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Dr. Bhimrao Ambedkar has been a recognized Great Hero of the past century. He is neither a Shudra by birth nor a Brahmin by birth. He is a Brahmin and a Shudra on the basis of Karma. The philosophy and way of life in the form of liberty, equality and fraternity can be found only in Buddhism. Teachings and characteristics of Buddha and Dhamma: Individual centre, truth-exploration, self-promotion, humanism, the spiritual state of religion, and the highest state of conscience Ambedkar Praneet Navayana was the cornerstone of Humanity. Abstract in Hindi Language: विगत शताब्दी के मान्य महानायक रहे हैं डॉ भीमराव अम्बेडकर। डॉ अम्बेडकर का चिंतन-व्यवहार का आधार बिंदु मानवता ही है जो नस्ल, रंग, जाति, लिंग भेद से परे है जिसमें समता समाज ही उनके नवयान का आधार बनी। यह धारणः प्रत्येक व्यक्ति न जन्म से शूद्र होता है और न ही जन्म से ब्राह्मण। वह कर्म के आधार पर ही ब्राह्मण और शूद्र होता है। डॉ भीमराव का धर्मांतरण अन्धानुकरण नहीं था। उन्होंने बौद्ध धर्म की प्रज्ञा, करुणा, और समता आधारित दृष्टि को देखा और पाया कि स्वतंत्रता, समानता और बन्धुत्त्व रूपी जीवन दर्शन और जीवन पद्धति बौद्ध धर्म में ही पाई जा सकती है। बुद्ध और धम्म की शिक्षाएँ और विशेषताएँः व्यक्ति केंद्र, सत्यान्वेषण, आत्मोन्नति, मानवतावाद, धर्म की साधनावत स्थिति, विवेक की सर्वोच्च स्थिति ही अम्बेडकर प्रणीत नवयान की आधारशिला रही। Keywords: महानायक, मानवतावाद, अछूत, धर्मांतरण, समतामूलक समाज, व्यक्ति केंद्र, सत्यावेषण, आत्मोन्नति, विवेक की सर्वोच्च स्थिति

The biggest criterion of a language is to maintain itself in the use and utility of common people for a long time. Hindi has tested and refined itself in the development of science and technology. That is why today Hindi has proved its usefulness. Words and expressions have also been included in its repository. This is the reason that Hindi has not only become a part of the life of the common man today, but is also advancing itself in the world of growing science and computer era. Abstract in Hindi Language: भाषा की सबसे बड़ी कसौटी अपने आप को लम्बे समय तक आम लोगों के प्रयोग में आने और उपयोगिता बनाए रखने की होती है |जिस भी भाषा ने अपने आप को समय के साथ अपना परिष्कार किया है वह भाषा दीर्घजीवी बनी है| हिंदी ने विज्ञान और प्रौद्योगिकी के विकास में अपने आप को परखा है और साधा है| इसी कारण आज हिंदी ने अपनी प्रयोजनीयता सिद्ध की है|हिंदी ने अपने कलेवर में न केवल स्थानीय बोलियों को अपने आप में समेटा है अपितु देश विदेश की अनेकानेक भाषाओँ की शब्द सम्पदा और अभिव्यक्तियों को भी अपने भंडार में शामिल किया है | यही कारण है कि हिंदी न केवल आज आम व्यक्ति के जीवन का अंग बनी है अपितु बढती विज्ञान और कंप्यूटर की दुनिया में भी अपने आपको नित प्रति आगे बढ़ा रही Keywords: भाषा, विज्ञान और प्रौद्योगिकी, कंप्यूटर युग

Big data has been utilized by numerous businesses to produce straightforward goods and offer improved customer insights via predictive technology like artificial intelligence. Big data is a discipline that primarily focuses on the extraction and systems analysis of enormous data sets to assist organizations in identifying trends. Big Data is now widely used by businesses to boost development across a range of functional areas and increase their capacity for managing sizable client databases. Due to the increased demand for information management specialists brought on by big data, a growing number of software corporations are making investments in companies that focus on data management and analytics. Nevertheless, big data management is threatened by the problem of data protection or privacy. This research paper presents security Challenges and Privacy Issues of Big Data. Further, we created a framework for assessing information security risks in the big data world.

The entire creation of Bhishma Sahni is an attempt to accept, a understand the presence of darkness and then realize the desire to come out of expections. Partisanship is also the act of shedding light on the life immersed in the deep darkness of his characters. The function of creation is the observance of duty. Sacrifice, and sacrifice have been considered as the test of love. Various dimensions of time, society and culture are found in Sahni's novels. His deep vision has gone on the subject of family relations, all classes, inter-caste marriage, love, and illegal children in society. Sahni's literature became the heart of the masses, the heartbeat of the hearts. He broke the stale traditions settled in illogical systems, and established relevant ideals for the new generation. Abstract in Hindi Language: भीष्म साहनी का सम्पूर्ण सृजन अँधेरों की उपस्थिति को स्वीकारना,समझना और फिर उन अंधेरों से निकलने की आकांक्षा को साकार करने की कोशिश है। भीष्म साहनी आजीवन तमस के पार जाने के रास्ते खोजते रहे। उनके सारे लेखन में गरीब, शोषित, दलित वर्ग की पक्षधरता भी उनके पात्रों के गहन अंधेरों में डूबे जीवन पर प्रकाश डालने का कृत्य है। सृजन का उत्स कर्तव्य पालन ही है। त्याग, बलिदान को प्रेम की कसौटी माना गया है। साहनी के उपन्यासों में समय,समाज और संस्कृति के विभिन्न आयाम मिलते हैं। पारिवारिक सम्बन्ध, सभी वर्ग, समाज में अंतरजातीय विवाह, प्रेम, अवैध संतान विषय पर उनकी गहरी दृष्टि गई है। समग्र वैश्विक साहित्य आस्था और विश्वास के उजाले को साथ लेकर अस्तित्व को सार्थक बनाने का यज्ञ है। साहनी का साहित्य जनमानस का ह्रदयग्राही, दिलों की धड़कन बना। उन्होंने विसंगत व्यवस्थाओं में बसी बासी परम्पराओं को तोड़ा, नई पीढ़ी के लिए प्रासंगिक आदर्श स्थापित किया। Keywords: प्रतिबद्धता, व्यवस्था-विसंगति, कर्तव्य पालन, समाज-संस्कृति, आस्था और विश्वास

Data visualisation entails presenting information in a graphical or pictorial format that is easy to understand. It aids in the explanation of facts and the formulation of action plans. It will be useful in any field of study that requires novel approaches to presenting large amounts of complex information. The introduction of computer graphics shaped modern visualisation. This paper provides an overview of data visualisation. Data visualisation, information visualisation, scientific visualisation, and big data are all keywords. Information visualisation and scientific visualisation are other terms for data visualisation. Humans have always used visualisations to make messages or information last longer. What cannot be touched, smelled, or tasted can be visually represented.

Data visualisation entails presenting information in a graphical or pictorial format that is easy to understand. It aids in the explanation of facts and the formulation of action plans. It will be useful in any field of study that requires novel approaches to presenting large amounts of complex information. The introduction of computer graphics shaped modern visualisation. This paper provides an overview of data visualisation. Data visualisation, information visualisation, scientific visualisation, and big data are all keywords. Information visualisation and scientific visualisation are other terms for data visualisation. Humans have always used visualisations to make messages or information last longer. What cannot be touched, smelled, or tasted can be visually represented.

Background and objective Despite significant gains and achieving progress in the last decade, maternal anemia remains a major public health concern in India. Both antenatal (AN) and postnatal (PN) women are populations adversely affected by anemia. Reducing anemia among AN and PN women is one of the national priorities of Anemia Mukt Bharat. The present study aimed at assessing the prevalence of anemia, utilization of AN and PN care (ANC and PNC) services, and drivers of anemia among pregnant and lactating women (PLWs) in Devbhumi Dwarka district, Gujarat. Methods A descriptive cross-sectional study was conducted in four blocks of Devbhumi Dwarka district, Gujarat. A total of 1,185 PLWs were interviewed. Anemia was determined based on the last Hb test record on the Mamta Card (Mother and Child Protection Card). Results The mean age of the study population was 25.19 ±3.91 years: 24.95 ±3.94 years for pregnant women and 25.45 ±4.01 years for lactating women. The prevalence of anemia among pregnant women (Hb:11 g/dL) was 72.92%; 33.91% had moderate and 0.83% had severe anemia. The prevalence of anemia among lactating women (Hb:12 g/dL) was 26%; 63.93% had moderate and 1.29% had severe anemia. The utilization of nutrition and health services was found to be limited. Of the other sociodemographic factors, age (p=0.045), birth spacing (p=0.014), and education (p=0.017) were significant determinants of anemia among pregnant women, whereas parity (p=0.002), birth spacing (p=0.003), religion (p=0.041), and receipt of take-home ration (THR) (p=0.018) were significantly associated with anemia among lactating women. Conclusion The study revealed a high prevalence of anemia among PLWs and sub-optimal utilization of nutritional and healthcare services in Devbhumi Dwarka. Implementing interventions such as comprehensive nutrition education and counseling can contribute toward improving maternal and child health outcomes.

Penile lichen sclerosis is a longstanding inflammatory disease of the skin with a controversial aetiology. Penile lichen sclerosis (PLS) is a growing, inflammatory dermatitis of the anogenital region, which involves the meatus, prepuce, penile shaft, and glans penis. Although the accurate aetiology of PLS is contentious, multiple factors including genetics, autoimmunity, infections of human papillomavirus, hepatitis C, Epstein-Barr virus, risk factors (hormonal and trauma), etc., can be considered to be a part of the etiopathogenesis of PLS. The initial clinical presentations of penile lichen sclerosis are white plaques, atrophied skin, erythema, erosions, and sclerosis in the anogenital region. When the disease advances, the following can occur, including meatal constraints, telangiectasia, petechiae, soreness, papular lesions, tightness of the foreskin, difficulties in passing urine, itching, tenderness on erections, pain, cracking, bleeding, redness, rashes, tightness at frenulum, and dysuria. This disease has a dangerous course of action and if untreated it may be linked with severe urologic and sexual morbidities. PLS is usually treated with medical and surgical interventions like topical or intralesional steroids and circumcision. The role of circumcision is very critical in the course of action and prognosis of PLS, and its treatment is dependent on the stage of the disease. This review brings up the knowledge regarding epidemiology, etiopathology, clinical presentation, and management of PLS with an emphasis on the role of circumcision.

BACKGROUND A podcast is an unconventional method of disseminating information through audio to the mass. Nowadays, the podcast is an emerging new portable technology and is a valuable resource that provides unlimited access for promoting health in participants. Podcast in relevance to healthcare has been used as a source of medical education but there is a dearth of studies for use of podcast as a source of Health Information. This study will provide new perspectives by implementing the SANGYAN podcast that contains information about COVID-19 and other health conditions. OBJECTIVE The study aims to determine the usefulness and effectiveness of the SANGYAN podcast as a digital health intervention to address misinformation related to COVID-19 and other health conditions among individuals in Chennai, Tamil Nadu. METHODS An intervention study will be conducted with 500 participants from PMCHRI and RHTC in Chennai, Tamil Nadu. Individuals, 18 years and older residing in the selected urban and rural settings visiting the OPD of PMCHRI and RHTC will be recruited. The consented participant’s socio-demographic details will be noted and their health literacy will be assessed using REALM-R. Once, the participants will have listened to the podcast the usability and acceptance, of participants' satisfaction will be assessed. RESULTS Descriptive analysis will be gathered to for continuous variables and frequency analysis of the categorical variables. Bivariate analysis was conducted to understand the correlation of socio-demographic features in response to their perception, usefulness, acceptance, and user satisfaction with the study. All analysis will be performed using SPSS v.24., and the results will be reported at 95% CIs and p CONCLUSIONS The finding from this research project will aid in the development and implementation of data-driven, evidence-based, and human-centered behavior change using podcasts to address public health challenges among populations living in diverse settings. This would also help in enhancing the acceptability of podcasts as a source of health-related information. CLINICALTRIAL

BACKGROUND Health inequity is interlinked with good health and well-being of an individual. Digital health interventions have the potential to reduce the existing health inequities. OBJECTIVE The study aims to identify determinants of social, economic, and health inequity in diverse settings to enhance Healthy Eating and Active Living. It further aims to design and develop a digital health intervention HEAL (Healthy Eating Active Living) incorporating framework of human centred design to enhance Healthy Eating and Active Living among rural and urban population groups of Chennai, Tamil Nadu. METHODS A prospective, 3-year cohort study will be conducted. The study aims to recruit 6350 individuals across rural and urban settings of Chennai, Tamil Nadu. A total of eleven sites have been selected for participation in the study. Data will be gathered for sociodemographic, economic inequity, HEAL profile, depression, anxiety and stress, well-being, sources of health information, perceived access to health care, health literacy, navigation of health literacy, satisfaction with health system. RESULTS The study would help to explore the determinants of social, economic and health inequity across multiple sites. Data analysis will be performed using SAS v9.3 and results will be reported as 95% CI and p-values. The findings of the study will inform the design and development of a tailored, interactive human centered digital intervention to enhance the health and well-being of the population groups of Chennai. CONCLUSIONS The proposed study will help in examining the role of the proposed HEAL intervention to enhance the good health and well-being of the population groups of Chennai.

Introduction: It is well established that medical practitioners experience high levels of job stress. This study assessed the perceived stress and stressors among medical practitioners from various specialities in Gandhinagar City of the Gujarat State.Materials and Methods: Using a cross-sectional research design, 72 medical practitioners, from different specialities and practice settings in Gandhinagar City of the Gujarat State, were briefly interviewed using the semi-structured interview protocol and Perceived Stress Scale (PSS), during the period from May to June 2019.Results: The study showed that the majority of medical practitioners had a moderate level of stress (70%), and 11% of them had a severe level of stress. A chi-square test of independence revealed that gender (p=.0029), speciality (p=.0021), practice setting (p=.021), and experience (p=.001) were statistically significant for stress levels. Further, the average PSS score differed significantly according to stressors, namely, work related (p=.004), financial (p=.008), family (p=.049), excessive workload (p=.001), night duty (p=.012) and long working hours (p=.000). However, most medical practitioners did not perceive stress as a problem, and perhaps for that reason necessary actions for stress management were not reported by them.Conclusion: Present study reported high level of stress among medical practitioners, indicating a critical need to address occupational stress among medical practitioners in Gandhinagar City. Immediate actions are warranted for preventing adverse effect of stress on overall health and wellbeing of medical practitioners.. Future study on a large sample that assesses stress, mental health risk, and coping responses would be important step.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has resulted in the current COVID-19 pandemic. Worldwide this disease has infected over 2.5 million individuals with a mortality rate ranging from 5 to 10%. There are several efforts going on in the drug discovery to control the SARS-CoV-2 viral infection. The main protease (MPro) plays a critical role in viral replication and maturation, thus can serve as the primary drug target. To understand the structural evolution of MPro, we have performed phylogenetic and Sequence Similarity Network analysis, that depicted divergence of Coronaviridae MPro in five clusters specific to viral hosts. This clustering was corroborated with the comparison of MPro structures. Furthermore, it has been observed that backbone and binding site conformations are conserved despite variation in some of the residues. These attributes can be exploited to repurpose available viral protease inhibitors against SARS-CoV-2 MPro. In agreement with this, we performed screening of ∼7100 molecules including active ingredients present in the Ayurvedic anti-tussive medicines, anti-viral phytochemicals and synthetic anti-virals against SARS-CoV-2 MPro as the primary target. We identified several natural molecules like δ-viniferin, myricitrin, taiwanhomoflavone A, lactucopicrin 15-oxalate, nympholide A, afzelin, biorobin, hesperidin and phyllaemblicin B that strongly binds to SARS-CoV-2 MPro. Intrestingly, these molecules also showed strong binding with other potential targets of SARS-CoV-2 infection like viral receptor human angiotensin-converting enzyme 2 (hACE-2) and RNA dependent RNA polymerase (RdRp). We anticipate that our approach for identification of multi-target-directed ligand will provide new avenues for drug discovery against SARS-CoV-2 infection. Communicated by Ramaswamy H. Sarma

BACKGROUND Background: Well-being is a multidimensional concept and has been extended to many areas. Student well-being has garnered attention over the last decade due to raised concerns. Digital health interventions have the potential to enhance and improve student well–being. OBJECTIVE Objective: The objective of the study is to design, develop and pilot test a digital health platform to enhance student well-being. METHODS Methods: A sample size of five thousand participants will be recruited across Gujarat and Tamil Nadu, India. Students will be enrolled from, Parul University, Vadodara, Gujarat, and Panimalar Medical College Hospital and Research Institute, (PMCHRI), Panimalar Engineering College, Panimalar Institute of Technology and Panimalar College of Nursing, Chennai, Tamil Nadu. Current undergraduate and graduate students consenting to participate will be recruited using convenience sampling from these institutes. The study will collect baseline data to construct the student well-being index. Based on the risk profile, a random subset of the population will be provided access to the digital health intervention that will deliver tailored interactive messages addressing the various dimensions of well-being among undergraduate and graduate students. The eligible study participants will be of 18 years and older, enrolled in these institutes, and willing to give their consent to participate in the study. RESULTS Results: The proposed research is an unfunded study. The enrolment of the individuals in study will begin from October 2022. Data gathered will be analyzed using SAS v9.3 and results will be reported as 95% CI and p-values. CONCLUSIONS Conclusions: The proposed study will help to determine the factors affecting well-being among college students and help in designing digital health intervention to improve the well-being of the undergraduate and graduate students. CLINICALTRIAL

Anserine and carnosine represent histidine-containing dipeptides that exert a pluripotent protective effect on human physiology. Anserine is known to protect against oxidative stress in diabetes and cardiovascular diseases. Human carnosinases (CN1 and CN2) are dipeptidases involved in the homeostasis of carnosine. In poikilothermic vertebrates, the anserinase enzyme is responsible for hydrolyzing anserine. However, there is no specific anserine hydrolyzing enzyme present in humans. In this study, we have systematically investigated the anserine hydrolyzing activity of human CN1 and CN2. A targeted multiple reaction monitoring (MRM) based approach was employed for studying the enzyme kinetics of CN1 and CN2 using carnosine and anserine as substrates. Surprisingly, both CN1 and CN2 can hydrolyze anserine effectively. The observed catalytic turnover rate (V

Introduction: Trochanteric fractures are among the most common injuries which are usually resulting from minimal to moderate physical trauma to areas of bone signicantly weakened by osteoporosis. The greatest problems for the orthopaedic surgeon to treating the unstable trochanteric fracture and the complications (implant failure, varus collapse, non-union) occur from xation that result of instability. The implants have evolved from extramedullary implants like dynamic hip screw to the intramedullary types of PFNA2 nail. 50 patients were included in our study from July 2019 to July 2021. 25 patient Material And Methods : s were treated with PFNA2 (Group I) and 25 patients with PFN (Group II) Nails. Mean blood loss during surgery was 153.8±10.92 Results : & 201.6±38.48 in group I & group II respectively and the mean duration of surgery in group I and group II was 65.24+6.57 min and 85.44 + 11.08 min respectively. Conclusion : We concluded that use of helical blade PFN is certainly better in 31A2.2 and 31A3.3 type of fracture than screw PFN.

The ability to ablate a gene in a given tissue by generating a conditional knockout (cKO) is crucial for determining its function in the targeted tissue. Such tissue-specific ablation is even more critical when the gene’s conventional knockout (KO) is lethal, which precludes studying the consequences of its deletion in other tissues. Therefore, here we describe a successful strategy that generated a Matrix Gla floxed mouse (Mgp.floxed) by the CRISPR/Cas9 system, that subsequently allowed the generation of cKOs by local viral delivery of the Cre-recombinase enzyme. MGP is a well-established inhibitor of calcification gene, highly expressed in arteries’ smooth muscle cells and chondrocytes. MGP is also one of the most abundant genes in the trabecular meshwork, the eye tissue responsible for maintenance of intraocular pressure (IOP) and development of Glaucoma. Our strategy entailed one-step injection of two gRNAs, Cas9 protein and a long-single-stranded-circular DNA donor vector (lsscDNA, 6.7 kb) containing two loxP sites in cis and 900–700 bp 5′/3′ homology arms. Ocular intracameral injection of Mgp.floxed mice with a Cre-adenovirus, led to an Mgp.TMcKO mouse which developed elevated IOP. Our study discovered a new role for the Mgp gene as a keeper of physiological IOP in the eye.

Iron deficiency anemia (IDA) seems to be a major public health challenge amongst pregnant females in India, despite having multiple national health programs, challenges across the management of IDA continues. The Government of India in recently announced Anemia Mukt Bharat Programme recommends parenteral iron across moderate and severe grades of anemia. In contrast to its clinical efficacy, the cost-effectiveness of Intravenous Iron Sucrose (IVIS) and Ferric Carboxy Maltose (FCM) is not yet established. This paper details the protocol for health technology assessment to assess the effect of intravenous therapy on the improvement of Haemoglobin concentration over oral therapy. The study will be conducted in two districts of the state. The study unit will be selected by a proportionate sampling method from the divided region of district i.e. rural, tribal, desert and coastal based on previous year registered pregnancy. A mixed-method approach will be adopted. Data will be collected over a period of 1 year along with the assessment of key outcome indicators. Additionally, key informant interviews will be conducted and cost data will be gathered to perform cost-effectiveness analysis.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has resulted in the current COVID-19 pandemic. Worldwide this disease has infected around 1.5 million individuals with a mortality rate ranging from 5 to 10%. It has also imposed extreme challenges on global health, economy, and social behavior. Due to the unavailability of therapeutics, several efforts are going on in the drug discovery to control the SARS-CoV-2 viral infection. The main protease (MPro) plays a critical role in viral replication and maturation, thus can serve as the primary drug target. To understand the structural evolution of MPro, we have performed phylogenetic and SSN analysis, that depicted divergence of Coronaviridae MPro in five clusters specific to viral hosts. This clustering was also corroborated with the comparison of MPro structures. Furthermore, it has been observed that backbone and binding site conformations are conserved despite variation in some of the residues. This conservation can be exploited to repurpose available viral protease inhibitors against SARS-CoV-2 MPro. In agreement with this, we performed screening of the custom-made library of ~7100 molecules including active ingredients present in the Ayurvedic anti-tussive medicines, anti-viral phytochemicals and synthetic anti-virals against SARS-CoV-2 MPro as the primary target. We identified several natural molecules that strongly binds to SARS-CoV-2 MPro among which top seven molecules are d-Viniferin, Myricitrin, Taiwanhomoflavone A, Lactucopicrin 15-oxalate, Nympholide A, Biorobin and Phyllaemblicin B. Most of the predicted lead molecules are from Vitis vinifera, also reported for anti-tussive and/or antiviral activities. These molecules also showed strong binding with other main targets RdRp and hACE-2. We anticipate that our approach for identification of multi-target-directed ligand will provide new avenues for drug discovery against SARS-CoV-2 infection.

The ongoing COVID-19 pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from the Coronaviridae family. The disease has infected more than 1 million individuals worldwide with a mortality rate ranging from 5 to 10 %. It has also imposed extreme challenges on global health, economy, and social behavior. Due to the unavailability of therapeutics, several efforts are going on in the drug discovery to control the SARS-CoV-2 viral infection. The main protease (MPro) plays a critical role in viral replication and maturation, thus can serve as the primary drug target. To understand the structural evolution of MPro, we have performed phylogenetic and SSN analysis, that depicted divergence of Coronaviridae MPro in five clusters specific to viral hosts. This clustering was also corroborated with the structural comparison of MPro structures. Furthermore, it has been observed that backbone and binding site conformations are conserved despite variation in some of the residues. This conservation can be exploited to repurpose available viral protease inhibitors against SARS-CoV-2 MPro. In agreement with this, we performed screening of the custom-made library of ~ 7000 molecules including active ingredients present in the Ayurvedic anti-tussive medicines, anti-viral phytochemicals and synthetic anti-virals against SARS-CoV-2 MPro as the primary target. It has been observed that natural molecules like d-Viniferin, Myricitrin, Taiwanhomoflavone A, Lactucopicrin 15-oxalate, Nympholide A, Biorobin, Phyllaemblicin B and other several molecules show strong binding with the SARS-CoV-2 MPro. Most of the predicted lead molecules are from Vitis vinifera, also reported for anti-tussive and/or antiviral activities. These molecules also showed strong binding with other main targets RdRp and hACE-2. We anticipate that our approach for identification of multi-target-directed ligand will provide new avenues for drug discovery against SARS-CoV-2 infection.

PurposeIn the past decade, entrepreneurship research has evolved with the contribution of different scholars, but there is a lack of studies available that focused on entrepreneurship with disabilities. The objective of the research is understanding differently abled entrepreneurs and their entrepreneurial journey. How challenges caused by disability contribute to motivate them to pursue entrepreneurship as a career. This study is based on “Underdog entrepreneurs: Challenge-based entrepreneurship model” theoretical model proposed by Miller and Breton-Miller (2017).Design/methodology/approachThis qualitative research includes case study methodology to study eight differently abled entrepreneurs. All the identified cases are located in the city of Ahmedabad, Gujarat, India. In-depth interviews and multiple visits were scheduled to collect the data. Transcripts of the interview and observation notes were developed for the analysis of the content according to the adopted theoretical model.FindingsDifferently abled entrepreneurs show similar traits as the non-disabled entrepreneurs. They are also found to be more resilient and persistent while dealing with the challenges of failure, stress and uncertainty. Difficult conditions and experiences of discrimination indirectly prepare them for tackling challenges while pursuing entrepreneurship. People close to differently abled entrepreneurs play a critical role in shaping and supporting their ventures.Research limitations/implicationsOwing to the lack of authentic information available on disabled entrepreneurs, the study does not include different entrepreneurs with more disabilities such as hearing impairment, speech impairment and mental illness. The study also focuses on the entrepreneurs of Ahmedabad City, Gujarat because of the similar reason.Originality/valueThis paper is an original submission and contributes towards understanding the differently abled entrepreneurs.

Carnosine, a histidine containing dipeptide, exerts beneficial effects by scavenging reactive carbonyl compounds (RCCs) that are implicated in pathogenesis of diabetes. However, the reduced carnosine levels may aggravate the severity of diabetes. The precise quantification of carnosine levels may serve as an indicator of pathophysiological state of diabetes. Therefore, we have developed a highly sensitive targeted multiple reaction monitoring (MRM) method for quantification of carnosine in human plasma samples. Various mass spectrometry parameters such as ionization of precursor, fragment abundance and stability, collision energy, tube lens offset voltage were optimized to develop a sensitive and robust assay. Using the optimized MRM assay, the lower limit of detection (LOD) and limit of quantification (LOQ) for carnosine were found to be 0.4 nM and 1.0 nM respectively. Standard curves were constructed ranging from 1.0 nM to 15.0 μM and the levels of carnosine in mice and human plasma were determined. Further, the MRM assay was extended to study carnosine hydrolyzing activity of human carnosinases, the serum carnosinase (CN1) and the cytosolic carnosinase (CN2). CN1 showed three folds higher activity than CN2. The MRM assay developed in this study is highly sensitive and can be used for basal plasma carnosine quantification, which can be developed as a novel marker for scavenging of RCCs in diabetes.

A one day male baby was brought with absent anal opening and meconuria. Clinically anal opening was absent.There was meconium in urine and left undescended testis. Postnatal X ray and Invertogram showed high anorectal malformation with air filled cystic area in the lower abdomen. Intraoperatively we found a pouch- like dilatation of colon with its distal end opening into the dome of bladder forming a fistula between pouch and bladder. Divided diversion colostomy proximal to pouch with preservation of pouch was done as first stage of operative procedure. Colonorraphy and pull through has been planned as second stage operation after 4 months.nbsp

India’s economy is considered to be among the most vibrant economies of the world in the twenty-first century. One of the pillars of its growth has been the information technology (IT) industry, which contributes 8.1% of the country’s GDP. The IT industry in India has grown at an average rate of 13.1% between 2008 and 2014, and such a rise augurs well for the development of innovative products and services. This growth has provided both sustainable infrastructure and a competitive environment for the development of entrepreneurship. With the expansion of the entrepreneurial ecosystem, there has been a rise in the number of venture capital (VC) firms and foreign investors. However, a considerable number of innovative IT-based new ventures have either been closed down or acquired by other investors. There are very few ventures which have expanded globally and have sustained innovativeness of their services and products over time. Entrepreneurial failure at the international level has been examined in the literature, and one of the major reasons identified for such phenomenon is the lack of skills among the managers or founders required to manage the innovations and growth. When an organization is expanding its operations and markets globally, it requires a holistic development of all the factors affecting the innovativeness of both the organization and its people. This study explores the organizational challenges faced by technology-based new ventures in the management of innovation and organizational growth during their phase of scaling-up toward becoming an international organization at a global level and their relationship with the development of organizational leadership, structure, culture, and change. The study also discusses the ‘Collaborative Internationalization’ model for the internationalizing technology ventures and discusses various organizational development-related aspects.

The two genes of the cardiac Myosin Heavy Chain (MHC) locus - alpha MHC (aMHC) and beta MHC (bMHC) - are reciprocally regulated in the mouse ventricle during development and in adult conditions such as hypothyroidism and pathological cardiac hypertrophy. Their expressions are under the control of thyroid hormone T3 levels. To gain insights into the epigenetic mechanisms that underlie this inducible and reversible switching of the aMHC and bMHC isoforms, we have investigated the histone modification patterns that occur over the two cardiac MHC promoters during T3-mediated reversible switching of gene expression. Mice fed a diet of propylthiouracil (PTU-an inhibitor of T3 synthesis) for 2 weeks dramatically reduce aMHC mRNA expression and increase bMHC mRNA levels to high levels, while a subsequent withdrawal of PTU diet for 2 weeks completely reverses the T3-mediated changes in MHC expression. Using hearts from mice treated in this way, we carried out chromatin immunoprecipitation-qPCR assays with antibodies against acetylated histone H3 (H3ac) and trimethylated histone (H3K4me3) - two well-documented markers of activation. Our results show that the re-expression of bMHC is associated at the bMHC promoter with increased H3ac but not H3K4me3. In contrast, the silencing of aMHC is associated at its promoter with decreased H3K4me3, but not decreased H3ac. The epigenetic changes at the two MHC promoters are completely reversed when the gene expression returns to initial levels. These data indicate that during reciprocal and inducible gene expression H3ac parallels bMHC isoform expression while H3K4me3 parallels expression of the tightly linked aMHC isoform.

of our daily activities are now routinely recorded and analysed by a variety of governmental and commercial organizations for the purpose of security and business related applications. From telephone calls to credit card purchases, from internet surfing to medical prescription refills, we generate data with almost every action we take. These data sets need to be analyzed for identifying patterns which can be used to predict future behaviour. However, data owners may not be willing to share the real values of their data due to privacy reason. Hence, some amount of privacy preservation needs to be done on data before it is released. Privacy preserving data mining (PPDM) tends to transform original data, so that sensitive data are preserved. In this paper we have proposed a new method CAMDP (Combination of Additive and Multiplicative Data Perturbation) for privacy preserving in data mining.

Chronic low-grade inflammation/meta-inflammation in adipose tissue leads to obesity-associated metabolic complications. Despite growing understanding, the roles of immune cell subsets, their interrelationship, and chronological events leading to progression of obesity-associated insulin resistance (IR) remains unclear.We carried out temporal immunometabolic profiling of adipose tissue from C57BL/6 mice fed a high-fat diet (HFD) for 4, 8, 12, 16, and 20 weeks. We used clodronate sodium liposomes (CLODs) to deplete macrophages and disodium cromoglycate sodium liposomes (DSCGs) to stabilize mast cells.In the temporal HFD settings, mice showed progressive glucose intolerance, insulin resistance, and adipose tissue senescence. Histochemistry analysis of epididymal white adipose tissue (eWAT) using picro-sirius red and Masson's trichrome staining showed extensive collagen deposition in the 16th and 20th weeks. Flow cytometry analysis of the stromal vascular fraction (SVF) from eWAT revealed T-cell subsets as early-phase components and pro-inflammatory macrophages, as well as mast cells as the later phase components during obesity progression. In our therapeutic strategies, macrophage depletion by CLOD and mast stabilization by DSCG attenuated obesity, adipose tissue fibrosis, and improved whole-body glucose homeostasis. In addition, mast cell stabilization also attenuated senescence (p53 and X-gal staining) in eWAT, signifying the role of mast cells over macrophages during obesity.New-generation mast cell stabilizers can be exploited for the treatment of obesity-associated metabolic complications.

By classical competitive antagonism, a substrate and competitive inhibitor must bind mutually exclusively to the active site. The competitive inhibition of O-acetyl serine sulfhydrylase (OASS) by the C-terminus of serine acetyltransferase (SAT) presents a paradox, because the C-terminus of SAT binds to the active site of OASS with an affinity that is 4–6 log-fold (104–106) greater than that of the substrate. Therefore, we employed multiple approaches to understand how the substrate gains access to the OASS active site under physiological conditions. Single-molecule and ensemble approaches showed that the active site-bound high-affinity competitive inhibitor is actively dissociated by the substrate, which is not consistent with classical views of competitive antagonism. We employed fast-flow kinetic approaches to demonstrate that substrate-mediated dissociation of full length SAT–OASS (cysteine regulatory complex) follows a noncanonical “facilitated dissociation” mechanism. To understand the mechanism by w...

Dug1p, a M20 family metallopeptidase and human orthologue of carnosinase, hydrolyzes Cys-Gly dipeptide, the last step of glutathione (GSH) degradation in Saccharomyces cerevisiae. Molecular bases of peptide recognition by Dug1p and other M20 family peptidases remain unclear in the absence of structural information about enzyme-peptide complexes. We report the crystal structure of Dug1p at 2.55 Å resolution in complex with a Gly-Cys dipeptide and two Zn(2+) ions. The dipeptide is trapped in the tunnel-like active site; its C-terminus is held by residues at the S1' binding pocket, whereas the S1 pocket coordinates Zn(2+) ions and the N-terminus of the peptide. Superposition with the carnosinase structure shows that peptide mimics the inhibitor bestatin, but active site features are altered upon peptide binding. The space occupied by the N-terminus of bestatin is left unoccupied in the Dug1p structure, suggesting that tripeptides could bind. Modeling of tripeptides into the Dug1p active site showed tripeptides fit well. Guided by the structure and modeling, we examined the ability of Dug1p to hydrolyze tripeptides, and results show that Dug1p hydrolyzes tripeptides selectively. Point mutations of catalytic residues do not abolish the peptide binding but abolish the hydrolytic activity, suggesting a noncooperative mode in peptide recognition. In summary, results reveal that peptides are recognized primarily through their amino and carboxyl termini, but hydrolysis depends on the properties of peptide substrates, dictated by their respective sequences. Structural similarity between the Dug1p-peptide complex and the bestatin-bound complex of CN2 suggests that the Dug1p-peptide structure can be used as a template for designing natural peptide inhibitors.

One of the challenges of writing a chapter on inter sectionality, with a specific emphasis on sexuality and ethnicity, is that specificity distorts the ideal of inter sectionality, which seeks to see people as multiple composites of identities, not simply two or three of these. However, the pragmatics of understanding some of these intersections in depth does require us to develop a focus. This chapter, therefore, seeks to investigate the experiences, dilemmas, challenges and triumphs of being a sexual and ethnic minority. Using case studies and narratives from clinical encounters, research studies, web-blogs, and mass media, I chronicle these accounts to understand the complex psychological and political journeys people caught between seemingly conflicting identities have to make on a daily basis.

Chemical modifications of substrate peptides are often necessary to monitor the hydrolysis of small bioactive peptides. We developed an electrospray ionization mass spectrometry (ESI-MS) assay for studying substrate distributions in reaction mixtures and determined steady-state kinetic parameters, the Michaelis-Menten constant (K(m)), and catalytic turnover rate (V(max)/[E](t)) for three metallodipeptidases: two carnosinases (CN1 and CN2) from human and Dug1p from yeast. The turnover rate (V(max)/[E](t)) of CN1 and CN2 determined at pH 8.0 (112.3 and 19.5s(-1), respectively) suggested that CN1 is approximately 6-fold more efficient. The turnover rate of Dug1p for Cys-Gly dipeptide at pH 8.0 was found to be slightly lower (73.8s(-1)). In addition, we determined kinetic parameters of CN2 at pH 9.2 and found that the turnover rate was increased by 4-fold with no significant change in the K(m). Kinetic parameters obtained by the ESI-MS method are consistent with results of a reverse-phase high-performance liquid chromatography (RP-HPLC)-based assay. Furthermore, we used tandem MS (MS/MS) analyses to characterize carnosine and measured its levels in CHO cell lines in a time-dependent manner. The ESI-MS method developed here obviates the need for substrate modification and provides a less laborious, accurate, and rapid assay for studying kinetic properties of dipeptidases in vitro as well as in vivo.

There has been an increasing recognition that mitochondrial perturbations play a central role in human heart failure. Mitochondrial networks, whose function is to maintain the regulation of mitochondrial biogenesis, autophagy ('mitophagy') and mitochondrial fusion/fission, are new potential therapeutic targets. Yet our understanding of the molecular underpinning of these processes is just emerging. We recently identified a role of the SWI/SNF ATP-dependent chromatin remodeling complexes in the metabolic homeostasis of the adult cardiomyocyte using cardiomyocyte-specific and inducible deletion of the SWI/SNF ATPases BRG1 and BRM in adult mice (Brg1/Brm double mutant mice). To build upon these observations in early altered metabolism, the present study looks at the subsequent alterations in mitochondrial quality control mechanisms in the impaired adult cardiomyocyte. We identified that Brg1/Brm double-mutant mice exhibited increased mitochondrial biogenesis, increases in 'mitophagy', and alterations in mitochondrial fission and fusion that led to small, fragmented mitochondria. Mechanistically, increases in the autophagy and mitophagy-regulated proteins Beclin1 and Bnip3 were identified, paralleling changes seen in human heart failure. Evidence for perturbed cardiac mitochondrial dynamics included decreased mitochondria size, reduced numbers of mitochondria, and an altered expression of genes regulating fusion (Mfn1, Opa1) and fission (Drp1). We also identified cardiac protein amyloid accumulation (aggregated fibrils) during disease progression along with an increase in pre-amyloid oligomers and an upregulated unfolded protein response including increased GRP78, CHOP, and IRE-1 signaling. Together, these findings described a role for BRG1 and BRM in mitochondrial quality control, by regulating mitochondrial number, mitophagy, and mitochondrial dynamics not previously recognized in the adult cardiomyocyte. As critical to the pathogenesis of heart failure, epigenetic mechanisms like SWI/SNF chromatin remodeling seem more intimately linked to cardiac function and mitochondrial quality control mechanisms than previously realized.

1) The Introduction lays out the background, the results of Poliseno et al. (2010), and consistent data reported in other studies. However, the Introduction should include other studies that lead to questions concerning the ceRNA hypothesis (e.g. PMID: 24793693 & PMID: 24905003 and references therein). The ceRNA hypothesis remains an active area for inquiry. The impression given by the Introduction is that the overwhelming balance of evidence favors the ceRNA hypothesis – this is not correct. We have updated the Introduction to include these references and reflect a more balanced view of the ceRNA hypothesis. 2) It is well established that over-expression of miRNA can repress target mRNA (Figure 1D) and that over-expression of a miRNA sponge can de-repress an miRNA target (Figure 2A-D and Figure 4A). While repetition of these figures panels is useful, none of these experiments tackle the central conclusions of Poliseno et al. (2010) – that expression of an endogenous miRNA competitor (e.g. a pseudogene) can de-repress miRNA function. This is because of the reliance on over-expression strategies in Figure 1D, Figure 2A-D, and Figure 4A. Studies of the endogenous genes are required to test reproducibility of the central conclusion of the original study. The major conclusion of Poliseno et al. (2010) is that decreased expression of PTEN or PTENP1 mRNA should cause mutual changes in miRNA-mediated repression. The only experimental test on endogenous PTEN and PTENP1 mRNA (rather than over-expression of cDNA or miRNA) is represented by Figure 2F-H. It is therefore essential that Figure 2F-H are the focus of the replication study because these are the only experiments that address the central conclusion of Poliseno et al. (2010). Thank you for the suggestion. We have included Figures 2F-H in the revised manuscript. We are also removing Figures 2A, 2C, and 2D from the replication attempt. 3) The normalization used for the mRNA measurements by Poliseno et al. (2010) and the Registered Report is ACTIN. This is a problem because Actin mRNA is dynamically regulated in cells and is a target of cell signaling pathway. Perhaps the ACTIN measurements can be retained in the replication study, but I would strongly recommend that an independent normalization control is also employed – for example 36B4 or another control gene. Thank you for the suggestion. We have included 36B4 as an additional control gene. We will maintain the original analysis (using ACTIN measurements for normalization) and include an additional exploratory analysis from the original, using 36B4 to normalize the gene expression. 4) The authors adopt two statistical procedures, ANOVA and 2 tailed t-test, in the analysis. However, neither of them is used correctly. Take Protocol 1 as an example. A) In ANOVA, for testing the main effect of siRNA, there are only 3 different levels: siLUC, 19b and 20a, but the numerator degree of freedom of the F-test is stated to be 4 by the authors. I also couldn't follow how the authors got the denominator d.f. to be 20 given there are only 18 observations in total. Thank you for catching this error. We have gone through and checked all calculations. Regarding Protocol 1 in particular, the error with d.f. was an error upon transferring the information to the manuscript. The d.f. of the F-test is indeed (2,12) (3 levels with 18 total measurements). We have corrected any other location where this might have occurred in error. B) In 2 tailed t-test, the effective size calculated by the authors assumed that the two populations have the same variance, which is apparently not true for many pairs according to the data. For example, in the comparison of siLuc PTENP1 and 19b PTENP1, the two sample standard deviations are 0.386 and 0.065, respectively. The former is almost 6-fold of the later. Moreover, given this small sample size, t-test is not recommended unless one is very confident that the two population distributions are normal. Thank you for the comment. We have gone through and included non-parametric comparisons (Wilcoxon-Mann-Whitney) along with the student’s t-tests whenever the standard deviations are large. We have also included a statement in the confirmatory analysis plan where the normality and homoscedasticity of the data will be assessed prior to performing the proposed statistical analysis. We have also included a link to the scripts, other files, and summary of calculations (https://osf.io/cd2yq/?view_only=c9c2497ac93c46b6a8ab786769fcff74). 5) Besides the paper that the authors intend to replicate, the oncosuppressive properties of PTEN 3'UTR have been shown also in Tay et al. (Cell, 2015), a paper which should be quoted. Analogously, the oncosuppressive properties of PTENP1 have been shown in other papers besides Yu et al. (2014), namely Chen et al. (2015) and Guo et al. (2015). We have updated the Introduction to reflect a more balanced view of the ceRNA hypothesis. 6) In more general terms, we suggest that the Introduction should include a comprehensive review of all the papers in which the ceRNA-based relationship between PTEN and PTENP1 has been confirmed: Poliseno, L., et al., Deletion of PTENP1 pseudogene in human melanoma. J Invest Dermatol, 2011. 131(12): p. 2497-500. Ioffe, Y.J., et al., Phosphatase and tensin homolog (PTEN) pseudogene expression in endometrial cancer: a conserved regulatory mechanism important in tumorigenesis? Gynecol Oncol, 2012. 124(2): p. 340-6. Johnsson, P., et al., A pseudogene long-noncoding-RNA network regulates PTEN transcription and translation in human cells. Nat Struct Mol Biol, 2013. 20(4): p. 440-6. Yu, G., et al., Pseudogene PTENP1 functions as a competing endogenous RNA to suppress clear cell renal cell carcinoma progression. Mol Cancer Ther, 2014. Chen, C.L., et al., Suppression of hepatocellular carcinoma by baculovirus-mediated expression of long non-coding RNA PTENP1 and MicroRNA regulation. Biomaterials, 2015. 44: p. 71-81. Guo, X., et al., Pseudogene PTENP1 Suppresses Gastric Cancer Progression by Modulating PTEN. Anticancer Agents Med Chem, 2015. We have included a more balanced discussion of the literature of the ceRNA hypothesis. In the Introduction, we try to focus on the experiments being replicated and any direct replications of them, instead of a comprehensive review of all the literature on this subject, which includes many conceptual or related experiments, which would be more appropriate for a review.

The relative utility of 3D, microscopic resolution assessments of fixed mouse myocardial structure via diffusion tensor imaging is demonstrated in this study. Isotropic 100-m resolution fiber orientation mapping within 5.5° accuracy was achieved in 9.1 hr scan time. Preliminary characterization of the diffusion tensor primary eigenvector reveals a smooth and largely linear angular rotation across the left ventricular wall. Moreover, a higher level of structural hierarchy is evident from the organized secondary and tertiary eigenvector fields. These findings are consistent with the known myocardial fiber and laminar structures reported in the literature and suggest an essential role of diffusion tensor microscopy in developing quantitative atlases for studying the structure–function relationships of mouse hearts. Magn Reson Med 52:453– 460, 2004. © 2004 Wiley-Liss, Inc.

Many bandages tend to be harmful when being removed from the human skin. This is a crucial issue, especially faced by burn victims. When bandages are removed from the burn wound, they tend to be harmful by peeling off the newly formed layer of skin over the burn wound. Such nature causes the patient to endure a longer recovery time with additional pain. The objective of this project is to 3D print artificial skin for the victims of burn wounds by using natural gelation. The main aim for creating the artificial skin will be used in place of the current burn wound treatment techniques of dressing the wounds in bandages. The inner layer of this skin was lined with a natural adhesive, a thin layer of agar-agar, which has been reinforced with crushed eggshells to increase its adhesive strength and durability. The synthesized gel contained non adhesive behavior, yet aids in wound healing abilities. Applying hydrocolloids ensures that the wound is kept cool and the gel also ensures efficient heat transfer. This was done so that less sweating occurs on the patient. Based on the experiments that were conducted, the results conclude that the best ratio of artificial skin layer would be 2:1 of agar gel: crushed eggshells. This golden ratio of gel: crushed eggshells for the longest period of time for attachment on the skin without sweating, is achieved. The skin will be printed using Acrylonitrile-Butadiene-Styrene (ABS). The colour of the skin and the shape of the skin was individually designed for each specific patient. The inner gel has the capabilities of reducing the rehabilitation time, without compromising the comfort of the patient. This approach has the potential to be used as a new method to treat burn wounds..

Erythroid Kruppel-like Factor (EKLF) is an erythroid-specific transcription factor that plays a critical role in gamma- to beta-globin gene switching during development. To identify essential domains required for EKLF transactivation function, we cotransfected a human erythroleukemia cell line (K562) with a locus control region gamma/Luc-beta/Cat reporter and an EKLF expression vector. In this assay EKLF mediates a 500-fold induction of beta/CAT expression compared with controls. To map essential transactivation domains, progressive NH(2)-terminal and internal deletion mutants of EKLF were constructed. All EKLF mutants were expressed at wild-type levels, localized to the nucleus, and bound DNA. When mutant EKLF proteins were tested for beta/CAT activation, a novel transactivation domain was identified. This novel domain, encompassing amino acids (aa) 140-358, is sufficient for maximal beta/CAT activation. An 85-amino acid subdomain within this region (aa 140-225) is essential for its activity. Interestingly, this central transactivation subdomain is functionally redundant with the amino-terminal domain (aa 1-139). Thus, EKLF possesses at least two potent transactivation domains that appear to function in a redundant manner.

Student performance in university courses is of great concern to the higher education where several factors may affect the performance. This paper is an attempt to apply the data mining processes, particularly classification, to help in enhancing the quality of the higher educational system by evaluating student data to study the main attributes that may affect the student performance in courses. For this purpose, we have used data obtained from Vikram University, Ujjain of course B.A. first year student. The classification rule generation process is based on the decision tree as a classification method where the generated rules are studied and evaluated. A system that facilitates the use of the generated rules is built which allows students to predict the final grade in a course under study.

Rationale: Repopulation of the injured heart with new, functional cardiomyocytes remains a daunting challenge for cardiac regenerative medicine. An ideal therapeutic approach would involve an effective method at achieving direct conversion of injured areas to functional tissue in situ. Objective: The aim of this study was to develop a strategy that identified and evaluated the potential of specific micro (mi)RNAs capable of inducing reprogramming of cardiac fibroblasts directly to cardiomyocytes in vitro and in vivo. Methods and Results: Using a combinatorial strategy, we identified a combination of miRNAs 1, 133, 208, and 499 capable of inducing direct cellular reprogramming of fibroblasts to cardiomyocyte-like cells in vitro. Detailed studies of the reprogrammed cells demonstrated that a single transient transfection of the miRNAs can direct a switch in cell fate as documented by expression of mature cardiomyocyte markers, sarcomeric organization, and exhibition of spontaneous calcium flux characteristic of a cardiomyocyte-like phenotype. Interestingly, we also found that miRNA-mediated reprogramming was enhanced 10-fold on JAK inhibitor I treatment. Importantly, administration of miRNAs into ischemic mouse myocardium resulted in evidence of direct conversion of cardiac fibroblasts to cardiomyocytes in situ. Genetic tracing analysis using Fsp1Cre-traced fibroblasts from both cardiac and noncardiac cell sources strongly suggests that induced cells are most likely of fibroblastic origin. Conclusions: The findings from this study provide proof-of-concept that miRNAs have the capability of directly converting fibroblasts to a cardiomyocyte-like phenotype in vitro. Also of significance is that this is the first report of direct cardiac reprogramming in vivo. Our approach may have broad and important implications for therapeutic tissue regeneration in general.

Expression in the adult heart of a number of cardiac genes, including the two genes comprising the cardiac Myosin heavy chain locus (Myh), is controlled by thyroid hormone (T3) levels, but there is minimal information concerning the epigenetic status of the genes when their expressions change. We fed mice normal chow or a Propyl thio uracil (PTU, an inhibitor of T3 production)-diet for 6 weeks, or the PTU diet for 6 weeks followed by normal chow for a further two weeks. Heart ventricles from these groups were then used for ChIP-seq analyses with an antibody to H3K4me3, a well documented epigenetic marker of gene activation. The resulting data show that, at the Myh7 locus, H3K4me3 modifications are induced primarily at 5’ transcribed region in parallel with increased expression of beta myosin heavy chain (MHC). At the Myh6 locus, decreases in H3K4me3 modifications occurred at the promoter and 5’ transcribed region. Extensive H3K4me3 modifications also occurred at the intergenic region between the two Myh genes which extended into the 3’ transcribed region of Myh7. The PTU-induced changes in H3K4me3 levels are, for the most part, reversible but are not invariably complete. We found full restoration of Myh6 gene expression upon PTU withdrawal, however the H3K4me3 pattern was only partially restored at Myh6, suggesting that full re-expression of Myh6 does not require that the H3K4me3 modifications return fully to the untreated conditions. Together, our data show that the H3K4me3 modification is an epigenetic marker closely associated with changes in Myh gene expression.

See related article, pages 629–638 In response to stress signals, the mammalian heart responds by an increase in its size.1 This is largely accomplished by an increase in the size of myocytes (hypertrophy) rather than by increasing their numbers (hyperplasia). At the molecular level, pathological stresses induce multiple changes, including genetic reprogramming—the reexpression of a battery of fetal genes and the downregulation of multiple adult genes.2 Together, the changes in gene expression result in substantial phenotypic changes, including changes in size, contractility, metabolic state, and electric conductance. Indeed, the reexpression of fetal genes, including β-myosin heavy chain (β-MyHC), atrial natriuretic factor (ANF), and alpha-skeletal actin, has for many years been looked on as an important molecular indicator of pathological hypertrophy. In spite of this use of β-MyHC reexpression as a marker of pathological hypertrophy, until recently there have been surprisingly few reports on β-MyHC reexpression at the level of individual myocytes within the hypertrophic heart. Nevertheless, previous work using in situ hybridization against β-MyHC mRNA has shown that reexpression of β-MyHC does not take place in all areas of the hypertrophic heart; rather, it is distributed in distinct regions.3 More recently, it has been possible to study individual cells from the heart using genetically altered mice in which a fluorescent protein (yellow fluorescent protein [YFP]) is tagged onto the native β-MyHC gene. This procedure confirmed that β-MyHC reexpression takes place in cells located in distinct regions, including regions adjacent to areas of …

Unknown molecular responses to sarcomere protein gene mutations account for pathologic remodeling in hypertrophic cardiomyopathy (HCM), producing myocyte growth and increased cardiac fibrosis. To determine if hypertrophic signals activated myocyte enhancer factor-2 (Mef2), we studied mice carrying the HCM mutation, myosin heavy-chain Arg403Gln, (MHC 403/+ ) and an Mef2-dependent β-galactosidase reporter transgene. In young, prehypertrophic MHC 403/+ mice the reporter was not activated. In hypertrophic hearts, activation of the Mef2-dependent reporter was remarkably heterogeneous and was observed consistently in myocytes that bordered fibrotic foci with necrotic cells, MHC 403/+ myocytes with Mef2-dependent reporter activation reexpressed the fetal myosin isoform (βMHC), a molecular marker of hypertrophy, although MHC 403/+ myocytes with or without βMHC expression were comparably enlarged over WT myocytes. To consider Mef2 roles in severe HCM, we studied homozygous MHC 403/403 mice, which have accelerated remodeling, widespread myocyte necrosis, and neonatal lethality. Levels of phosphorylated class II histone deacetylases that activate Mef2 were substantially increased in MHC 403/403 hearts, but Mef2-dependent reporter activation was patchy. Sequential analyses showed myocytes increased Mef2-dependent reporter activity before death. Our data dissociate myocyte hypertrophy, a consistent response in HCM, from heterogeneous Mef2 activation and reexpression of a fetal gene program. The temporal and spatial relationship of Mef2-dependent gene activation with myocyte necrosis and fibrosis in MHC 403/+ and MHC 403/403 hearts defines Mef2 activation as a molecular signature of stressed HCM myocytes that are poised to die.

Aims Cardiac hypertrophy is associated with a reduction in the contractile response to beta-adrenergic stimulation, and with re-expression of foetal genes such as beta-myosin heavy chain (MHC). However, whether these two markers of pathology develop concordantly in the same individual cells or independently in different cells is not known. Methods and results To answer this question, we examined the beta-adrenergic response of individual beta-MHC expressing and non-expressing myocytes from hypertrophic hearts, using a previously generated mouse model (YFP/beta-MHC) in which a yellow fluorescent protein (YFP) is fused to the native beta-MHC protein allowing easy identification of beta-MHC expressing cells. Yellow fluorescent protein/beta-MHC mice were submitted to 4 weeks of transverse aortic constriction (TAC), and the contractile parameters of isolated individual myocytes in response to the beta-adrenergic agonist isoproterenol were assessed. Our results demonstrate that the decrease in isoproterenol-induced cell shortening that develops in TAC hearts occurs only in those hypertrophic myocytes that re-express beta-MHC. Hypertrophic myocytes that do not express beta-MHC have contractility indices indistinguishable from non-TAC controls. Conclusion These data show that the reduction of beta-adrenergic response occurs only in subsets, rather than in all myocytes, and is coincident with re-expression of beta-MHC.

To determine whether the expression of cardiac genes changes in a graded manner or by on/off switching when cardiac myocytes change genetic programs in living animals, we have studied two indicator genes that change their expression oppositely in mouse binucleate ventricular cardiomyocytes during development and in response to cardiac hypertrophy. One is a single-copy transgene controlled by an α-myosin heavy chain (aMHC) promoter and coding for CFP. The other is the endogenous β-myosin heavy chain (bMHC) gene modified to code for a YFP–bMHC fusion protein. Using high-resolution confocal microscopy, we determined the expression of the two indicator genes in individual cardiomyocytes perinatally and after inducing cardiac hypertrophy by transverse aortic constriction. Our results provide strong evidence that the cardiac genes respond by switching their expression in an on/off rather than graded manner, and that responding genes within a single cell and within the two nuclei of cardiomyocytes do not necessarily switch concordantly.

MicroRNAs (miRNAs) are a class of small noncoding RNAs that have gained status as important regulators of gene expression. Here, we investigated the function and molecular mechanisms of the miR-208 family of miRNAs in adult mouse heart physiology. We found that miR-208a, which is encoded within an intron of alpha-cardiac muscle myosin heavy chain gene (Myh6), was actually a member of a miRNA family that also included miR-208b, which was determined to be encoded within an intron of beta-cardiac muscle myosin heavy chain gene (Myh7). These miRNAs were differentially expressed in the mouse heart, paralleling the expression of their host genes. Transgenic overexpression of miR-208a in the heart was sufficient to induce hypertrophic growth in mice, which resulted in pronounced repression of the miR-208 regulatory targets thyroid hormone-associated protein 1 and myostatin, 2 negative regulators of muscle growth and hypertrophy. Studies of the miR-208a Tg mice indicated that miR-208a expression was sufficient to induce arrhythmias. Furthermore, analysis of mice lacking miR-208a indicated that miR-208a was required for proper cardiac conduction and expression of the cardiac transcription factors homeodomain-only protein and GATA4 and the gap junction protein connexin 40. Together, our studies uncover what we believe are novel miRNA-dependent mechanisms that modulate cardiac hypertrophy and electrical conduction.