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Eyes have a unique position among all the sense organs. Primary importance has been given for the protection of eyes by ancient Indian scholar because without eyes whole world is full of darkness. With the advancement of science development of many electronic gadgets like computer mobile etc, which has been produce new diseases. Among those diseases computer vision Syndrome is one of them. In modern system there is no permanent cure of CVS. In Ayurvedic text there is no description of Computer Vision Syndrome but it can be a Anukta vyadhi as describe by a Charak. Some of ocular symptoms of computer vision syndrome can be correlated with Sushkaksipaka as describe by Acharya sushrut and A.Hridya. Modern medicine do not have a definite treatment for this problem except lubricating eye drops whereas Ayurvedic kriyakalpas that is Ayurvedic local ocular therapies could suggest definite treatment to cure the computer vision syndrome along with the change in working style and correction of refractive error. Drugs which are describe in the treatment of Suskhakshipaka have potent property to reduce the clinical features of CVS. So in this clinical trial, I have used different dosage form of Saindhavadiyoga in the management of CVS. Study was conducted among the BHU computer professionals and all the patient were divide in the three group. In the group A eye drop and Anjana of Saindhavadi yoga was given, in the Group B only Carboxy Methyl Cellulose eye drop was given while in the group C only Tarpan therapy was given with Saindhavadighrita All the group have positive result results in reduction of signs and symptoms of CVS without any side effect which are produce by the preservative of Artificial tear drops. But in the group C have better results in comparison to other two group, it was occur due to oleation, nutritive antioxidant property of Ghrita and Yogvahi property of Ghrita which increase the bioavailbilty of drug along with other content of the drug.

Standardization is the present time need for acceptance of Ayurvedic polyherbal formulation. Standard polyherbal formulations have potency to check the pathogenesis of disease process without producing another disease. The upgrade the standard operating procedure can give new identity to the present era. An objective of the study is standardizing Saindhavadi Ghrita on various parameters, in order to assure its safety and efficacy on various basis. The Ghrita was analyzed for its organoleptic, physico-chemical features and screened for its phyto-constituents. In the present work, the obtained results were found within prescribed limits. The standard analytical methodologies provide safety and efficacy to the product, to avoid complication because eye is very sensitive and precious gift of god. So we cannot use Ghrita for Tarpan therapy without standardization.

Differentiation of T and B cells to effector and memory cell fates are associated with extensive metabolic changes which are accompanied by altered mitochondrial dynamics. However, whether alterations in mitochondrial structure and function plays an active role in regulating effector versus memory cell fate decisions during immune responses remains unclear. Our studies here characterize changes in mitochondrial dynamics in activated B cells and show that increased mitochondrial mass and activity is a distinct feature of memory B cell lineage commitment in vivo. Using a directed screen of mitochondrial modulators, we identify mitochondrial fission inhibitor, Mdivi-1 as an agent that could enhance mitochondrial mass and function leading to augmented memory B cell differentiation. The enhanced memory B cell responses mediated by Mdivi-1, translated to more robust recall responses upon secondary antigen exposures. Moreover, Mdivi-1 when used in combination with subunit (SARS-CoV2) and inactivated (H1N1 influenza) vaccines led to remarkably improved vaccine efficacies and protection from lethal viral (H1N1) challenge. Single-cell transcriptomics revealed enhanced commitment to memory lineage differentiation in B cells following Mdivi-1 treatment. We propose that mitochondrial modulators such as Mdivi-1 are a novel class of “immune enhancers” that specifically reinforces immunological memory and could be broadly applied to improve the fidelity of immune responses and vaccine efficacies.

Eyes hold a unique position among all the sense organs. Eyes are the most valuable gift of God to the living beings. Primary importance has been given for the protection of eyes by ancient Indian scholar because without eyes whole world is full of darkness. Acharya Vagbhata says that as long as there is desire for liv-ing, so long all efforts should be made always by men to protect the eyes; because for the blind man, night and day are the same; all the things of this world are useless though he might have plenty of money. In this present time, the human lifestyle has been radically changed. When the human race evolved, hunting was very crucial for survival; hence, they were focused for distant vision works that could allow the man to hunt. As the passage of time, science become more developed more and more vehicle develop for liveli-hood. These days, no longer he requires hunting in the forests. Rather, he has to track the internet on com-puter in a closed compartment. With over 560 million internet users, India is the second largest online mar-ket1 in the world, ranked only behind China. By 2021, there will be over 600 million internet users in India. But in modern system of medicine there is no permanent cure except temporary relieve by artificial tears. Ayurveda is good treatment of the choice.

Somatic hypermutation (SHM) produces point mutations in immunoglobulin (Ig) genes in B cells when uracils created by the activation-induced deaminase are processed in a mutagenic manner by enzymes of the base excision repair (BER) and mismatch repair (MMR) pathways. Such uracil processing creates DNA strand breaks and is susceptible to the generation of deleterious deletions. Here, we demonstrate that the DNA repair factor HMCES strongly suppresses deletions without significantly affecting other parameters of SHM in mouse and human B cells, thereby facilitating the production of antigen-specific antibodies. The deletion-prone repair pathway suppressed by HMCES operates downstream from the uracil glycosylase UNG and is mediated by the combined action of BER factor APE2 and MMR factors MSH2, MSH6, and EXO1. HMCES's ability to shield against deletions during SHM requires its capacity to form covalent cross-links with abasic sites, in sharp contrast to its DNA end-joining role in class switch recombination but analogous to its genome-stabilizing role during DNA replication. Our findings lead to a novel model for the protection of Ig gene integrity during SHM in which abasic site cross-linking by HMCES intercedes at a critical juncture during processing of vulnerable gapped DNA intermediates by BER and MMR enzymes.

Enzymes of the TET family are methylcytosine dioxygenases that undergo frequent mutational or functional inactivation in human cancers. Recurrent loss-of-function mutations in TET proteins are frequent in human diffuse large B cell lymphoma (DLBCL). Here, we investigate the role of TET proteins in B cell homeostasis and development of B cell lymphomas with features of DLBCL. We show that deletion of Tet2 and Tet3 genes in mature B cells in mice perturbs B cell homeostasis and results in spontaneous development of germinal center (GC)-derived B cell lymphomas with increased G-quadruplexes and R-loops. At a genome-wide level, G-quadruplexes and R-loops were associated with increased DNA double-strand breaks (DSBs) at immunoglobulin switch regions. Deletion of the DNA methyltransferase DNMT1 in TET-deficient B cells prevented expansion of GC B cells, diminished the accumulation of G-quadruplexes and R-loops and delayed B lymphoma development, consistent with the opposing functions of DNMT and TET enzymes in DNA methylation and demethylation. Clustered regularly interspaced short palindromic repeats (CRISPR)-mediated depletion of nucleases and helicases that regulate G-quadruplexes and R-loops decreased the viability of TET-deficient B cells. Our studies suggest a molecular mechanism by which TET loss of function might predispose to the development of B cell malignancies.

The three members of TET family of Fe(II) and alpha-ketoglutarate-dependent dioxygenases mediate DNA demethylation by sequentially oxidizing 5-methylcytosine (5mC) to 5-hydroxymethyl- (5hmC), 5-formyl- (5fC) and 5-carboxyl-cytosine (5caC). TET enzymes are required for normal development, and loss of TET function due to mutations, metabolic perturbations and hypoxia, among other mechanisms, occurs frequently in many hematological malignancies and solid tumors. Recent studies have identified mutations in TET proteins (TET2, most commonly) and metabolic enzymes which regulate TET catalytic activity in a large cohort of patients with Diffuse Large B-cell Lymphoma (DLBCL). However, the clinical significance of these mutations in DLBCL and the molecular mechanisms through which TET proteins suppress development of malignancies in general, are not fully-understood. To investigate the role of TET loss-of-function in the pathogenesis of DLBCL, we generated mice with B-cell-specific deletion of TET2 and TET3, the major TET homologs expressed in mature B cells. TET deficiency in B cells perturbed mature B cell homeostasis resulting in spontaneous development of Germinal Center-derived B cell lymphomas. Moreover, B cells with TET deficiency demonstrated increased genomic instability, a feature previously associated with TET loss-of-function in other hematopoietic lineages. Transcriptional profiling of TET-deficient expanded B cells revealed altered expression of genes and proteins involved in modulating the levels of secondary DNA structures, G-quadruplexes and DNA:RNA hybrids (R-loops) which have been linked to genomic instability and transcriptional perturbations in many different cancers. Using previously described methods and newer approaches, we observed a substantial increase in the levels of G-quadruplex and R-loop structures in TET-deficient B cells compared with control B cells. The increase in G-quadruplex and R-loop structures was evident in naïve, activated and GC B cells following acute TET deletion as well as in TET-deficient myeloid cells and T cells. Genome-wide mapping studies and high-throughput genome-wide translocation sequencing (HTGTS) showed a strong correlation of increased G-quadruplex and R-loop structures with increased DNA DSBs in switch regions of immunoglobulin heavy chain locus in TET-deficient B cells. Deletion of the DNA methyltransferase DNMT1 in TET-deficient B cells prevented the expansion of germinal center B cells, diminished the accumulation of G-quadruplexes and R-loops, and caused a notable delay in lymphoma development, consistent with the opposing functions of DNMT and TET enzymes in DNA methylation and demethylation. CRISPR-mediated depletion of nucleases and helicases that regulate G-quadruplexes and R-loops decreased the viability of TET-deficient B cells. Our studies suggest a molecular mechanism by which TET loss-of-function might predispose to development of B cell-derived and other malignancies, and highlight novel therapeutic avenues that could be further explored. Disclosures Rao: Cambridge Epigenetix: Membership on an entity's Board of Directors or advisory committees.

Chronic lymphocytic leukemia (CLL) is a heterogeneous B cell malignancy that still remains incurable. Recent studies have highlighted cellular and non-cellular components of the tissue microenvironment in CLL that help nurture the growth of leukemic cells by providing the necessary stimuli for their proliferation and survival. The diverse stimuli in the specialized tissue microenvironment of CLL lead to constitutive activation of several signaling pathways that includes B cell receptor signaling and the associated mitogen-activated protein kinase (MAPK) signaling. Recent findings have described aberrant activation of MAPK signaling and its interactions with other cellular signaling pathways in the pathogenesis of CLL. These studies have shed light on the deregulated molecular mechanisms contributing to hyperactivation of MAPK signaling and provided avenues for therapeutic options for aggressive CLL. In this review, we describe and discuss the current status of our understanding into the role of MAPK signaling in the pathogenesis of CLL.

Background : This study assessed health care trainees perceptions of their education regarding transgender health care issues and personal and professional comfort level with the provision of transgender health care. An online 20 question survey (2013) about trainee experience and education on transgender health care was administered to medical (MD), nurse practitioner (NP), and physician assistant (PA) students at a single institution. Methods : The survey instrument consisted of seven demographic questions and 13 non-demographic questions about education in transgender health, hours of education received, and personal and professional comfort with transgender medical care. Non-demographic questions were written as even-point Likert scale questions with four rating options (very comfortable, somewhat comfortable, somewhat uncomfortable, and very uncomfortable). Results : A total of 484 MD, NP, and PA students responded to the online survey out of 1,035 eligible students (47% response rate). Only 74 (15%) respondents had provided medical care to a transgender patient. Provision of medical care to a transgender patient was associated with increased hours of education on transgender health issues (P

HMCES (5hmC binding, embryonic stem cell-specific-protein), originally identified as a protein capable of binding 5-hydroxymethylcytosine (5hmC), an epigenetic modification generated by TET proteins, was previously reported to covalently crosslink to DNA at abasic sites via a conserved cysteine. We show here that Hmces-deficient mice display normal hematopoiesis without global alterations in 5hmC. HMCES specifically enables DNA double-strand break repair through the microhomology-mediated alternative-end-joining (Alt-EJ) pathway during class switch recombination (CSR) in B cells, and HMCES deficiency leads to a significant defect in CSR. HMCES mediates Alt-EJ through its SOS-response-associated-peptidase domain (SRAPd), a function that requires DNA binding but is independent of its autopeptidase and DNA-crosslinking activities. We show that HMCES is recruited to switch regions of the immunoglobulin locus and provide a potential structural basis for the interaction of HMCES with long DNA overhangs generated by Alt-EJ during CSR. Our studies provide further evidence for a specialized role for HMCES in DNA repair.

TET enzymes are dioxygenases that promote DNA demethylation by oxidizing the methyl group of 5-methylcytosine to 5-hydroxymethylcytosine (5hmC). Here, we report a close correspondence between 5hmC-marked regions, chromatin accessibility and enhancer activity in B cells, and a strong enrichment for consensus binding motifs for basic region-leucine zipper (bZIP) transcription factors at TET-responsive genomic regions. Functionally, Tet2 and Tet3 regulate class switch recombination (CSR) in murine B cells by enhancing expression of Aicda, which encodes the activation-induced cytidine deaminase (AID) enzyme essential for CSR. TET enzymes deposit 5hmC, facilitate DNA demethylation, and maintain chromatin accessibility at two TET-responsive enhancer elements, TetE1 and TetE2, located within a superenhancer in the Aicda locus. Our data identify the bZIP transcription factor, ATF-like (BATF) as a key transcription factor involved in TET-dependent Aicda expression. 5hmC is not deposited at TetE1 in activated Batf-deficient B cells, indicating that BATF facilitates TET recruitment to this Aicda enhancer. Our study emphasizes the importance of TET enzymes for bolstering AID expression and highlights 5hmC as an epigenetic mark that captures enhancer dynamics during cell activation.

In India Doctorate of Medicine (MD) pharmacology is primarily knowledge oriented based on teaching, seminars, lectures and research related activities including animals and paper-based experiments and day to day management of undergraduate classes. MD pharmacology student should be competent of both clinical and experimental pharmacology. So, the postgraduate pharmacology curriculum should be competent to meet all the job requirements. Therefore, medical council of India (MCI) has introduced new post graduate curriculum which is based on knowledge, practical, clinical skills, thesis skills, and attitudes including communication and training in research. In India demand for skilled clinical research professionals is increasing day by day for growing pharma industries and good academician. So, there is an urgent need for the experienced and skilled pharmacologist to fulfil the requirements. MD pharmacology students should get posting in different clinical departments and observatory posting in industry, clinical research organization (CRO), regulatory body and research organisations. The course of MD Pharmacology should be like that fulfil all the skills that a pharmacologist must have.

// Vipul Shukla 1 , Ashima Shukla 1 , Shantaram S. Joshi 1 and Runqing Lu 1,* 1 Department of Genetics Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE, USA * Has deceased Correspondence to: Vipul Shukla, email: // Keywords : IRF4, Notch signaling, CLL, B1 cells Received : May 03, 2016 Accepted : May 14, 2016 Published : May 25, 2016 Abstract Molecular pathogenesis of Chronic Lymphocytic Leukemia (CLL) is not fully elucidated. Genome wide association studies have linked Interferon Regulatory Factor 4 (IRF4) to the development of CLL. We recently established a causal relationship between low levels of IRF4 and development of CLL. However, the molecular mechanism through which IRF4 suppresses CLL development remains unclear. Deregulation of Notch signaling pathway has been identified as one of the most recurrent molecular anomalies in the pathogenesis of CLL. Yet, the role of Notch signaling as well as its regulation during CLL development remains poorly understood. Previously, we demonstrated that IRF4 deficient mice expressing immunoglobulin heavy chain Vh11 (IRF4 -/- Vh11) developed spontaneous CLL with complete penetrance. In this study, we show that elevated Notch2 expression and the resulting hyperactivation of Notch signaling are common features of IRF4 -/- Vh11 CLL cells. Our studies further reveal that Notch signaling is indispensable for CLL development in the IRF4 -/- Vh11 mice. Moreover, we identify E3 ubiquitin ligase Nedd4, which targets Notch for degradation, as a direct target of IRF4 in CLL cells and their precursors. Collectively, our studies provide the first in vivo evidence for an essential role of Notch signaling in the development of CLL and establish IRF4 as a critical regulator of Notch signaling during CLL development.

DNA methyltransferase 3a (DNMT3A) catalyzes the formation of 5-methyl-cytosine in mammalian genomic DNA, and it is frequently mutated in human hematologic malignancies. Bi-allelic loss of Dnmt3a in mice results in leukemia and lymphoma, including chronic lymphocytic leukemia (CLL). Here, we investigate whether mono-allelic loss of Dnmt3a is sufficient to induce disease. We show that, by 16 months of age, 65% of Dnmt3a(+/-) mice develop a CLL-like disease, and 15% of mice develop non-malignant myeloproliferation. Genome-wide methylation analysis reveals that reduced Dnmt3a levels induce promoter hypomethylation at similar loci in Dnmt3a(+/-) and Dnmt3a(Δ/Δ) CLL, suggesting that promoters are particularly sensitive to Dnmt3a levels. Gene expression analysis identified 26 hypomethylated and overexpressed genes common to both Dnmt3a(+/-) and Dnmt3a(Δ/Δ) CLL as putative oncogenic drivers. Our data provide evidence that Dnmt3a is a haplo-insufficient tumor suppressor in CLL and highlights the importance of deregulated molecular events in disease pathogenesis.

Increasing evidence suggests that posttraumatic stress disorder (PTSD) symptoms are highly prevalent and pervasive among National Guard members who served in recent wars. Previous meta-analyses report a lack of social support as one of the strongest risk factors for the development of PTSD symptoms. Social support among military members is typically categorized into two types: assistance and support which is received from military leaders and fellow members of one’s unit and civilian social support which is obtained from civilian family and friends. Prior research has demonstrated that unit support is associated with less severe PTSD symptoms. In addition to unit support, the influence of civilian social support was also considered a potent buffer for PTSD symptoms. Civilian social support is important to National Guard members because their experiences integrate military and civilian life more than active duty soldiers. Unlike intensive studies in active duty military personnel, fewer studies have examined the role of social support in National Guard members, and civilian social support is rarely investigated in these limited studies. This review article examines the role of civilian social support in National Guard members as a potential protective factor against the development of PTSD symptoms.

TET enzymes are dioxygenases that promote DNA demethylation by oxidizing the methyl group of 5-methylcytosine to 5-hydroxymethylcytosine (5hmC). Here, we report a close correspondence between 5hmC-marked regions, chromatin accessibility and enhancer activity in B cells, and a strong enrichment for consensus binding motifs for basic region-leucine zipper (bZIP) transcription factors at TET-responsive genomic regions. Functionally, Tet2 and Tet3 regulate class switch recombination (CSR) in murine B cells by enhancing expression of Aicda, which encodes the activation-induced cytidine deaminase (AID) enzyme essential for CSR. TET enzymes deposit 5hmC, facilitate DNA demethylation, and maintain chromatin accessibility at two TET-responsive enhancer elements, TetE1 and TetE2, located within a superenhancer in the Aicda locus. Our data identify the bZIP transcription factor, ATF-like (BATF) as a key transcription factor involved in TET-dependent Aicda expression. 5hmC is not deposited at TetE1 in activated Batf-deficient B cells, indicating that BATF facilitates TET recruitment to this Aicda enhancer. Our study emphasizes the importance of TET enzymes for bolstering AID expression and highlights 5hmC as an epigenetic mark that captures enhancer dynamics during cell activation.

Medical students matriculating in the coming years will be faced with treating an expansive increase in the population of older lesbian, gay, bisexual, and transgender (LGBT) patients. While these patients face healthcare concerns similar to their non-LGBT aging peers, the older LGBT community has distinct healthcare needs and faces well-documented healthcare disparities. In order to reduce these healthcare barriers, medical school curricula must prepare and educate future physicians to treat this population while providing high quality, culturally-competent care. This article addresses some of the unique healthcare needs of the aging LGBT population with an emphasis on social concerns and healthcare disparities. It provides additional curricular recommendations to aid in the progressive augmentation of medical school curricula. Abbreviations: Liaison Committee on Medical Education (LCME); LGBT: Lesbian, gay, bisexual, transgender

In the present scenario of all over the world like develops and undeveloped countries are fastly used Distributed Generations (DGs) and Flexible Alternating Current Transmission Systems (FACTS) controllers in power systems for reactive power supports so that the overall power system performances are improved such as minimization of real and reactive power losses, environmental pollutions and maximization of loadability of system, power system stability, the short circuit capacity of the line and also enhancement of voltage profile, available power transfer capacity of the system, reliability as well as security of the system and more flexible operations of the system. This survey paper presents a taxonomical review on impact assessment of DGs and FACTS controllers in power systems from different power system performance viewpoints and reveals the current status of research work in this field.

The transgender community has many unique health needs that are often inadequately addressed by the current medical system. Healthcare workers may lack adequate training to deal with these unique issues, compromising care. Healthcare workers must understand the major barriers to healthcare and heath disparities experienced by the transgender community in order to work to overcome them. This article presents a case illustrating some of the issues that may compromise healthcare for the transgender community as well as measures taken by the providers in response to this case to improve caring for this population in the future.

Interferon Regulatory Factor 4 (IRF4) and IRF8 are critical regulators of immune system development and function. In B lymphocytes, IRF4 and IRF8 have been shown to control important events during their development and maturation including pre-B cell differentiation, induction of B cell tolerance pathways, marginal zone B cell development, germinal center reaction and plasma cell differentiation. Mechanistically, IRF4 and IRF8 are found to function redundantly to control certain stages of B cell development, but in other stages, they function nonredundantly to play distinct roles in B cell biology. In line with their essential roles in B cell development, deregulated expressions of IRF4 and IRF8 have been associated to the pathogenesis of several B cell malignancies and diseases. Recent studies have elucidated diverse transcriptional networks regulated by IRF4 and IRF8 at distinct B cell developmental stages and related malignancies. In this review we will discuss the recent advances for the roles of IRF4 and IRF8 during B cell development and associated diseases.

TET enzymes are dioxygenases that promote DNA demethylation by oxidizing the methyl group of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC). Here we report a close correspondence between 5hmC-marked regions, chromatin accessibility and enhancer activity in B cells, and a strong enrichment for consensus binding motifs for basic region-leucine zipper (bZIP) transcription factors at TET-responsive genomic regions. Functionally, Tet2 and Tet3 regulate class switch recombination (CSR) in murine B cells by enhancing expression of Aicda, encoding the cytidine deaminase AID essential for CSR. TET enzymes deposit 5hmC, demethylate and maintain chromatin accessibility at two TET-responsive elements, TetE1 and TetE2, located within a superenhancer in the Aicda locus. Our data identify BATF as a key bZIP transcription factor involved in TET-dependent Aicda expression. 5hmC is not deposited at TetE1 in activated Batf-deficient B cells, indicating that BATF facilitates TET recruitment to this Aicda enhancer. Our study emphasizes the importance of TET enzymes for bolstering AID expression, and highlight 5hmC as an epigenetic mark that captures enhancer dynamics during cell activation.

Interferon regulatory factor 4 (IRF4) is a critical transcriptional regulator of B-cell development and function. A recent genome-wide single-nucleotide polymorphism (SNP) association study identified IRF4 as a major susceptibility gene in chronic lymphocytic leukemia (CLL). Although the SNPs located in the IRF4 gene were linked to a downregulation of IRF4 in CLL patients, whether a low level of IRF4 is critical for CLL development remains unclear. In rodents, CLL cells are derived from B1 cells whose population is dramatically expanded in immunoglobulin heavy chain Vh11 knock-in mice. We bred a Vh11 knock-in allele into IRF4-deficient mice (IRF4(-/-)Vh11). Here, we report that IRF4(-/-)Vh11 mice develop spontaneous early-onset CLL with 100% penetrance. Further analysis shows that IRF4(-/-)Vh11 CLL cells proliferate predominantly in spleen and express high levels of Mcl-1. IRF4(-/-)Vh11 CLL cells are resistant to apoptosis but reconstitution of IRF4 expression in the IRF4(-/-)Vh11 CLL cells inhibits their survival. Thus, our study demonstrates for the first time a causal relationship between low levels of IRF4 and the development of CLL. Moreover, our findings establish IRF4(-/-)Vh11 mice as a novel mouse model of CLL that not only is valuable for dissecting molecular pathogenesis of CLL but could also be used for therapeutic purposes.

A recent genome-wide SNP association study identified IRF4 as a major susceptibility gene for chronic lymphocytic leukemia (CLL). Moreover, the SNPs located in the 3' UTR of the IRF4 gene have been linked to a down-regulation of IRF4. However, whether a low level of IRF4 is critical for CLL development remains unclear. New Zealand Black (NZB) mice are a naturally occurring, late-onset mouse model of CLL. To examine the role of a reduced level of IRF4 in CLL development, we generated, through breeding, IRF4 heterozygous mutant mice in the NZB background (NZB IRF4(+/-)). Our results show that CLL development is accelerated dramatically in the NZB IRF4(+/-) mice. The average onset of CLL in NZB mice is 12 months, but CLL cells can be detected in NZB IRF4(+/-) mice at 3 months of age. By 5 months of age, 80% of NZB IRF4(+/-) mice developed CLL. CLL cells are derived from B1 cells in mice. Interestingly, NZB IRF4(+/-) B1 cells exhibit prolonged survival, accelerated self-renewal, and defects in differentiation. Although NZB IRF4(+/-) CLL cells are resistant to apoptosis, high levels of IRF4 inhibit their survival. High levels of IRF4 also reduce the survival of MEC-1 human CLL cells. Our analysis further reveals that high levels of IRF4 suppress Akt activity and can do so without the IRF4 DNA binding domain. Thus, our findings reveal a causal relationship between a low level of IRF4 and the development of CLL and establish IRF4 as a novel regulator in the pathogenesis of CLL.

Violent video game (VVG) use has repeatedly been found to be associated with hostile expectations about others, desensitization to violence, decreased empathy and prosocial behavior, and aggressive thoughts and behaviors. Although these research findings have been widely publicized, VVGs remain the most extensively played games and represent a multi-billion dollar industry. Although VVGs are typically rated "mature," indicating they are not suitable for youths, they are often purchased for youths. This may be in part because there is currently no system available to consumers that thoroughly describes the content of video games, and much of the public is unaware of the types of violence that characterize game play. The purpose of this paper is to describe the violent content of some of the top VVGs, based on sales. For the purposes of this issue, acts of senseless, unprovoked violence will be described in detail.

Purpose Evidence-based guidance is presented to assist clinicians in selecting alternative formulations of antiretroviral (ARV) agents for patients with human immunodeficiency virus (HIV) infection who are unable to swallow tablets or capsules. Summary The inability to take medications in standard oral dosage forms can be associated with nonadherence or the use of alternative administration strategies such as capsule or tablet breaking, crushing, or chewing. Patients with HIV infection require long-term ARV therapy to maintain viral suppression; ARV agents are predominately available as tablets and capsules that may pose swallowing difficulties for some patients. Using a variety of sources (the primary literature, pharmaceutical package inserts, and requests for unpublished data from drug manufacturers), available evidence on the bioavailability of ARV medications after disruption of the capsule or tablet matrix was reviewed; information on alternative formulations of ARV agents was also assessed. With several ARV agents, disruption of the solid oral dosage form by crushing, chewing, or breaking tablets or opening capsules prior to ingestion has been shown to result in altered bioavailability or pharmacokinetics and thus the potential for incomplete virological suppression, increased adverse effects, and suboptimal health outcomes. Conclusion Of the 33 single-agent ARV medications and combination ARV products in five classes available at the time of review, approximately half exist as powders, liquids, injectables, or chewable or dissolvable tablets. If alternative ARV formulations or administration methods are used, close monitoring for achievement of virological and immunologic success and potential toxicities is recommended.

Neuroanatomy is an essential course for healthcare students and imparts knowledge regarding the structure of the human nervous system. Its study requires calling upon many skills from students as well as educators. Particularly, challenging is to understand 3-dimensional structures and their relative positions and interfaces from primarily 2-dimensional images and MRI scans. Use of multiple modalities in teaching has been proposed, particularly supplementation of existing teaching methods with plastinated versions of actual brains. While the benefits of using plastinated specimens are many, it is a labor-intensive process that results in a relatively fragile specimen. Furthermore, any time idiosyncrasies of a specimen make it especially valuable, plastination is limited, because it results in only one specimen. To alleviate these issues, the authors propose scanning and reproduction of these samples using digital modeling and manufacturing techniques focused particularly on 3-D printing. In the context of converting from a purely preservation process to one of replication, it becomes immediately clear that the problem of proper mass reproduction, takes on a Design for Manufacture (DFM) construct, particularly, a design for assembly/disassembly/modularity shape. We show how this problem can be approached within the context of DFM, posed as a mathematical optimization problem and present preliminary results from our experiments.Copyright © 2015 by ASME

Summary The zebra mussel, Dreissena polymorpha, is a highly prolific biofouling freshwater mollusk accidentally introduced into North America in the 1980s. Understanding its reproductive mechanisms may give insight into its dispersion, adaptive mechanisms, and possible control methods. The spread of zebra mussels into tidal regions and estuaries could be affected by the responses of zebra mussel gametes to seawater. This study investigated the effect of dilute seawater (Instant Ocean, IO) and several of its components on zebra mussel oocyte viability. Zebra mussels were induced to spawn with serotonin. Oocytes exposed to various concentrations of IO or its component ions began to break down within 3 h, usually after a latency of 1–2 h. The percentages of oocytes fragmenting after 3-h exposure to low IO concentration were: control, 11±3%; 2% IO, 18±2%; 5% IO, 47±4%; and 10% IO, 53±4% (ANOVA, p

Review on IRF4, with data on DNA/RNA, on the protein encoded and where the gene is implicated.

Due to the complexity of the central nervous system, neuroanatomy is cited as one of the most challenging portions of the anatomical sciences to study by first year medical students, and junior physicians alike. Fiber paths are challenging to visualize to their full extent, and it is difficult for a novice learner to clearly dissect them. Interpretation of a two-dimensional image to a three-dimensional anatomical structure is a daunting task, and creates palpable dread to many students. Since its inception, plastination has served as an adjunct in teaching anatomy. If used properly, plastinated specimens can augment students' understanding of neural architecture. Brains dissected following the preparation method by Klingler can be plastinated and utilized for visualization of fiber paths, and used to accent complex structures. This technique can isolate neuroanatomical structures that students often have trouble visualizing in three dimensions, such as the corpus callosum. This poster will discuss the spe...

B cell central tolerance is a process through which self-reactive B cells are removed from the B cell repertoire. Self-reactive B cells are generally removed by receptor editing in the bone marrow and by anergy induction in the periphery. IRF8 is a critical transcriptional regulator of immune system development and function. A recent study showed that marginal zone B cell and B1 B cell populations are dramatically increased in IRF8-deficient mice, indicating that there are B cell–developmental defects in the absence of IRF8. In this article, we report that mice deficient for IRF8 produced anti-dsDNA Abs. Using a hen egg lysozyme double-transgenic model, we further demonstrate that B cell anergy was breached in IRF8-deficient mice. Although anergic B cells in the IRF8-proficient background were blocked at the transitional stage of development, anergic B cells in the IRF8-deficient background were able to mature further, which allowed them to regain responses to Ag stimulation. Interestingly, our results show that IRF8-deficient B cells were more sensitive to Ag stimulation and were resistant to Ag-induced cell death. Moreover, our results show that IRF8 was expressed at a high level in the anergic B cells, and an elevated level of IRF8 promoted apoptosis in the transitional B cells. Thus, our findings reveal a previously unrecognized function of IRF8 in B cell anergy induction.

Chronic Lymphocytic Leukemia (CLL) is the most common adult leukemia in the western world. Despite considerable progress in our current understanding of CLL, the molecular events underlying the complex pathogenesis and clinical behavior of CLL have not been fully elucidated. IRF4 belongs to the IRF superfamily of transcription factors and plays critical roles at multiple stages of B cell development. Genome Wide Association Studies had linked Single Nucleotide Polymorphism (SNP) mediated IRF4 down regulation to the pathogenesis of human CLL (Di Bernardo et al. Nature Genetics, 2008). Moreover, low levels of IRF4 were found to correlate with poorer disease outcomes in human CLL (Chang et al. Blood, 2002). More recently, the studies from our group have established a causal relationship between low levels of IRF4 and development of CLL. We used Vh11 knock-in (Vh11) mice that express a prearranged immunoglobulin variable heavy chain (Vh11) leading to preferential generation of B1 cells (CLL precursors). Interestingly, we have shown that mice expressing the Vh11 transgene in the IRF4 deficient background (IRF4-/-Vh11) spontaneously develop CLL at complete penetrance (Shukla et al. Blood, 2013). Additionally, we have also shown that low levels of IRF4 dramatically accelerate CLL development in the New Zealand Black (NZB) mice (Ma et al. JBC, 2013). Although our studies have demonstrated a critical role for IRF4 in the pathogenesis of CLL, the molecular mechanism through which IRF4 controls CLL development remains unclear. In an attempt to identify the molecular networks regulated by IRF4 in CLL cells we performed gene expression profiling of CLL cells isolated from IRF4-/-Vh11 mice. Intriguingly, our analysis revealed that Notch signaling was hyperactive in the IRF4-/-Vh11 CLL cells compared to the IRF4+/+Vh11 B1 cells. This is particularly important because recent high depth sequencing studies have identified mutations in Notch1 protein leading to its stabilization and activation in ~10% of all CLL patients (Puente et al. Nature, 2011, Fabbri et al. JEM, 2011). Furthermore, the individuals carrying these mutations have an aggressive disease course and poor overall survival (Fabbri et al. JEM, 2011). Recently, studies by Simonetti et al have shown that the IRF4 deficient B cells express high levels of Notch receptors and display increased Notch activity (Simonetti et al. JEM, 2013). We also detected high levels of Notch2 but not Notch1 on the cell surface of IRF4-/-Vh11 CLL cells. To examine the contribution of Notch signaling in the pathogenesis of CLL, we crossed IRF4-/-Vh11 mice with Notch2 conditional mutant mice and with mice expressing cre recombinase in B cells (CD19cre). This approach allowed us to genetically delete Notch2 receptor specifically in B cells of IRF4-/-Vh11 mice (CD19creNotch2fl/flIRF4-/-Vh11). Surprisingly, CLL cells could still emerge from the majority of CD19creNotch2fl/flIRF4-/-Vh11 mice. However, further analysis revealed that those CLL cells expressed high levels of Notch2 on their surface and were apparently derived from IRF4-/-Vh11 B1 cells that escaped cre-mediated deletion. Importantly, our analysis also showed that the majority of B1 and B2 cells in the CD19creNotch2fl/flIRF4-/-Vh11 mice had lost the expression of Notch2, indicating that Notch2 is dispensable for their generation. We also performed a series of analysis to determine how Notch signaling affects B1 and CLL cells in vitro. We used a co-culture system to engage Notch receptors to their ligand (Delta like 1 or DL1) expressed on S17 stromal cells (S17-DL1). Co-culture of wild type B1 cells with S17-DL1 stromal cells led to enhanced proliferation and survival of B1 cells when compared with cells co-cultured on control vector transfected S17 stromal cells (S17-R1). Importantly, Notch2 receptor deficient B1 cells failed to respond to DL1 expressing S17 stromal cells. Similarly, IRF4-/-Vh11 CLL cells also showed enhanced survival on S17-DL1 stromal cells compared to S17-R1 stromal cells. In summary, our studies show that Notch2 is absolutely required for CLL development in the IRF4-/-Vh11 mice, thereby supporting a critical role for Notch signaling in the pathogenesis of CLL. Disclosures No relevant conflicts of interest to declare.

Abstract 714 Chronic Lymphocytic Leukemia (CLL) representing 30% of all the adult leukemia is a heterogeneous B cell malignancy with complex clinical behavior and pathophysiology. Even though an increasing body of literature has helped in understanding the biology of CLL the precise molecular events that underlie the pathogenesis of this disease are still not fully understood. Recent Whole Genome Sequencing and Genome Wide Association Studies have linked different genetic loci to the origin and evolution of the disease. A Genome Wide Association Study (GWAS) by Di Bernardo et al identified Single Nucleotide Polymorphisms in the 3'UTR of Interferon Regulatory Factor 4 (IRF4) as a familial risk allele for CLL. These SNPs are associated with lower mRNA levels of IRF4 in individuals carrying the risk allele. Another study by Havelange V. et al identified heterozygous mutations in the DNA binding domain of IRF4 in patients with CLL. These studies have linked lower levels of IRF4 to the pathogenesis of CLL. Even though studies have linked low levels of IRF4 to the pathogenesis of CLL, whether low levels of IRF4 indeed contribute to CLL development remains to be determined. In rodent, CLL cells are presumably derived from B1 cells. Like CLL cells, B1 cells are B220+CD5+ and possess self-reactive B cell receptors (BCR). B1 cells normally reside in peritoneal and pleural cavities and constitute only about 5% of total B lymphocytes in mice. However, B1 cell population is dramatically elevated in the VH11 knock-in mice. VH11 family of variable IgH chain is expressed exclusively on B1 cells where it pairs with Vk9 to form BCR that recognizes phosphatidylcholine on senescent RBCs. We did not observe an overt increase of CLL development in IRF4 deficient mice. However, when we crossed the VH11 knock-in mice to an IRF4 deficient mice it resulted in a CLL like lymphoproliferative disorder with marked expansion of CD5+ B220low/int population in blood, spleen and other lymphoid organs. To monitor pathogenesis of CLL, we collected blood from saphenous vein of IRF4−/−VH11 and IRF4+/+VH11 littermate control mice monthly. A total of ten mice from each group were used for this study. The appearance of a monoclonal B220lowCD5+ population and its percentage among peripheral blood mononuclear cells (PBMC) were used for the initial CLL diagnosis. The diagnostic criterion for CLL in VH11 mice is the appearance of monoclonal B220lowCD5+ cells constituting over 20% of PBMCs. If the percentage of the B220lowCD5+ clone is under 20% of the PBMC, the diagnosis would be monoclonal B cell lymphocytosis (MBL), a disease which can progress into CLL. Interestingly, 6 out of 10 IRF4−/−VH11 mice developed CLL after just 5 months while the four remaining IRF4−/−VH11 mice developed MBL. In contrast, none of the IRF4+/+VH11 control mice developed CLL or MBL. Two of the mice developed aggressive CLL where leukemic cells infiltrated liver and other non-lymphoid organs. BrdU incorporation assay identified spleen as the primary site of CLL cells expansion although proliferating CLL cells were also identified in lymph nodes and bone marrow. We further isolated CLL cells from IRF4−/−VH11 mice and transplanted them into immunodeficient mice (Rag2−/−gamma−/−). Our result shows that the disease was transplantable as all the transplanted mice succumbed to CLL within 1 month post-transplantation. Treatment of IRF4−/−VH11 CLL cells with specific Syk and Btk inhibitors targeting BCR signaling showed that like human CLL, the leukemic cells were depended on active BCR signaling for their survival and proliferation. In summary, we provide evidence for a causal relationship between low levels of IRF4 and development of CLL. Further studies are needed to provide mechanistic insights into how IRF4 is involved in the development and progression of this disease. The fine dissection of this relationship will allow for development of new therapeutic strategies and identification of new prognostic markers for diagnosis and treatment of CLL. Disclosures: No relevant conflicts of interest to declare. Disclosures: No relevant conflicts of interest to declare.

After tick transmission, the spirochete, Borrelia burgdorferi (Bb), remains at the skin inoculation site for up to 48 hours before spreading and causing Lyme disease. Bb surface agonists should elicit potent inflammation and clearance in skin. However, the ID50 is 10x faster than any immune cell velocities observed in our model. This suggests that Bb motility is a primary mechanism for host cell evasion and indicates our imaging techniques will allow true delineation of host-Bb interactions critical for Lyme disease development.