Proteomic-miRNA Biomics Profile Reveals 2D Cultures of Human iPSC-Derived Neural Progenitor Cells More Sensitive than 3D Spheroid System Against the Experimental Exposure to Arsenic.

The iPSC-derived 3D models are considered to be a connective link between 2D culture and in vivo studies. However, the sensitivity of such 3D models is yet to be established. We assessed the sensitivity of the hiPSC-derived 3D spheroids against 2D cultures of neural progenitor cells. The sub-toxic dose of Sodium Arsenite (SA) was used to investigate the alterations in miRNA-proteins in both systems. Though SA exposure induced significant alterations in the proteins in both 2D and 3D systems, these proteins were uncommon except for 20 proteins. The number and magnitude of altered proteins were higher in the 2D system compared to 3D. The association of dysregulated miRNAs with the target proteins showed their involvement primarily in mitochondrial bioenergetics, oxidative and ER stress, transcription and translation mechanism, cytostructure, etc., in both culture systems. Further, the impact of dysregulated miRNAs and associated proteins on these functions and ultrastructural changes was compared in both culture systems. The ultrastructural studies revealed a similar pattern of mitochondrial damage, while the cellular bioenergetics studies confirm a significantly higher energy failure in the 2D system than to 3D. Such a higher magnitude of changes could be correlated with a higher amount of internalization of SA in 2D cultures than in 3D spheroids. Our findings demonstrate that a 2D culture system seems better responsive than a 3D spheroid system against SA exposure.

Proteome architecture of human-induced pluripotent stem cell-derived three-dimensional organoids as a tool for early diagnosis of neuronal disorders.

BACKGROUND AND OBJECTIVES: Induced pluripotent stem cells (iPSCs) derived three-dimensional (3D) model for rare neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS) is emerging as a novel alternative to human diseased tissue to explore the disease etiology and potential drug discovery. In the interest of the same, we have generated a TDP-43-mutated human iPSCs (hiPSCs) derived 3D organoid model of ALS disease. The high-resolution mass spectrometry (MS)-based proteomic approach is used to explore the differential mechanism under disease conditions and the suitability of a 3D model to study the disease. MATERIALS AND METHODS: The hiPSCs cell line was procured from a commercial source, grown, and characterized following standard protocols. The mutation in hiPSCs was accomplished using CRISPR/Cas-9 technology and predesigned gRNA. The two groups of organoids were produced by normal and mutated hiPSCs and subjected to the whole proteomic profiling by high-resolution MS in two biological replicates with three technical replicas of each. RESULTS: The proteomic analysis of normal and mutated organoids revealed the proteins associated with pathways of neurodegenerative disorders, proteasomes, autophagy, and hypoxia-inducible factor-1 signaling. Differential proteomic analysis revealed that the mutation in TDP-43 gene caused proteomic deregulation, which impaired protein quality mechanisms. Furthermore, this impairment may contribute to the generation of stress conditions that may ultimately lead to the development of ALS pathology. CONCLUSION: The developed 3D model represents the majority of candidate proteins and associated biological mechanisms altered in ALS disease. The study also offers novel protein targets that may uncloud the precise disease pathological mechanism and be considered for future diagnostic and therapeutic purposes for various neurodegenerative disorders.

Insights of Extracellular Vesicles of Mesenchymal Stem Cells: a Prospective Cell-Free Regenerative Medicine for Neurodegenerative Disorders.

Mesenchymal stem cells (MSCs) are multipotent, adult stem cells which are found in numerous tissues like the umbilical cord, Wharton's jelly, bone marrow, and adipose tissue. They possess the capacity of self-renewal by dividing and differentiating into various cellular lineages. Their characteristic therapeutic potential exploited so far has made them a desirable candidate in regenerative medicine. Neurodegenerative diseases (NDs) like Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), and ischemic stroke have been treated with MSCs and MSC-derived products. Over the past few decades, we have witnessed significant contributions in discovering the etiology of various NDs and their possible therapeutic solutions. One of the MSC-based therapeutics is extracellular vesicles (EVs), which contain multiple biologically active molecules like nucleic acids and proteins. The contents of EVs are ferried between cells for intercellular communication which then leads to regulation of the homeostasis of recipient cells. EVs serve as a considerable means of cell-free therapies like for tissue repair or regeneration as EVs can maintain therapeutically effective cargo of parent cells and are free of various ethical issues in cell-based therapies. Due to paucity of standard protocols in extraction procedures of EVs and their pharmacological properties and mechanisms, the development of new EV dependent therapies is challenging. With this review, an attempt has been made to annotate these mechanisms, which can help advance the novel therapeutic approaches towards the treat and define a more narrowed down approach for each ND to devise effective MSC-based therapies to cure and avert these diseases.

Socio-ecological vulnerability and resilience of mountain communities residing in capital-constrained environments.

The study evaluated perceived reactions and counter-actions of Himalayan communities to climate change. The evaluation was conducted through identification and characterization of 62 socio-environment-specific indicators in three altitude zones (< 1200 m asl (zone A), 1200-1800 m asl (zone B), and > 1800 m asl (zone C)) in Pauri district, Uttarakhand, India, using a bottom-up, indicator-based approach. Indicators with higher significance for the local economy, livelihoods, or conservation were selected and assimilated into dimensions of vulnerability and resilience. Finally, these were integrated into a sustainable livelihood framework in an approach intended to calculate vulnerability and resilience jointly. The results indicated that the vulnerability and resilience of the mountain communities studied varied widely along the altitude gradient, due to variations in socioeconomic profile, livelihood requirements, resource availability, accessibility, and utilization pattern, and climate risk. The overall values for vulnerability (exposure + sensitivity-adaptive capacity) and resilience (exposure + sensitivity-restorative capacity) were, respectively, 0.34 and 0.28 in zone A, 0.54 and 0.37 in zone B, and 0.65 and 0.59 in zone C. There was a significant difference in contribution of indicators to vulnerability and resilience along the altitudinal gradient was recorded. Strategies for dealing with site-specific vulnerability are required and should address bottlenecks in accessibility and availability of food, water, and healthcare; sustainable utilization of forest resources; educational attainment and skill enhancement; and migration. These results extend current knowledge among the research community and policymakers on socio-ecological changes affecting mountain communities. To reduce the policy level gap between bottom-up and top-down approaches, we suggest precautionary and ongoing site-specific traditional practices and modern adaptation practices, leading to effective and efficient handling of local issues in the context of climate change.

To study the awareness of osteoporosis in postmenopausal Indian women in a Northeast part of India: An evaluation of the Osteoporosis Health Belief Scale.

AIM AND OBJECTIVE: Our study's objective is to determine the level of awareness in postmenopausal women in Guwahati, Northeast part of India, using the Osteoporosis Health Belief Scale (OHBS). SETTINGS AND DESIGN: Osteoporosis is the most common silent health problem in postmenopausal Indian women. An awareness level among this subset of women needs to study. The present study is based on a self-explanatory questionnaire. METHODS: We conducted a self-explanatory questionnaire study on 2000 postmenopausal women in Guwahati, North East region of India. The bone mineral density (BMD) was measured using calcaneum Quantitative ultrasound (QUS) to assess BMD. Baseline characteristics were noted and analyzed. RESULTS: The awareness level was noted only in757 (37.85%). Women with education level 12th standard and above have some awareness regarding osteoporosis. CONCLUSIONS: Based on the present study, we can conclude that there is a lack of awareness in postmenopausal women regarding osteoporosis in India's North-East region. This subset of women is unaware of the condition that can leads to fragility fracture if not address in time. The study emphasizes that health care professionals should conduct frequent awareness programs in the community to prevent this silent disease, and morbidities so arise from osteoporosis can be minimized.

Assessment of climate change pattern in the Pauri Garhwal of the Western Himalayan Region: based on climate parameters and perceptions of forest-dependent communities.

This study assessed the climate change in Pauri district, Uttarakhand, India, a region highly vulnerable to climate change with potentially high loss of livelihoods and lives. The scale of change in the district's climate was analyzed using meteorological station data (1901-2000) and grid data (1985-2015). Perceptions of climate change among forest-dependent communities in three altitude zones (< 1200 m asl (zone A); 1200-1800 m asl (zone B), and > 1800 m asl (zone C)) in the study region were surveyed with respect to 14 climate-specific indicators. Annual mean, maximum, and minimum temperature of seasonal data indicated increasing trends except monsoon. Percentage cloud cover showed an increase, of approximately 3%, while diurnal temperature displayed decreasing trends. Rainfall in the district showed a decreasing trend, with more than 50% of years 1985-2015 receiving less rainfall than the annual average. More than 90% of respondents in zones A and B, and around 65-70% respondents in zone C, reported changes in climate parameters. These findings confirm the long-term observable changes in climate in the region and demonstrate the utility of station data, grid data, and surveys of local communities' perceptions when analyzing climate change. The analysis provided important clues about the nature of climate changes in the district. The results can be used to reduce the gap between bottom-up understanding and top-down policies and to formulate precautionary and ongoing site-specific adaptation practices for communities in different altitude zones in the study region, leading to effective and efficient mitigation of climate change impacts.

Morphological and molecular characterization of Neogastrostyla aqua nov. gen., nov. spec. (Ciliophora, Hypotrichia) from River Yamuna, Delhi; comparison with Gastrostyla-like genera.

A novel hypotrichous ciliate, Neogastrostyla aqua nov. gen., nov. spec. isolated from River Yamuna, Delhi, India, is described using morphological, morphometric and morphogenetic characters from live and stained cells supplemented with derivation of phylogenetic relationships using SSU rRNA gene. Neogastrostyla aqua nov. gen., nov. spec., is characterized by light yellow cortical granules present singly or in clusters of two to five, distributed randomly; parental adoral zone of membranelles retained unchanged for proter; Oxytricha-pattern of paroral and endoral; 25-27 frontal-ventral-transverse cirri including a distinct bipartite frontoventral row of composite origin; three dorsal kineties with no fragmentation of the third kinety, two dorsomarginal kineties, three caudal cirri, two macronuclear nodules and two or three micronuclei. Though Neogastrostyla aqua nov. gen., nov. spec. has a frontoventral row like Gastrostyla species, it differs distinctly in the combination of characters from Gastrostyla and other genera with Gastrostyla-like ventral ciliature (Protogastrostyla, Hemigastrostyla, Apogastrostyla and Pseudogastrostyla) particularly in the colour and distribution of cortical granules as well as arrangement and formation of ciliature. According to the phylogenetic analyses, Neogastrostyla aqua nov. gen., nov. spec. clustered consistently with Gastrostyla sp. Y2 (no description available), Oxytricha granulifera and Architricha indica as close neighbours.

MR Geyser Sign in chronic rotator cuff tears.

Acromio-clavicular (AC) joint cysts are rare presentation of chronic shoulder pathology. These cysts may be observed secondary to either degenerative changes in the AC joint with an intact rotator cuff (type 1 cyst) or following a chronic rotator cuff tear (type 2 cyst). The latter phenomenon is known as Geyser Sign and is described by ultrasound, conventional arthrogram and magnetic resonance imaging (MRI). We present a case of chronic rotator cuff tear presenting with a large type 2 cyst and Geyser Sign on MRI.

Phylogeography of freshwater fish Puntius sophore in India.

Phylogeography and evolutionary history of the freshwater species are poorly known. We document the phylogeography of widely distributed Puntius sophore using cytochrome oxidase subunit I (COI) gene of 650 bp. In the present study, we used 61 individual sequences from known geographic locations across India whereas data are lacking from other parts of its distribution range. Total 20 haplotypes with the intra-species sequence divergence ranging from 0.004 to 0.025 were observed and they were split into two major clades (North and Northeastern to Central India). Two distant geographic (North and Northeastern to Central India) regions shared haplotype suggesting ancient river connectivity or introduction of species from Northeast and Central India. Overall nucleotide and haplotype diversities were 0.00971 and 0.915. The Tajima's D and Fu's Fs values were found negative but non-significant thus rejecting the population expansion model followed by the multimodal mode of mismatch distribution. Bayesian skyline plots from both the clade showed steady population history over time; and start of decline in recent years in the clade B (∼1000-1500 years). The present finding is in support to the 'Satpura hypothesis' proposed to explain species movement patterns from Southeast Asian countries to Indian subcontinent, seconded by P. sophore showing high genetic diversity within Northern India clade (high genetic splits) because of presence of high river network in comparison to other parts of the country.

Askin's tumour in pregnancy with dismal prognosis.

Askin's tumour is a rare malignant neoplasm of the chest wall. A 23-year-old primigravida patient was admitted to the hospital with complaints of chest wall swelling, cough and dyspnoea. Radiological examination showed a chest wall mass in right hemithorax with erosion of ribs and moderate pleural effusion. Fine-needle aspiration cytology was suggestive of malignant small round cell tumour. Immunohistochemical analysis together with the clinical and radiological findings led to the definitive diagnosis of Askin's tumour. Despite receiving multimodality therapy, she had residual soft tissue mass and died after 6 months. Rarity of this malignancy during pregnancy renders the case unique.

Spectrum of Clinical Presentation of Pulmonary Alveolar Microlithiasis: Case Series from North India.

We describe five cases of pulmonary alveolar microlithiasis (PAM) from North India with characteristic radiological and histopathological features. All patients were symptomatic with variable severity and duration of the symptoms and one patient developed cor-pulmonale during the course of follow-up. Diagnosis of PAM was suspected on the basis of classical sand-storm appearance on radiological examination and confirmed by transbronchial lung biopsy in four of the five cases. Apart from other features, presence of pleural and pericardial calcification in one case makes this discussion valuable. Awareness of this specific entity among the clinicians is essential to avoid unnecessary investigations.

Seasonal variation of benthic macro invertebrates from Tons River of Garhwal Himalaya Uttarakhand.

Present investigation was carried out to assess the seasonal variation of benthic macro-invertebrates from the Tons river, a tributary of Yamuna River in Garhwal Himalaya, Uttrakhand during December, 2007 to November, 2009. The seasonal benthic diversity was correlated with various physic-chemical parameters which documented that the macrobenthic diversity is mostly regulated by the dissolved oxygen in the water while temperature and free CO2 were found to be inversely correlated with the benthic fauna. Maximum diversity of benthos was reported at the upstream site ('H' 0.204) during the winter season while it was recorded minimum during the rainy season at all the sites. Maximum diversity is reported during the winter season at all the sites. The benthic fauna is represented by three phylum, 4 classes and 10 orders with Insecta emerging as the most dominant class. Maximum genera were reported from midstream site as it acts as ecotone between upstream and downstream.

Atrial myxoma: a rare cause of ischemic stroke.

Arial myxoma can present as stroke and should be considered as a differential diagnosis of stroke in young individuals. We present here a 42 years female who presented with sudden loss of conciousness. After extensive work up for young stroke, left atrial myxoma was detected and tumor was removed surgically and histopathological report was consistent with the atrial myxoma.

Vitamin B12 deficiency presenting as pyrexia.

Vitamin B12 deficiency is an uncommon cause of pyrexia. We report the case of a patient who presented with pyrexia and anaemia, which after exclusion of infective, inflammatory or endocrine causes was attributed to megaloblastic anaemia secondary to vitamin B12 deficiency. The patient's fever responded to treatment of vitamin B12 deficiency.

Concurrent chemoradiation in locally advanced carcinoma cervix patients.

PURPOSE: To investigate the feasibility of concurrent chemo radiation in locally advanced carcinoma cervix patients in our clinical setting. MATERIALS AND METHODS: From Sept. 1st 2005 to Aug. 31st 2006, 102 patients of carcinoma cervix belonging to stage IIA to IV A were enrolled in the study. External beam radiation therapy was administered using Cobalt 60 teletherapy machine. Cisplatinum (40 mg/m 2) and 5 Fluorouracil (500 mg/m 2 ) continuous infusions with radiotherapy on D2-D5 in first and last 5 # of radiation therapy were administered. RESULTS: Response to treatment and toxicities were monitored and analyzed in 102 patients (50 study group and 52 control group). All 102 patients completed treatment. Out of 50 patients in the study group, 30, 10 and 4 patients had complete, partial and progressive disease, respectively. While out of 52 patients in the control group, 26 had complete and 12 showed partial response. No difference in overall renal, hematological and cutaneous toxicity was seen between two groups. CONCLUSION: This study did not show any benefit of concurrent chemo radiation as compared to radiotherapy alone in locally advanced cervical cancer patients. This could be due to more bulk of tumor stage per stage, poor nutritional status, less number of patients in both arms, not enough to pick up statistically significant small difference in outcome.

Transfusion transmitted malaria in a non-endemic area.

Transfusion transmitted malaria in non-endemic areas is a rare and alarming diagnosis. It deserves a special mention because of its rarity, delay in diagnosis, treatment and serious complications. Shimla, though nonendemic, but being a tourist place, can get malaria transported from other parts of India. We present here a case of transfusion transmitted falciparum malaria in IGMC Shimla. We have discussed the strategies for diagnosis and prevention of transfusion transmitted malaria in endemic and non-endemic regions.

Unilateral blepharospasm and blepharoptosis with bilateral vertical gaze palsy in thalamic hemorrhage.

We present here case of a 49-year-old female with unilateral left sided blepharospasm and blepharoptosis, bilateral vertical gaze palsy, right hemiparesis and right seventh nerve supranuclear palsy as a manifestation of left thalamic hemorrhage. To our knowledge this is the first reported case of a combination of unilateral blepharospasm and blepharoptosis with bilateral vertical gaze palsy.