Updates on the role of epigenetics in familial mediterranean fever (FMF).

Familial Mediterranean Fever (FMF) is an autosomal recessive autoinflammatory disease caused by mutations in the MEFV (MEditerranean FeVer) gene that affects people originating from the Mediterranean Sea. The high variability in severity and clinical manifestations observed not only between ethnic groups but also between and within families is mainly related to MEFV allelic heterogeneity and to some modifying genes. In addition to the genetic factors underlying FMF, the environment plays a significant role in the development and manifestation of this disease through various epigenetic mechanisms, including DNA methylation, histone modification, and noncoding RNAs. Indeed, epigenetic events have been identified as an important pathophysiological determinant of FMF and co-factors shaping the clinical picture and outcome of the disease. Therefore, it is essential to better understand the contribution of epigenetic factors to autoinflammatory diseases, namely, FMF, to improve disease prognosis and potentially develop effective targeted therapies. In this review, we highlight the latest updates on the role of epigenetics in FMF.

Experimental models in Familial Mediterranean Fever (FMF): Insights into pathophysiology and therapeutic strategies.

Familial Mediterranean Fever (FMF) is a recurrent polyserositis characterized by self-limiting episodes or attacks of fever along with serosal inflammation. It mainly impacts people of the Mediterranean and Middle Eastern basin. FMF is a recessive autoinflammatory condition caused by mutation in the MEFV gene located on chromosome 16p13. MEFV mutations lead to the activation of the pyrin inflammasome resulting in an uncontrolled release of IL-1ß. Various in vitro, in vivo and ex vivo experimental models have been developed to further comprehend the etiology and pathogenesis of FMF. These models have been proven to be clinically relevant to human FMF and can provide significant information about biological systems with respect to this condition. Additionally, these models have provided pertinent contributions to the development of potent therapeutic strategies against FMF. In this review, we describe the different experimental models utilized in FMF and we focus primarily on the most widely used models that have produced prominent insights into the pathophysiology of the disease.

Relationship between emotional expressions and lifestyle changes among university students during COVID-19 lockdown in Lebanon.

INTRODUCTION: Emotional expressivity plays an important role in terms of communication and interpersonal relationships in the context of society. Our research aims at assessing the perception of emotional expressivity and its association with lifestyle changes during COVID-19 lockdown among university students in Lebanon. METHODOLOGY: A total of 742 participants completed an anonymous online questionnaire including socio-demographic characteristics, lifestyle habits, and emotional expression evaluated through the Berkeley Expressivity Questionnaire (BEQ). Participants also reported their perception of the relationship between emotional expressivity and lifestyle changes experienced during lockdown. RESULTS: The expression of positive emotions was decreased among students who have a weight loss from decreased eating during lockdown. Moreover, significantly lower negative emotions and increased positive emotions were found to be associated respectively with the increase in quality and quantity of distance learning, which emphasizes the psychological investment in education. In terms of extensive time spent on TV and social media, results point towards increased emotional expressivity, particularly positive emotions and impulse facets. Finally, changes in social interaction during this period impacted all facets of emotional expressivity. CONCLUSIONS: In the context of lockdown due to the pandemic, emotional aspects were associated by university students with lifestyle changes. Our research highlights the beneficial role of social networking, distance learning, physical activity, and well-balanced nutrition on emotional regulation in this particularly stressful situation, thus the importance of a global approach to emotional expressivity including the social aspects and biological ones.

Spectrum of MEFV Variants and Genotypes among Clinically Diagnosed FMF Patients from Southern Lebanon.

Background: Familial Mediterranean Fever (FMF) is an autosomal recessive auto-inflammatory disease characterized by pathogenic variants in the MEFV gene, with allele frequencies greatly varying between countries, populations and ethnic groups. Materials and methods: In order to analyze the spectrum of MEFV variants and genotypes among clinically diagnosed FMF patients from South Lebanon, data were collected from 332 participants and 23 MEFV variants were screened using a Real-Time PCR Kit. Results: The mean age at symptom onset was 17.31 ± 13.82 years. The most prevalent symptoms were abdominal pain, fever and myalgia. MEFV molecular analysis showed that 111 patients (63.79%) were heterozygous, 16 (9.20%) were homozygous, and 47 (27.01%) carried two variants or more. E148Q was the most encountered variant among heterozygous subjects. E148Q/M694V was the most frequent in the compound heterozygous/complex genotype group, while M694I was the most common among homozygous patients. Regarding allele frequencies, M694V was the most common variant (20.7%), followed by E148Q (17.1%), V726A (15.7%) and M694I (13.2%). Conclusion: The high percentage of heterozygous patients clinically diagnosed as FMF highlights the pseudo-dominant transmission of the disease in Lebanon and emphasizes the importance of molecular testing for a more accurate diagnosis and better management and treatment of FMF.

Comparative effect of sodium butyrate and sodium propionate on proliferation, cell cycle and apoptosis in human breast cancer cells MCF-7.

INTRODUCTION: Short-chain fatty acids (SCFAs) are ubiquitous lipids produced as a result of bacterial fermentation of dietary fiber. While their role in colorectal cancer is well known, the effect of SCFAs in breast cancer is poorly defined. OBJECTIVE: To understand the various effects of SCFAs on breast carcinogenesis, we investigated the effect of sodium butyrate (NaB) and sodium propionate (NaP) in MCF-7 cell line. MATERIALS AND METHODS: Cells were incubated with different concentrations of NaB or NaP for 24, 48, 72 or 96 h. Cell proliferation was assayed using MTT kit. Cell cycle was performed using propidium iodide staining then analyzed with a flow cytometer. Apoptosis was assessed by Hoechst technique and cell-cycle sub-G1 phase. RESULTS: NaB and NaP inhibited MCF-7 cell proliferation in a dose-dependent manner with respective IC50 of 1.26 mM and 4.5 mM, thus indicating that NaB is more potent than NaP. Low and medium levels of both SCFAs induced morphology changes which are characteristic of a differentiated phenotype. Flow cytometry analysis revealed a blockage in G1 growth phase. Interestingly, removing NaB or NaP from culture media after few days of treatment showed a reversible effect on cell morphology and proliferation where cells reentered the cycle after 24 h of drug wash-out. Finally, treatment with medium levels of these molecules induced low MCF-7 apoptosis, while higher doses led to massive apoptosis. CONCLUSION: Our results show that SCFAs may be considered as an interesting inhibitor for breast cancer progression.

Improvement of Human Sperm Vacuolization and DNA Fragmentation Co-Cultured with Adipose-Derived Mesenchymal Stem Cell Secretome: In Vitro Effect.

BACKGROUND AND OBJECTIVES: Oxidative stress (OS) is known to be an important factor of male infertility. Adipose-derived mesenchymal stem cells (AD-MSCs) are known to have immune-modulatory and anti-oxidant effects through their secretions, hence raising the idea of their potential benefit to improve sperm parameters. This study aims at investigating the effect of AD-MSCs conditioned medium (CM) on human sperm parameters in the presence and absence of H2O2-induced OS. METHODS AND RESULTS: Sperm samples were collected from 30 healthy men and divided into two groups: non-stressed and H2O2-stressed. Isolated AD-MSCs from healthy donors undergoing liposuction were cultured and CM was collected at 24, 48 and 72 h. Both sperm groups were cultured with CM and a time course was performed followed by an evaluation of sperm parameters. The incubation of non-stressed and stressed sperm samples with AD-MSCs-CM for 24 h was found to have the optimum impact on sperm vacuolization, DNA fragmentation and OS levels in comparison to other incubation timings, while preserving motility, viability and morphology of cells. Incubation with CM improved all sperm parameters except morphology in comparison to the non-treated group, with the best effect noted with CM collected at 24 h rather than 48 or 72 h for sperm vacuolization and DNA fragmentation. When compared to fresh semen parameters (T0), samples cultured with CM 24 h showed a significant decrease in sperm vacuolization and DNA fragmentation while keeping other parameters stable. CONCLUSIONS: AD-MSCSs-CM improves sperm quality, and hence can be used in treating infertility and subsequently enhancing IVF outcomes.

Emergence of Vulvovaginal Candidiasis among Lebanese Pregnant Women: Prevalence, Risk Factors, and Species Distribution.

Objective: Candida species colonize the vagina in at least 20% of women, with rates rising to 30% during pregnancy. This study aimed at determining the prevalence and risk factors of vulvovaginal candidiasis (VVC) in pregnant women at 35-37 weeks of gestation. It also aims at finding possible correlations between VVC and vaginal colonization by other agents, such as Group B Streptococcus (GBS) and bacterial vaginosis. Methodology: Over a one-year period, high vaginal swabs were collected from pregnant women during their regular antenatal checkup in different polyclinics in Beirut and South Lebanon. Swabs were examined microscopically, cultured on Sabouraud Dextrose Agar, and Candida isolates were identified using Chromatic Candida medium and Germ Tube Test. Results: VVC was detected in 44.8% of samples, with C. glabrata (44.4%) and C. albicans (43.4%) being the most isolated species. Approximately, half of pregnant women (57.7%) were coinfected with Candida and bacterial vaginosis, while 26% of them carried simultaneously Candida spp. and GBS. No significant correlation was found between the occurrence of VVC and demographic, clinical, medical, and reproductive health characteristics of pregnant women. In contrast, participants with previous miscarriages and those being hospitalized during the past 12 months were more susceptible to develop vaginal C. krusei infection in comparison to other Candida species (p=0.0316 and p=0.0042, respectively). Conclusion: The prevalence of VVC in pregnant women is an increasing trend in our community. Therefore, routine medical examination and regular screening for candidiasis in the antenatal care program is highly recommended to manage the disease and its complications.

Impact of alexithymia on glycemic control among Lebanese adults with type 2 diabetes.

PURPOSE: Alexithymia, defined as the inability of a person to identify, describe and express emotions, has been found to influence glycemic control in type 2 diabetes patients (D2). The characteristics and influencing factors of alexithymia and the association of this psychological construct with D2 has not yet been studied in Lebanon where 14.6% of adults are diagnosed with the disease. This study aims at evaluating the prevalence of alexithymia and its relationship with glycemic control among Lebanese adults with D2. METHODS: Alexithymia was assessed in 104 patients diagnosed with D2 and 100 healthy controls using the 20-item Toronto Alexithymia Scale (TAS-20). The impact of alexithymia on glycemic control was evaluated using HbA1c values, fasting blood glucose levels, number of severe hyperglycemic episodes and hospitalizations for hyperglycemia within the past months. RESULTS: Alexithymia prevalence was significantly higher in D2 patients compared to controls (35.5% vs 15%). Patients with alexithymia showed higher levels of HbA1c and glucose in comparison to those without alexithymia. Consistently, significant positive correlations were found between the TAS-20 total and subscale scores and both HbA1c and glucose levels. Alexithymic patients had three times more severe hyperglycemic episodes and five times more hospitalizations for hyperglycemia compared to those without alexithymia. According to multivariate regression analysis, lifestyle factors alone were not found predictive of alexithymia in D2 patients. CONCLUSION: Given the impact of alexithymia on D2 regulation, screening of alexithymia in case of D2 and appropriate psychological follow-up are important for a better prognosis, management and treatment of the disease.

Prevalence, antimicrobial resistance and risk factors for campylobacteriosis in Lebanon.

INTRODUCTION: The rapid increase in Campylobacter strains resistant to antibiotics represents a major problem for public health. In Lebanon, campylobacteriosis is underdiagnosed since bacteria detection in stool samples is not performed routinely. This study aims to evaluate the prevalence, sources and routes of transmission, risk factors and antimicrobial susceptibility patterns of Campylobacter spp. in Lebanon. METHODOLOGY: Stool samples collected from 1000 Lebanese patients with diarrhea, and 150 meat samples taken from supermarkets and slaughterhouses were subjected to Campylobacter detection. Colonies were identified by Gram staining, oxidase and catalase activities. They were then differentiated at the species level by hippurate test and PCR. Susceptibility of Campylobacter spp. to antibiotics was studied by the disc diffusion standard method. RESULTS: Campylobacter spp. were detected in 21.5% of stool samples; the main isolated species being C. jejuni (83.2%) and C. coli (13.9%). The highest Campylobacter infection rates were detected among children (41.8%) and during summer (31.6%). Consumption of contaminated meat and salads, and contact with animals represented the major risk factors for campylobacteriosis, with poultry carcasses and bovine cuts identified as the main bacteria reservoirs. Neither demographic determinants nor season had a major effect on the prevalence of campylobacteriosis. Erythromycin was the most active agent against Campylobacter spp. A multi-resistance rate was observed in 35.9% of isolates. CONCLUSIONS: Campylobacteriosis is a major public health concern in Lebanon. Bacteria detection in stool culture should be performed routinely to allow an early diagnosis and a better monitoring of the disease and its burden.

RAC1 expression and role in IL-1ß production and oxidative stress generation in familial Mediterranean fever (FMF) patients.

Familial Mediterranean fever (FMF) is a recessively inherited autoinflammatory disorder. The caspase-1-dependent cytokine, IL-1ß, plays an important role in FMF pathogenesis, and RAC1 protein has been recently involved in IL-1ß secretion. This study aims to investigate RAC1 expression and role in IL-1ß and caspase-1 production and oxidative stress generation in FMF. The study included 25 FMF patients (nine during attack and remission, and 16 during remission only), and 25 controls. RAC1 expression levels were analyzed by real-time PCR. Ex vivo production of caspase-1, IL-1ß, IL-6 and markers of oxidative stress (malondialdehyde, catalase, and glutathione system) were evaluated respectively in supernatants of patients' and controls' PBMC and PMN cultures, in the presence and absence of RAC1 inhibitor. RAC1 gene expression and IL-1ß levels were increased in patients in crises compared to those in remission or controls. RAC1 expression levels were correlated with MEFV genotypes, patients carrying the M694V/M694V genotype having a two-fold increase in the expression levels compared to those carrying other genotypes. Caspase-1 levels were higher in LPS-induced PBMC of patients in remission than controls. Spontaneous and LPS-induced IL-1ß production were comparable in patients in remission and controls, whereas LPS-induced IL-6 production was enhanced in patients, compared to controls. RAC1 inhibition resulted in a decrease in caspase-1 and IL-1ß, but not IL-6, levels. Malondialdehyde levels produced by LPS-stimulated PMNs were increased in patients in remission compared to those in controls, but decreased following RAC1 inhibition. Catalase and GSH activities were reduced in unstimulated PMN culture supernatants of patients in remission compared to controls and were increased in the presence of RAC1 inhibitor. These results show the involvement of RAC1 in the inflammatory process of FMF by enhancing IL-1ß production, through caspase-1 activation, and generating oxidative stress, even during asymptomatic periods.

Cytokine signatures in hereditary fever syndromes (HFS).

Hereditary fever syndromes (HFS) include a group of disorders characterized by recurrent self-limited episodes of fever accompanied by inflammatory manifestations occurring in the absence of infection or autoimmune reaction. Advances in the genetics of HFS have led to the identification of new gene families and pathways involved in the regulation of inflammation and innate immunity. The key role of several cytokine networks in the pathogenesis of HFS has been underlined by several groups, and supported by the rapid response of patients to targeted cytokine blocking therapies. This can be due to the direct effect of cytokine overproduction or to an absence of receptor antagonist resulting in dysbalance of downstream pro- and anti-inflammatory cytokine networks. The aim of this study was to present an overview and to discuss the major concepts regarding the cellular and molecular immunology of HFS, with a particular focus on their specific cytokine signatures and physiopathological implications. Based on their molecular and cellular mechanisms, HFS have been classified into intrinsic and extrinsic IL-1ß activation disorders or inflammasomopathies, and protein misfolding disorders. This review integrates all recent data in an updated classification of HFS.

Study of the association of IL-1ß and IL-1RA gene polymorphisms with occurrence and severity of Familial Mediterranean fever.

Familial Mediterranean fever (FMF) is a recessive autoinflammatory disorder. The balance between the pro-inflammatory cytokine IL-1ß and its receptor antagonist IL-1RA plays an important role in the development of FMF. In order to determine a possible association of polymorphisms in IL-1ß and IL-1RA genes with occurrence and/or severity of the disease, 42 genetically confirmed FMF patients and 42 controls were genotyped for IL-1ß(-511C/T), IL-1ß(-31T/C), IL1-1ß(+3954T/C) and IL-1RA VNTR polymorphisms. IL-1ß and IL-1RA levels were evaluated by multiplex ELISA in supernatants of PBMC cultures of 30 FMF patients with and without 24h stimulation of monocytes by LPS. The CC genotype and C allele at positions -31 and + 3954 of IL-1ß gene were more frequent in FMF patients than in controls. FMF patients carriers of IL-1ß(-31) CC genotype were associated with a 2-fold increase in LPS-induced IL-1ß secretion as well as a higher disease severity score (11.2 ± 2.9) when compared to patients carrying the TC and TT genotypes (6.1 ± 2.1 and 4.5 ± 2.4, respectively). These results indicate that IL-1ß gene polymorphisms at positions -31 and + 3954 may be associated with an increased risk for FMF. IL-1ß(-31) contributes also to the severity of the disease, probably by modulating IL-1ß synthesis.

Hypovitaminosis D in a Young Lebanese Population: Effect of GC Gene Polymorphisms on Vitamin D and Vitamin D Binding Protein Levels.

Bioactive vitamin D is a steroid hormone transported in blood via the vitamin D binding protein (DBP). Our study aimed to investigate the vitamin D status in a young Lebanese population and study the association of hypovitaminosis with levels of DBP. Polymorphisms in the GC gene that encodes DBP were also screened. Blood samples were collected from 179 university students. Vitamin D status and DBP levels were assayed by enzyme-linked immunosorbent assay (ELISA). DNA was extracted from 128 participants, and genotyping of the two GC gene SNPs, rs7041, and rs4588, was carried out by restriction fragment length polymorphism. Forty-seven percent of participants had hypovitaminosis D (<20 ng/ml). A significant positive correlation was observed between vitamin D status and DBP. Genotyping data showed that participants carrying the rs7041 GG and rs4588 AA genotypes had higher concentrations of DBP than those carrying other genotypes. Four allelic versions of the GC gene were observed, one of which, GC*3, was encountered for the first time in this study, and was found to be associated with both normal vitamin D and high DBP levels. Modifying genes such as GC could therefore affect DBP levels, and contribute, along with environmental factors, to the hypovitaminosis D observed in sunny countries.

Ex vivo PBMC cytokine profile in familial Mediterranean fever patients: Involvement of IL-1ß, IL-1α and Th17-associated cytokines and decrease of Th1 and Th2 cytokines.

In order to clarify the inflammatory mechanism underlying familial Mediterranean fever (FMF), we aimed to evaluate the ex vivo cytokine profile of FMF patients during acute attacks and attack-free periods, and compare it with that of healthy controls. The study included 34 FMF patients, of whom 9 were studied during attack and remission and 24 healthy controls. Cytokine levels were evaluated by Luminex technology in serum and supernatants of PBMC (Peripheral Blood Mononuclear Cells) cultures with and without 24h stimulation of monocytes by LPS and T lymphocytes by anti-CD3/CD28 beads. Levels of IL-6 and TNF-α were higher in unstimulated and LPS-stimulated PBMC supernatants of FMF patients in crises compared to controls. In response to LPS stimulation, higher levels of IL-1ß and IL-1α were found in PBMC supernatants of patients during crises compared to those in remission and to controls. IFN-γ and IL-4 levels were the lowest in unstimulated and anti-CD3/CD28 stimulated PBMCs supernatants of patients during crises compared to remission and controls. The Th17 cytokines IL-17 and IL-22 were respectively higher in anti-CD3/CD28 stimulated PBMC supernatants of FMF patients during and between crises compared to controls. Amongst cytokines tested in serum, only IL-6 and TNFα were enhanced in FMF patients. The ex vivo study represents an interesting approach to evaluate cytokines' involvement in FMF. Our results suggest an ongoing subclinical inflammation and define an elevated inflammatory cytokine signature, distinctly for M694V homozygous patients. The absence of spontaneous IL-1ß release by PBMCs reflects no constitutive activation of the inflammasome in FMF physiopathology.