Awareness of the Effect of Vitamin B12 Deficiency on the Nervous System Among the General Population in Taif, Saudi Arabia.

BACKGROUND: Vitamin B12 is important for the health of the nervous system, its deficiency leads to various neurological manifestations such as visual problems, ataxia, peripheral neuropathy, dementia, etc. The deficiency can be caused by malnutrition, malabsorption, or increased demand. Early detection is important for the control and prevention of complications. METHOD: In December 2021, a population-based cross-sectional survey was carried out among Saudi males and women at least 18 years old. There were 383 participants in the sample. An electronic survey distributed over social media was used to collect the data. SPSS version 28 was used to analyze the data. RESULTS: The majority of the respondents were female (88%). Most participants were aged 18-25 years (44%). Regarding participants' awareness and knowledge of vitamin deficiency, 64% were aware of vitamin B12 deficiency. 41.7% of participants knew about food sources of vitamin B12, 29.0% knew how to prevent it, and 30.0% took vitamin B12 supplements. (92.2%) of the participants were not following vegetarian or vegan diets. Difficulty concentrating accounted for the highest number of reported symptoms by the respondents (53.8%). CONCLUSION: This study recommends a scientific approach encouraging patients to self-report their B12 deficiency in medical institutions. In addition, a study about the relationship between B12 deficiency and other neurodegenerative disorders is also a recommendation of this study.

Successful Use of Virtual Microscopy in the Assessment of Practical Histology during Pandemic COVID-19: A Descriptive Study.

BACKGROUND: Practical knowledge and skills of microscopy has classically been delivered for medical students using conventional microscopes (CMs). Using virtual microscopy (VM) in teaching practical histology was established during distance learning for Taif medical students during COVID 19 pandemic period. However, the suitable assessment methods for student performance during distance learning are still debatable. We focused on how to ensure the learner's achievement of course practical outcomes and learning domain. AIMS AND OBJECTIVES: This study aimed to ascertain whether using VM to evaluate student learning in practical Histology during distance education programs and if moving to VM affecting students' scores. MATERIALS AND METHODS: For the first time, we used VM during online objective structural practical examination (OSPE) of 3rd year medical students. Different sets of virtual slides were given for students at the time of assessment, then different tasks were described and each student was asked to finalize his/her task during the designed time. A specific rubric was designed for the evaluation of student work. Moreover, student perceptions of VM as teaching and assessment method were assessed using online survey. Post examination psychometric analysis of VM OSPE was done and compared with previous OSPE results of the same batch of students. RESULTS: The average student score was 4.63 ± 0.51 with no significant difference from previous student's scores. The net students' feedback was positive. Their average satisfaction on all items ranged from 3.7 to 4.25 on Likert scale. Students recorded the easy image access at any time and place with VM as the most distinctive feature. CONCLUSION: Our results indicated that VM is not only an effective method in teaching histology but also it is an assessment method for measuring student performance during online assessment.

Morphological and Biochemical Features of Cerebellar Cortex After Exposure to Zinc Oxide Nanoparticles: Possible Protective Role of Curcumin.

Zinc oxide nanoparticles (ZnONPs) are widely used in the last decades. Therefore, investigation of its neurotoxic effect is important. This work aimed to investigate the potential adverse effects of ZnONPs on rat's cerebellar cortex and the possible neuroprotective role of curcumin (Cur). Forty male albino rats were randomly divided into four equal groups. Two groups were injected with ZnONPs and one group was previously received Cur before ZnONPs. At the end of the experiment, cerebellar tissue samples were prepared for histological, morphometric, immunohistochemical study, and tissue levels of oxidative stress markers and cytokine analysis. cerebellar damage is clearly visible with ZnONPs. Degeneration, loss, disorganization of cerebellar neurons was observed. Histopathological degeneration of Purkinje and granular cells together with loss of Nissl substance, astrocyte gliosis, and affection of cerebellar blood brain barrier were detected. Moreover, an apoptotic marker (caspase-3) was significantly expressed in Purkinje and granular layers together with elevated gene expression of P53 and COX-2 in cerebellar tissue of ZnONPs intoxicated group. Astrocyte gliosis and inflammatory markers IL-1, IL-6, and TNF-α were expressed significantly in ZnONPs intoxicated cerebellum. These changes were associated with evidence of cerebellar oxidative stress. Strikingly, treatment with Cur together with ZnONPs recorded morphological improvement, with increased number of Purkinje cells and decreased caspase +ve cells. These findings were confirmed by morphometric and statistical analysis. Cur ameliorates the deterious effect of ZnONPs on the cerebellar cortex through its antioxidant, antiapoptotic, and anti-inflammatory efficacies. Anat Rec, 2018. © 2018 Wiley Periodicals, Inc.

Role of adipose tissue derived stem cells differentiated into insulin producing cells in the treatment of type I diabetes mellitus.

Generation of new ß cells is an important approach in the treatment of type 1 diabetes mellitus (type 1 DM). Adipose tissue-derived stem cells (ADSCs) might be one of the best sources for cell replacement therapy for diabetes. Therefore, this work aimed to test the possible role of transplanted insulin-producing cells (IPCs) differentiated from ADSCs in treatment of streptozotocin (STZ) induced type I DM in rats. Type 1 DM was induced by single intra peritoneal injection with STZ (50 mg/kg BW). Half of the diabetic rats were left without treatment and the other half were injected with differentiated IPCs directly into the pancreas. ADSCs were harvested, cultured and identified by testing their phenotypes through flow cytometry. They were further subjected to differentiation into IPCs using differentiation medium. mRNA expression of pancreatic transcription factors (pdx1), insulin and glucose transporter-2 genes by real time PCR was done to detect the cellular differentiation and confirmed by stimulated insulin secretion. The pancreatic tissues from all groups were examined 2 months after IPC transplantation and were subjected to histological, Immunohistochemical and morphometric study. The differentiated IPCs showed significant expression of pancreatic ß cell markers and insulin secretion in glucose dependent manner. Treatment with IPCs induced apparent regeneration, diffused proliferated islet cells and significant increase in C-peptide immune reaction. We concluded that transplantation of differentiated IPCs improved function and morphology of Islet cells in diabetic rats. Consequently, this therapy option may be a promising therapeutic approach to patient with type 1 DM if proven to be effective and safe.

Pomegranate protective effect on experimental ischemia/reperfusion retinal injury in rats (histological and biochemical study).

Oxidative stress is one mechanism involved in the pathogenesis of ischemia/reperfusion (/R) retinal injury. The histological, biochemical, and functional changes associated with pomegranate (PMG) treatment prior to retinal I/R were analyzed using 40 adult male albino rats. Rats were divided into four groups: Groups I and II (sham operated and received saline or PMG, respectively); Groups III and IV (I/R rat models with prior administration of saline or 250 mg/kg/day PMG, respectively). Electroretinogram (ERG) results were recorded and eye specimens were taken and processed for light and electron microscopic examinations and for assessment of oxidative status in retinal homogenate. I/R lead to degenerative changes in retinal layers with a significant reduction in nuclear factor erythroid 2-related factor 2 (Nrf2) immunoreactivity in concomitant with significant oxidant-antioxidant disturbance and decreased a- and b-wave amplitude in the ERG. These alterations were ameliorated with prior PMG treatment. In conclusion, PMG treatment, as an antioxidant, attenuated retinal structural and functional I/R injury through activation of Nrf2 which could be a base for future therapy designs.

Caffeine intake decreases oxidative stress and inflammatory biomarkers in experimental liver diseases induced by thioacetamide: Biochemical and histological study.

Liver disease remains a significant global health problem. Increased caffeine consumption has been associated with a lower prevalence of chronic liver disease. This study aimed to investigate the modifying effects of caffeine on liver injury induced by thioacetamide (TAA) administration in male rats and the possible underlying mechanisms. Forty adult male rats were equally classified into four groups: control group, received only tap water; caffeine-treated group, received caffeine (37.5 mg/kg per day); TAA-treated group, received intraperitoneal (i.p.) TAA (200 mg/kg b.w.) twice a week; and caffeine + TAA-treated group, received combined TAA and caffeine in the same previous doses. After eight weeks of treatment, blood samples were collected for biochemical analysis and liver specimens were prepared for histological and immunohistochemical studies and for assessment of oxidative stress. TAA induced liver toxicity with elevated liver enzymes and histological alterations, fatty changes, apoptosis, and fibrosis evidenced by increased immunohistochemical reaction to matrix metalloproteinase-9 (MMP-9) and collagen type IV in hepatocytes. Also, the levels of pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6) in serum were significantly elevated. Co-treatment with caffeine and TAA restored normal liver structure and function. Caffeine provided an anti-fibrogenic, anti-inflammatory, and antioxidant effect that was associated with recovery of hepatic histological and functional alterations from TAA-induced hepatotoxicity.

Gestational protein restriction: study of the probable effects on cardiac muscle structure and function in adult rats.

Intrauterine growth restriction (IUGR) has been linked to heart disease in adulthood. This study aimed to examine the effect of gestational protein restriction during fetal and early postnatal life on the cardiac muscle structure and function in adult offspring. Pregnant female rats were randomly divided into two dietary groups: normal-protein diet (NP) and low-protein diet (LP). Fifteen male offspring from each group were included in the study. Offspring body weights were recorded at birth and monthly from weaning until 24 weeks of age while systolic blood pressure was measured weekly. At the end of the experiment, hearts were weighed and processed for light and electron microscopy and immunohistochemical study. Immunohistochemical staining for localization of inducible nitric oxide synthase (iNOS) and connexin 43 proteins was performed. The gestational protein restriction induced deleterious effects on adult offspring including decreased birth weight, heart weight, and heart rate, and increased systolic blood pressure. Histologically, the number of cardiomyocytes decreased and cardiac fibrosis increased. Signs of degeneration at both structural and ultra-structural levels of cardiomyocytes were also seen. The iNOS was up regulated in LP offspring which was a promoter for apoptosis, while connexin 43 was down regulated which would affect heart conductivity and contractility. Our results demonstrate that adult offspring body weight and cardiac muscle structure and function can be programmed by maternal gestational nutrition. These adverse outcomes suggest the criticality of dietary behavior during pregnancy on long-term offspring cardiac health.

DNA repair genes polymorphisms and risk of colorectal cancer in Saudi patients.

BACKGROUND AND STUDY AIMS: Polymorphisms in the DNA repair genes may influence individual capacity to repair DNA damage, which may be associated with increased genetic instability and carcinogenesis. Our aim was to evaluate the relation of genetic polymorphisms in 2 DNA repair genes, XPD Lys751Gln and XRCC1 (A399G), with colorectal cancer (CRC) susceptibility. We further investigated the potential effect of these DNA repair variants on clinicopathological parameters of CRC patients. PATIENTS AND METHODS: Both XPD and XRCC1 polymorphisms were characterised in one hundred CRC patients and one hundred healthy controls who had no history of any malignancy by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method and PCR with confronting two-pair primers (PCR-CTPP), using DNA from peripheral blood in a case control study. RESULTS: Our results revealed that the frequencies of GG genotype of XRCC1 399 polymorphism were significantly higher in the CRC patients than in the normal individuals (p⩽0.03), and did not observe any association between the XPD Lys751Gln polymorphism and CRC risk. We found association between both XRCC1 A399G polymorphisms and histological grading of disease. CONCLUSION: Our results suggested that, XRCC1 gene is an important candidate gene for susceptibility to colorectal carcinoma.

Cilostazol attenuates gentamicin-induced nephrotoxicity in rats.

INTRODUCTION: Gentamycin is a widely used antibiotic. The nephrotoxic adverse effects of the drug may limit its use. Cilostazol, a phosphodiesterase III inhibitor, was reported to protect from renal oxidative stress. This work aimed to investigate the possible protective effect of cilostazol on gentamicin-induced nephrotoxicity and the possible underlying mechanisms. MATERIALS AND METHODS: 40 male albino rats were divided into 4 equal groups: (1) Control; (2) Cilostazol, 10mg/kg, p.o.; (3) Gentamicin, 80 mg/kg, i.p.; (4) Gentamicin 80 mg/kg, i.p. along with cilostazol 10mg/kg, p.o. All drugs were administered once daily for 8 days. On 9th day blood samples were collected for the estimation of creatinine, urea and uric acid in serum. Then the rats were sacrificed and kidneys were removed for light and electron microscope studies. Moreover, reduced glutathione (GSH) and malondialdehyde (MDA) levels as well as catalase (CAT) and superoxide dismutase (SOD) activities were determined in renal tissues. RESULTS: Gentamicin elevated the serum levels of creatinine, urea and uric acid as well as the MDA level in the renal tissue, while it decreased CAT, SOD activities and GSH levels as well as produced degenerative changes in glomeruli and tubules associated with increased expression of apoptotic markers and decreased expression of anti-apoptotic markers. Administration of cilostazol decreased urea, creatinine, uric acid and MDA levels while increased CAT and SOD activities and GSH levels as well as ameliorated the histopathological changes in relation to gentamicin group. CONCLUSION: Cilostazol protected rats from gentamicin-induced nephrotoxicity possibly, in part through its antioxidant and anti-apoptotic activity.

Anti-apoptotic effect of spermatogonial stem cells on doxorubicin-induced testicular toxicity in rats.

The present study was designed to investigate whether spermatogonial stem cells (SSCs) have possible effect on doxorubicin (DOX)-induced testicular apoptosis and damaged oxidant/antioxidant balance in rats. Sixty male Albino rats were divided into 3 groups: the saline control group, the testicular toxicity group (2mg/kg DOX once a week for 8 weeks) and the third group is a donor stem cells transplanted following pre-treatment with DOX. After the 8th week, the rats were sacrificed and tissues were collected and examined for CD95, CD95L, Caspase 3, and Caspase 8 gene expression using RT-PCR. While malondialdehyde (MDA), glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD) were determined using colorimetric kits. Biochemical, histopathological and PCR results showed improvement of the SSCs' group compared to the DOX-group. It was observed that spermatogonial stem cell affected DOX-induced activation of intrinsic apoptotic signaling pathway via preventing DOX-induced increases in CD95 and CD95L levels as well as cleaved Caspase-8 and Caspase-3 levels in testicular tissues, however, spermatogonial stem cell decreased Dox-induced NF-κB activation as well. It can be concluded that SSCs may be utilized to develop new cell-based therapies, and to advance germline gene therapy.

Vitamin D receptor gene polymorphisms and steroid receptor status among Saudi women with breast cancer.

The vitamin D receptor (VDR) is a mediator for the cellular effects of vitamin D and interacts with other cell signaling pathways that influence cancer development. We evaluated the associations of the FOK1 and Taq1 VDR polymorphisms and breast cancer risk and possible effect modification by steroid receptor status of the tumor. This case-control study includes 95 breast cancer patients and 100 age-matched controls. Genotyping for VDR FOK1 and Taq1 polymorphisms was performed using polymerase chain reaction-based restriction fragment length polymorphism. Level of 25(OH)D in serum was determined using ELISA. Immunohistochemical studies were performed for estrogen receptors (ER) and progesterone receptors (PR). The frequencies of ff genotype were significantly increased in the breast cancer group compared to the control group. Carriers of the f allele were significantly more likely to develop BC. We observed a statistically significant interaction for the Fok1 polymorphism and ER status. Our results demonstrated that FOK1 f. genotype and f allele have an important role in breast cancer risk in Saudi patients.

Role of calorie restriction in alleviation of age-related morphological and biochemical changes in sciatic nerve.

BACKGROUND: Aging is associated with structural, functional and biochemical alterations in the nervous system. Calorie restriction (CR) was found to retard most physiological indices of aging. OBJECTIVES: This work aimed to investigate the effect of CR on age-related changes in sciatic nerves. MATERIALS AND METHODS: Thirty male albino rats aged 1 month were equally divided into three groups; Group I [control adult-ad libitum AL]: fed a regular diet and sacrificed at the age of 6 months, group II (aged-AL group): fed a regular diet AL and sacrificed at the age of 18 months, and group III (aged CR) fed a 40% calorie restricted diet and sacrificed at the age of 18 months. Rats were anesthetized and sciatic nerves were processed for light, electron microscope and morphometric studies. Oxidative stress in sciatic nerves was investigated by estimation of lipid perioxidation by product malondialdehyde (MDA) tissue level and antioxidant enzyme; superoxide dismutase activity (SOD). RESULTS: The aged (AL) sciatic nerves appeared disorganized, with thick perineurium and increased collagen fibers associated with decreased g-ratio. Abnormal myelin forms were seen as outfolded myelin loops, thin denuded myelin, splitting of myelin into myelin figures and interlamellar vacuoles. Schwann cells revealed vacuolated cytoplasm. There was also significant increase in MDA level and a significant decrease in SOD activity in comparison to control adult (AL). Apparent structural and histomorphological improvement were noticed after CR in aged rats. CONCLUSION: Aging caused structural and biochemical alterations in sciatic nerves with alleviating effect of calorie restriction on such effects.

Mechanistic action of mesenchymal stem cell injection in the treatment of chemically induced ovarian failure in rabbits.

BACKGROUND: No curative treatment is known for primary ovarian failure; however, mesenchymal stem cells (MSCs), through self-renewal and regeneration, push the trial to evaluate their role in the treatment of ovarian failure. The aim of this study was to explore the impact of MSCs on cyclophosphamide (CTX)-induced ovarian failure in rabbits and to clarify the mechanism(s) by which MSCs exert their action. METHODS: Thirty-five adult female rabbits were injected with CTX to induce ovarian failure. Five rabbits were euthanized after the last injection of CTX for histological examination. The others (30 rabbits) were further subdivided into two groups: group 1 (ovarian failure group, 15 rabbits) received no treatment; group 2 (ovarian failure and MSC recipient group, 15 rabbits) received MSCs isolated from extracted bone marrow of male rabbits. RESULTS: A decrease of follicle-stimulating hormone and an increase of estrogen and vascular endothelial growth factor (VEGF) levels in the MSC recipient group versus the ovarian failure group were found. Weak caspase-3 expression and +ve proliferating cell nuclear antigen staining after MSC injection were detected. Cytological and histological examinations showed increased follicle numbers with apparent normal structure of ovarian follicles in the MSC recipient group. Moreover, Y chromosome-containing cells from male donors were present within the ovarian tissues in group 2. CONCLUSIONS: The current study suggests that intravenous injection of MSCs into rabbits with chemotherapy-induced ovarian damage improved ovarian function. MSCs accomplish this function by direct differentiation into specific cellular phenotypes and by secretion of VEGF, which influence the regeneration of the ovary.

Epicatechin attenuates doxorubicin-induced brain toxicity: critical role of TNF-α, iNOS and NF-κB.

Doxorubicin (DOX) is considered one of the most important chemotherapeutic agents that is used for the treatment of solid tumors. Its long-term use can cause neurodegenerative disorders due to its prolonged activation of microglia. The present study proved that the use of epicatechin prior to DOX treatment significantly attenuated not only the increase in TNF-α, iNOS and NF-κB expressions but also the increase in TNF-α and total nitrite levels in brain tissue when compared with rats treated with DOX-only. Thus, our study revealed that epicatechin can be used for the treatment of neuroinflammation and also for preventing the development of neurodegenerative disease during antineoplastic therapy because of its protective role in attenuation of neurotoxic pro-inflammatory mediators including TNF-α, NF-κB, and iNOS.