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TSC renal cystic disease is poorly understood and has no approved treatment. In a new principal cell-targeted murine model of Tsc cystic disease, the renal cystic epithelium is mostly composed of type A intercalated cells with an intact Tsc2 gene confirmed by sequencing, although these cells exhibit a Tsc-mutant disease phenotype. We used a newly derived targeted murine model in lineage tracing and extracellular vesicle (EV) characterization experiments and a cell culture model in EV characterization and cellular induction experiments to understand TSC cystogenesis. Using lineage tracing experiments, we found principal cells undergo clonal expansion but contribute very few cells to the cyst. We determined that cystic kidneys contain more interstitial EVs than noncystic kidneys, excrete fewer EVs in urine, and contain EVs in cyst fluid. Moreover, the loss of the Tsc2 gene in EV-producing cells greatly changes the effect of EVs on renal tubular epithelium, such that the epithelium develops increased secretory and proliferative pathway activity. We demonstate that the mTORC1 pathway activity is independent form the EV production, and that the EV effects for a single cell line can vary significantly. TSC cystogenesis involves significant contribution from genetically intact cells conscripted to the mutant phenotype by mutant cell derived EVs.

Ulcerative colitis is a chronic inflammation of the colonic mucosa. Gum Arabic (GA) has been reported to exert anti-inflammatory and antifibrotic activity. This study aimed to evaluate the effect o...

During the last decade electronic cigarettes (e-cigarettes) have been studied as an alternative devices to the tobacco cigarette, but with better safety for the health of smokers, so as to create a new approach to smoking addiction, such as the “smoking harm reduction”. This new approach, suggested by a part of the scientific world, aroused interest and debates in the regulatory field, involving all the major regulatory bodies and often creating divergences from nation to nation on the rules driving the production, distribution and consumption of these alternative products. Many studies have been conducted both in vitro and in vivo, to clarify the effects of the e-cigarette compared to the classic one. In this context, the Center of Excellence for the Acceleration of HArm Reduction (CoEHAR) was established within the University of Catania (Italy) and the multi-center project, created under its leadership, the REPLICA project, which aims to replicate in vitro studies originally conducted by tobacco and e-cigarette manufacturers, in order to verify the robustness and replicability of the data. In this work the REPLICA Team replicated part of the work published by Rudd and colleagues in 2020, which aims to establish the aerosol-induced cytotoxicity, mutagenesis and genotoxicity of a pod system e-cigarette aerosol compared to tobacco cigarette smoke. As in the original paper, we performed Neutral Red Test (NRU) for the evaluation of cytotoxicity, AMES test for the evaluation of mutagenesis and In Vitro Micronuclei (IVM) assay for the evaluation of genotoxicity on cells treated with cigarette smoke or e-cigarette aerosol. The results obtained showed high cytotoxicity, mutagenicity and genotoxicity induced by cigarette smoke, but slight or no cytotoxic, mutagenic and genotoxic effects induced by the e-cigarette aerosol. The data obtained support those previously presented by Rudd and colleagues, although we have highlighted some methodological flaws of their work. Overall, we can affirm that the results obtained by Rudd and colleagues have been established and our data also confirm the idea that e-cigarette aerosol is much safer and less harmful than cigarette smoking, making it a useful device in smoking harm reduction.

Next-generation sequencing, such as whole-exome sequencing (WES), is increasingly used in the study of Mendelian disorders, yet many are reported as “negative.” Inappropriate variant annotation and filtering steps are reasons for missing the molecular diagnosis. Noncoding variants, including splicing mutations, are examples of variants that can be overlooked. Herein, we report a family of four affected newborns, and all presented with severe congenital microcephaly. Initial research WES analysis identified a damaging homozygous variant in NME1 gene as a possible cause of primary microcephaly phenotype in these patients. However, reanalysis of the exome data uncovered a biallelic splice site variant in asparagine synthetase gene which seems to be the possible cause of the phenotype in these patients. This study highlights the importance of revisiting the exome data and the issue of “negative” exome and the afterward approaches to identify and prove new candidate genes.

Tuberous sclerosis complex (TSC) is associated with TSC1 or TSC2 gene mutations resulting in hyperactivation of the mTORC1 pathway. This mTORC1 activation is associated with abnormal tissue development and proliferation such that in the kidney there are both solid tumors and cystic lesions. This review summarizes recent advances in tuberous sclerosis complex nephrology and focuses on the genetics and cell biology of tuberous sclerosis complex renal disease, highlighting a role of extracellular vesicles and the innate immune system in disease pathogenesis.

Background Sleep patterns have changed continuously worldwide and it can be influenced by social, cultural, and environmental factors. These patterns may be associated with poor sleep quality and daytime sleepiness. The aim of the study was to investigate sleep patterns and quality in Omani adults using actigraphy. Subjects and methods This was a cross-sectional study conducted between June 2015 and February 2017. Four hundred subjects agreed to participate in the study (52% male, 48% female). Subjects were randomly selected and enrolled in the study among young adults and middle aged individuals living in the City of Muscat. Subjects were asked to fill-in Epworth sleepiness scale (ESS) and Pittsburgh sleep quality index (PSQI). Actigraphy was used to measure their sleep patterns for one week. Results The mean age of participants was 32.80±11.50 years. Four sleep patterns were identified: monophasic, bi-phasic (post-dawn), bi-phasic (afternoon siesta), and polyphasic (three sleep periods/24 hours). The study revealed that 35% of participants had biphasic-siesta sleep pattern, 28% polyphasic, 26% monophasic, and 11% biphasic-dawn. The biphasic siesta pattern was found to be associated with younger age group (25-34 years) (P=0.001). Polyphasic sleep was associated with higher ESS score (P=0.001) but not with poor sleep quality (P=0.24). There was no significant difference in night sleep duration among all the sleep patterns (P=0.07) but the polyphasic sleep pattern had higher total 24-hour day sleep duration (P=0.03). Nearly 90% of participants practiced afternoon siestas with mean duration of 45±43 minutes. Conclusion The predominant sleep pattern among Omanis was biphasic-siesta and majority of people practiced afternoon siesta. Polyphasic sleep pattern is associated with daytime sleepiness.

Background: Serum creatinine and estimated glomerular filtration rate (eGFR) are keys to assess kidney function and suggest a propensity for development of renal failure. Genetic factors are reported in different ethnic groups to play a role in the variability of these phenotypes. In this study, we investigate the heritability and quantitative trait loci (QTL) of serum creatinine and eGFR. Multiple phenotypes were collected from a multi-generation pedigree of 281 subjects form Oman family study. Genotype analysis was used to calculate heritability and linkage was performed using variance components decomposition-based methods implemented in SOLAR. Results: The multivariate-adjusted heritability estimates for serum creatinine and eGFR were 0.70 and 0.63 (p-value 2.5x10-11 and 1.8x10-11), respectively. Genome-wide linkage analysis showed significant loci with LOD score ≥2 at chromosomes 9p21.1, 9p21.3, 15p26.3 and 16.p13.3. Functional annotation identified angiopoietin 1-TEK pathway as a candidate pathway where TEK is associated with chronic kidney disease and expressed in renal glomeruli. Conclusion: This study showed high heritability of serum creatinine and eGFR with significant QTLs in chromosomes 9, 15 and 16. Further research is needed to study the genetic association studies of angiopoietin 1-TEK axis with glomerulopathies like type 2 diabetes and to predict and diagnose progression of renal diseases.

The incidence of connective tissue diseases such as systemic lupus erythematous (SLE), in adult patients with sickle cell disease (SCD), appears to be increasing. The exact causes underlying this increased risk are still unknown, but a link with B regulatory (Breg) cells is possible as these cells suppress inflammatory responses, and maintain tolerance. Quantitative and qualitative analyses of circulating Breg cells were performed in a cohort of SCD patients with SLE, and their levels were correlated with key soluble mediators promoting autoreactive B cells. We demonstrated that levels of Breg cells were significantly decreased in SCD patients with SLE compared to patients with SCD only or healthy controls. Functional analysis of Breg cells from SCD patients with SLE revealed impairments in IL-10 production that correlated with lower levels of STAT3 phosphorylation, without abnormal expression of IL-10 receptor on Breg cells. On the other hand, BAFF levels were substantially elevated in SCD patients with SLE, but not significantly associated with Breg cell levels. Collectively, these results indicated numerical and functional deficits of Breg cells in SCD patients with SLE and their capacity to maintain tolerance and control inflammation is imbalanced, which leads to the development of autoimmune responses.

Mucolipidosis Type IV (MLIV) is caused by a deficiency of the mucolipin cation channel encoded by Mucolipin TRP Cation Channel 1 gene (

To identify the biochemical, hematological, and immune biomarkers in COVID-19 patients on admission that are predictive of eventual admission to the intensive care unit (ICU).This retrospective cohort study was conducted on all confirmed COVID-19 cases hospitalized at Royal Hospital, Oman from 24 February to 30 July 2020. The demographic, clinical, and laboratory data were collected from the hospital information system. Patients were divided into two groups: non-ICU admitted group and ICU admitted group.Out of 445 patients, 276 (62.0%) were male and 169 (38.0%) were female; 259 (58.2%) patients were admitted to COVID-19 general wards whereas 186 (41.8%) were admitted to ICU. Admission to ICU was more likely when patient had the following comorbidities: diabetes (OR = 1.8; 95% CI: 1.3-2.7), liver diseases (OR = 2.1; 95% CI: 1.1-4.3), and respiratory diseases (OR = 2.0; 95% CI: 1.1-3.7). Between ICU and non-ICU patients, there were significant differences in on-admission laboratory blood/serum parameters: total white blood cells (WBCs) count, lymphocytes count, C-reactive protein (CRP), ferritin, corrected calcium, interleukin 6 (IL-6), D-dimer, alanine transaminase (ALT), lactate dehydrogenase (LDH), albumin, and troponin.The current study identified the presence of the comorbidities (i.e., diabetes, liver diseases, and respiratory diseases) and on-admission laboratory blood and serum test results (i.e., WBC, lymphocytes, CRP, ferritin, corrected calcium, IL-6, D-dimer, ALT, LDH, albumin, and troponin) that are associated with ICU admission.

To describe the effect of seasonal variations on sleep patterns in a hot climate Arab region.This is a cross-sectional study that included healthy Omani subjects of both genders between ages 18 and 59 years. Data for sleep pattern identification in summer and winter were collected from participants using an actigraphy wristband.Among 321 participants, in summer seasons, a polyphasic sleep pattern (40%) prevailed over other sleep patterns (P0.001). While in the winter season, monophasic sleep (31%) was the dominant pattern (P0.001). Subjects slept longer during the winter seasons with total hours of sleep during the day 48 min longer than in the summer, though the difference was not statistically significant (P0.05), while siesta duration in the summer was significantly longer (13 min, P0.01). In summer, the sleep quality was good (PSQI ≤ 5); however, it was poor (PSQI5) in winter (P0.05). Night sleep duration, daytime sleepiness, and sleep latency were not statistically different between the summer and winter seasons.Sleep patterns may be influenced by seasonal changes. A polyphasic sleep pattern prevailed in summer while a monophasic pattern was the predominant sleep pattern in winter. In summer, the sleep quality was good and the siesta duration was longer compared to the winter.

Objectives: To identify the biochemical, hematological, and immune biomarkers in COVID-19 patients on admission that are predictive of eventual admission to the intensive care unit (ICU). Methods: This retrospective cohort study was conducted on all confirmed COVID-19 cases hospitalized at Royal Hospital, Oman from 24 February to 30 July 2020. The demographic, clinical, and laboratory data were collected from the hospital information system. Patients were divided into two groups: non-ICU admitted group and ICU admitted group. Results: Out of 445 patients, 276 (62.0%) were male and 169 (38.0%) were female; 259 (58.2%) patients were admitted to COVID-19 general wards whereas 186 (41.8%) were admitted to ICU. Admission to ICU was more likely when patient had the following comorbidities: diabetes (OR = 1.8; 95% CI: 1.3–2.7), liver diseases (OR = 2.1; 95% CI: 1.1–4.3), and respiratory diseases (OR = 2.0; 95% CI: 1.1–3.7). Between ICU and non-ICU patients, there were significant differences in on-admission laboratory blood/serum parameters: total white blood cells (WBCs) count, lymphocytes count, C-reactive protein (CRP), ferritin, corrected calcium, interleukin 6 (IL-6), D-dimer, alanine transaminase (ALT), lactate dehydrogenase (LDH), albumin, and troponin. Conclusions: The current study identified the presence of the comorbidities (i.e., diabetes, liver diseases, and respiratory diseases) and on-admission laboratory blood and serum test results (i.e., WBC, lymphocytes, CRP, ferritin, corrected calcium, IL-6, D-dimer, ALT, LDH, albumin, and troponin) that are associated with ICU admission.

BACKGROUND: PTRHD1 was proposed as a disease‐causing gene of intellectual disability, spasticity, and parkinsonism. OBJECTIVES: To characterize the clinical phenotype and the molecular cause of intellectual disability in four affected individuals of a consanguineous family. METHODS: Clinical evaluation, whole‐exome sequencing, Sanger sequencing, reverse transcription polymerase chain reaction (PCR), real‐time PCR, immunoblot, and isoelectric focusing. RESULTS: A homozygous 28‐nucleotide frameshift deletion introducing a premature stop codon in the PTRHD1 exon 1 was identified in the four affected members. We further confirmed the apparent transcript escape of the nonsense‐mediated messenger RNA (mRNA) decay pathway. Real‐time PCR showed that mRNA expression of the mutant PTRHD1 is higher compared to the wild‐type. Western blotting and isoelectric focusing identified a truncated, but stable mutant PTRHD1 protein expressed in the patient's primary cells. CONCLUSIONS: We provide further evidence that PTRHD1 mutations are associated with autosomal‐recessive childhood‐onset intellectual disability associated with spasticity and parkinsonism.

Objective: Studies from the past decades have shown that mood disorders are common during childhood and adolescence. This study aimed to estimate the point prevalence of depression in Omani children and adolescents during social distancing and lockdown and identify the risk factors for developing depressive symptoms during the COVID-19 pandemic.Methods: This is an analytical cross-sectional study conducted in May 2020, in which all young Omani people attending a mainstream school aged 8–18 years old were eligible to participate. Parents were asked to complete the online survey, which consisted of the parent version of the Mood and Feelings Questionnaire (MFQ-Parent). In addition, the option of a self-reported version (MFQ-Self) was provided in cases where children preferred to fill out the survey themselves. Logistic regression was used to identify the contributing socio-demographic variables associated with depressive symptoms.Results: A total of 445 participants completed the MFQ, out of which 72.1% were parents, and 27.9% were children, adolescents and young people. 13.9% of children and adolescents exhibited depressive symptoms during the COVID-19 pandemic in Oman. The presence of depressive symptoms was associated with increased food intake (OR 1.81, 95% CI 1.00–3.29, p-value p-value p-value Conclusion: This study from Oman concurs with recent reports of depression being common during the COVID-19 pandemic. Concerted efforts are needed to mitigate this trend and identify high-risk groups during the lockdown period.

Aim: To determine the prevalence, genetic characteristics, current management and outcomes of familial hypercholesterolaemia (FH) in the Gulf region. Methods: Adult (18-70 years) FH patients were recruited from 9 hospitals and centres across 5 Arabian Gulf countries. The study was divided into 4 phases and included patients from 3 different categories. In phase 1, suspected FH patients (category 1) were collected according to the lipid profile and clinical data obtained through hospital record systems. In phase 2, patients from category 2 (patients with a previous clinical diagnosis of FH) and category 1 were stratified into definitive, probable and possible FH according to the Dutch Lipid Clinic Network criteria. In phase 3, 500 patients with definitive and probable FH from categories 1 and 2 will undergo genetic testing for 4 common FH genes. In phase 4, these 500 patients with another 100 patients from category 3 (patients with previous genetic diagnosis of FH) will be followed for 1 year to evaluate clinical management and cardiovascular outcomes. The Gulf FH cohort was screened from a total of 34,366 patients attending out-patient clinics. Results: The final Gulf FH cohort consisted of 3,317 patients (mean age: 47±12 years, 54% females). The number of patients with definitive FH is 203. In this initial phase of the study, the prevalence of (probable and definite) FH is 1/232. Conclusion: The prevalence of FH in the adult population of the Arabian Gulf region is high. The Gulf FH registry, a first-of-a-kind multi-national study in the Middle East region, will help in improving underdiagnosis and undertreatment of FH in the region.

The endothelium contributes to the maintenance of vasodilator tone by releasing endothelium-derived relaxing factors, including nitric oxide (NO). In hypertension, endothelial nitric oxide synthase (eNOS) produces less NO and could be one of the contributing factors to the increased peripheral vascular resistance. Agonist-induced Ca(2+) entry is essential for the activation of eNOS. The transient receptor potential vanilloid type 4 (TRPV4) channel, a Ca(2+)-permeant cation channel, is expressed in the endothelial cells and involved in the regulation of vascular tone. The present study aimed to investigate the role of TRPV4 channel in endothelium-dependent NO-mediated relaxation of the resistance artery in hypertensive rats. Using a wire myograph, relaxation response to the TRPV4 activator, 4alpha-phorbol-12,13-didecanoate (4alphaPDD) was assessed in mesenteric arteries obtained from Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHRs). Compared to WKY, SHR demonstrated a significantly attenuated 4alphaPDD-induced endothelium-dependent NO-mediated relaxation. Immunohistochemical analysis revealed positive staining for TRPV4 in the endothelium of mesenteric artery sections in both WKY and SHR. Furthermore, TRPV4 mRNA and protein expressions in SHR were significantly lower than their expression levels in WKY rats. We conclude that 4alphaPDD-induced endothelium-dependent NO-mediated vasorelaxation is reduced in SHR and downregulation of TRPV4 could be one of the contributing mechanisms.

In tuberous sclerosis complex (TSC), Tsc2 mutations are associated with more severe disease manifestations than Tsc1 mutations and the role of extracellular vesicles (EVs) in this context is not yet studied. We report a comparative analysis of EVs derived from isogenic renal cells except for Tsc1 or Tsc2 gene status and hypothesized that in spite of having similar physical characteristics, EVs modulate signaling pathways differently, thus leading to TSC heterogenicity. We used mouse inner medullary collecting duct (mIMCD3) cells with the Tsc1 (T1G cells) or Tsc2 (T2J cells) gene disrupted by CRISPR/CAS9. EVs were isolated from the cell culture media by size-exclusion column chromatography followed by detailed physical and chemical characterization. Physical characterization of EVs was accessed by tunable resistive pulse sensing and dynamic light scattering, revealing similar average sizes and zeta potentials (at pH 7.4) for EVs from mIMCD3 (123.5 ± 5.7 nm and −16.3 ± 2.1 mV), T1G cells (131.5 ± 8.3 nm and −19.8 ± 2.7 mV), and T2J cells (127.3 ± 4.9 nm and −20.2 ± 2.1 mV). EVs derived from parental mIMCD3 cells and both mutated cell lines were heterogeneous (>90% of EVs < 150 nm) in nature. Immunoblotting detected cilial Hedgehog signaling protein Arl13b; intercellular proteins TSG101 and Alix; and transmembrane proteins CD63, CD9, and CD81. Compared to Tsc2 deletion, Tsc1 deletion cells had reduced EV production and release rates. EVs from Tsc1 mutant cells altered mTORC1, autophagy, and β-catenin pathways differently than EVs from Tsc2-mutated cells. Quantitative PCR analysis revealed the down regulation of miR-212a-3p and miR-99a-5p in EVs from Tsc2-mutated cells compared to EVs from Tsc1-mutant cells. Thus, EV-derived miR-212-3p and mIR-99a-5p axes may represent therapeutic targets or biomarkers for TSC disease.

Objectives: Although typically considered a condition affecting children, scientific evidence has shown that 30–50% of those diagnosed with ADHD in childhood continue to suffer from this condition after the age of 18. This study sought to address the gap in the literature, and to describe the socio-demographic and clinical characteristics of ADHD patients in a sample of adult patients attending an outpatient clinic at a tertiary care hospital in Oman, and to evaluate their association with the different subtypes of the disorder. Methods: Data from adult patients with ADHD from the outpatient clinic at Sultan Qaboos University Hospital (SQUH), Muscat, Oman, were retrospectively collected from medical records from January 2018 to April 2020. Socio-demographic characteristics, clinical profiles and psychiatric co morbidities were examined. Results: This study included 100 adults who fulfilled the standard diagnosis of ADHD, with 54% (n=54), and 46% (n=46) from the inattentive and combined subtypes, respectively. It was found that ADHD was more prevalent among males (64.0%) compared to females (36%), with the inattentive subtype being more predominant among females. The ADHD patients with the inattentive subtype were associated with co-morbid substance use disorders (OR=11.29, P = 0.049), personality disorders (OR=7.96, P = 0.017), and major depressive disorder (OR=15.94, P = 0.002) compared to patients with the predominantly combined subtype. Conclusion: This study from Oman echoes the findings from the current literature, that adult patients with ADHD commonly have co-morbid psychiatric disorders, leading to significant functional impairment. Psychiatric co-morbidities must be identified and urgently treated, to allow for better clinical and functional outcomes in adult patients with ADHD. Keywords: ADHD, co-morbidity, hyperactivity, Oman, psychiatry

Cigarette smoking is the leading cause of preventable deaths worldwide. Electronic Nicotine Delivery Systems (ENDS) may reduce health risks associated with chronic exposure to smoke and their potential benefits have been the matter of intense scientific debate. Here we replicated three key published studies from the Tobacco Industry on cytotoxic and inflammatory effects of cigarette smoke and ENDS aerosol in an independent multicentric study. We aimed to establish the reliability of results and the robustness of conclusions by replicating the authors’ experimental protocols and further validating them with different techniques. We exposed human bronchial epithelial cell (NCI-H292) to cigarette smoke and to aerosol from ENDS. All the exposure were conducted at air-liquid interface to assess cytotoxicity effects of smoke and aerosol. Moreover, we aimed to assess different inflammatory mediators release (IL-6, IL-8 and MMP-1) from cells exposed to whole smoke and to smoke without particulate matter (vapor phase). We were able to replicate the results obtained in the original studies on cytotoxicity confirming that almost 80% of the cytotoxic effect of smoke is due to the vapor phase of smoke. Moreover, our results substantiated the reduced cytotoxic effects of ENDS aerosol in respect to cigarette smoke. However, our data are significantly different from the original ones in terms of inflammatory and remodeling activity triggered by smoke. Taken all together, the data obtained independently in different laboratories clearly demonstrate the reduced toxicity of ENDS products compared to cigarettes and thus providing a valuable tool to the harm reduction strategies in smokers. Competing Interest Statement Riccardo Polosa is full tenured professor of Internal Medicine at the University of Catania (Italy) and Medical Director of the Institute for Internal Medicine and Clinical Immunology at the same University. In relation to his recent work in the area of respiratory diseases, clinical immunology, and tobacco control, RP has received has received lecture fees and research funding from Pfizer, GlaxoSmithKline, CV Therapeutics, NeuroSearch A/S, Sandoz, MSD, Boehringer Ingelheim, Novartis, Duska Therapeutics, and Forest Laboratories. Lecture fees from a number of European EC industry and trade associations (including FIVAPE in France and FIESEL in Italy) were directly donated to vaper advocacy no-profit organizations. RP has also received grants from European Commission initiatives (U-BIOPRED and AIRPROM) and from the Integral Rheumatology & Immunology Specialists Network (IRIS) initiative. He has also served as a consultant for Pfizer, Global Health Alliance for treatment of tobacco dependence, CV Therapeutics, Boehringer Ingelheim, Novartis, Duska Therapeutics, ECITA (Electronic Cigarette Industry Trade Association, in the UK), Arbi Group Srl., and Health Diplomats. RP has served on the Medical and Scientific Advisory Board of Cordex Pharma, Inc., CV Therapeutics, Duska Therapeutics Inc, Pfizer, and PharmaCielo. RP is also founder of the Center for Tobacco prevention and treatment (CPCT) at the University of Catania and of the Center of Excellence for the acceleration of HArm Reduction (CoEHAR) at the same University, which has received support from Foundation for a Smoke Free World to conduct 8 independent investigator-initiated research projects on harm reduction. RP is also currently involved in the following pro bono activities: scientific advisor for LIAF, Lega Italiana Anti Fumo (Italian acronym for Italian Anti-Smoking League), the Consumer Advocates for Smoke-free Alternatives (CASAA) and the International Network of Nicotine Consumers Organizations (INNCO); Chair of the European Technical Committee for standardization on -Requirements and test methods for emissions of electronic cigarettes- (CEN/TC 437; WG4). Giovanni Li Volti is currently elected Director of the Center of Excellence for the acceleration of HArm Reduction. Konstantinos Poulas has received service grants and research funding from a number of Vaping Companies. He is the Head of the Institute of Research and Innovations, which has received a grant from the Foundation for a Smoke Free World. All the other authors declare no conflicts of interest., This investigator-initiated experimental research was produced with the help of a grant from the Foundation for a Smoke Free World. The funder had no role in the study design, or the writing of the experimental research. The contents, selection and presentation of facts, as well as any opinions expressed in the experimental research are the sole responsibility of the author and under no circumstances shall be regarded as reflecting the positions of the funder.

Ovarian cancer (OC) is characterized by a high morbidity and mortality, highlighting a great need for a better understanding of biological mechanisms that affect OC progression and improving its early detection methods. This study investigates effects of prolactin (PRL) on ovarian cancer cells, analyzes PRL receptors (PRLR) in tissue micro arrays and relates PRLR expression to survival of ovarian cancer. A database, composed of transcript profiles from OC, was searched for PRLR expression and results were put in relation to survival. Expression of PRLR in OC tissue sections and OC cell lines SKOV3, OV2008 and OVSAHO was assessed using immunohistochemistry, western blots and quantitative real-time PCR. The biological function of PRLR was evaluated by proliferation, colony formation and wound healing assays. Levels of PRLR mRNA are related to survival; in epithelial OC a high PRLR mRNA expression is related to a shorter survival. Analysis of a tissue micro array consisting of 84 OC showed that 72% were positive for PRLR immuno-staining. PRLR staining tended to be higher in OC of high grade tumors compared to lower grades. PRLR mRNA and protein can further be detected in OC cell lines. Moreover, in vitro treatment with PRL significantly activated the JAK/STAT pathway. PRLR expression is associated with OC survivals. PRL and its receptor may play an onco-modulatory role and promote tumor aggressiveness in OC. Alternatively, increased PRLR levels may form a base for the development of PRLR antagonist or PRLR antagonist-drug conjugate to increase selective uptake of anti-cancer drugs.

Background and aims Familial hypercholesterolemia (FH) is a common autosomal dominant disorder that can result in premature atherosclerotic cardiovascular disease (ASCVD). Limited data are available worldwide about the prevalence and management of FH. Here, we aimed to estimate the prevalence and management of patients with FH in five Arabian Gulf countries (Saudi Arabia, Oman, United Arab Emirates, Kuwait, and Bahrain). Methods The multicentre, multinational Gulf FH registry included adults (≥18 years old) recruited from outpatient clinics in 14 tertiary-care centres across five Arabian Gulf countries over the last five years. The Gulf FH registry had four phases: 1- screening, 2- classification based on the Dutch Lipid Clinic Network, 3- genetic testing, and 4- follow-up. Results Among 34,366 screened patient records, 3713 patients had suspected FH (mean age: 49±15 years; 52% women) and 306 patients had definite or probable FH. Thus, the estimated FH prevalence was 0.9% (1:112). Treatments included high-intensity statin therapy (34%), ezetimibe (10%), and proprotein convertase subtilisin/kexin type 9 inhibitors (0.4%). Targets for low-density lipoprotein cholesterol (LDL-C) and non-high-density lipoprotein cholesterol were achieved by 12% and 30%, respectively, of patients at high ASCVD risk, and by 3% and 6%, respectively, of patients at very high ASCVD risk (p Conclusions This snap-shot study was the first to show the high estimated prevalence of FH in the Arabian Gulf region (about 3-fold the estimated prevalence worldwide), and is a “call-to-action” for further confirmation in future population studies. The small proportions of patients that achieved target LDL-C values implied that health care policies need to implement nation-wide screening, raise FH awareness, and improve management strategies for FH.

To investigate the agreement in sleep pattern recording by self-reported sleep questionnaires and actigraphy in adults. This is a cross-sectional study. Men and women who met inclusion criteria were recruited for this study. The inclusion criteria were apparently healthy Omani nationals ages 19 to 50 years. Sleep questionnaires were randomly distributed in Muscat either directly or via electronic and paper announcements. Data were collected from the participants using the self-reported questionnaires with four piloted questions for sleep pattern identification and through the actigraphy wristband given to subjects to wear for a week. Cohen’s kappa test was performed for agreement analysis. A total of 964 Omani subjects between ages 18 and 59 years of both genders were recruited and completed the questionnaires successfully. Out of these, only 321 subjects wore the actigraphy wristband for 1 week (response rate = 33%). Agreement analysis reported a mild level of agreement for the monophasic (41%), moderate level for biphasic (59%), and good level for polyphasic (70%) sleep patterns. The overall agreement level of sleep patterns between the two methods was 57%. There is a low specificity of self-reported assessment in reporting sleep pattern. The average agreement level of subjective versus objective assessments of sleep patterns was moderate at 57% and self-reported sleep pattern is not specific. The study recommends the use of actigraphy along with sleep questionnaires for accurate assessment of sleep patterns in cohort studies.

The tuberous sclerosis complex (Tsc) proteins regulate the conserved mTORC1 growth regulation pathway. We identified that loss of the Tsc2 gene in mouse inner medullary collecting duct (mIMCD) cells induced a greater than two-fold increase in extracellular vesicle (EV) production compared to the same cells having an intact Tsc axis. We optimized EV isolation using a well-established size exclusion chromatography method to produce high purity EVs. Electron microscopy confirmed the purity and spherical shape of EVs. Both tunable resistive pulse sensing (TRPS) and dynamic light scattering (DLS) demonstrated that the isolated EVs possessed a heterogenous size distribution. Approximately 90% of the EVs were in the 100&ndash, 250 nm size range, while approximately 10% had a size greater than 250 nm. Western blot analysis using proteins isolated from the EVs revealed the cellular proteins Alix and TSG101, the transmembrane proteins CD63, CD81, and CD9, and the primary cilia Hedgehog signaling-related protein Arl13b. Proteomic analysis of EVs identified a significant difference between the Tsc2-intact and Tsc2-deleted cell that correlated well with the increased production. The EVs may be involved in tissue homeostasis and cause disease by overproduction and altered protein content. The EVs released by renal cyst epithelia in TSC complex may serve as a tool to discover the mechanism of TSC cystogenesis and in developing potential therapeutic strategies.

Several bioactive compounds from microalgae have demonstrated diverse biological activities with positive effects on human health. However, the potential of bioactive peptides as functional foods is still undervalued. Therefore, the exploration of microalgae strains as sources of bioactive peptides could reveal strong and unique bioactivities, especially when these marine sources have never been explored before. For this aim, protein extracts from six indigenous marine diatoms were subjected to enzymatic hydrolysis using four proteases (flavourzyme, pepsin, papain and trypsin). The hydrolysates were then tested for angiotensin converting enzyme (ACE)‐inhibitory, antioxidant and antihypertensive properties. Results showed that papain hydrolysates from all microalgae strains exhibited strong ACE‐inhibitory activities and antioxidant properties. In particular, protein hydrolysates from Bellerochea malleus were found to reduce blood pressure properties of 17 mmHg after 5 days of oral administration to SHR animals. These results revealed the potential of bioactive peptides from indigenous marine diatoms for use as functional foods or nutraceuticals.

Background The development of anti-cancer drugs with the ability to inhibit brain metastasis through the blood-brain barrier (BBB) is substantially limited due to the lack of reliable in vitro models. Main methods In this study, the Geltrex™-based Transwell® and microfluidic BBB models were applied to screen the effect of β-boswellic acid (β-BA) on the metastasis of MDA-MB-231 cells through the BBB in static and dynamic conditions, respectively. Major results The toxicity assay revealed that β-BA deteriorates MDA-MB-231 cells, while β-BA had no detectable toxic effects on human umbilical vein endothelial cells (HUVECs) and astrocytes. Trans-endothelial electrical resistance evaluation showed sustainable barrier integrity upon treatment with β-BA. Vimentin expression in HUVECs, evaluated using western blot, confirmed superior barrier integrity in the presence of β-BA. The obtained results were confirmed using an invasion study with a cell tracker and a scanning electron microscope. β-BA significantly inhibited metastasis by 85%, while cisplatin (Cis), a positive control, inhibited cancer cell migration by 12% under static conditions. Upon applying a dynamic BBB model, it was revealed that β-BA-mediated metastasis inhibition was significantly higher than that mediated by Cis. Conclusions and implications In summary, the current study proved the anti-metastatic potential of β-BA in both static and dynamic BBB models. This article is protected by copyright. All rights reserved.

For the successful, large-scale cultivation of microalgae, which for our work refers to diatoms, it is important to select the appropriate species for the right environment. Intensive research on the growth of different strains of microalgae from several regions worldwide is an ongoing effort. The aim of our research in this context was to select the best isolates of diatoms from different coastal sites in Oman, a region whose flora is poorly known, and identify these species using molecular (nuclear-encoded small subunit ribosomal RNA and chloroplast-encoded rbcL and psbC) and morphologic data. We aimed also to investigate and measure the growth rate, biomass production and fatty acid composition under ambient environmental conditions. Our results showed that in contrast to most of the research reported in the literature, the production of biomass from six isolates has been successfully carried out with reduced amounts of nutrients and without CO2 addition. Growth rates for species cultivated outdoors were high and ranged from 0.37 to 0.92 day−1. These findings are economically very promising as fast growth rates, associated with reduced operational costs, could remarkably improve the production efficiency and thereby justify their use as biomass feedstocks. Molecular data revealed that one isolate was Bellerochea malleus, four isolates represented an unidentified species of Bellerochea and the remaining strain studied represented an unidentified species of the genus Nitzschia. These six strains were rich in polyunsaturated fatty acids, with an average of 47% of the total fatty acids, confirming their potential use as aquaculture feed supplements.