The assessment of liver function test and fertility hormones in Saudi athletes using anabolic androgenic steroids.

Background: A growing number of athletes are using synthetic anabolic-androgenic steroids (AAS), comprised of testosterone and other derivatives, to enhance athletic performance and muscle mass. Over the years, numerous reports elucidated the side effects of the illegal use of AAS, such as infertility, and liver disorders. The effect of AAS on the hepatic and reproductive systems in Saudi athletes has not yet been studied. Therefore, this study examined the liver function and sex hormone parameters of AAS users as compared to non-users. Methods: Fasting blood samples were collected from 16 male Saudi athletes, 10 AAS-users (cases) and 6 non-users (controls) to measure liver function tests (ALT, AST, GGT, ALP, total protein, albumin, direct and total bilirubin) and muscle enzymes (CK, LDH), Fertility hormones (LH, FSH, total testosterone, estradiol, and prolactin) were included also. Furthermore, a self-reported questionnaire was obtained to identify the type of AAS used, the dosage, and the length of the course before sample collection. Results: The results show a statistically significant increase in ALT (P < 0.001), AST (P < 0.001), CK (P < 0.05), and a significant decrease (P < 0.05) in albumin (P < 0.001) and total bilirubin levels (P < 0.01) in AAS-users. Total testosterone increased significantly among AAS (P < 0.05), along with a significant decrease in LH (P < 0.01), and FSH (P < 0.001) levels, while serum prolactin and estradiol levels were significantly increased (P < 0.05). Conclusion: AAS can enhance physical performance and appearance, its potential adverse effects on the hepatic and reproductive systems necessitate careful consideration. Our research demonstrates an increase in the liver-specific enzyme ALT in AAS users relative to non-users and the possibility that short-term AAS usage increases the risk of liver injury.

Mouth rinses efficacy on salivary SARS-CoV-2 viral load: A randomized clinical trial.

Considering the global trend to confine the COVID-19 pandemic by applying various preventive health measures, preprocedural mouth rinsing has been proposed to mitigate the transmission risk of SARS-CoV-2 in dental clinics. The study aimed to investigate the effect of different mouth rinses on salivary viral load in COVID-19 patients. This study was a single-center, randomized, double-blind, six-parallel-group, placebo-controlled clinical trial that investigated the effect of four mouth rinses (1% povidone-iodine, 1.5% hydrogen peroxide, 0.075% cetylpyridinium chloride, and 80 ppm hypochlorous acid) on salivary SARS-CoV-2 viral load relative to the distilled water and no-rinse control groups. The viral load was measured by quantitative reverse transcription PCR (RT-qPCR) at baseline and 5, 30, and 60 min post rinsing. The viral load pattern within each mouth rinse group showed a reduction overtime; however, this reduction was only statistically significant in the hydrogen peroxide group. Further, a significant reduction in the viral load was observed between povidone-iodine, hydrogen peroxide, and cetylpyridinium chloride compared to the no-rinse group at 60 min, indicating their late antiviral potential. Interestingly, a similar statistically significant reduction was also observed in the distilled water control group compared to the no-rinse group at 60 min, proposing mechanical washing of the viral particles through the rinsing procedure. Therefore, results suggest using preprocedural mouth rinses, particularly hydrogen peroxide, as a risk-mitigation step before dental procedures, along with strict adherence to other infection control measures.

Study of trioleoylglycerol two-layer and adiposome cross-section mimicking four-layer systems through atomic-level simulations.

Adiposomes are artificially prepared lipid droplet (LD)-mimetic structures, which, unlike LDs, do not harbor proteins. The dynamics of interaction between triacylglycerols (TAGs), drug molecule, and phospholipids in adiposomes is currently not well-established. Trioleoylglycerol (TOG) molecule was divided into three parts: two oleoyl tails and one 2-monooleoylglycerol (MOG). Forcefield parameters for two oleoyl tails were adopted from the AMBER18 repository while that of the MOG forcefield was taken from the literature. Charge correction was performed on the MOG forcefield before its utilization. After charge correction, the resulting TOG molecule had zero charge. TOG bilayer (2L) and tetralayer (4L) systems were prepared and simulated. TOG bilayer (2L) systems-modeled from two different initial conformations, the TOG3 conformation and the TOG2:1 conformation-showed that TOG2:1 conformation was more prevailing irrespective of the starting conformation and was subsequently used in further simulations. The hydrated TOG 2L system showed TOG-water solution solubility of 0.051 mol L-1 which is near experimental values. This validated the correct parameterization of the TOG molecule. The simulations of 4L systems showed stable membrane behaviors toward the end of simulations. It was also observed that in the 4L system, the TOG molecules showed the formation of micelles with the drug molecule. Almost six TOGs remained continuously in contact with the drug molecule throughout the simulation. The availability of charge-corrected TOG parameterization is expected to equip future studies with a framework for molecular dynamics simulations of adiposomes and/or LDs at the atomic level.

Sugiol Suppresses the Proliferation of Human U87 Glioma Cells via Induction of Apoptosis and Cell Cycle Arrest.

The diterpenoid, sugiol, has been reported to exert anticancer effects against a number of human cancers. However, the anticancer effects of sugiol have not been evaluated against the human glioma cells. The present study was designed to examine the effects of sugiol on the proliferation of human U87 glioma cells. The results showed that sugiol significantly (P < 0.05) suppressed the viability of the U87 cells in a concentration dependent manner and exhibited an IC50 value of 15 µM. On the other hand, the growth inhibitory effects of sugiol were minimal on the normal human astrocytes. Acridine orange and ethidium bromide staining (AO/EB) staining revealed that sugiol induces apoptosis which was further confirmed by Western blot analysis, wherein upregulation of Bax and downregulation of Bcl-2 were observed in U87 cells. Flow cytometry showed that sugiol causes cell cycle arrest at the G 0/G 1 stage. The relative percentage of G1 phase was found to be increased from 26.58% at 0 µM to 70.96% at 30 µM sugiol. Taken together, the results suggest sugiol inhibits the growth of glioma cells and may prove to be a lead molecule in the management of human glioma.

A global report on the dynamics of COVID-19 with quarantine and hospitalization: A fractional order model with non-local kernel.

In this paper, a compartmental mathematical model has been utilized to gain a better insight about the future dynamics of COVID-19. The total human population is divided into eight various compartments including susceptible, exposed, pre-asymptomatic, asymptomatic, symptomatic, quarantined, hospitalized and recovered or removed individuals. The problem was modeled in terms of highly nonlinear coupled system of classical order ordinary differential equations (ODEs) which was further generalized with the Atangana-Balaeanu (ABC) fractional derivative in Caputo sense with nonlocal kernel. Furthermore, some theoretical analyses have been done such as boundedness, positivity, existence and uniqueness of the considered. Disease-free and endemic equilibrium points were also assessed. The basic reproduction was calculated through next generation technique. Due to high risk of infection, in the present study, we have considered the reported cases from three continents namely Americas, Europe, and south-east Asia. The reported cases were considered between 1st May 2021 and 31st July 2021 and on the basis of this data, the spread of infection is predicted for the next 200 days. The graphical solution of the considered nonlinear fractional model was obtained via numerical scheme by implementing the MATLAB software. Based on the fitted values of parameters, the basic reproduction number ℜ0 for the case of America, Asia and Europe were calculated as ℜ0≈2.92819, ℜ0≈2.87970 and ℜ0≈2.23507 respectively. It is also observed that the spread of infection in America is comparatively high followed by Asia and Europe. Moreover, the effect of fractional parameter is shown on the dynamics of spread of infection among different classes. Additionally, the effect of quarantined and treatment of infected individuals is also shown graphically. From the present analysis it is observed that awareness of being quarantine and proper treatment can reduce the infection rate dramatically and a minimal variation in quarantine and treatment rates of infected individuals can lead us to decrease the rate of infection.

Comparative proteomics reveals that lipid droplet-anchored mitochondria are more sensitive to cold in brown adipocytes.

Brown adipose tissue (BAT) is specialized for uncoupled heat production through mitochondrion fueled majorly from fatty acids (FAs) of lipid droplets (LDs). How the interaction between the two organelles contributes the generation of heat remains elusive. Here, we report that LD-anchored mitochondria (LDAM) were observed in the BAT of mice raised at three different temperatures, 30 °C, 23 °C, and 6 °C. The biochemical analyses including Western blotting of electron transport chain subunits showed that LDAM were functional. Comparative proteomics analysis was conducted, which revealed differential expressions of proteins between LDAM and cytoplasmic mitochondria (CM) at different temperatures. Higher expressions of proteins at low temperature were observed for i) FA ß-oxidation in LDAM including FA synthesis and uncoupling, ii) pseudo-futile cycle in CM, and iii) two shuttle systems: glycerol 3-phosphate in both CM and LDAM and citrate malate in CM. Together, these results suggest that LDs and LDAM form a preorganized and functional organelle complex that permits the rapid response to cold.

Stability and validity of intact parathyroid hormone levels in different sample types and storage conditions.

BACKGROUND: Several pre-analytical factors can affect the measurement of intact Parathyroid Hormone (IPTH). In this study, we have investigated the effects of using different types of tubes, time elapsed before separation, and storage conditions over time on the measured values of IPTH. METHOD: Blood samples from 30 subjects were collected into plain, SST, and EDTA tubes. All serum and plasma were separated immediately (first set) and after 2 hrs delay (second set). The first set of samples were aliquoted and stored at RT (25°C), at fridge (4°C), and freezer (-20°C). IPTH was measured in all the stored aliquots at 2,4, and 8 days after collection using Architect analyzer. RESULTS: Paired T test and ANOVA repeated measures showed no significant difference between IPTH levels in all tubes. The second set of serum and plasma were significantly lower (3.8% and 7.4%, p < 0.001, respectively) when compared to samples measured initially. Serum samples stored at RT were significantly lower (by 45%,59%, and 77%) on days 2,4, and 8 when compared to the initial time (p < 0.001 in all cases). Plasma samples stored at RT, were significantly lower on day 8 after collection, by 30.8% (p < 0.001). These differences would be clinically important. CONCLUSION: Plasma IPTH can be stored at RT for up to four days. Both plasma and serum IPTH are not affected by a delay in the separation of up to two h and they can be stored for up to 8 days in a fridge or freezer without any clinically significant changes in their values.

Rab18 binds PLIN2 and ACSL3 to mediate lipid droplet dynamics.

Lipid droplet (LD) is a vital organelle governing lipid homeostasis and Rab18 has been linked to lipid metabolism. However, the mechanisms of Rab18-mediated LD dynamics in myoblast cells remain elusive. Here, we report that Rab18 plays an important role in oleic acid (OA)-induced LD accumulation in mouse myoblast C2C12 cells. Rab18 was translocated from the endoplasmic reticulum (ER) to LDs during LD accumulation, which was regulated by perilipin 2 (PLIN2), a major LD protein. LD-associated Rab18 bound with the C terminus of PLIN2 and the LD localization of Rab18 was diminished when PLIN2 was depleted. Moreover, loss of function of Rab18 led to reduced triacylglycerol (TAG) level and fewer but larger LDs. In contrast, overexpression of Rab18 resulted in elevated TAG content and LD number. Furthermore, LD-associated Rab18 interacted with acyl-CoA synthetase long-chain family member 3 (ACSL3), which in turn promoted the LD localization of this protein. These data show that Rab18 interacts with PLIN2 and forms a complex with PLIN2 and ACSL3, which plays a critical role in LD accumulation and dynamics of myoblast cells.

Detection of Giardia lamblia by Microscopic Examination, Rapid Chromatographic Immunoassay Test, and Molecular Technique.

Background Giardia lamblia is a pathogenic intestinal flagellate transmitted by the ingestion of contaminated water or food with the cyst stage of the parasite. Giardiasis can cause severe acute diarrhea and malabsorption or may persist as a chronic infection. Effective treatment and control measures depend on proper laboratory diagnosis using diagnostic methods with high sensitivity and specificity. Objective To compare the sensitivity and specificity of direct smear, Ritchie sedimentation technique, two brands of rapid chromatographic immunoassay test, and real-time polymerase chain reaction (PCR) for the detection of G. lamblia in clinical human fecal samples. Materials and methods Unpreserved 100 stool specimens were collected in clean plastic containers and labeled with the patient's information and examined through light microscopy, immunochromatographic test (ICTs), and real-time PCR. Results Out of 100 fresh stool samples obtained from workers analyzed, real-time PCR targeting the SSU rRNA gene was able to detect Giardia deoxyribonucleic acid (DNA) in (42) samples followed by ImmunoCard STAT! (31) samples (Meridian Bioscience, Germany), direct smear (23) samples, CerTest (19) samples (Biotec, Zaragoza, Spain), and Ritchie technique (17) samples. Real-time PCR was the most sensitive for the diagnosis of G. lamblia in comparison to the other techniques. Conclusions All the techniques investigated were sensitive for the detection of G. lamblia in stool samples. Further studies are recommended using multiplex real-time PCR assay in order to increase the possibility of the presence or absence of the infection.

3D Printing of Metal/Metal Oxide Incorporated Thermoplastic Nanocomposites With Antimicrobial Properties.

Three-dimensional (3D) printing has experienced a steady increase in popularity for direct manufacturing, where complex geometric items can be produced without the aid of templating tools, and manufacturing waste can be remarkably reduced. While customized medical devices and daily life items can be made by 3D printing of thermoplastics, microbial contamination has been a serious obstacle during their usage. A very clever approaches to overcome this challenge is to incorporate antimicrobial metal or metal oxide (M/MO) nanoparticles within the thermoplastics during or prior to 3D printing. Many M/MO nanoparticles can prevent contamination from a wide range of microorganism, including antibiotic-resistant bacteria via various antimicrobial mechanisms. Additionally, they can be easily printed with thermoplastic without losing their integrity and functionality. In this mini review, we summarize recent advancements and discuss future trends related to the development of 3D printed antimicrobial thermoplastic nanocomposites by addition of M/MO nanoparticles.

Impact of Sequential Passaging on Protein Expression of E. coli Using Proteomics Analysis.

Urinary tract infection (UTI) is one of the most prevalent bacterial infections in the world affecting the bladder and the kidney. Escherichia coli (E. coli) is the main causative agent of 80-90% of community-acquired UTIs, about 40% of nosocomial UTIs, and 25% of recurrent UTIs. The field of proteomics has emerged as a great tool to analyze expressed proteins to identify possible biomarkers associated with many pathological states and, to the same extent, those associated with bacterial pathogenesis and their ability to cause recurrent infections. Here, in a descriptive cross-sectional pilot study, we employed proteomic techniques to investigate the effects of environmental stress on protein profiles of E. coli simulated by sequential passaging of samples from patients with UTIs to screen for unique proteins that arise under stressful environment and could aid in the early detection of UTIs. Four urine samples were collected from individuals with recurrent UTI and sequentially subcultured; protein samples were extracted from bacterial pellets and analyzed using 2-dimensional gel electrophoresis (2DGE). Protein spots of interest arising from changes in the protein profile were analyzed using liquid chromatography-mass spectrometry (LC-MS/MS) and matched against known databases to identify related proteins. We identified ATPB_ECOBW, ASPA ECOLI, DPS ECOL6, and DCEB ECOLI as proteins associated with higher passaging. We concluded that passaging resulted in identifiable changes in the protein profile of E. coli, namely, proteins that are associated with survival and possible adaptation of bacteria, suggestive of factors contributing to antibiotic resistance and recurrent UTIs. Furthermore, our method could be further used to identify indicator-protein candidates that could be a part of a growing protein database to diagnose and identify causative agents in UTIs.

Cross-sectional prevalence study of MERS-CoV in local and imported dromedary camels in Saudi Arabia, 2016-2018.

The Middle East Respiratory Syndrome-Coronavirus (MERS-CoV) is an endemic virus in dromedaries. Annually, Saudi Arabia imports thousands of camels from the Horn of Africa, yet the epidemiology of MERS-CoV in these animals is largely unknown. Here, MERS-CoV prevalence was compared in imported African camels and their local counterparts. A total of 1399 paired sera and nasal swabs were collected from camels between 2016 and 2018. Imported animals from Sudan (n = 829) and Djibouti (n = 328) were sampled on incoming ships at Jeddah Islamic seaport before unloading, and local camels were sampled from Jeddah (n = 242). Samples were screened for neutralizing antibodies (nAbs) and MERS-CoV viral RNA. The overall seroprevalence was 92.7% and RNA detection rate was 17.2%. Imported camels had higher seroprevalence compared to resident herds (93.8% vs 87.6%, p <0.01) in contrast to RNA detection (13.3% vs 35.5%, p <0.0001). Seroprevalence significantly increased with age (p<0.0001) and viral RNA detection rate was ~2-folds higher in camels <2-year-old compared to older animals. RNA detection was higher in males verses females (24.3% vs 12.6%, p<0.0001) but seroprevalence was similar. Concurrent positivity for viral RNA and nAbs was found in >87% of the RNA positive animals, increased with age and was sex-dependent. Importantly, reduced viral RNA load was positively correlated with nAb titers. Our data confirm the widespread of MERS-CoV in imported and domestic camels in Saudi Arabia and highlight the need for continuous active surveillance and better prevention measures. Further studies are also warranted to understand camels correlates of protection for proper vaccine development.

Impact of smoking cessation, coffee and bread consumption on the intestinal microbial composition among Saudis: A cross-sectional study.

The gut microbiota is often affected by the dietary and lifestyle habits of the host, resulting in a better efficacy that favors energy harvesting from the consumed food. Our objective was to characterize the composition of gut microbiota in adult Saudis and investigate possible association with lifestyle and dietary practices. Feces from 104 Saudi volunteers (48% males) were tested for microbiota by sequencing the V3-V4 region of bacterial 16S ribosomal RNA (rRNA). For all participants, data were collected related to their lifestyle habits and dietary practices. The relative abundance (RA) of Fusobacteria was significantly higher in normal weight Saudis (P = 0.005, false discovery rate-FDR = 0.014). Individuals who consumed more coffee presented marginally significant more RA of Fusobacteria (P = 0.02, FDR = 0.20) in their gut microbiota compared to those reporting low or no coffee intake, but the RA of Fusobacteria was significantly higher in smokers compared to non-smokers (P = 0.009, FDR = 0.027). The RA of Fusobacteria was also significantly higher in those reporting daily consumption of bread (P = 0.005, FDR = 0.015). At the species level, the gut microbiota of people who consumed coffee was dominated by Bacteroides thetaiotaomicron followed by Phascolarctobacterium faecium and Eubacterium rectale. Similarly, the gut microbiota of smokers was also enriched by B. thetaiotaomicron and Lactobacillus amylovorus. Smoking cessation, bread and coffee consumption induce changes in the intestinal microbial composition of Saudis. This indicates the significance of diet and lifestyle practices in the determination of the composition of the gut microbiota, which could possibly lead later to changes in metabolic profile and weight.

Seroprevalence of MERS-CoV in healthy adults in western Saudi Arabia, 2011-2016.

BACKGROUND: The Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly recognized zoonotic coronavirus. Current evidence confirms the role of dromedaries in primary human infections but does not explain the sporadic community cases. However, asymptomatic or subclinical cases could represent a possible source of infection in the community. METHODS: Archived human sera (7461) collected between 2011 and 2016 from healthy adult blood donors from 50 different nationalities in the western part of Saudi Arabia were obtained for MERS-CoV seroprevalence investigation. Samples were tested for MERS-CoV S1-specific antibodies (Abs) by ELISA and confirmed by testing for neutralizing Abs (nAbs) using both pseudotyped and live virus neutralization assays. RESULTS: Out of 7461 samples, 174 sera from individuals with 18 different nationalities were ELISA positive (2.3%, 95% CI 2.0-2.7). Presence of nAbs was confirmed in 17 samples (0.23%, 95% CI 0.1-0.4) of which one sample exhibited positivity in both neutralization assays. Confirmed seropositivity was identified in young (15-44 years) men and women from Saudi Arabia, Egypt, Yemen, Pakistan, Palestine, Sudan, and India without significant preference. CONCLUSIONS: An increasing trend of MERS-CoV seroprevalence was observed in the general population in western Saudi Arabia, suggesting that asymptomatic or mild infections might exist and act as an unrecognized source of infection. Seropositivity of individuals from different nationalities underscores the potential MERS exportation outside of the Arabian Peninsula. Thus, enhanced and continuous surveillance is highly warranted.

Chemokines and their association with body mass index among healthy Saudis.

Obesity is a chronic disorder that is associated with body mass index (BMI) of greater or equal to 30 kg/m2. The prevalence of obesity in the Kingdom of Saudi Arabia (KSA) is increasing at an alarming rate and is expected to reach 41% in men and up to 78% in women by 2022. Since chemokines are associated with involuntary weight loss, the objective of this study was to elucidate their association with BMI among Saudis. A questionnaire was used to collect information about diet, health conditions, and demographics from 15 men and 16 women who participated in the study. BMI was calculated based on clinical measurements and participants were classified according to their BMI category as: normal, underweight, overweight, or obese. Serum samples were collected for a multiplex assay using the Human Chemokine Magnetic 30-plex panel. The serum concentration of either the monokine induced by gamma interferon (MIG) or the CXC-motif chemokine ligand 9 (CXCL-9) was significantly increased in obese men (P = 0.0194) and women (P = 0.043) as compared to underweight men and women, respectively. However, the serum levels of other chemokines were not significantly different among the groups. We found that MIG levels are differentially regulated in serum, based on individuals' BMI.

Prevalence of human papillomavirus in Jeddah, Saudi Arabia.

BACKGROUND: Human papillomaviruses (HPVs) are small, non-enveloped, double-stranded DNA viruses that consist of more than 200 genotypes. Low-risk genotypes are associated with warts or benign lesions, whereas high-risk genotypes are usually associated with malignancies and cancers including cervical cancer. However, the real prevalence and incidence of HPV in Saudi Arabia may be understated due to a lack of comprehensive data reporting. OBJECTIVES: Determine the positivity rate of HPV in men and women in Jeddah, Saudi Arabia. DESIGN: Cross-sectional. SETTING: Tertiary care center in Jeddah. SUBJECTS AND METHODS: Self-collected vaginal swab samples were obtained from females attending the gynecological clinic in the period between October 2017 and April 2018 at a tertiary care center, Jeddah, Saudi Arabia. PCR-positive HPV samples were sequenced to determine genotype. Additionally, serum samples were collected from healthy male and female blood donors and screened for HPV IgG antibodies by ELISA. MAIN OUTCOME MEASURES: Molecular and serological positivity for HPV. SAMPLE SIZE: 119 self-collected vaginal swabs from females at a gynecology clinic and 966 serum samples from healthy blood donors. RESULTS: Of the 119 tested vaginal swabs, 7 samples (5.9%) were positive for HPV DNA. Several genotypes were identified. Most of the positive samples were from Saudi females in the age range of 31-50 years seeking care for infertility. Of the 966 serum samples, only 16 samples (1.7%) were positive for HPV IgG antibodies. CONCLUSION: While the prevalence of HPV in men and women in our sample from the western region of Saudi Arabia was low, our data clearly show that it is not uncommon among high-risk groups and people are still exposed to the risk of HPV infection. Most importantly, these data provide valuable information that could aid in enhancing national awareness about HPV and in introducing an HPV vaccination program. LIMITATIONS: Single hospital and a convenience sample CONFLICT OF INTEREST: None.