MicroRNAs in equine Endometritis: A review of pathophysiology and molecular insights for diagnostic and therapeutic strategies.

Endometritis plays an important role in mare infertility. Certain infectious agents interfere with the innate immune system of endometrium, causing a systemic inflammatory response that lasts for a long time and circulates via the blood or cellular degeneration, leading to endometritis due to bacterial endotoxins. Different small, non-coding RNA molecules are involved in many biological functions. For instance, microRNAs (miRNAs) are involved in the post-transcriptional regulation of gene expression. These miRNAs are important regulators of gene expression, primarily via inhibiting transcription and translation processes. This manuscript reviews: (1) pathomorphological findings in equine endometritis, (2) the expression and effects of eca-miR-17, eca-miR-223, eca-miR-200a, eca-miR-155, and eca-miR-205 in endometritis and (3) the therapeutic role of miRNA in equine endometritis. The miRNAs have a vital regulatory role in a wide range of inflammatory diseases by regulating the molecular mechanism of cytokines that cause inflammation through signal pathways. This review emphasizes the demand for cutting-edge genetic technologies and the development of novel pharmaceutical preparations to improve our understanding of the genes encoding by these miRNAs. It also focuses on the efficacy of miRNAs for control, early diagnosis, and prevention of endometritis.

6-Gingerol via overexpression of miR-322-5p impede lipopolysaccharide-caused inflammatory response in RAW264.7 cells.

Acute lung injury (ALI) and sepsis are complicated syndromes that are often left untreated in critically ill patients. 6-Gingerol is a phenolic phytochemical compound that is found in fresh ginger, has pharmacological effects against inflammation. This study explored the roles of 6-gingerol in a mouse model of acute lung injury caused by lipopolysaccharide (LPS) and RAW-264.7 cells inflammation. The LPS-induced animal model underwent histopathological examinations, and RAW-264.7 cells viability was determined by Cell counting Kit-8 (CCk-8) assay. Additionally, qRT-PCR, Immunofluorescence, Western blot, and ELISA were used in vivo and in vitro to identify inflammatory factors and proteins associated with NF-κB and MAPK signaling pathways. In a histological examination 6-gingerol exhibited protective effects. Moreover, 6-gingerol elevated cell viability and downregulated inflammatory factors Interlukin-1ß (IL-1ß), Interlukin-6 (IL-6) and Tumor necrosis factor-α (TNF-α) in LPS-treated RAW-264.7 cells. Furthermore, 6-gingerol decreased phosphorylation of P65, P38 and the level of JNK in NF-κB and MAPK pathways. Importantly, 6-gingerol increased transcript abundance of miR-322-5p which suppressed by LPS and miR-322-5p downregulation negated the protective functions of 6-gingerol. The protective activity of 6-gingerol was mediated by miR-322-5p up-regulation.

Seasonal changes in the scrotal skin histology in relation to thermoregulation and testosterone level in camel and buffalo bull.

This study was conducted to compare the scrotal skin dynamics of camel and buffalo bulls regarding thermoregulation, season and testosterone. Scrotal skin and blood samples (n = 8 each) were collected during breeding (October to March) and non-breeding seasons (April to August). Tissue slides were prepared by the paraffin embedding technique followed by hematoxylin and eosin (H&E) staining and were analysed by ImageJ®. Relative humidity and temperature were recorded in different seasons to calculate the thermal humidity index (THI). Serum testosterone level was estimated through radioimmunoassay (RIA). The data were analysed by applying ANOVA, the THS test was used as a post-ANOVA interface and the correlation coefficient was also calculated among season, testosterone and skin parameters. The results showed that skin thickness, season and THI were negatively correlated with testosterone levels. In the neck, body and apex, the papillary layer was thicker in buffalo bull compared to camel bull during the breeding season; however, the reticular layer followed a reverse trend in the apex when THI was low. In the scrotal neck during the non-breeding season, hair follicle (HF) length and density were recorded high in camel bull compared to buffalo bull when THI was elevated. The density of HF followed the otherwise trend in the apex during the breeding season. The density of SG was found highest in all regions of camel bull as compared to buffalo bull and showed direct relation with THI. To conclude, this study delineates how the histo-dynamics of scrotal skin varies and accommodate itself with testosterone and THI over the different season in camel and buffalo.

Effects of COVID-19 pandemic on anatomy education of medical and dental students of Pakistan; a reality check.

PURPOSE: Virtual anatomy education was the only way that supported the learning process of the students during the forced lockdown time of COVID-19 pandemic. The intention of the current study was to apprehend the expected challenges experienced by the medical and dental students during their cyber anatomy classes. METHODS: This study was carried on 300 1st and 2nd year medical and dental students who joined their respective college in January 2020, and consented to participate in the study. A multiple choice questionnaire regarding their stance about these online classes was formed and student's feedback was taken. RESULTS: Most (80%) of the students longed for their traditional anatomy learning i.e., dissection courses, didactic lectures, interaction and motivation from their mentors and peer. The students blamed the learning without live dissections, models and microscopic slides for their lack of confidence. More than 80% students blamed inappropriate gadgets, absence of high-band internet connections, as possible barriers in their digital learning. Lack of self-motivation was considered by 66% students. CONCLUSIONS: This situation of anatomy education while lockdown was not intentional and should not be taken as the silver bullet solution for a subject like anatomy. Although students had to face a lot of challenges but this shift to online mode went swiftly at the time of health crisis. This digital learning may extend for an indefinite period, the students' feedback will be helpful in bringing appropriate and timely modifications in digital anatomy education.

miR-424-5p overexpression inhibits LPS-stimulated inflammatory response in bovine endometrial epithelial cells by targeting IRAK2.

Endometritis is inflammation of endometrium due to various factors and is a common cause of infertility. Several remedies used for endometritis like antibiotics, hormones, and herbs. Studies confirm that microRNAs play a significant role in various inflammatory diseases. However, the role of miR-424-5p in endometritis is not clear. In our study, histopathology, real-time quantitative polymerase chain reaction, Western blot analysis, immunofluorescence, ELISA, and dual-luciferase reporter assay were used to elucidate the effect of miR-424-5p in lipopolysaccharide (LPS)-primed inflammatory response in bovine endometrial epithelial cells (BEECs) and clarify the potential mechanism. Our results revealed that miR-424-5p mimics noticeably decrease the production of proinflammatory cytokines (IL-1ß, IL-6, and TNF-α), while miR-424-5p inhibitors have inverse effects in BEECs. Moreover, overexpression of miR-424-5p on BEECs cells also suppressed NF-κB p65 activation. Afterwards, we verified that miR-424-5p inhibited Interleukin 1 Receptor Associated Kinase 2 (IRAK2) expression by binding to the 3'-UTR of IRAK2 mRNA. Further, co-transfection of miR-424-5p inhibitors and siRNA-IRAK2 revealed that negative regulation of miR-424-5p on LPS-induced inflammatory response in BEECs was mediated by IRAK2.Mutually, miR-424-5p pharmacologic stabilization represents an entirely unique medical aid for cow endometritis and other inflammation-related diseases.

Andrograpanin mitigates lipopolysaccharides induced endometritis via TLR4/NF-κB pathway.

Endometritis is an inflammatory disease that is caused by various pathogenic organisms. Andrograpanin is a compound of Andrographis paniculata, which has an important role in many inflammatory diseases, but the molecular mechanism of andrograpanin to combat inflammation is unclear. This study shows the anti-inflammatory effect of andrograpanin on Lipopolysaccharides (LPS) stimulated bovine endometrial epithelial cells (bEECs) and LPS-induced mouse model. We investigated the cytotoxic effect of bEECs by using CCK-8 analysis. Quantification of pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α) protein levels and mRNA was carried out using RT-qPCR and ELISA, respectively. The protein expressions of p65 and IκBα were assessed by western blot and immunofluorescence to check the inhibition of p65 translocation into the nucleus. The treatment effect of andrograpanin on mouse uterine tissues was determined by histopathology. in vivo, curative effect experiments showed that andrograpanin significantly reduced the endometrial injury in a mouse model. Our studies first confirmed that andrograpanin had no cytotoxic effect at 7.5,15 and 30 µg/mL concentration on bEECs. Following, Andrograpanin significantly reduced the mRNA and protein level of IL-1ß, IL-6, and TNF-α both in vivo and in vitro. Finally, Andrograpanin inhibited the IκBα degradation and p65 phosphorylation in LPS-stimulated bEECs and LPS-induced endometrial injury. Our results showed that andrograpanin might have therapeutic effects against endometritis.

MicroRNA: Could It Play a Role in Bovine Endometritis?

Endometritis in dairy cows is a major economic problem worldwide; without advances in lifestyle management and drug treatment, it causes high morbidity and death. Micro ribonucleic acid (miRNAs) these days is seen as an important part of gene control networks. It is a class of small nucleotides 20-25, single-stranded RNA molecules. In endometritis, the inflammatory response caused by the gram-negative bacteria Escherichia coli (E. coli) alters the expression of miRNA which can regulate the innate immune system. This manuscript reviews (1) the interaction of miRNAs with the signaling of NF-κB and dysregulation of miRNAs and NF-κB activity in endometritis and (2) the activity of miR-let-7c, miR-148a, and miR-488 in NF-κB activation and their effect on endometritis. Cows with reduced immunity are more vulnerable to transition diseases, such as endometritis. During post-partum, cows undergo stress, metabolic disorders, hormonal imbalance, negative energy balance, and changes in diet. One of the many categories of regulatory molecules, which explain its natural function and pathological impact on NF-κB dysregulation, is important to inform the complexity of the immune system and to develop treatments for endometritis. It shows that miRNAs could have multiple applications in veterinary medicine. Nevertheless, a comprehensive study of is essential which should be aimed at exploring the role of microRNA at physiological level and its effect due to dysfunction and dysregulation.

Effect of ethanolic preparations of cinnamon (Cinnamomum zeylanicum) extract on hematologic and histometric parameters of selected organs in Alloxan® induced diabetic female albino rats.

PURPOSE: Assessment of the antidiabetic effect of cinnamon bark extract in histologic damages and some hematologic parameters in Alloxan® induced diabetic female albino rats. METHOD: Thirty female albino rats weighing 150-230 g were divided into five groups (n = 6): normal (G1) and diabetic groups (intraperitoneally Alloxan®-injected) including diabetic control (G2), Getformin @ 0.25 (G3), CE @ 0.10 (G4), and CE @ 0.20 g/kg b.wt. (G5) for 49 days. Blood glucose level and weight were measured on weekly interval for the period of seven weeks (49th day). Blood samples were collected for hematologic analysis. Tissue samples from uterus, liver and kidneys were processed by routine paraffine technique. Histologic sections of uterus were studied to measure endometrial glands area and thickness of endo- and myometrium. Liver and kidneys were evaluated for diabetes-induced degenerative changes and antidiabetic effect of cinnamon extract (CE). One-way analysis of variance followed by Tukey test were used to compare the group means for each parameter. RESULTS: Statistical analysis revealed significant (P < 0.05) deleterious effects of diabetes on all parameters studied, however, CE recovered hematological parameters significantly (P < 0.05) as seen in G3 and G5 groups which showed significant (P < 0.05) improvement in uterus, liver and kidneys' histology. G4 significantly (P < 0.05) reduced the blood glucose at the 4th week which was maintained in subsequent weeks while G3 and G5 had significantly (P < 0.05) lowered the blood glucose from 1st week, although highly significant (P < 0.01) effect was observed during last two weeks of the study. CONCLUSION: Anti-diabetic activity of cinnamon extract was found significant in Alloxan® induced hyperglycemic rats in dose-dependent manners. CE has potential to restore diabetes induced hematological disturbances and histological damages in uterus, liver and kidney due to the presence of cinnamic acid, anhydride tannin and methyl-hydroxy chalcone polymer. Hence, CE can be recommended for the management of glucose homeostasis to avoid diabetes-associated disturbances in female rats.

Effects of oral administration of black seed (Nigella sativa) oil on histomorphometric dynamics of testes and testosterone profile in rabbits.

The present study was conducted to evaluate the influence of oral administration of black seed (Nigella sativa) oil on histomorphometrical characteristics of testes and testosterone profile in adult rabbits. Twenty adult male rabbits aged seven months were divided into two groups: control and treated. Black seed oil was administrated orally for 60 days at 5ml/kg body weight/day on daily basis in addition to the food and water ad lib to the treated group. Biometric parameters of the testes were recorded immediately after their removal. Tissue samples of testes were processed with paraffin tissue preparation technique. Histometrical parameters of testes were measured with the help of automated image analysis software Image J®. Serum testosterone concentration was determined with Radioimmunoassay technique. Statistical analysis revealed significant (P<0.05) rise in weight, length, circumference and volume of testis in treated group than control group. The values of histometrical parameters studied viz., thickness of spermatogenic epithelium, diameter and area of seminiferous tubules, diameter of lumen of seminiferous tubules, number of spermatogenic layers of testes and serum testosterone concentration were found significantly (P<0.05) higher in treated group than control group. Based on the data it is conceivable that the oral administration of black seed oil has potential to stimulate testicular function in adult rabbits.