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Patients with chronic bullous disease of childhood (CBDC) generally respond well to treatment with drugs like Dapsone and oral and topical steroids. Refractory cases are additionally treated with immunosuppressants like azathioprine, methotrexate, ciclosporin, colchicine, tetracycline, and IVIG (Intravenous Immunoglobulin). However, there are few patients who fail to respond to these therapies and pose a challenge both to the patient and the treating physician. Rituximab has been used widely to treat pemphigus cases in adults, but its use in children has not been much reported. We hereby report a case of a 5-year-old child with CBDC, refractory to other treatments, who responded favorably to Rituximab.

Magnesium acetyltaurate (MgAT) has been shown to have a protective effect against N-methyl-D-aspartate (NMDA)-induced retinal cell apoptosis. The current study investigated the involvement of nuclear factor kappa-B (NF-κB), p53 and AP-1 family members (c-Jun/c-Fos) in neuroprotection by MgAT against NMDA-induced retinal damage. In this study, Sprague-Dawley rats were randomized to undergo intravitreal injection of vehicle, NMDA or MgAT as pre-treatment to NMDA. Seven days after injections, retinal ganglion cells survival was detected using retrograde labelling with fluorogold and BRN3A immunostaining. Functional outcome of retinal damage was assessed using electroretinography, and the mechanisms underlying antiapoptotic effect of MgAT were investigated through assessment of retinal gene expression of NF-κB, p53 and AP-1 family members (c-Jun/c-Fos) using reverse transcription-polymerase chain reaction. Retinal phospho-NF-κB, phospho-p53 and AP-1 levels were evaluated using western blot assay. Rat visual functions were evaluated using visual object recognition tests. Both retrograde labelling and BRN3A immunostaining revealed a significant increase in the number of retinal ganglion cells in rats receiving intravitreal injection of MgAT compared with the rats receiving intravitreal injection of NMDA. Electroretinography indicated that pre-treatment with MgAT partially preserved the functional activity of NMDA-exposed retinas. MgAT abolished NMDA-induced increase of retinal phospho-NF-κB, phospho-p53 and AP-1 expression and suppressed NMDA-induced transcriptional activity of NF-κB, p53 and AP-1 family members (c-Jun/c-Fos). Visual object recognition tests showed that MgAT reduced difficulties in recognizing the visual cues (i.e. objects with different shapes) after NMDA exposure, suggesting that visual functions of rats were relatively preserved by pre-treatment with MgAT. In conclusion, pre-treatment with MgAT prevents NMDA induced retinal injury by inhibiting NMDA-induced neuronal apoptosis via downregulation of transcriptional activity of NF-κB, p53 and AP-1-mediated c-Jun/c-Fos. The experiments were approved by the Animal Ethics Committee of Universiti Teknologi MARA (UiTM), Malaysia, UiTM CARE No 118/2015 on December 4, 2015 and UiTM CARE No 220/7/2017 on December 8, 2017 and Ethics Committee of Belgorod State National Research University, Russia, No 02/20 on January 10, 2020.

Brain-derived neurotrophic factor (BDNF) could be considered a potential neuroprotective therapy in amyloid beta (Aβ)-associated retinal and optic nerve degeneration. Hence, in this study we investigated the neuroprotective effect of BDNF against Aβ1-40-induced retinal and optic nerve injury. In this study, exposure to Aβ1-40 was associated with retinal and optic nerve injury. TUNEL staining showed significant reduction in the apoptotic cell count in the BDNF-treated group compared with Aβ1-40 group. HE-stained retinal sections also showed a striking reduction in neuronal cells in the ganglion cell layer (GCL) of retinas fourteen days after Aβ1-40 exposure. By contrast, number of retinal cells was preserved in the retinas of BDNF-treated animals. After Aβ1-40 exposure, visible axonal swelling was observed in optic nerve sections. However, the BDNF-treated group showed fewer changes in optic nerve; axonal swelling was less frequent and less marked. In the present study, exposure to Aβ was associated with oxidative stress, whereas levels of retinal glutathione (GSH), superoxide dismutase (SOD) and catalase were significantly increased in BDNF-treated than in Aβ1-40-treated rats. Both visual object recognition tests using an open-field arena and a Morris water maze showed that BDNF improved rats' ability to recognise visual cues (objects with different shapes) after Aβ1-40 exposure, thus demonstrating that the visual performance of rats was relatively preserved following BDNF treatment. In conclusion, intravitreal treatment with BDNF prevents Aβ1-40-induced retinal cell apoptosis and axon loss in the optic nerve of rats by reducing retinal oxidative stress and restoring retinal BDNF levels.

The role of adenosine receptors as therapeutic targets for neuroprotection is now widely recognized. Their role, however, in protection against retinal ganglion cell (RGC) apoptosis in glaucoma needs further investigation. Hence, in this review, we look into the possibility of adenosine receptors as potential therapeutic targets by exploring their role in modulating various pathophysiological mechanisms underlying glaucomatous RGC loss.This review presents a summary of the adenosine receptor distribution in retina and the cellular functions mediated by them. The major pathophysiological mechanisms such as excitotoxicity, vascular dysregulation, loss of neurotrophic signaling, and inflammatory responses involved in glaucomatous RGC loss are discussed. The literature showing the role of adenosine receptors in modulating these pathophysiological mechanisms is discussed. The literature search was conducted using Pubmed search engine using key words such as 'RGC apoptosis,' 'adenosine,' adenosine receptors' 'retina' 'excitotoxicity,' 'neurotrophins,' 'ischemia', and 'cytokines' individually and in various combinations.Use of adenosine receptor agonists and antagonists, for preservation of the RGCs in glaucomatous eyes independent of the level of intraocular pressure seems a very useful strategy. Future application of this strategy would require appropriate designing of drug formulation for tissue and disease-specific receptor targeting. Furthermore, the modulation of physiological functions and potential adverse effects need further investigations.

Chronic myeloid leukemia (CML) results from hematopoietic stem cell transformation by the BCR-ABL kinase. Despite the success of BCR-ABL tyrosine kinase inhibitors (TKIs) in treating CML patients, leukemia stem cells (LSCs) resist elimination and persist as a major barrier to cure. Previous studies suggest that overexpression of the sirtuin 1 (SIRT1) deacetylase may contribute to LSC maintenance in CML. Here, by genetically deleting SIRT1 in transgenic CML mice, we definitively demonstrated an important role for SIRT1 in leukemia development. We identified a previously unrecognized role for SIRT1 in mediating increased mitochondrial oxidative phosphorylation in CML LSCs. We showed that mitochondrial alterations were kinase independent and that TKI treatment enhanced inhibition of CML hematopoiesis in SIRT1-deleted mice. We further showed that the SIRT1 substrate PGC-1α contributed to increased oxidative phosphorylation and TKI resistance in CML LSCs. These results reveal an important role for SIRT1 and downstream signaling mechanisms in altered mitochondrial respiration in CML LSCs.

Objective N-methyl-D-aspartate (NMDA) excitotoxicity has been proposed to mediate apoptosis of retinal ganglion cells (RGCs) in glaucoma. Taurine (TAU) has been shown to have neuroprotective properties, thus we examined anti-apoptotic effect of TAU against retinal damage after NMDA exposure. Methodology Sprague-Dawley rats were divided into 5 groups of 33 each. Group 1 was administered intravitreally with PBS and group 2 was similarly injected with NMDA (160 nmol). Groups 3, 4 and 5 were injected with TAU (320 nmol) 24 hours before (pre-treatment), in combination (co-treatment) and 24 hours after (post-treatment) NMDA exposure respectively. Seven days after injection, rats were sacrificed; eyes were enucleated, fixed and processed for morphometric analysis, TUNEL and caspase-3 staining. Optic nerve morphology assessment was done using toluidine blue staining. The estimation of BDNF, pro/anti-apoptotic factors (Bax/Bcl-2) and caspase-3 activity in retina was done using ELISA technique. Results Severe degenerative changes were observed in retinae after intravitreal NMDA exposure. The retinal morphology in the TAU pre-treated group appeared more similar to the control retinae and demonstrated a higher number of nuclei than the NMDA group both per 100 μm length (by 1.5-fold, p In conclusion our study revealed that pre-treatment with TAU prevents NMDA-induced retinal cell apoptosis more effectively than co- and post-treatment with TAU.

ET1 induced retinal and optic nerve injury is widely used as an experimental model to study unexplained pathophysiology and therapeutic targets involved in several ophthalmic diseases such as glaucoma and diabetic retinopathy. Although a dose-dependent effect of ET1 on retinal vascular tone and retinal blood flow has been studied, the dose-dependent effects of ET1 on retinal and optic nerve morphology remain unexplored. Current study, therefore investigated the dose-dependent effects of ET1 on retinal and optic nerve morphology. Sprague Dawley rats were injected with ET1 in the dose range of 0.025–2.5 nM, intravitreally. Seven days post-injection, eyes were enucleated and retinal sections were examined for morphological changes using H&E staining and extent of retinal cell apoptosis was determined using TUNEL staining. Optic nerve morphology was studied using toluidine blue staining. It was observed that ET1 induces significant changes in retinal and optic nerve morphology at 0.25 and 2.5 nM doses but not at 0.025 nM. The outcomes of in vivo studies using different doses of ET1, therefore, may not be comparable. Furthermore, while designing experimental studies using ET1 induced retinal ischemia, it is important to carefully make the dose selection based on the study objectives.

Time-varying neurophysiological activity has been classically explored using correlation based sliding window analysis. However, this method employs only lower order statistics to track dynamic functional connectivity of the brain. We introduce recursive dynamic functional connectivity (rdFC) that incorporates higher order statistics to generate a multi-order connectivity pattern by analyzing neurophysiological data at multiple time scales. The technique builds a hierarchical graph between various temporal scales as opposed to traditional approaches that analyze each scale independently. We examined more than a million rdFC patterns obtained from morphologically diverse EEGs of 2378 subjects of varied age and neurological health. Spatiotemporal evaluation of these patterns revealed three dominant connectivity patterns that represent a universal underlying correlation structure seen across subjects and scalp locations. The three patterns are both mathematically equivalent and observed with equal prevalence in the data. The patterns were observed across a range of distances on the scalp indicating that they represent a spatially scale-invariant correlation structure. Moreover, the number of patterns representing the correlation structure has been shown to be linked with the number of nodes used to generate them. We also show evidence that temporal changes in the rdFC patterns are linked with seizure dynamics.

SUMMARY Chronic myeloid leukemia (CML) is propagated by leukemia stem cells (LSCs) that are not eradicated by tyrosine kinase inhibitor (TKI) treatment and persist as a source of disease recurrence. Bone marrow (BM) mesenchymal niches play an essential role in hematopoietic stem cell (HSC) and LSC maintenance. Using a murine CML model, we examine leukemia-induced alterations in mesenchymal cell populations. We show that 6C3+ stromal progenitors expand in CML BM and exhibit increased LSC but reduced HSC supportive capacity. Tumor necrosis factor alpha (TNF-α) signaling mediates expansion and higher expression of CXCL1 in CML BM 6C3+ cells and higher expression of the CXCL1 receptor CXCR2 in LSCs. CXCL1 enhances LSC proliferation and self-renewal, whereas CXCR2 inhibition reduces LSC growth and enhances LSC targeting in combination with tyrosine kinase inhibitors (TKIs). We find that TNF-α-mediated alterations in CML BM stromal niches enhance support of LSC maintenance and growth via CXCL1-CXCR2 signaling and that CXCR2 inhibition effectively depletes CML LSCs., Graphical abstract, In brief Chronic myeloid leukemia (CML) leukemia stem cells (LSCs) resist elimination by treatment. Agarwal et al. show that CML 6C3+ bone marrow stromal progenitors support LSC maintenance. TNF-α signaling mediates increased CXCL1 expression in 6C3+ cells, which support LSC proliferation and self-renewal. Targeting the CXCL1 receptor CXCR2 inhibits LSC maintenance.

Introduction:irritable bowel syndrome (IBS) is one of the most common functional gastrointestinal disorder. The medical programme is among the most challenging and stressful undergraduate programmes which may predispose to higher rates of IBS. This study sought to determine the prevalence of undiagnosed IBS and the factors associated with IBS among medical students in a Malaysian private university. Methods:a cross sectional study was conducted among the medical students from semester 6 to semester 9 (year 3, year 4 and year 5) of a Malaysian private university. The questionnaire consisted of 2 main sections. Section A was on demographic data and section B consisted of the Hospital Anxiety and Depression Scale (HADS), the Pittsburgh Sleep Quality Index (PSQI) and the Rome IV Questionnaire for IBS. Association between the factors gathered and IBS was assessed using the Chi-Square test. Variables with a p-value of less than 0.2 in the univariate analysis were entered into a multivariate analysis model. Results:number of students who responded were 190 (56.3%) were females, 66.3% were Chinese and 37.9% were from semester 9. Twenty-eight (14.7%) students had symptoms consistent with a diagnosis of IBS. Depression was found to be significantly associated with IBS (Adjusted OR: 4.749, CI: 1.809-12.470). Conclusion:this study suggests that IBS is common among Malaysian medical students. There was a significant association between IBS and depression.

During recent decades, stress-related neuropsychiatric disorders such as anxiety, depression, chronic tension headache, and migraine have established their stronghold in the lives of a vast number of people worldwide. In order to address this global phenomenon, intensive studies have been carried out leading to the advancement of drugs like anti-depressants, anxiolytics, and analgesics which although help in combating the symptoms of such disorders but also create long-term side effects. Thus, as an alternative to such clinical practices, various complementary therapies such as yoga and meditation have been proved to be effective in alleviating the causes and symptoms of different neuropsychiatric disorders. The role of altered brain waves in this context has been recognized and needs to be pursued at the highest level. Thus, the current study provides a review focused on describing the effects of yoga and meditation on anxiety and depression as well as exploring brain waves as a tool for assessing the potential of these complementary therapies for such disorders.

Sigmoid volvulus in pregnancy is a very rare clinical situation responsible for high rates of mortality and morbidity in both mother and fetus. It is due to the gravid uterus which itself clouds the clinical picture of sigmoid volvulus.A long and redundant sigmoid colon compressed by enlarged gravid uterus, might explain increased incidence of sigmoid volvulus during pregnancy especially during third trimester. This article describes a case with 36 weeks of gestation where sigmoid volvulus was encountered during caesarean section. Keywords: Sigmoid, volvulus, pregnancy

Physics is everywhere in our daily life. It helps in understanding concepts of surgery easily. Application of physics in surgical field is very helpful especially for a surgeon. In this article we have summarized some common laws of physics, many of them we see in our operation theatre like Poiseuille’s Law, Pascal’s Law, Laplace’s Law, Bernoulli Equation or Theorem, Ohm’s Law, Floating Ball Valve Mechanism, Newton’s Law of Cooling, Laws of Vector and Air-Water Seal System. The application and understanding of these laws would make a surgeon think more like a scientist. Doppler Effect is used in ultrasonography to evaluate the direction and velocity of blood flow. Reynolds number (NR) is used to predict whether the flow of blood will be laminar or turbulent. The laws of Thermodynamics, Mechanics, and Vectors as they apply to soft and bony tissues. These include the Laplace’s Law as applied to colonic perforation and esophageal varices etc. In Electrocautery, alternating current is passed through a resistant metallic wire electrode and generates heat. The heated electrode is used to achieve hemostasis and dissection. The Pascal’s Law finds use in hernia repair Keywords: Physics, Surgery, Fundamentals.

Identifying the patients with acute heart failure (HF) at high risk for rehospitalization after hospital discharge will enable proper optimization of treatment. This study is aimed to evaluate the rehospitalization rate at 60 days of discharge and their predictors in patients of chronic heart failure with reduced ejection fraction (HFrEF).This prospective observational study enrolled patients with left ventricle ejection fraction (LVEF)40%, who were admitted because of acute decompensation. Patients were followed for 60 days to analyze rehospitalization rate and its predictors.Of 103 HFrEF patients (74% male; mean age 55.8 years) enrolled, 7 patients died during index admission and 3 patients lost to follow up. The 60-day rehospitalization rate was 37% (34/93). We studied 23 clinical and 9 biochemical predictors of rehospitalization. Out of 34 events of rehospitalization, 79.41% (n=28) was due to cardiac cause followed by respiratory 5.8% (n=2), renal 5.8% (n=2) and others 5.8% (n=2). Among all the parameters, on logistic regression analysis having longer length of index hospital stay (7 days) (52.8% vs. 28.8%; odds ratio [OR], 1.79; confidence interval [CI], 1.2-7.25; p=0.040) and chronic kidney disease (CKD) (26.5% vs. 8.5%; OR, 3.06; CI, 1.1-57.04; p=0.050) independently increased the risk of rehospitalization at 60 days of discharge. Further higher haemoglobin level (11.3 vs. 9.9 gm/dL; OR, 0.71; CI, 0.48-0.97; p=0.050) and higher LVEF at index admission (30.4% vs. 26.5%; OR, 0.87; CI, 0.75-0.99; p=0.049) were associated with decreased the risk of rehospitalization.Our study reveals that patients with HFrEF have significantly higher rehospitalization rate (37%) and in-hospital mortality rates (6.78%) of any chronic cardiac disease conditions. Correction of low hemoglobin and special care in those who are having very low LVEF, CKD and longer length of stay, including tailored therapy and frequent visits may play an important role in preventing future rehospitalization in these patients.

Retinal ganglion cell apoptosis in glaucoma is associated with elevated levels of endothelin-1 (ET1), a potent vasoconstrictor. ET1-induced retinal ischemia leads to altered expression of nitric oxide synthase (NOS) isoforms leading to increased formation of nitric oxide (NO) and retinal nitrosative stress. Since magnesium (Mg) is known to improve endothelial functions and reduce oxidative stress and taurine (TAU) possesses potent antioxidant properties, we investigated the protective effects of magnesium acetyltaurate (MgAT) against ET1-induced nitrosative stress and retinal damage in rats. We also compared the effects of MgAT with that of TAU alone.Sprague Dawley rats were intravitreally injected with ET1. MgAT and TAU were administered as pre-, co-, or posttreatment. Subsequently, the expression of NOS isoforms was detected in retina by immunohistochemistry, retinal nitrotyrosine level was estimated using ELISA, and retinal cell apoptosis was detected by TUNEL staining.Intravitreal ET1 caused a significant increase in the expressions of nNOS and iNOS while eNOS expression was significantly reduced compared to vehicle treated group. Administration of both MgAT and TAU restored the altered levels of NOS isoform expression, reduced retinal nitrosative stress and retinal cell apoptosis. The effect of MgAT, however, was greater than that of TAU alone.MgAT and TAU prevent ET1-induced retinal cell apoptosis by reducing retinal nitrosative stress in Sprague Dawley rats. Addition of TAU to Mg seems to enhance the efficacy of TAU compared to when given alone. Moreover, the pretreatment with MgAT/TAU showed higher efficacy compared to co- or posttreatment.

Background: Helicobacter pylori infection is a major health ailment in developing countries. The infection is related with high morbidity and mortality ranging from chronic gastritis to gastric malignancies. The prevalence of H. pylori infection greatly varies from country to country and region to region. The aim is to study the prevalence of H. pylori infection in upper gastrointestinal (GI) tract disorders (dyspepsia) patients. Methods: The study was carried out in an outpatient department, India. Patients presenting with dyspeptic symptoms were subjected to serological investigation and upper GI endoscopy, histopathological examination, and rapid urease test of specimen. Diagnosis of H. pylori infection was made if one or both diagnostic test results were positive. Results: H. pylori infection was diagnosed in 85% of patients. There was no significant difference in sex- and age-related distribution (50 years' age group) of H. pylori infection. However, association of H. pylori infection was positive in 83.3% (45) of patients with endoscopic abnormalities (54). Conclusion: The prevalence of H. pylori infection is high in the study population. Early detection and prompt treatment are essential for prevention of serious complications.

Tyrosine kinase inhibitor (TKI) treatment of chronic myeloid leukemia (CML) has limited efficacy against leukemia stem cells (LSC) responsible for disease propagation, and most CML patients require continued TKI treatment to maintain remission. LSC maintenance is related, at least in part, to signals from the bone marrow microenvironment (BMM). Our previous studies have shown that Wnt signaling from the BMM contributes to preservation of CML LSC following TKI treatment. Secretion of Wnt ligands requires their modification by the O-acyl transferase Porcupine (PORCN). Here we investigated the activity of a potent and selective PORCN inhibitor, WNT974, against CML stem and progenitor cells. WNT974 efficiently antagonized Wnt signaling in human CML CD34+ cells, and in combination with the TKI nilotinib (NIL) significantly enhanced inhibition of proliferation and colony-forming potential of CML stem and progenitor cells and reduced their growth in immunodeficient mice in vivo, in comparison with NIL alone. Treatment of transgenic CML mice in vivo with NIL in combination with WNT974 significantly reduced leukemic stem and progenitor cell numbers, reduced regeneration of leukemic long-term hematopoietic stem cells in secondary transplant recipients, and enhanced survival of mice after discontinuation of treatment, in comparison with NIL alone. CML progenitors demonstrated enhanced sensitivity to Wnt stimulation, associated with increased expression of the FZD4 receptor. FZD4 knockdown inhibited CML progenitor growth. These results support further investigation of PORCN targeting to inhibit Wnt secretion and signaling and enhance targeting of CML stem cells while sparing their normal counterparts.

Rodents have widely been used to represent glaucomatous changes both in the presence and absence of elevated intraocular pressure (IOP) as they offer clear advantages over other animal species. IOP elevation is commonly achieved by creating an obstruction in the aqueous outflow pathways, consequently leading to retinal ganglion cell and optic nerve (ON) damage, the hallmark of glaucoma. These changes may also be achieved in the absence of elevated IOP by directly inflicting injury to retina or ON. Areas covered: This paper presents a summary of currently used rodent models of glaucoma. The characteristics of these models from several studies are summarized. The benefits and shortcomings of these models are also discussed. Expert opinion: The choice of animal model that closely represents human disease is key for successful translational of preclinical research to clinical practice. Rodent models of rapid IOP elevation are likely to be least representative, whereas models such as steroid-induced glaucoma models more closely resemble the trabecular meshwork changes seen in glaucomatous human eyes. However, this model needs further characterization. Rodent models based on direct retinal and ON injury are also useful tools to investigate molecular mechanisms involved at the site of final common pathology and neuroprotective strategies.

Molluscum contagiosum is a very common skin and mucosal disease of viral origin, caused by molluscum contagiosum virus (MCV) 0 of poxvirus family. With the eradication of smallpox, MCV is the only member of the poxvirus family that causes substantial disease in humans. Although frequently reported, its unusual clinical presentation makes its diagnosis a challenging task.

Several electroencephalogram (EEG)-based predictive models for automated epilepsy diagnosis have been proposed over more than a decade. However, to the best of our knowledge, none have been evaluated on a holdout/test set. A vast majority of these studies have reported accuracies above 95% on a benchmark EEG dataset, but the dataset has been shown here to have certain limitations when used for building classifiers for epilepsy diagnosis. We implemented two previously reported classifiers trained on the benchmark dataset whose accuracies were observed to drop sharply when evaluated on a test set. We propose a feature, engineered specifically, for epilepsy diagnosis that attempts to characterize the neuronal synchronization using scalp EEG by extending the concept of the impulse response of linear time-invariant systems to matrices. This feature was tested on the EEG of 50 epileptics and 50 healthy subjects and yielded an area under the curve (AUC) of 0.87. It outperforms the existing models implemented by us that gave the AUC of 0.80 when trained and tested on scalp EEG data, thereby, setting the new benchmark for automated epilepsy diagnosis on test set evaluation. The feature has also been shown to have statistical consistency across time and vigilance states with robustness against EEG artifacts.

Chronic myeloid leukemia (CML) originates in a hematopoietic stem cell (HSC) transformed by the breakpoint cluster region (BCR)-abelson (ABL) oncogene and is effectively treated with tyrosine kinase inhibitors (TKIs). TKIs do not eliminate disease-propagating leukemic stem cells (LSCs), suggesting a deeper understanding of niche-dependent regulation of CML LSCs is required to eradicate disease. Cxcl12 is expressed in bone marrow niches and controls HSC maintenance, and here, we show that targeted deletion of Cxcl12 from mesenchymal stromal cells (MSCs) reduces normal HSC numbers but promotes LSC expansion by increasing self-renewing cell divisions, possibly through enhanced Ezh2 activity. In contrast, endothelial cell-specific Cxcl12 deletion decreases LSC proliferation, suggesting niche-specific effects. During CML development, abnormal clusters of colocalized MSCs and LSCs form but disappear upon Cxcl12 deletion. Moreover, MSC-specific deletion of Cxcl12 increases LSC elimination by TKI treatment. These findings highlight a critical role of niche-specific effects of Cxcl12 expression in maintaining quiescence of TKI-resistant LSC populations.

Summary: The bone marrow microenvironment is composed of heterogeneous cell populations of non-hematopoietic cells with complex phenotypes and undefined trajectories of maturation. Among them, mesenchymal cells maintain the production of stromal, bone, fat, and cartilage cells. Resolving these unique cellular subsets within the bone marrow remains challenging. Here, we used single-cell RNA sequencing of non-hematopoietic bone marrow cells to define specific subpopulations. Furthermore, by combining computational prediction of the cell state hierarchy with the known expression of key transcription factors, we mapped differentiation paths to the osteocyte, chondrocyte, and adipocyte lineages. Finally, we validated our findings using lineage-specific reporter strains and targeted knockdowns. Our analysis reveals differentiation hierarchies for maturing stromal cells, determines key transcription factors along these trajectories, and provides an understanding of the complexity of the bone marrow microenvironment. : Using single-cell RNA sequencing, Wolock et al. reconstruct the transcriptional hierarchy of mouse bone marrow stromal cell states and infer differentiation paths to fat, bone, and cartilage. These cell state relations were validated using lineage-specific reporter strains and targeted knockdowns of transcription factors that mediate fate decisions. Keywords: stromal cell, differentiation, scRNA-seq, bone marrow, transcription factor

Endothelin-1 (ET-1), a potent vasoconstrictor, plays a significant role in the pathophysiology of ocular conditions like glaucoma. Glaucoma is characterized by apoptotic loss of retinal ganglion cells (RGCs) and loss of visual fields and is a leading cause of irreversible blindness. In glaucomatous eyes, retinal ischemia causes release of pro-inflammatory mediators such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α and promotes activation of transcription factors such as nuclear factor kappa B (NFKB) and c-Jun. Magnesium acetyltaurate (MgAT) has previously been shown to protect against ET-1 induced retinal and optic nerve damage. Current study investigated the mechanisms underlying these effects of MgAT, which so far remain unknown. Sprague dawley rats were intravitreally injected with ET-1 with or without pretreatment with MgAT. Seven days post-injection, retinal expression of IL-1β, IL-6, TNF-α, NFKB and c-Jun protein and genes was determined using multiplex assay, Western blot and PCR. Animals were subjected to retrograde labeling of RGCs to determine the extent of RGC survival. RGC survival was also examined using Brn3A staining. Furthermore, visual functions of rats were determined using Morris water maze. It was observed that pre-treatment with MgAT protects against ET-1 induced increase in the retinal expression of IL-1β, IL-6 and TNF-α proteins and genes. It also protected against ET-1 induced activation of NFKB and c-Jun. These effects of MgAT were associated with greater RGC survival and preservation of visual functions in rats. In conclusion, MgAT prevents ET-1 induced RGC loss and loss of visual functions by suppressing neuroinflammatory reaction in rat retinas.

Aim To investigate dose-dependent effects of N-methyl-D-aspartate (NMDA) on retinal and optic nerve morphology in rats. Methods Sprague Dawley rats, 180-250 g in weight were divided into four groups. Groups 1, 2, 3 and 4 were intravitreally administered with vehicle and NMDA at the doses 80, 160 and 320 nmol respectively. Seven days after injection, rats were euthanized, and their eyes were taken for optic nerve toluidine blue and retinal hematoxylin and eosin stainings. The TUNEL assay was done for detecting apoptotic cells. Results All groups treated with NMDA showed significantly reduced ganglion cell layer (GCL) thickness within inner retina, as compared to control group. Group NMDA 160 nmol showed a significantly greater GCL thickness than the group NMDA 320 nmol. Administration of NMDA also resulted in a dose-dependent decrease in the number of nuclei both per 100 µm GCL length and per 100 µm2 of GCL. Intravitreal NMDA injection caused dose-dependent damage to the optic nerve. The degeneration of nerve fibres with increased clearing of cytoplasm was observed more prominently as the NMDA dose increased. In accordance with the results of retinal morphometry analysis and optic nerve grading, TUNEL staining demonstrated NMDA-induced excitotoxic retinal injury in a dose-dependent manner. Conclusion Our results demonstrate dose-dependent effects of NMDA on retinal and optic nerve morphology in rats that may be attributed to differences in the severity of excitotoxicity and oxidative stress. Our results also suggest that care should be taken while making dose selections experimentally so that the choice might best uphold study objectives.

Amyloid beta (Aβ) is known to contribute to the pathophysiology of retinal neurodegenerative diseases such as glaucoma. Effects of intravitreal Aβ(1-42) on retinal and optic nerve morphology in animal models have widely been studied but not those of Aβ(1-40). Hence, we evaluated the time- and dose-related effects of intravitreal Aβ(1-40) on retinal and optic nerve morphology. Since oxidative stress and brain derived neurotrophic factor (BDNF) are associated with Aβ-induced neuronal damage, we also studied dose and time-related effects of Aβ(1-40) on retinal oxidative stress and BDNF levels.Five groups of rats were intravitreally administered with vehicle or Aβ(1-40) in doses of 1.0, 2.5, 5 and 10 nmol. Animals were sacrificed and eyes were enucleated at weeks 1, 2 and 4 post-injection. The retinae were subjected to morphometric analysis and TUNEL staining. Optic nerve sections were stained with toluidine blue and were graded for neurodegenerative effects. The estimation of BDNF and markers of oxidative stress in retina were done using ELISA technique.It was observed that intravitreal Aβ(1-40) causes significant retinal and optic nerve damage up to day 14 post-injection and there was increasing damage with increase in dose. However, on day 30 post-injection both the retinal and optic nerve morphology showed a trend towards normalization. The observations made for retinal cell apoptosis, retinal glutathione, superoxide dismutase activity and BDNF were in accordance with those of morphological changes with deterioration till day 14 and recovery by day 30 post-injection. The findings of this study may provide a guide for selection of appropriate experimental conditions for future studies.

Topical administration is the preferred route of drug delivery for ophthalmic ailments. However, poor permeation through ocular surface and significant systemic absorption, makes the topical drug delivery challenging. Furthermore, distribution of topically delivered drugs varies with their physicochemical properties and the type of formulation used. Hence, this study was done to understand the pattern of ocular drug distribution of topically applied hydrophilic and lipophilic substances in two different formulations.5-Carboxyfluorescein and 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate were used as representative candidates for hydrophilic and lipophilic substances, respectively. They were formulated in solution and liposomes. Single drop of either formulation containing hydrophilic or lipophilic substance was instilled topically, unilaterally to rat eyes. Subsequently, rats were sacrificed at 10, 30 and 120 min post-instillation. Eyes were cryosectioned and examined under confocal microscope to determine the fluorescence intensity in ocular tissues.Corneal permeation of hydrophilic and lipophilic substances in both formulations peaked at 30 min post-instillation. Liposomal-lipophilic dye and non-liposomal-hydrophilic dye showed better corneal distribution. Fluorescence was absent in contralateral eyes of non-liposomal-hydrophilic dye-treated animals but was present in contralateral eyes of liposomal-hydrophilic dye-treated animals. Fluorescence in contralateral eyes of liposomal-lipophilic dye-treated animals was significantly higher compared to non-liposomal-lipophilic dye-treated animals.Topically applied liposomal formulation of lipophilic substance provides higher corneal concentration of drug with lesser systemic absorption compared to its solution. For hydrophilic substance, topical use of solution provides greater corneal concentration compared to liposomes which is more likely to be absorbed systemically.

Hematopoietic stem cells (HSCs) reside in regulatory niches in the bone marrow (BM). Although HSC niches have been extensively characterized, the role of endosteal osteoblasts (OBs) in HSC regulation requires further clarification, and the role of OBs in regulating leukemic stem cells (LSCs) is not well studied. We used an OB visualization and ablation mouse model to study the role of OBs in regulating normal HSCs and chronic myelogenous leukemia (CML) LSCs. OB ablation resulted in increase in cells with a LSK Flt3(-)CD150(+)CD48(-) long-term HSC (LTHSC) phenotype but reduction of a more highly selected LSK Flt3(-)CD34(-)CD49b(-)CD229(-) LTHSC subpopulation. LTHSCs from OB-ablated mice demonstrated loss of quiescence and reduced long-term engraftment and self-renewal capacity. Ablation of OB in a transgenic CML mouse model resulted in accelerated leukemia development with reduced survival compared with control mice. The notch ligand Jagged-1 was overexpressed on CML OBs. Normal and CML LTHSCs cultured with Jagged-1 demonstrated reduced cell cycling, consistent with a possible role for loss of Jagged-1 signals in altered HSC and LSC function after OB ablation. These studies support an important role for OBs in regulating quiescence and self-renewal of LTHSCs and a previously unrecognized role in modulating leukemia development in CML.

Background Steroid-induced ocular hypertension is currently treated in the same way as primary open-angle glaucoma. However, the treatment is often suboptimal and is associated with adverse effects. We evaluated the oculohypotensive effects of topical trans-resveratrol in rats with steroid-induced ocular hypertension and involvement of adenosine receptors (AR) in intraocular pressure (IOP) lowering effect of trans-resveratrol. Methods The oculohypotensive effect of unilateral single-drop application of various concentrations of trans-resveratrol was first studied in oculonormotensive rats. Concentration with maximum effect was similarly studied in rats with steroid-induced ocular hypertension. Involvement of AR was studied by observing the alterations of IOP in response to trans-resveratrol after pretreating animals with AR subtype-specific antagonists. Additionally, we used computational methods, including 3D modelling, 3D structure generation and protein–ligand interaction, to determine the AR–trans-resveratrol interaction. Results All concentrations of trans-resveratrol produced significant IOP reduction in normotensive rat eyes. Maximum mean IOP reduction of 15.1% was achieved with trans-resveratrol 0.2%. In oculohypertensive rats, trans-resveratrol 0.2% produced peak IOP reduction of 25.2%. Pretreatment with A1 antagonist abolished the oculohypotensive effect of trans-resveratrol. Pretreatment with A3 and A2A AR antagonists produced significant IOP reduction in both treated and control eyes, which was further augmented by trans-resveratrol application in treated eyes. Computational studies showed that trans-resveratrol has highest affinity for A2B and A1, followed by A2A and A3 AR. Conclusion Topically applied trans-resveratrol reduces IOP in rats with steroid-induced ocular hypertension. Trans-resveratrol-induced oculohypotension involves its agonistic activity at the A1 AR.

SummaryThe FLT3-ITD mutation is frequently observed in acute myeloid leukemia (AML) and is associated with poor prognosis. In such patients, FLT3 tyrosine kinase inhibitors (TKIs) are only partially effective and do not eliminate the leukemia stem cells (LSCs) that are assumed to be the source of treatment failure. Here, we show that the NAD-dependent SIRT1 deacetylase is selectively overexpressed in primary human FLT3-ITD AML LSCs. This SIRT1 overexpression is related to enhanced expression of the USP22 deubiquitinase induced by c-MYC, leading to reduced SIRT1 ubiquitination and enhanced stability. Inhibition of SIRT1 expression or activity reduced the growth of FLT3-ITD AML LSCs and significantly enhanced TKI-mediated killing of the cells. Therefore, these results identify a c-MYC-related network that enhances SIRT1 protein expression in human FLT3-ITD AML LSCs and contributes to their maintenance. Inhibition of this oncogenic network could be an attractive approach for targeting FLT3-ITD AML LSCs to improve treatment outcomes.

The present study was undertaken with the purpose to compare the sensitivity of various methods for detection of lymph node metastases:Intra-operative frozen sections H & E staining, Conventional H & E staining on formalin fixed tissue, Serial -step sectioning by conventional H & E staining & Immunohistochemical staining by Pancytokeratin antibody. METHOD: The study included 80 consecutive cases of oral squamous cell carcinoma, who underwent radical neck dissection. The various level of lymph nodes in these cases were checked for metastases by 4 techniques i.e. intra-operative frozen sections H & E staining, conventional H & E staining on formalin fixed tissue, serial -step sectioning by conventional H & E staining &immunohistochemical staining by Pancytokeratin antibody. RESULTS: Considering IHC as a gold standard, we observed highest sensitivity & specificity for serial sectioning at 53.7%, & 98.9% when compared to intraoperative frozen section and conventional H&E which were 32.5%, 97.1% & 44.7%, 98.2% respectively. CONCLUSION: Thus we conclude that the most sensitive method to detect lymph node metastasis in case of Oral Squamous cell carcinoma is step serial section when considering IHC as a gold standard.

Angiotensin converting enzyme inhibitors (ACEIs) have been shown to lower intraocular pressure (IOP). Since, the ACEIs cause increased tissue prostaglandin levels, we hypothesized that the mechanisms of ACEI-induced IOP reduction have similarity with those of prostaglandin analogs. The present study investigated the involvement of matrix metalloproteinases (MMPs) and cytokine activity modulation as the underlying mechanisms of ACEI-induced ocular hypotension. The IOP lowering effect of single drop of enalaprilat dehydrate 1% was evaluated in rats pretreated with a broad spectrum MMP inhibitor or a cytokine inhibitor. Effect of angiotensin receptor blocker, losartan potassium 2%, was also studied to evaluate involvement of angiotensin II receptor type 1 (AT1) in IOP lowering effect of ACEI. Topical treatment with single drop of enalaprilat resulted in significant IOP reduction in treated eye with mean peak reduction 20.3% at 3h post-instillation. Treatment with losartan resulted in a peak IOP reduction of 13.3%, which was significantly lower than enalaprilat, indicating involvement of mechanisms in addition to AT1 blockade. Pretreatment with a broad spectrum MMP inhibitor or a cytokine inhibitor significantly attenuated the enalprilat-induced IOP reduction with mean peak IOP reduction of 11.2% and 13.6% respectively. The IOP-lowering effect of enalaprilat seems to be attributed to reduced angiotensin II type 1 receptor stimulation and modulation of MMP and cytokines activities.

Skin is the largest organ of human body and stands as a guard for our internal organs. It can be regarded as a mirror giving a reflection of metabolic, biochemical and functional status of our internal organs. Dermatologists/Gastroenterologist should be aware of the dermatological manifestations as these change may be the first clue that a patient has underlying gastrointestinal (GI) or liver disease. Recognizing these signs is important in early and appropriate diagnosis. This article reviews the important dermatological manifestation of various GI and liver diseases.

The FLT3-ITD mutation occurs in 25-30% of AML patients, and is associated with increased relapse rate and poor survival. FLT3 tyrosine kinase inhibitors (TKI) have shown encouraging results, but responses are not sustained. Therefore, there is considerable interest in understanding mechanisms of resistance of FLT3-ITD AML stem cells after TKI treatment. Autophagy is an important cellular response that maintains cell survival under stress. Here we investigated the regulation of autophagy in resistance of FLT3-ITD+ AML stem cells to FLT3 TKI treatment. MV4-11 and Molm13 FLT3-ITD AML cells were transduced with GFP-LC3-RFP-LC3 ΔG constructs to measure autophagic flux. Autophagyhigh AML cells showed slower growth(p We tested the effect of Lys05 in vivo by treating NSG mice engrafted with primary FLT3-ITD AML cells with vehicle, AC220 (10mg/kg), Lys05 (20mg/kg) or combination for 3 weeks. Lys05 treatment reduced human CD45+ cell engraftment (8.7% vs 17%, p=0.03), and reduced CD45+CD33+ myeloid cells in BM and spleen (BM: 42% vs 81%, p We conclude that autophagy inhibition enhances sensitivity of FLT3-ITD+ AML stem cells to TKI treatment both in vitro and in vivo. We demonstrate important roles for autophagy in regulating p53 levels and cell cycle in FL3-ITD AML cells. These studies support further development of strategies to target autophagy and related pathways to enhance efficacy of TKI in eliminating AML stem cells. Disclosures No relevant conflicts of interest to declare.

Background: Item analysis is a quality assurance of examining the performance of the individual test items that measures the validity and reliability of exams. This study was performed to evaluate the quality of the test items with respect to their performance on difficulty index (DFI), Discriminatory index (DI) and assessment of functional and non-functional distractors (FD and NFD).Methods: This study was performed on the summative examination undertaken by 113 students. The analyses include 120 one best answers (OBAs) and 360 distractors.Results: Out of the 360 distractors, 85 distractors were chosen by less than 5% with the distractor efficiency of 23.6%. About 47 (13%) items had no NFDs while 51 (14%), 30 (8.3%), and 4 (1.1%) items contained 1, 2, and 3 NFDs respectively. Majority of the items showed excellent difficulty index (50.4%, n=42) and fair discrimination (37%, n=33). The questions with excellent difficulty index and discriminatory index showed statistical significance with 1NFD and 2 NFD (p=0.03).Conclusions: The post evaluation of item performance in any exam in one of the quality assurance method of identifying the best performing item for quality question bank. The distractor efficiency gives information on the overall quality of item.

Eczema herpeticum is a potentially life threatening viral infection with pre-existing skin disease. It occurs most commonly in patients with atopic dermatitis. An extensive rash with small vesicles filled with yellow pus and high fever is the commonest presentation of eczema herpeticum. Children are more commonly affected and primary episode is more severe than the recurrence. We are reporting a case of Kaposi's varicelliform eruption in 30 year old atopic female who presented with an abrupt onset of umbilicated vesicular eruptions with pustular crusting over the face. The mortality of Eczema Herpeticum is approximately 10% in which most fatalities occurring in infants and children with primary type of herpetic infection and in adults with inadequate cellular

Cerebriform intradermal nevus is a rare form of cutis verticis gyrata. Clinically it manifests as a scalp deformity resembling the surface of the brain, with cerebriform morphologic characteristics these lesions may become malignant or infected. Here we report a 20 year old boy, with cerebriform and soft fold on right temporo-parietal area present since birth. The clinical and histopathological (solitary or clusters of nevus cells) presentations of cerebriform intradermal nevus are described and the therapeutic possibilities discussed. INTRODUCTION: Cutis verticis gyrata (CVG) is a skin deformity characterized by excessive growth of the skin of the scalp, resulting in furrows and folds which are similar to gyri of the brain cortex. CVG can be classified into two forms: 'primary' (essential and non-essential) and 'secondary'. The primary non-essential type is associated with the neurological and ophthalmological changes, while the primary essential form does not present any other disorder except for the cutaneous alterations. Secondary forms of CVG are much more frequent and accompany other pathologies. The treatment consists of local hygiene and surgical resection of skin excess. Intradermal cerebriform nevus is a rare entity, predominantly affecting females at birth or during the first years of life. The condition usually begins with a macular lesion that develops into parietal and occipital symmetric convolutions, occupying one half to three quarters of the scalp6, 7. It has a growth peak at puberty. The mechanism for the cerebriform pattern on the surface of the intradermal cerebriform nevus is unknown. 12

INTRODUCTION-Reproductive tract infections (RTIs) and sexually transmitted infections (STIs) represent a major public health problem in developing countries, so their quantification and study is very important. METHODOLOGY-Case records of 231 patients with RTIs and STIs who attended the Department of Dermatology and Venereology, Mahatma Gandhi Medical College and Hospital, Jaipur for a period of seven months from March 2011 to April 2013 were analysed. RESULTS- The total number of cases studied were 231, out of which 137 cases ( 59%) were males and 94 cases (41%) were females , with a ratio of males to females of 1.4 : 1. CONCLUSION -Male to female sex ratio came out almost equal (1.4:1). Majority of females were married 85 cases (90.42%) and almost half of the males were also married 66 cases (48.17%). Viral STIs constitute the major burden of the STI clinic and enhance the susceptibility of an individual to acquire or transmit HIV through sexual contact. Syndromic algorithms have some shortcomings, and they need to be periodically reviewed and adapted to the epidemiological patterns of STI in a given setting.

Aims: Enhancement of cholinergic activity and reduction of oxidative stress by scavenging free radicals such as nitric oxide are well recognized therapeutic approaches in several pathological conditions. We evaluated the anticholinesterase, antioxidant and nitric oxide scavenging activity of the aqueous extracts of Ocimum basilicum, Curcuma

Senile cataract is the most common cause of bilateral blindness and results from the loss of transparency of the lens. Maintenance of the unique tissue architecture of the lens is vital for keeping the lens transparent. Membrane transport mechanisms utilizing several magnesium (Mg)-dependent ATPases, play an important role in maintaining lens homeostasis. Therefore, in Mg-deficiency states, ATPase dysfunctions lead to intracellular depletion of K(+) and accumulation of Na(+) and Ca(2+). High intracellular Ca(2+) causes activation of the enzyme calpain II, which leads to the denaturation of crystallin, the soluble lens protein required for maintaining the transparency of the lens. Mg deficiency also interferes with ATPase functions by causing cellular ATP depletion. Furthermore, Mg deficiency enhances lenticular oxidative stress by increased production of free radicals and depletion of antioxidant defenses. Therefore, Mg supplementation may be of therapeutic value in preventing the onset and progression of cataracts in conditions associated with Mg deficiency.