Comparing the effect of using 2-Mercaptoethanol in the cell culture medium of different cell passages of human mesenchymal stem cells.

Preparing a suitable cell culture medium that supports the biological needs of the growing cells is crucial to enhancing the success rate of any in vitro and in vivo experiments and minimizing undesirable interferences.  Mesenchymal stem cells ( MSCs) which are powerful regenerative stem cells require being grown in proper culture media to preserve their stemness and therapeutic properties. MSCs are usually grown in Dulbecco's Modified Eagle low glucose Medium (DMEM low glucose) which contains 5.6 mmol/L of glucose and is supplemented with Fetal Bovine Serum (FBS), antibiotics, and 2-Mercaptoethanol. The addition of 2-Mercaptoethanol to the cell culture medium was proposed long ago and has continued to be used until now. Despite the positive effects of adding 2-Mercaptoethanol in the cell culture medium, its use is still controversial and needs continuous updates to limit its interference with experimental treatments. Herein, we found that 2-Mercaptoethanol is beneficial to enhancing the proliferation and survival of MSCs at higher passage numbers while its effect is negligible for earlier passages. This concise study provides updates regarding the suitable time to add 2-Mercaptoethanol which can minimize its intermeddling with the experimental design and treatments.

The toxic profile of tramadol combined with nicotine on the liver and testicles: evidence from endoplasmic reticulum stress.

BACKGROUND: Tramadol is one of the most commonly abused substances in the Middle East. Furthermore, smoking is extremely common among the population. METHODS: An experimental study was performed on Sprague-Dawley rats to explore the effects of both nicotine and tramadol on the liver and testes. The tramadol was administered at 10 and 20 mg/kg, respectively, while the nicotine was administered at 125 mg/kg. Histological examination and androgen receptor ELISA assay showed mild effects on the liver and proofed safety on the testis. Western blot analysis of BIP (immunoglobulin heavy-chain binding protein) and CHOP (CCAAT-enhancer-binding protein homologous protein) revealed that fewer problems were induced by adding nicotine to tramadol. Autophagy marker LCIII and apoptosis marker caspase-8 showed similar effects to CHOP and BIP on liver samples. The real-time PCR of BIP expression showed similar but not identical results. CONCLUSIONS: The results showed mild endoplasmic reticulum stress, autophagy, and apoptosis in the liver samples. Histological examination revealed stable spermatogenesis with average androgen receptor blood levels in the different groups.

Neurological Effects of Combining Low Toxic Dose of Tramadol and Nicotine: An Animal Model Evidence of Endoplasmic Reticulum Stress.

Tramadol abuse is a common problem in the Middle East in conjunction with smoking. The current study applied immunohistochemistry, western blot, real-time PCR, and ELISA to test the combination toxicity. Low toxic doses of tramadol induced animal brain cortex inflammation and hippocampus injury. Adding nicotine reverted hippocampus pathological changes without triggering marked brain injury. The expression of CHOP protein with real-time PCR showed mild endoplasmic reticulum stress (ER) in rat's brain. Histological, immunohistochemical, and western blotting analysis of CHOP (CCAAT-enhancer-binding protein homologous protein) and BIP (immunoglobulin heavy chain-binding protein) chaperones demonstrated endoplasmic reticulum stress in the brains of animals. Furthermore, the levels of apoptosis and autophagy markers demonstrated a mild reaction. The blood level of serotonin was high in all study groups, with a marked increase in the combined one. The high serotonin levels in the blood can be critical and associated with a high risk of serious withdrawal and pathological consequences. Serotonin receptor blockers such as olanzapine may increase systemic serotonin levels and need further investigation to utterly pinpoint their roles in managing mood disorders. In conclusion, the combination of tramadol and nicotine is less harmful than expected. However, serious withdrawal effects can occur as a result of high systemic serotonin effects.

The impact of multiple sclerosis on the size and morphology of corpus callosum: an MRI-based retrospective study / Impacto de la esclerosis múltiple en el tamaño y la morfología del cuerpo calloso: un estudio retrospectivo basado en resonancia magnética

SUMMARY: The corpus callosum (CC) includes the majority of fibers linking the two brain hemispheres. Several cross sectional studies showed an association between callosal atrophy and malfunction and neurodegenerative diseases, which may play a role in their pathological manifestations. As a result, the accurate quantification of the corpus callosum is important to have normative values according to sex, age and ethnicity. The purpose of this study is to determine the size of CC in patients suffering from multiple sclerosis, and compare it to CC size in healthyindividuals. Midsagittal size of CC were recorded prospectively from 404 routine MR brain examinations in normal individuals. The internal skull surface was measured to calculate CC/ internal skull surface ratio. Two groups of patients were studied: 200 (100 male /100 female) healthy individuals and 204 (101 males/103 females) with multiple sclerosis (MS). Mean surface area of CC in controls was 6.58±1.04 cm2 and there was no significant difference between males and females (P< 0.627). CC/ internal skull surface ratio was 4.44±0.77 %. MS patients showed a significant decrease in CC size compared to normal controls. Using MR imaging, we measured the mean sizes of the various portions of the CC in normal individuals, in addition to MS patients; these values may provide a useful basis to determine changes occurring in CC structures.

El cuerpo calloso (CC) incluye la mayoría de las fibras que unen los dos hemisferios cerebrales. Varios estudios transversales mostraron una asociación entre la atrofia y el mal funcionamiento calloso y las enfermedades neurodegenerativas, lo que puede desempeñar un papel en sus manifestaciones patológicas. En consecuencia, la cuantificación precisa del cuerpo calloso es importante para tener valores normativos según sexo, edad y etnia. El propósito de este estudio fue determinar el tamaño de CC en pacientes que padecen esclerosis múltiple y compararlo con el tamaño de CC en individuos sanos. El tamaño sagital medio del CC se registró prospectivamente a partir de 404 exámenes cerebrales de RM de rutina en individuos normales. Se midió la superficie interna del cráneo para calcular la relación CC/superficie interna del cráneo. Se estudiaron dos grupos de pacientes: 200 (100 hombres/100 mujeres) sanos y 204 (101 hombres/103 mujeres) con esclerosis múltiple (EM). El área superficial media de CC en los controles fue de 6,58±1,04 cm2 y no hubo diferencia significativa entre hombres y mujeres (P< 0,627). La relación CC/superficie interna del cráneo fue de 4,44±0,77 %. Los pacientes con EM mostraron una disminución significativa en el tamaño de CC en comparación con los controles normales. Usando imágenes de RM, medimos los tamaños medios de las diversas porciones del CC en individuos normales, además de pacientes con EM; estos valores pueden proporcionar una base útil para determinar los cambios que ocurren en las estructuras CC.

Hypoxia disturbs the Migration and Adhesion characteristics of Mesenchymal Stem Cells.

Mesenchymal stem cells (MSCs) have been successfully used in treating many diseases which being verified by many preclinical and clinical trials.  Despite the exciting therapeutic potential of MSCs, multiple challenges encountered which hinder researchers from achieving successful clinical translations. Many studies have showed that moderate hypoxia (1-7% O2) considered as an important regulator of MSCs homing, migration, and differentiation. Additionally, low oxygen tension levels have been implicated in the maintenance of MSCs quiescence and plasticity in general. On the other hand, severe hypoxia (<1% O2) negatively affects the in vitro therapeutic potential of MSCs and causing their poor survival. Using Elisa assay we assessed some major adhesion markers that are known to be secreted by MSCs and play a role in cell-cell and cell-matrix adhesion under normoxia (21% O2) and severe hypoxia (0.5% O2). These markers including: SDF1-α, CXCR4, FAK, VEGF and ICAM-1. The results showed a significant drop in the adhesion markers in MSCs under severe hypoxia in comparing to normoxia, which causes a disruption in the cell-cell adhesion abilities of MSCs and ultimately can affect the incorporation of MSCs at the host site. These Findings can open new avenue to improve the attachment of MSCs at the transplantation site by targeting the adhesion and chemokines markers.

Assessing the racial differences in Q angle measurements in relevance to articular cartilage of the femoral condyle: a retrospective ultrasound study / Evaluación de las diferencias raciales en las mediciones del ángulo Q en relación con el cartílago articular del cóndilo femoral: un estudio ecográfico retrospectivo

SUMMARY: The Q-angle is defined as the acute angle formed by the vectors for combined pull of the quadriceps femoris muscle and the patellar tendon. This study aimed to investigate the variations in Q angle with respect to race. Moreover, this study performed ultrasound to evaluate of the thickness of articular cartilage covering the medial and lateral femoral condyle in volunteers with an increased Q angle. The study included 487 Jordanian and 402 Malaysians with age range 18-23 years. Moreover, the study included 30 participants aged between 18 and 22 years, with a total of 15 volunteers with Q >14° and 15 patients with Q ≤14°. Both Q angle and condylar distance were measured by well-trained medical practitioners according to a well-established protocol. The thickness of articular cartilage covering the medial and lateral femoral condyle of the femoral bone was measured using ultrasound. Regardless of race, Q angle was greater in females. Furthermore, Q angle was significantly greater in Arab volunteers compared to Malay volunteers. Q angle significantly increase with increasing condylar distance in both races. Finally, the statistical analysis showed a significantly reduced thickness of articular cartilage on both medial and lateral femoral condyle (P = 0.05) in the Q >14° group. Multiple factors including race and condylar distance and even the articular cartilage of femoral condyle should be considered during the examination and management of knee fractures and condylar diseases.

RESUMEN: El ángulo Q se define como el ángulo agudo formado por los vectores de tracción combinada del músculo cuádriceps femoral y el tendón patelar. Este estudio tuvo como objetivo investigar las variaciones en el ángulo Q con respecto a la raza. Además, se realizó una ecografía para evaluar el grosor del cartílago articular que cubre los cóndilos femorales medial y lateral en voluntarios con un ángulo Q aumentado. El estudio incluyó a 487 jordanos y 402 malayos con un rango de edad de 18 a 23 años. Además, el estudio incluyó a 30 participantes con edades comprendidas entre 18 y 22 años, 15 voluntarios con Q> 14 ° y 15 pacientes con Q ≤ 14 °. Tanto el ángulo Q como la distancia condilar fueron medidos por médicos bien entrenados de acuerdo con un protocolo establecido. El grosor del cartílago articular que cubre los cóndilos femorales medial y lateral del fémur se midió mediante ecografía. Independientemente del grupo racial, el ángulo Q fue mayor en las mujeres. Además, el ángulo Q fue significativamente mayor en los voluntarios árabes en comparación con los voluntarios malayos. El ángulo Q se aumenta significativamente al incrementarse la distancia condilar en ambas grupos raciales. Finalmente, el análisis estadístico mostró una reducción significativa del grosor del cartílago articular en los cóndilos femorales medial y lateral (P = 0,05) en el grupo Q> 14. Durante la exploración y el tratamiento de las fracturas de rodilla y de las enfermedades condilares, se deben considerar múltiples factores, incluida la raza y la distancia condilar e incluso el cartílago articular del cóndilo femoral.

Anatomy education of medical students during the COVID 19 pandemic / Educación en anatomía de estudiantes de medicina durante la pandemia de COVID-19

SUMMARY: COVID-19 created extraordinary challenges to anatomy education and teaching practices, as the anatomists try to achieve best knowledge delivery level for their discipline, without the use of traditional teaching aids such as the cadavers and microscopic slides. The present study was conducted to collect medical students' response and opinions regarding the pros and cons of online teaching vs traditional teaching. 2263 medical studentswere recruited from the first three academic years. A multiple choice close-ended questionnaire regarding their opinion about virtual teaching mode for the anatomy discipline during COVID-19 pandemic was designed and circulated via emails.The majority (78.12 %) of the students agreed that they missed their traditional anatomy learning mode. Moreover, (92.92 %) of the students missed their campus and the college social life. The students strongly felt there is a gap and difficulty in understanding the topics that required practicing and visual orientation such as dissections, models, microscopic slides which help them in better memorizing and recalling the anatomical terms. The lack of proper devices and the absence of high quality internet were among the top reported issues that negatively affect online learning. These results indicated that, compared with traditional methods of teachings, the online learning in the medical schools had relatively poor planning and required continuous and combined efforts in order to improve the quality of online teaching specially for anatomy discipline, which may be an essential response for any unforeseen situation such as the COVID-19 pandemic. We should look at the current situation as an opportunity to apply modern anatomy education approaches which may be a necessity at the present time, with huge accomplishments achieved in the information and online technology field.

RESUMEN: La pandemia de COVID-19 creó desafíos extraordinarios para la educación y las prácticas de enseñanza de la anatomía, debido al objetivo del anatomista de lograr el mejor nivel de educación para su disciplina sin el uso materiales didácticos tradicionales, tal como los cadáveres y las láminas microscópicas. En este studio se analizó la respuesta y las opiniones de los estudiantes sobre los pros y los contras de la enseñanza en línea frente a la enseñanza tradicional. Se reclutaron 2263 estudiantes de medicina de los tres primeros años académicos. Se diseñó un cuestionario cerrado de opción múltiple con respecto a su opinión sobre el método de enseñanza virtual para la disciplina de anatomía durante la pandemia de COVID-19 y se distribuyó por correo electrónico. Además, (92,92 %) de los estudiantes dejaron de participar en la vida social universitaria y se ausentaron del campus. Los estudiantes sintieron fuertemente que hay una brecha y dificultad para comprender los temas que requieren práctica y orientación visual, como disecciones, modelos, diapositivas microscópicas que les ayudan a memorizar y recordar mejor los términos anatómicos. La falta de internet de calidad, y de dispositivos adecuados se encuentran entre los principales problemas reportados que afectan negativamente el aprendizaje en línea. Estos resultados indicaron que, en comparación con los métodos tradicionales de enseñanza, el aprendizaje en línea en las escuelas de medicina tenía una planificación relativamente deficiente y requería una planificación continua, además de esfuerzos para mejorar la calidad de la enseñanza en línea de anatomía, lo que que podría ser de apoyo fundamental ante cualquier situación imprevista como la pandemia de COVID-19. Actualmente, se debe considerar esta situación como una oportunidad para aplicar enfoques modernos de educación en anatomía con importantes logros en el campo de la tecnología informática y en línea.

Cross talk between 26S proteasome and mitochondria in human mesenchymal stem cells' ability to survive under hypoxia stress.

Mesenchymal stem cells (MSCs) are regarded as unique cells which play an imperative role in the field of regenerative medicine. They are characterized by the self-renewal capacity, multi-lineage differentiation abilities and immunomodulation properties which render them perfectly ideal cell type for treating a wide range of chronic diseases. Despite these enchanted features, there are many hurdles that need to be circumvented to ensure their long-term survival and viability after transplantation. Recently, hypoxia has been indicated as one of the most baffling stress conditions that can affect the survival rate of MSCs either positively or negatively depending on the level of hypoxia. MSCs can survive well under moderate hypoxia, but die shortly if they were exposed to severe hypoxia without clearly convincing explanation for this enigma. The current study reveals a novel mechanism of 26S proteasome in controlling the ability of BM-MSCs to withstand hypoxic stress by maintaining proper mitochondrial function. The results indicated that 26S proteasome remains functioning once BM- MSCs are exposed to moderate hypoxia (2.5%O2) and preserves their survival and proliferation mediated by intact mitochondrial performance, whereas 26S proteasome becomes inactive when BM-MSCs faces severe hypoxia that lead to poor mitochondrial function and less chance to survive longer. The outcomes of this study demonstrated the importance of 26S proteasome machinery in enhancing the resistance of BM-MSCs to hypoxic stress condition which may help in better planning future studies that target this system. Schematic representation summarizing the findings of the current study. 26S proteasome function preservation in normoxia and moderate hypoxia leads to maintain appropriate proliferation and mitochondrial activity in human BM-MSCs and promote their survival. On the opposite side, severe hypoxia disrupts the 26S proteasome function leading to significant reduction in the proliferation, survival and mitochondrial dynamics in human BM-MSCs causing their death.

Addressing the impact of different fetal bovine serum percentages on mesenchymal stem cells biological performance.

Human mesenchymal stem cells (MSCs) are presently on the top of hierarchy in the field of stem cell therapy, due to their miraculous therapeutic abilities in diminishing the symptoms of many chronic diseases and initiating regeneration and repair for various damaged tissues and organs. The foremost initial step to reach high success rate in any MSCs based study is the optimization of culture growth media by establishing a suitable fetal bovine serum (FBS) percentage that suits the purpose of MSCs based experiment. Choosing the suitable FBS percentage is a controversial issue and merely depends on the researchers experience and suggested recommendations by the suppliers. Despite the huge improvements in overall MSCs investigating approaches, there are no definite protocols that set up a range of FBS percentages that can be followed. Toward achieving this objective, we evaluate in the present report the effect of using various FBS percentages (5-20%) added to DMEM low glucose media, on the biological behaviour of MSCs. Growing MSCs in high FBS percentages containing culture media (15% and 20% FBS) increase the proliferation and expansion rate of MSCs, although it decreases the immunosuppressive properties. On the other hand, adding low FBS percentage (7% FBS) to MSCs culture media enhanced the immunosuppression characteristics of MSCs, even though the proliferation rate was moderately reduced. 7% FBS is the cut off percentage that can be used without negatively altering major MSCs biological properties in which using 5% FBS will cause a tremendous decrease in the proliferation capacity and immunosuppressive properties. This report may assist other researchers in choosing appropriate FBS percentage when preparing MSCs culture media that serve the purpose of their MSCs based studies.