Sterol O-acyltransferase (SOAT/ACAT) activity is required to form cholesterol crystals in hepatocyte lipid droplets.

OBJECTIVE: Excess cholesterol storage can induce the formation of cholesterol crystals in hepatocyte lipid droplets. Such crystals distinguish metabolic dysfunction associated steatohepatitis (MASH) from simple steatosis and may underlie its pathogenesis by causing cell damage that triggers liver inflammation. The mechanism linking cholesterol excess to its crystallization in lipid droplets is unclear. As cholesteryl esters localize to and accumulate in lipid droplets more readily than unesterified free cholesterol, we investigated whether cholesterol esterification by sterol O-acyltransferase (SOAT), also known as acyl co-A cholesterol acyltransferase (ACAT), is required for hepatocyte lipid droplet crystal formation. METHOD: Cholesterol crystals were measured in cholesterol loaded Hep3B hepatocytes, RAW264.7 macrophages, and mouse liver using polarizing light microscopy. We examined the effect of blocking SOAT activity on crystal formation and compared these results to features of cholesterol metabolism and the progression to intracellular crystal deposits. RESULTS: Cholesterol loading of Hep3B cells caused robust levels of lipid droplet localized crystal formation in a dose- and time-dependent manner. Co-treatment with SOAT inhibitors and genetic ablation of SOAT1 blocked crystal formation. SOAT inhibitor also blocked crystal formation in low density lipoprotein (LDL) treated Hep3B cells, acetylated LDL treated RAW 264.7 macrophages, and in the liver of mice genetically predisposed to hepatic cholesterol overload and in mice with cholesterol enriched diet-induced MASH. CONCLUSION: SOAT1-mediated esterification may underlie cholesterol crystals associated with MASH by concentrating it in lipid droplets. These findings imply that inhibiting hepatocyte SOAT1 may be able to alleviate cholesterol associated MASH. Moreover, that either a lipid droplet localized cholesteryl ester hydrolase is required for cholesterol crystal formation, or the crystals are composed of cholesteryl ester.

An eight-week single-blind randomised controlled trial on dual task balance training and motor imagery practice in ischaemic stroke survivors.

The purpose of this study was to find the effects of motor imagery on balance and mobility of stroke patients.This was a single-blind randomised controlled trial which included a total of 30 patients. These patients were assigned into two group: experimental (n=15) and control (n=15) by sealed envelope method. Functional Reach Test (FRT), Time Up and Go Test (TUGT), Berg Balance Scale (BBS), and Movement Imagery Questionnaire (MIQ) were used as assessment tools. The overall mean age of the participants was 52.63 ±8.78 years. Between groups analysis showed non-statistically significant difference (p>0.05) in FRT but significant difference in TUGT, BBS, and MIQ-3 (p<0.009) after eight weeks of training. Within group analysis showed statistically significant difference (p<0.05) for all outcome measures. This study concluded that motor imagery is an effective treatment strategy for improving balance and mobility in stroke patients and brings better results as compared to conventional treatments.

Frequency of migraine and its associated triggers and relievers among medical students of Lahore: a cross-sectional study.

Introduction: Headache is a widely prevalent illness that negatively impacts people's lives, leaving them functionally incapable of performing regular everyday tasks. The global burden of headache is 40%, of which migraine accounts for 47%. Objective: This study aims to determine the frequency of migraine among medical students of Lahore and its characteristics, associated triggers, and relievers among medical students. Methodology: A cross-sectional study was conducted in medical colleges after obtaining ethical approval. Migraine was diagnosed using the criteria provided in the International Classification of Headache Disorders, Third Edition (ICHD-3), and data on triggers, relief, and demographics were collected. The information is analyzed using SPSS 22. Result: The data were collected from 522 medical students. The average age of the students was 21.3 ± 2.0 SD (in years). About 146 (28.0%) of the medical students have migraine according to ICHD-3 criteria and were diagnosed initially. Gender was insignificantly associated with migraine (P-value=0.32). Students with psoriasis, hypertension, and polycystic ovarian syndrome were statistically significantly related to migraines with a P-value=0.002. Dehydration is the most frequent aggravator, and adequate sleep is the most frequent reliever of migraine. Conclusion: The findings show a high frequency of migraine. They are similar in both genders, depicting that stressful lifestyles, inadequate sleep patterns, and bizarre dietary habits make them more prone to migraine episodes. So, further, detailed studies should be done on evaluating triggers and relievers of migraine and their interrelations with migraine so we can focus on preventive strategies, diagnosis, and treatment of migraine.

Acid sphingomyelinase deactivation post-ischemia promotes brain angiogenesis and remodeling by small extracellular vesicles.

Antidepressants have been reported to enhance stroke recovery independent of the presence of depressive symptoms. They have recently been proposed to exert their mood-stabilizing actions by inhibition of acid sphingomyelinase (ASM), which catalyzes the hydrolysis of sphingomyelin to ceramide. Their restorative action post-ischemia/reperfusion (I/R) still had to be defined. Mice subjected to middle cerebral artery occlusion or cerebral microvascular endothelial cells exposed to oxygen-glucose deprivation were treated with vehicle or with the chemically and pharmacologically distinct antidepressants amitriptyline, fluoxetine or desipramine. Brain ASM activity significantly increased post-I/R, in line with elevated ceramide levels in microvessels. ASM inhibition by amitriptyline reduced ceramide levels, and increased microvascular length and branching point density in wildtype, but not sphingomyelinase phosphodiesterase-1 ([Smpd1]-/-) (i.e., ASM-deficient) mice, as assessed by 3D light sheet microscopy. In cell culture, amitriptyline, fluoxetine, and desipramine increased endothelial tube formation, migration, VEGFR2 abundance and VEGF release. This effect was abolished by Smpd1 knockdown. Mechanistically, the promotion of angiogenesis by ASM inhibitors was mediated by small extracellular vesicles (sEVs) released from endothelial cells, which exhibited enhanced uptake in target cells. Proteomic analysis of sEVs revealed that ASM deactivation differentially regulated proteins implicated in protein export, focal adhesion, and extracellular matrix interaction. In vivo, the increased angiogenesis was accompanied by a profound brain remodeling response with increased blood-brain barrier integrity, reduced leukocyte infiltrates and increased neuronal survival. Antidepressive drugs potently boost angiogenesis in an ASM-dependent way. The release of sEVs by ASM inhibitors disclosed an elegant target, via which brain remodeling post-I/R can be amplified.

Tocilizumab and Cytokine Release Syndrome in COVID-19 Pneumonia: Experience From a Single Center in Pakistan.

Background Tocilizumab (TCZ), an interleukin-6 (IL-6) receptor blocker, emerged as a treatment for cytokine release syndrome (CRS) in patients with severe COVID-19 pneumonia. The main objective of the study is to discuss the treatment response of TCZ in severe and critically ill patients with COVID-19 pneumonia. Patient demographics, laboratory parameters before and after TCZ therapy, and clinical outcomes in 20 patients in a single center were prospectively reviewed. Results Out of 120 patients, 96 (80%) were males and 24 (20%) were females. Only eight (10%) patients did not have any previously known comorbidity. There were 78 (65%) patients with severe disease, while 42 (35%) have critically severe disease. Of the 120 patients, only 36 required a second dose of TCZ in our study based on clinical background. Neutrophils and C-reactive protein (CRP) levels were observed to be raised in all patients, while lymphopenia was observed in 114/120, and D-dimer levels were elevated in 102 (85%) patients. After the second dose of tocilizumab, 102 (85%) patients reduced oxygen requirement within four days, and 14 patients were removed on the second dose of tocilizumab on clinical grounds. Of these 120 patients, in two weeks, 30 (25%) were discharged. Within three weeks, 60 of them were discharged, while 12 were discharged after three weeks, and 18 patients died in our study despite treatment. Conclusion TCZ appeared to be a good treatment option in patients with CRS and severe and critical pneumonia, and for patients with raised IL-6 levels despite single TCZ therapy, a repeat dose is recommended.

Correlation between Internet addiction, depression, anxiety and stress among undergraduate medical students in Azad Kashmir.

OBJECTIVE: To find out the correlation between internet addiction and depression, anxiety, and stress among undergraduate medical students in Azad Kashmir. METHODS: A cross-sectional study including 210 undergraduate medical students (first to the fifth year) was done in Poonch Medical College, Azad Kashmir. The data collection tools were DASS21 questionnaire and Young's internet addiction questionnaire. Spearman rank correlation test was done to see the correlation between internet addiction and depression, anxiety, and stress. Data were analyzed by SPSS v23 at a 95% confidence interval. RESULTS: A very high prevalence (52.4%) of moderate to extremely severe internet addiction was observed among the respondents. The mild positive correlation between internet addiction and depression was identified (p <.001) and similar type of correlation was observed between internet addiction and stress (p .003). However, anxiety and internet addiction were not significantly correlated. The prevalence of anxiety and depression among the males were higher than the females, whilst the stress level was almost the same across gender. CONCLUSION: Internet addiction has been found to be associated with various psychiatric diseases. In this study, we also observed such correlation. We have also observed a very high level of internet addiction among medical students. The prevalence of internet addiction may further increase in the coming years as the internet will become more cheap, available and include more high quality psychologically addictive contents.

Review: Oncogenic Insertional Mutagenesis as a Consequence of Retroviral Gene Therapy for X-Linked Severe Combined Immunodeficiency Disease.

For X-linked severe combined immunodeficiency (SCID-X1), the practice of gene therapy has revealed an unusual effect: insertional mutagenesis that can lead to leukemia. Even though incorporation of the retrovirus close to the oncogene for T-cell acute lymphoblastic leukemia (T-ALL), LIM-only protein 2 (LMO2) is observed frequently, but it is not clear why LMO2 expression is affected. It was demonstrated that in all the typical T-ALL oncogenes, there is mainly transcription of LMO2 in CD34+ progenitor cells. LYL1, TAN1, and TAL1 are very important intensification factors that are classically used in the gene therapy for copying LMO2 when they are stimulated. For this reason, oncogenes are susceptible to amalgamation with viruses. The IL-2R-gamma (IL-2 receptor γ chain) was found to be a supporting oncogene to LMO2. Nevertheless it was illustrated that excessive expression of IL-2R-gamma did not affect T-cell growth. In comparison to it, the excessive expression of LMO2 in CD34+ cells can cause ongoing increases in the development of T cells. Conversely, there is no effect on the development of B cells and myeloid cells. This information helps explain why LMO2 is mostly affected by various identified T-ALL oncogenes. In addition, throughout the process of T-cell development, expression of IL2R-gamma mediated by retrovirus may not always be oncogenic. As an alternative, replacement of signals of common IL-7 receptors may increase development of T cells wherever LMO2 was expressed and caused abnormal thymocyte development.

TSG101: Tumor Susceptibility Gene 101 (tsg101) Product-Role in Therapy Against HIV/AIDS.

HIV infection presents a major community health hazard, partially because the HIV virus is capable of evading antiretroviral therapies. Most anti-HIV drugs were intended to target virus-encoded mechanisms; however, some host-encoded molecules comparatively execute a vital role in the life cycle of virus. Thus, these might be considered as target sites for antiviral agents. TSG101 is important among these antiviral therapies because, as a cytoplasmic molecule, it facilitates viral budding and release. In this review, HIV-infected cells have TSG101 on their surface and thus can be used in antibody-based therapies. The development of a monoclonal antibody CB8-2 lessens the assembly of viruses from infected cells. This mechanism represents the potential use of TSG101-directed antibodies to fight against AIDS.

Availability of expanded programme of immunization services provided to children in a rural Pakistani village.

OBJECTIVES: To evaluate the immunization status of children under 5 years age, living in the Nurpur Shahan area and to identify various factors which influenced the rate of immunization. METHODS: A cross sectional study was conducted in which mothers of children under 5 years of age were questioned about the immunization of their children and the immunization services provided in the area. Systematic random sampling was used and the collected data was analyzed using SPSS v. 10.0. RESULTS: The overall immunization rate in the children of Nurpur Shahan was 77.4%. This study found strong correlations to immunization rates in children. Increased educational status of the mother, greater maternal immunization during pregnancy, and home delivery of vaccines, were all directly linked to higher rates of immunization amongst children. Lack of awareness, followed by inadequate facilities were the most commonly cited reasons for discontinuing the programme or abstaining from it altogether. CONCLUSION: The EPI coverage in Nurpur Shahan is quite well established, but still left room for improvement. Factors that encouraged higher immunization rates were more educated mothers, better awareness and availability of door-to-door services.