Challenges Faced by Behavioral Genetic Studies: Researchers Perspective from the MENA Region.

Background: Behavioral genetic studies are important for the understanding of the contribution of genetic variations to human behavior. However, such studies might be associated with some ethical concerns. Methods: In the current study, ethical challenges related to studies of genetic variations contributing to human behavior were examined among researchers. To achieve the study purpose, the Middle East and North Africa (MENA) region researchers were taken as an example, where the after- mentioned ethical challenges were discussed among a group of researchers, who were the participants of an online forum. Discussions and responses of the participants were monitored and were later qualitatively analyzed. Results: Discussions revealed that several ethical challenges, including subjects' recruitment, the difficulty of obtaining informed consents, and issues of privacy and confidentiality of obtained data as information leakage, in this case, will lead to social stigma and isolation of the participants and their immediate family members. Jordanian social and cultural norms, faith, and the tribal nature of the population were raised as a major challenge that might face conducting behavioral genetic studies in the Arab populations of the MENA. The lack of regulation related to the conduction of genetic studies, misunderstanding, and misuse of genetic information are other challenges. A full explanation of genetic research and the current and future possible benefits/risks of such research could be potential solutions. Conclusion: In conclusion, the MENA populations are tackled with major challenges in relation to conducting research studies in genetics/antisocial behavior field/s. Establishment of guidelines related to genetic studies, capacity building, increasing public awareness about the importance of genetic testing, and enhancing responsible conduct of research will facilitate the conduct of such sensitive studies in the future in the region.

Anticancer and antimutagenic activity of Silybum marianum L. and Eucalyptus camaldulensis Dehnh. against skin cancer induced by DMBA: In vitro and in vivo models.

The current study investigated the prospective effect of Silybum marianum L. and Eucalyptus camaldulensis Dehnh extracts against skin cancer. Skin cancer was induced by 7,12-dimethylbenz(a) anthracene (DMBA) in young Balb/c mice. Plant extracts were administered to animals orally, once/day (100mg/kg, 5 days/week) for the 20 weeks. Anticancer activity was examined via tumor progression, where antimutagenic activity was measured using 8-OHdG and sister chromatid exchange (SCE) levels. Eucalyptus camaldulensis Dehnh. leaves extract and Silybum marianum L. leaves extract significantly reduced 8-OHdG in cultured human lymphocytes in a dose-response manner (P<0.05). Similarly, the leave extracts of both plants significantly reduced chromosomal damage as measured by SCE levels (P<0.05). In the skin painting assay, the leave extracts of both plants significantly delayed the onset of tumors compared to DMBA treated group (P<0.05). The Silybum marianum leaves extract significantly reduced tumor incidence (P<0.01) and papilloma frequency (P<0.01) induced by DMBA. The Eucalyptus camaldulensis leaves extract significantly reduced the number of tumors per animal (P<0.05) and incidence of tumors (P<0.001). The in vitro and in vivo findings showed that leaves of Silybum marianum L. and Eucalyptus camaldulensis Dehnh. extracts might be a promising source for anticancer and antimutagenic agents against human cancer.

The aPKC/Par3/Par6 Polarity Complex and Membrane Order Are Functionally Interdependent in Epithelia During Vertebrate Organogenesis.

The differential distribution of lipids between apical and basolateral membranes is necessary for many epithelial cell functions, but how this characteristic membrane organization is integrated within the polarity network during ductal organ development is poorly understood. Here we quantified membrane order in the gut, kidney and liver ductal epithelia in zebrafish larvae at 3-11 days post fertilization (dpf) with Laurdan 2-photon microscopy. We then applied a combination of Laurdan imaging, antisense knock-down and analysis of polarity markers to understand the relationship between membrane order and apical-basal polarity. We found a reciprocal relationship between membrane order and the cell polarity network. Reducing membrane condensation by exogenously added oxysterol or depletion of cholesterol reduced apical targeting of the polarity protein, aPKC. Conversely, using morpholino knock down in zebrafish, we found that membrane order was dependent upon the Crb3 and Par3 polarity protein expression in ductal epithelia. Hence our data suggest that the biophysical property of membrane lipid packing is a regulatory element in apical basal polarity.

Attitude towards donation of the excised foreskin after circumcision surgery for research: A study from Madinah, Saudi Arabia.

Studies have shown the possibility of using the part of the foreskin removed after circumcision in the field of scientific and therapeutic research. Donations of tissues and organs are always associated with ethical challenges posed by bioethicists and societies to ensure the appropriate use of these tissues/organs. The purpose of this study was to understand the attitudes and awareness of parents/guardians regarding donation of excised foreskin to research and medical use. The study was based on a questionnaire and included 133 parents/guardians who visited Uhud Children's Hospital in Madinah, Saudi Arabia for newborn male circumcision. The results showed a high willingness (61.7%) to donate the extracted foreskin to research. The willingness to donate the extracted foreskin to research associated with undergraduate degree (P = 0.018), male sex (P = 0.011), high income (P = 0.029), and participation in previous research studies (P = 0.002). About 41.8% were convinced that written informed consent should be obtained before circumcision surgery, 38.1% (n = 51) were convinced that written informed consent should be taken after surgery, while the remaining 19.4% reported that the timing of written informed consent is unimportant. Finally, fear of excision of excess tissue (74.5%), lack of confidence in the research (68.6%), and potential for commercial use (64.7%) were the main barriers to unwillingness to donate the excised foreskin for research. In conclusion, a reasonable portion of Saudis agreed to donate their foreskin for research purposes. There is an urgent need to enhance awareness and attitudes towards tissue donation for research and therapeutic use.

Safety in Medical Laboratories: Perception and Practice of University Students and Laboratory Workers.

Background: Several safety-related accidents occur in the laboratories because of insufficient regulations, inappropriate implementation of safety measures, or unawareness attitude and practices toward safety precautions. Thus, establishment of efficient regulations and safe habits toward workplace safety is crucial to prevent or minimize such accidents. Here we investigate the levels of laboratory safety awareness among undergraduate medical science students and laboratory workers at major hospitals in Taif, Saudi Arabia. Methods: An anonymous cross-sectional survey was conducted on a random sample of 185 students and workers. The survey was categorized into five sections: (1) variable demographic items, (2) orientation around the warning signage of medical laboratory picograms, (3) assess participants' attitudes toward laboratory safety, (4) assess participants' safety awareness and practices in medical laboratories, and (5) assess participants' knowledge in dealing with emergency equipment and related procedures. Results: Data analyses indicate that workers are generally more aware of the key aspects of laboratory safety. Although students demonstrated moderate to good knowledge of major laboratory signage, there are some areas of deficiency, particularly regarding the use of fire extinguishers. Of interest is that female students are more familiar with laboratory signage than male students, although general laboratory safety awareness among students is poor. Students also showed poor awareness of certain critical aspects of safety practices and emergency procedures. Conclusion: The study data reveal inadequate awareness and knowledge of certain laboratory safety aspects among undergraduate students. Therefore, academic institutions should introduce effective safety regulations, education, and training to improve student safety.

Highlights on selected microscopy techniques to study zebrafish developmental biology.

Bio-imaging is a tedious task when it concerns exploring cell functions, developmental mechanisms, and other vital processes in vivo. Single-cell resolution is challenging due to different issues such as sample size, the scattering of intact and opaque tissue, pigmentation in untreated animals, the movement of living organs, and maintaining the sample under physiological conditions. These factors might lead researchers to implement microscopy techniques with a suitable animal model to mimic the nature of the living cells. Zebrafish acquired its prestigious reputation in the biomedical research field due to its transparency under advanced microscopes. Therefore, various microscopy techniques, including Multi-Photon, Light-Sheet Microscopy, and Second Harmonic Generation, simplify the discovery of different types of internal functions in zebrafish. In this review, we briefly discuss three recent microscopy techniques that are being utilized because they are non-invasive in investigating developmental events in zebrafish embryo and larvae.

The Association between Adiponectin Single Nucleotide Polymorphisms and Side Effects of Isotretinoin in Acne Patients.

BACKGROUND: Acne is a common condition of pilosebaceous follicle especially among young. Clinically, the most used medication in the treatment of moderate to severe acne is oral isotretinoin. However, interindividual variability in therapeutic response to isotretinoin and many side effects such as musculoskeletal pain, headache, and alteration in lipid profile can be seen with this treatment. AIM: In this study, the effect of genetic polymorphisms, rs2241766 and rs1501299, of the adiponectin gene was investigated in relation to the side effects of isotretinoin-treated young adult acne patients (n = 230). METHODS: Several biochemical parameters were measured at baseline and after treatments with isotretinoin. The ADIPOQ gene SNPs, rs2241766 and rs1501299, were genotyped in 230 patients. RESULTS: Alterations in lipid profile with a significant increase of ALT (P=0.007) were detected after isotretinoin treatment. Moreover, percentage change in HDL following isotretinoin treatment was significantly associated with rs1501299 (P=0.008). On the other hand, no associations between examined SNPs and side effects of isotretinoin and other lipid parameters (total cholesterol, LDL, and triglycerides) or liver function enzymes (ALT and AST) were detected. CONCLUSIONS: Current findings showed that rs1501299 of the ADIPOQ gene might be associated with changes in HDL level in acne patients following treatment with isotretinoin.

The effect of genetic variations in the choline acetyltransferase gene (ChAT) on waterpipe tobacco smoking dependence.

INTRODUCTION: Waterpipe tobacco smoking (WS) is a popular form of tobacco use, globally. While the impact of genetic variations on smoking behavior has been well-investigated, few studies have examined this issue with regard to WS. In the current study, associations between choline acetyltransferase gene (ChAT) rs1917810 and rs7094248 polymorphisms and WS dependence were examined. METHODS: Genotyping of rs1917810 and rs7094248 in 266 Jordanian waterpipe smokers was performed using RFLP-PCR. Dependence on WS was measured using the LWDS-10J scale. RESULTS: The frequency of rs1917810 G allele was 38.5% and that of the rs7094248 G allele was 40.8%. The rs1917810 was significantly associated with the WS dependence level (p<0.05). Carriers of the AA genotype of rs1917810 were more dependent on WS than those with GG genotype (p<0.05). However, no association between rs7094248 single-nucleotide polymorphism (SNP) and dependence on WS was observed (p>0.05). CONCLUSIONS: The rs1917810 in the ChAT gene might be associated with dependence on WS.

The CHRNA5 Polymorphism (rs16969968) and its Association with Waterpipe Smoking Addiction among Jordanians.

Waterpipe smoking is a form of tobacco use that causes nicotine/tobacco dependence and has become a global health problem. In the current study, the association of rs16969968 SNP in the CHRNA5 gene with waterpipe dependence was investigated. A total of 386 men and women who used a waterpipe to smoke tobacco were recruited and divided into less dependent and more dependent smokers based on their score on the Lebanon Waterpipe Dependence Scale (LWDS). Results showed a significant difference in the distribution of GG, GA, and AA genotypes by waterpipe dependence status (P<0.001). The more dependent group showed a higher frequency of the AA genotype than the less dependent smokers' group (38% versus 23% respectively). In addition, the more dependent smokers exhibited more A allele than less dependent smokers (53% versus 37% respectively, P<0.001). In conclusion, there is an association between the rs16969968 SNP and waterpipe dependence as assessed by the LWDS.

The Influence of IL-1B Gene Polymorphisms on H. pylori Infection and Triple Treatment Response Among Jordanian Population.

BACKGROUND: Helicobacter pylori (H. pylori) is considered the main cause of gastritis, peptic ulcer and gastric carcinoma in the human populations. H. pylori infection influences the secretion level of several proinflammatory cytokines including IL-1ß, which encoded by the IL-1B gene. OBJECTIVE: The current study aimed to investigate whether IL-1B gene polymorphisms are associated with H. pylori infection among the Jordanian population and responses to triple therapy. SUBJECTS AND METHODS: The gastroscopic examination was performed on 412 subjects for H. pylori infection diagnosis, 257 subjects were found to be infected by H. Pylori (positive cases), whereas 155 subjects were uninfected (negative controls). The IL-1B gene T-31C and C3954T polymorphisms were genotyped by PCR-RFLP. RESULTS: It was found that the T-31C polymorphism has a significant association with H. pylori infection (P<0.05), and the TT genotype frequency was significantly higher in infected subjects (50.2%) compared to controls (38.7%). On the other hand, no significant association was detected between C3954T SNPs and H. pylori infection among the Jordanian population. In addition, none of the examined polymorphisms were found to influence the responses to triple therapy. CONCLUSION: The IL-1B gene T-31C SNP might be associated with an enhanced risk of H. pylori infection among the Jordanian population.

Challenges that face the establishment of diabetes biobank in Jordan: a qualitative analysis of an online discussion forum.

Introduction: Diabetes is common in Jordan with a prevalence of about 13% of the population. Establishment of a diabetes biobank in Jordan could have an enormous impact on the management and prevention of the disease. Methods: In the current study, ethical challenges that might face the establishment of a biobank were examined by 28 researchers from the Middle East and North Africa region using an online discussion forum. Results: All participants agreed on the importance of the establishment of a diabetes biobank in Jordan. The possible challenges that were discussed included confidentiality and privacy, informed consent, specimen ownership and participants' rights, data sharing, returning of research results and incidental findings, lack of legislations, and importance of social awareness and public engagements with biobanks. Conclusion: In conclusion, participants support the establishment of a diabetes biobank in Jordan; however, some ethical issues should be considered to ensure the success of the biobank.

Salivary Microbiome and Cigarette Smoking: A First of Its Kind Investigation in Jordan.

There is accumulating evidence in the biomedical literature suggesting the role of smoking in increasing the risk of oral diseases including some oral cancers. Smoking alters microbial attributes of the oral cavity by decreasing the commensal microbial population and increasing the pathogenic microbes. This study aims to investigate the shift in the salivary microbiota between smokers and non-smokers in Jordan. Our methods relied on high-throughput next-generation sequencing (NGS) experiments for V3-V4 hypervariable regions of the 16S rRNA gene, followed by comprehensive bioinformatics analysis including advanced multidimensional data visualization methods and statistical analysis approaches. Six genera-Streptococcus, Prevotella, Vellionella, Rothia, Neisseria, and Haemophilus-predominated the salivary microbiota of all samples with different percentages suggesting the possibility for the salivary microbiome to restored after quitting smoking. Three genera-Streptococcus, Prevotella, and Veillonella-showed significantly elevated levels among smokers at the expense of Neisseria in non-smokers. In conclusion, smoking has a definite impact on shifting the salivary microbiota in smokers. We can suggest that there is microbial signature at the genera level that can be used to classify smokers and non-smokers by Linear Discriminant Analysis Effect Size (LEfSe) based on the salivary abundance of genera. Proteomics and metabolomics studies are highly recommended to fully understand the effect of bacterial endotoxin release and xenobiotic metabolism on the bacterial interrelationships in the salivary microbiome and how they affect the growth of each other in the saliva of smokers.

Evaluation of antioxidant status and oxidative stress markers in thermal sulfurous springs residents.

Sulfurous springs have been traditionally used in medical treatment for different purposes. These beneficial effects of sulfurous water have been attributed to the presence of sulfurous compounds mainly in the form of hydrogen sulfide (H2S). The purpose of the present study is to explore the effects of long-term exposure to sulfurous springs on oxidative stress and antioxidant biomarkers responses in individuals who lived nearby the sulfurous springs. The studied area was Al- Hammah sulfurous springs, which is located in the northern part of the Jordan Rift Valley and host many sulfurous springs. Residents in sulfurous springs area are continuously exposed to water and gases emission more than the overall population. We have found that the sulphate levels were 7 times higher in sulfurous springs water samples than control water samples. The majority of the volunteers involved in the present study were more than ten years long residence and lived in range distance between one to five kilometers (less than 3 miles) away from main sulfurous spring, and visited the sulfurous spring at least once a month. We did not find any noticeable symptoms in sulfur spring residents such as headaches, nausea, breathing problems. The total oxidative stress (TOS) and oxidative stress index (OSI) in sulfurous spring residents were lower than control individuals. The total antioxidant capacity (TAC) and total nitric oxide (NOX) levels were higher in sulfurous spring residents compared to control group. Furthermore, we have highlighted that living nearby the sulfurous springs does not affect oxygen saturation levels (SPO2) or heart pulse rate . These findings suggest that long-term exposure to sulfurous springs boost the antioxidant capacity and reduce oxidative stress levels in the human body. Hence, visiting sulfurous springs can act as natural remedies to diminish oxidative stress as they show promising potential in several-oxidative stress-related diseases treatment.

The role of PAX9 promoter gene polymorphisms in causing hypodontia: a study in the Jordanian population.

BACKGROUND: The congenital absence of one or few teeth, hypodontia, is considered one of the utmost dental ageneses in human beings. Several genes have been shown to be involved in the development of hypodontia such as paired box gene 9 (PAX9). The expression of PAX9 is controlled by several polymorphic elements in the promoter region of the gene on 14q13.3 locus. The aim of this study was to find any association between PAX9 c.-912T>C (rs2073247) and c.-1031G>A (rs2073244) promoter polymorphisms and the development of hypodontia among the Jordanian population. METHODS: Genotyping of the polymorphisms in 72 unrelated subjects with hypodontia was performed using PCR-restriction fragment length polymorphism (RFLP) technique and compared with that of 72 normal healthy unrelated control individuals. RESULTS: The hypodontia group had a significantly higher -1031GG genotype (P<0.01) and a significantly lower -912TC genotype (P<0.01) compared with the control group. The results suggest that the transcriptional activity of PAX9 gene is affected by polymorphisms in the promoter region of this gene and is associated with hypodontia phenotype. CONCLUSION: The rs2073247) and rs2073244 promoter polymorphisms of PAX9 might play a role in the development of hypodontia in the Jordanian population.

Characterization of a new series of fluorescent probes for imaging membrane order.

Visualization and quantification of lipid order is an important tool in membrane biophysics and cell biology, but the availability of environmentally sensitive fluorescent membrane probes is limited. Here, we present the characterization of the novel fluorescent dyes PY3304, PY3174 and PY3184, whose fluorescence properties are sensitive to membrane lipid order. In artificial bilayers, the fluorescence emission spectra are red-shifted between the liquid-ordered and liquid-disordered phases. Using ratiometric imaging we demonstrate that the degree of membrane order can be quantitatively determined in artificial liposomes as well as live cells and intact, live zebrafish embryos. Finally, we show that the fluorescence lifetime of the dyes is also dependent on bilayer order. These probes expand the current palate of lipid order-sensing fluorophores affording greater flexibility in the excitation/emission wavelengths and possibly new opportunities in membrane biology.

Quantitative imaging of membrane lipid order in cells and organisms.

It is now recognized that lipids and proteins in cellular membranes are not homogenously distributed. A high degree of membrane order is the biophysical hallmark of cholesterol-enriched lipid rafts, which may induce the lateral sorting of proteins within the membrane. Here we describe a quantitative fluorescence microscopy technique for imaging localized lipid environments and measuring membrane lipid order in live and fixed cells, as well as in intact tissues. The method is based on the spectral ratiometric imaging of the polarity-sensitive membrane dyes Laurdan and di-4-ANEPPDHQ. Laurdan typically requires multiphoton excitation, making it suitable for the imaging of tissues such as whole, living zebrafish embryos, whereas di-4-ANEPPDHQ imaging can be achieved with standard confocal microscopes. This approach, which takes around 4 h, directly examines the organization of cellular membranes and is distinct from alternative approaches that infer membrane order by measuring probe partitioning or dynamics.