors arising from other organs of the body. Not too long ago, liver tumors were left untreated because the liver was considered a complex and mysterious organ inaccessible to surgery. Advances in imaging procedures and surgical techniques over the past 40 years have revolutionized the approaches to the treatment of benign and malignant liver tumors. Subsegmentectomy, segmentectomy, lobectomy, and transplantation are routinely performed for the treatment of primary and metastatic liver tumors with minimal morbidity and mortality. Since accurate diagnosis remains the key to clinical and surgical management, the emphasis of this chapter is on classification, morphological features and differential diagnosis of malignant neoplasms of the liver.

Genetic modification is an indispensable tool to study gene function in normal development and disease. The recent breakthrough of creating human induced pluripotent stem cells (iPSCs) by defined factors (Takahashi et al., Cell 131:861-872, 2007) provides a renewable source of patient autologous cells that not only retain identical genetic information but also give rise to many cell types of the body including neurons and glia. Meanwhile, the rapid advancement of genome modification tools such as gene targeting by homologous recombination (Capecchi, Nat Rev Genet 6:507-512, 2005) and genome editing tools such as CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas (CRISPR-associated) system, TALENs (Transcription activator-like effector nucleases), and ZFNs (Zinc finger nucleases) (Wang et al., Cell 153:910-918, 2013; Mali et al., Science 339:823-826, 2013; Hwang et al., Nat Biotechnol 31:227-229, 2013; Friedland et al., Nat Methods 10(8):741-743, 2013; DiCarlo et al., Nucleic Acids Res 41:4336-4343, 2013; Cong et al., Science 339:819-823, 2013) has greatly accelerated the development of human genome manipulation at the molecular level. This chapter describes the protocols for making neural lineage reporter lines using homologous recombination and the CRISPR/Cas system-mediated genome editing, including construction of targeting vectors, guide RNAs, transfection into hPSCs, and selection and verification of successfully targeted clones. This method can be applied to various needs of hPSC genetic engineering at high efficiency and high reliability.

Human induced pluripotent stem cell (hiPSC) technologies are powerful tools for modeling development and disease, drug screening, and regenerative medicine. Faithful gene targeting in hiPSCs greatly facilitates these applications. We have developed a fast and precise clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) technology-based method and obtained fluorescent protein and antibiotic resistance dual knockin reporters in hiPSC lines for neurogenin2 (NEUROG2), an important proneural transcription factor. Gene targeting efficiency was greatly improved in CRISPR/Cas9-mediated homology directed recombination (∼ 33% correctly targeted clones) compared to conventional targeting protocol (∼ 3%) at the same locus. No off-target events were detected. In addition, taking the advantage of the versatile applications of the CRISPR/Cas9 system, we designed transactivation components to transiently induce NEUROG2 expression, which helps identify transcription factor binding sites and trans-regulation regions of human NEUROG2. The strategy of using CRISPR/Cas9 genome editing coupled with fluorescence-activated cell sorting of neural progenitor cells in a knockin lineage hiPSC reporter platform might be broadly applicable in other stem cell derivatives and subpopulations.

Background: Atrial fibrillation (AF) is one of the most common arrhythmias accounting for significant mortality and morbidity, especially in elderly. Though oral anticoagulation (OAC) is an effective mode of prevention of stroke in patients of AF, bleeding complication remains a major concern. Because of these issues, a significant proportion of patients either does not receive or receive suboptimal doses of OAC., Methods: In such patients, percutaneous left atrial appendage (LAA) closure remains an interesting option. Experience and literature of this procedure from India have been sparse. We report the first single center experience, from India, of percutaneous LAA closure with Amplatzer Cardiac Plug in 10 patients of non-valvular AF. These patients had contraindications for OAC or had high risk of bleeding or labile international normalized ratio (INR) on therapy., Results: We successfully deployed the devices in all of the cases with no major complications perioperatively and on short-term follow-up. We also report a comprehensive review on the technique of percutaneous LAA closure using Amplatzer Cardiac Plug, including some novel modification with our experience of doing percutaneous mitral balloon valvuloplasty., (Copyright © 2015 Cardiological Society of India. Published by Elsevier B.V. All rights reserved.)

The discovery of induced pluripotent stem cells (iPSCs) and the concurrent development of protocols for their cell-type-specific differentiation have revolutionized our approach to cell therapy. It has now become critical to address the challenges related to the generation of iPSCs under current good manufacturing practice (cGMP) compliant conditions, including tissue sourcing, manufacturing, testing, and storage. Furthermore, regarding the technical challenges, it is very important to keep the costs of manufacturing and testing reasonable and solve logistic hurdles that permit the global distribution of these products. Here we describe our efforts to develop a process for the manufacturing of iPSC master cell banks (MCBs) under cGMPs and announce the availability of such banks., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

Mitochondrial genome can provide information for genomic structure as well as for phylogenetic analysis and evolutionary biology. The complete 15,935 bp mitochondrial genome of Bactrocera zonata (Diptera: Tephritidae), is assembled from Illumina MiSeq read data. The mitogenome information for B. zonata was compared to the homologous sequences of other tephritids. Annotation indicated that the structure and orientation of 13 protein coding genes (PCGs), 22 tRNA and 2 rRNA sequences were typical of, and similar to, the ten closely related tephritid species. The nucleotide composition shows heavily biased toward As and Ts accounting 73.34% and exhibits a slightly positive AT skew, which is similar to other known tephritid species. All PCGs are initiated by ATN codons, except for cox1 with TCG and atp8 with GTG. Nine PCGs use a common stop codon of TAA or TAG, whereas the remaining four use an incomplete termination codon T or TA likely to be completed by adenylation. All tRNAs have the typical clover-leaf structure, with an exception for trnS((AGN)). Four short intergenic spacers showed high degree of conservation among B. zonata and other ten tephritids. A poly(T) stretch at the 5' end followed by [TA(A)]n-like stretch and a tandem repeats of 39 bp has been observed in CR. The analysis of gene evolutionary rate revealed that the cox1 and atp6 exhibits lowest and highest gene substitution rates, respectively than other genes. The phylogenetic relationships based on Maximum Likelihood method using all protein-coding genes and two ribosomal RNA genes confirmed that B. zonata is closely related to Bactrocera correcta, Bactrocera carambolae, Bactrocera papayae, and Bactrocera philippinensis and Bactrocera dorsalis belonging to B. dorsalis species complex forms a monophyletic clade, which is in accordance with the traditional morphological classification and recent molecular works., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Mesenchymal stem/stromal cells (MSCs) have been extensively investigated for their regenerative, immune-modulatory, and wound healing properties. While the laboratory studies have suggested that MSC's have a unique potential for modulating the etiopathology of multiple diseases, the results from clinical trials have not been encouraging or reproducible. One of the explanations for such variability is explained by the "art" of isolating and propagating MSCs. Therefore, establishing more than minimal criteria to define MSC would help understand best protocols to isolate, propagate and deliver MSCs. Developing a calibration standard, a database and a set of functional tests would be a better quality metric for MSCs. In this review, we discuss the importance of selecting a standard, issues associated with coming up with such a standard and how these issues can be mitigated., (Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)

The NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines) for Colorectal Cancer Screening provide recommendations for selecting individuals for colorectal cancer screening, and for evaluation and follow-up of colon polyps. These NCCN Guidelines Insights summarize major discussion points of the 2015 NCCN Colorectal Cancer Screening panel meeting. Major discussion topics this year were the state of evidence for CT colonography and stool DNA testing, bowel preparation procedures for colonoscopy, and guidelines for patients with a positive family history of colorectal cancer., (Copyright © 2015 by the National Comprehensive Cancer Network.)

It has been recently recognized that adenovirus is a pathogen with high morbidity and mortality among immunocompromised patients, particularly after solid organ or stem cell transplant. Confluent necrotizing hepatitis secondary to adenovirus infection alone or together with other organ involvement is extremely rare. There are only 32 cases of confluent necrotizing hepatitis reported in adults since 1960 and most occur after iatrogenic immunosuppression for bone marrow or solid organ transplantation or in other states of immunosuppression, including acquired immunodeficiency syndrome or chemotherapy treatment. We present the first case of concurrent adenovirus-induced necrotizing hepatitis and enterocolitis in an adult patient after double cord stem cell transplant for refractory Crohn's disease. Additionally, we report the imaging and morphologic findings and discuss the potential significance of morphology and immunohistochemistry as a practical approach for identifying adenovirus., (© The Author(s) 2015.)

Malignant melanoma has a variety of morphologic patterns and can metastasize and mimic any type of neoplastic process creating significant diagnostic difficulty. When metastasis to the gastrointestinal system is identified, it is most commonly associated with widely metastatic disease. We report a rare case of isolated gallbladder intramucosal metastatic melanoma with features mimicking lymphoepithelial carcinoma in an adult patient who presented with cholecystitis. Additionally, we report the imaging and morphologic features and discuss the importance of these findings along with a clear clinical history and immunohistochemical profile to make a definitive diagnosis., (© The Author(s) 2015.)

The discovery of induced pluripotent stem cells (iPSCs) and the rapid evolution of clinically compliant protocols to generate such lines from a variety of tissue sources has raised the possibility that personalized medicine may be achievable in the near future. Several strategies to deliver iPSCs for iPSC-derived cell-based therapy have been proposed: one such model has been the cell-banking model, using processes developed by the cord blood industry. The cord blood industry has evolved primarily as a banking model in which units of cord blood harvested from discarded placenta are stored either in a public or a private cord blood bank for future use. The consideration of a cord blood--like banking model has been further spurred by the realization that this population of cells is an ideal starting sample to generate pluripotent cells. Spurred by these technological advances, major efforts are underway to develop a current Good Manufacturing Practice--compliant protocol to generate iPSCs from cord blood and to develop a haplobanking strategy. In this article, we discuss the issues that may affect such an effort., (Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)