Calreticulin regulates Src kinase in osteogenic differentiation from embryonic stem cells

Calreticulin, the major Ca2+ buffer of the endoplasmic reticulum plays an important role in the choice of fate by embryonic stem cells. Using the embryoid body method of organogenesis, we showed impaired osteogenesis in crt−/− cells vis-à-vis calreticulin-containing osteogenic WT cells. In the non-osteogenic crt−/− cells, c-Src- a non-receptor tyrosine kinase- was activated and its inhibition rescued osteogenesis. Most importantly, we demonstrated that calreticulin-containing cells had lower c-Src kinase activity, and this was accomplished via the Ca2+-homeostatic function of calreticulin. Specifically, lowering cytosolic [Ca2+] in calreticulin-containing osteogenic WT cells with BAPTA-AM, activated c-Src and impaired osteogenic differentiation. Conversely, increasing cytosolic [Ca2+] in crt−/− cells with ionomycin deactivated c-Src kinase and restored osteogenesis. The immediate effector of calreticulin, the Ser/Thr phosphatase calcineurin, was less active in crt−/− cells, however, its activity was rescued upon inhibition of c-Src activity by small molecule inhibitors. Finally, we showed that higher activity of calcineurin correlated with increased level of nuclear Runx2, a transcription factor that is the master regulator of osteogenesis. Collectively, our work has identified a novel pathway involving calreticulin regulated Ca2+ signalling via c-Src in osteogenic differentiation of embryonic stem cells

c-Src Controls Mouse Embryonic Osteogenic Differentiation Through Regulation of Stat1 Stability

The protein tyrosine kinase Src is expressed ubiquitously and is involved in differentiation. However, the role of Src in osteogenic differentiation is complex since it has been reported to function both as repressor and activator. Using the small molecule inhibitor PP2, c-Src specific siRNAs, and tet-inducible lentiviral vectors overexpressing constitutively active c-Src, I confirmed c-Src inhibitory role in mouse embryonic stem cells (mESCs) and mouse preosteoblast MC3T3-E1s osteogenic differentiation. I further predicted that the increased level of osteogenic differentiation is regulated through Runt-related transcription factor 2 (Runx2). Additional analysis revealed that active c-Src (p-Y416-c-Src) lowers Runx2 nuclear localization and transcriptional activity in mESCs without having any effect on Runx2 expression level. I provide the first evidence linking inhibitory role of c-Src to Runx2 subcellular localization through signal transducer and activator of transcription 1 (Stat1), the cytoplasmic anchoring partner of Runx2. I discovered that higher level of active c-Src increases Stat1 protein half-life by inhibiting Stat1 proteasomal degradation. Therefore, by inhibition of c-Src activity, Stat1 protein level is reduced resulting in less of interaction with Runx2. This in turn increases nuclear localization and transcriptional activity of Runx2. Collectively, my study has defined a new mechanism, by which c-Src activity inhibits transcriptional regulation of osteogenesis.Ph.D.2019-07-10 00:00:0

c-Src kinase inhibits osteogenic differentiation via enhancing STAT1 stability.

The proto-oncogene Src is ubiquitously expressed and is involved in cellular differentiation. However, the role of Src in embryonic stem (ES) cell osteogenic differentiation is largely unknown. Using the small molecule inhibitor PP2, c-Src specific siRNAs, and tet-inducible lentiviral vectors overexpressing active c-Src, we delineated an inhibitory role of c-Src in osteogenic differentiation of mouse embryonic stem cells (mESCs) and mouse MC3T3-E1s preosteoblasts. Active c-Src was shown to restrict the nuclear residency of Runt-related transcription factor 2 (Runx2) and its transcriptional activity with no detectable effect on Runx2 expression level. Furthermore, we showed Signal Transducer and Activator of Transcription 1 (STAT1) was indispensable to the inhibitory role of c-Src on Runx2 nuclear localization. Specifically, higher levels of active c-Src increased STAT1 half-life by inhibiting its proteasomal degradation, thereby increasing the cytoplasmic abundance of STAT1. More abundant cytoplasmic STAT1 bound and anchored Runx2, which restricted its nucleocytoplasmic shuttling and ultimately reduced Runx2 transcriptional activity. Collectively, this study has defined a new mechanism by which c-Src inhibits the transcriptional regulation of osteogenesis from mESCs in vitro

Survey on Ionizing Radiation Including Gamma-Ray Exposure among the Medical Staff Working in the Non-Intensive Ward of Farshchian Cardiovascular Hospital in Hamadan, Iran

Background and Objective: With the increase of cardiovascular diseases worldwide, diagnostic and therapeutic measures, particularly nuclear medicine technology, have also increased. A multiplicity of diagnostic procedures are used for treating many patients with cardiovascular diseases in Farshchian Cardiovascular Hospital, Hamadan, Iran; therefore, the staff of this hospital, especially non-intensive care workers, have encountered a large number of these patients and have fear and concerns caused by this exposure. In this regard, this study aimed to determine the exposure of staff working in non-intensive care units of this Cardiovascular Hospital to ionizing radiation, including gamma-ray. Materials and Methods: This cross-sectional study was conducted in which 40 film badges (for radiation measurement) were attached to staff working in two separated inpatient wards (Ward A and Ward B) from non-intensive wards of Farshchian Cardiovascular Hospital in three separate shifts (morning, evening, and night) and in three occupational categories, including nurses, health workers, and service provider staff.[A1]  The amount of radiation exposure of people (in mSv) was measured in two separate sections for four mounths (the first two months, the second two months) and SPSS version 26 software was used to analyze the data. Results: Exposure to ionizing radiation, including gamma-ray, in all three occupational categories of nurses, health workers, and service providers was less than 0.05 mSv in both wards, all below the risk threshold. Conclusion: The results of this study showed that the exposure of nurses, health workers, and service providers working in the studied wards to ionizing radiation (e.g., gamma-ray) was within the permissible limit; therefore, the working environment of these employees is safe to take care of patients  [A1]There is one sentence in Persian abstract which was not translated here. Please check

Dietary acid load and its interaction with CETP TaqB1 polymorphisms on lipid profile among patients with Type 2 diabetes mellitus

Abstract Objective Gene-diet interaction plays a key role in the inter-individual differences in lipid abnormalities as a major risk factor for cardiovascular diseases (CVDs). Thus, we explored the interaction between CETP TaqB1 polymorphism with dietary acid load (DAL) on lipid profile among type 2 diabetes mellitus (T2DM). Method This cross-sectional study conducted on 220 Iranian patients with T2DM. Dietary acid load (PRAL and NEAP) was calculated via a validated food-frequency questionnaire (FFQ). The polymerase chain reaction (PCR) used for genotyping Taq1B polymorphism. Biochemical markers were measured by standard protocol. The interaction between CETP Taq1B polymorphism and DAL (PRAL and NEAP) on lipid profile was performed by a generalized linear regression model (GLM). Results The overall prevalence of rs708272 genotypes was 8.6%, 72.7% and 18.6% for B1B1, B1B2 and B2B2 genotype respectively. This study showed that people with the B1B1 genotype had greater LDL, TC, LDL/HDL, and TG when they consumed diets that scored higher on the NEAP and PRAL indexes than those with the B1B2 and B2B2 genotypes. Besides, carriers of the B1B1 allele who were in the highest tertile of NEAP, had lower HDL (P Interaction < 0.05). Conclusions In summary, the lipid profile might be improved in B1B1 homozygotes by less adherence to DAL indexes, however, the findings should be validated in high-quality interventional studies

Optimization of Antioxidant Activities and Intracellular Polysaccharide Contents Using Agaricus bisporus Extract as Elicitor in Submerged Fermenting Ganoderma lucidum: Optimization of IPS production from ganoderma lucidum

Background and Objective: Ganoderma lucidum is one of the medicinal fungi frequently used as supplement. The intracellular polysaccharides of this fungus include high molecular weights and help strengthen the immune system. Furthermore, these polysaccharides act as antioxidants by inhibiting free radicals and enhancing activity of the enzymes. Addition of various elicitors to the fungi submerged culture media affects the cell growth and metabolite production. Fungal extracts are one of these elicitors. Material and Methods: In this study, Ganoderma lucidum was first cultured in various culture media to investigate the base media. Using three various methods (soaking the fruit body in water, disintegrating the fruit body with a blender and boiling the fruit body), Agaricus bisporus fruit body extract was prepared as elicitor and the extract with the highest sugar content was used. For the optimization of growth and antioxidant activity of the intracellular polysaccharides, effects of six independent factors were investigated using Placket-Burman method, including Agaricus bisporus extract, peptone, maltose, pH, vitamin B1 and CaCl2. Response surface method was used to optimize three factors of vitamin B1, Agaricus bisporus extract and maltose. Then, stirred tank bioreactor was used to culture Ganoderma lucidum. Results and Conclusion: The YPG culture medium was selected as the base medium based on mycelial growth and antioxidant activity of the intracellular polysaccharides (IC50). Sugar content of the Agaricus bisporus extract was 30.66 µg.ml-1. Placket-Burman method revealed that the extracts of Agaricus bisporus, maltose and vitamin B1 significantly increased antioxidant activity of the intracellular polysaccharides. After optimizing these factors using RSM, the IC50 was reported as 1.047 mg.ml-1. Ganoderma lucidum cultivation in bioreactor significantly increased the cell growth (5.29 g.l-1). Intracellular polysaccharides included an IC50 of 1.14 mg.ml-1, which was significantly higher than that the intracellular polysaccharides included in YPG culture media. Conflict of interest: The authors declare no conflict of interest

Medicinal mushrooms as potential therapeutic agents in the treatment of diabetes mellitus: a review with focusing on in vivo and clinical studies

Diabetes is a metabolic disorder that affects 1 in 11 people. More than 1.5 million deaths worldwide each year are related to the disease. Many medications are used for diabetes, but these medications only prevent the disease from progressing and its symptoms. Medicinal mushrooms have been used to treat diabetes since 2,000 years ago in the traditional medicine of different countries. This study reviews the research conducted on the anti-diabetic effect of several medicinal mushrooms reducing blood sugar level by various mechanisms. Mushrooms affect carbohydrate metabolism by inhibiting α-amylase and α-glycosidase enzymes, affect cellular pathways involved in fat metabolism, insulin secretion, and cellular apoptosis. The antioxidant activity of mushrooms prevents oxidative damage to pancreatic beta cells, reduce blood sugar, cholesterol, and triglycerides and normalize liver enzymes levels. The anti-diabetic activity of medicinal mushrooms has been proven by in vitro and in vivo experiments, as well as by clinical trials. They were suggested as dietary food supplement in the treatment of diabetes

Optimization of Soy-based Media for the Production of Biologically Active Exopolysaccharides by Medicinal Mushroom Trametes versicolor

Background and Objective: Diabetes mellitus is the fifth leading cause of death in the world. Damaging effects of diabetes include advanced metabolic complications and various organ lesions. More than 90% of diabetic cases belong to type 2 diabetes. For decades, medicinal fungi such as Trametes versicolor have been considered as treatments for diabetes. The fungi exopolysaccharides show α-glucosidase activity in the intestinal membrane. Material and Methods: In this study, Iranian Trametes versicolor was cultured in various media based on soy milk, cow milk and soy protein extraction. After selecting appropriate culture media for biomass and exopolysaccharide production, response surface method with Box-Behnken design was used to investigate effects of independent variables of glucose (g l-1), soy milk (v v-1) and pH on biomass and exopolysaccharide production and optimization of these products. An animal (rat) model of streptozotocin-induced diabetic rats was used to investigate effects of exopolysaccharides on diabetes. Rats were treated with exopolysaccharides for 21 days. Results and Conclusion: Results showed that the culture media containing soy milk was appropriate for the growth and production of fungal products. Productions of biomass and exopolysaccharides increased to 2.87 g l-1 and 1.37 g l-1 respectively. Based on the analysis of response surface method, pH with p-value of 0.0004 and pH and soybean interaction with p-value of 0.0129 included significant effects on exopolysaccharide production. In optimum condition of the culture media with pH 4.67, 13.01 g l-1 glucose and 75% v v-1 soybean milk, biomass and exopolysaccharides reached 21.80 g l-1 and 9.6 g l-1. In diabetic rat model treated with exopolysaccharides, a 50.38% decrease in blood glucose was seen while triglycerides, cholesterol and low-density lipoproteins included 89, 20 and 21.67%, respectively. Furthermore, high-density lipoproteins increased by nearly 2.5%

The roles of the nurses in family physician team: A qualitative systematic review

Background: There are various approaches in primary health care regarding the key role of nurses. This systematic review investigates the roles of nurses, as part of the family medicine team. Materials and Methods: All published articles related to the role of nurses in the family medicine team from January 2000 to March 2022 were the subjects of this study. The eligibility criteria included original articles published in English or Persian in the last two decades. International credible scholarly databases (PubMed, Scopus, Magiran, IranMedex, and SID) were searched using keywords and syntax. Some of the keywords included “Family Health Nurse,” “Nurses in Primary Care,” “Family Medicine,” “Family Physician Care Program,” “General Physician Program,” “Role,” and “Nurse.” Data were extracted based on Sample, Phenomenon of Interest, Design, Evaluation, Research type (SPIDER) technique and reported based on the structure of Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Results: Of 147 identified publications by the search strategy, eight eligible empirical studies were included. The results showed that besides providing nursing care, nurses play vital roles in communication and teamwork, assessment, securing health services relevant to communities' needs, education, empowerment, clinical practice, health promotion, prevention, reflective research practice, and counseling. Conclusions: This study identified different roles nurses could play in providing primary health care. These findings are helpful for the replanning or reform of primary health care in countries, which aimed to expand the Family Physician Program (FPP) in their countries, including Iran