Optimal human ovarian follicle isolation: A review focused on enzymatic digestion.

The damage or depletion of ovarian reserves due to aging or cancer treatment can increase the need for fertility preservation techniques. One of the most common ways of supporting fertility in prepubertal girls and women who require immediate cancer treatment is through ovarian tissue cryopreservation and re-transplantation following cancer treatment. However, a more appropriate method should be employed in diseases such as leukemia, where malignant cells may be present in cryopreserved tissue, instead of ovarian tissue transplantation. Human ovarian follicle isolation for in vitro culture or the use of artificial ovaries for their growth can decrease the risk of reintroducing cancer cells into these individuals. Here we review the methods for the isolation of human ovarian follicles.

Fabrication of affinity-based drug delivery systems based on electrospun chitosan sulfate/poly(vinyl alcohol) nanofibrous mats.

Benign electrospinning of chitosan in aqueous medium is an open challenge mainly due to its insolubility in neutral pH and inter- and intramolecular hydrogen bonding interactions. Here, we developed a simple and widely-used methodology to improve the chitosan electrospinnability through the sulfation of chitosan and its further mixing with poly(vinyl alcohol) for the first time. The FTIR, 1H NMR and elemental analyses showed the successful sulfation of chitosan. Furthermore, the viscosity and electrical conductivity measurements revealed the high solubility of chitosan sulfate (CS) in aqueous media. In the next step, a uniform electrospun nanofibrous mat of CS/PVA was fabricated with a fiber diameter ranging from 90 to 340 nm. The crosslinked CS/PVA (50/50) nanofibrous mat as the optimum sample showed a swelling ratio of 290 ± 4 % and a high Young's modulus of 3.75 ± 0.10 GPa. Finally, malachite green (MG) as a cationic drug model was loaded into different samples of chitosan film, CS film, and CS/PVA (50/50) nanofibrous mat and its release behavior was studied. The results of these analyses revealed that the CS/PVA (50/50) nanofibrous mat can successfully load higher contents of the MG and also release it in a sustained manner.

Systematic levels of IL-29 and microRNA185-5p were not associated with severe COVID-19 in the Iranian population.

BACKGROUND: Increased systematic pro-inflammatory cytokines is the main cause of the inflammatory conditions of the hospitalized severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infected patients. In this project, serum levels of IL-29 and whole blood levels of microRNA-185-5p (miR-185-5p) were evaluated in the hospitalized SARS-CoV-2 infected patients. METHODS: This project was performed on the 60 hospitalized SARS-CoV-2 infected patients and 60 healthy controls to evaluate IL-29 and miR185-5p expression levels. IL-29 expression was explored using enzyme linked immunoassay (ELISA), while miR185-5p was evaluated using Real-Time PCR techniques. RESULTS: The results demonstrated that neither IL-29 serum levels nor relative expressions of miR-185-5p were significantly different between patients and healthy controls. CONCLUSION: Due to the results that are presented here, systematic levels of IL-29 and miR-185-5p cannot be considered as the main risk factors for induction of inflammation in the hospitalized SARS-CoV-2 infected patients.

5-Azacytidine incorporated skeletal muscle-derived hydrogel promotes rat skeletal muscle regeneration.

Decellularized skeletal muscle is a promising biomaterial for muscle regeneration due to the mimicking of the natural microenvironment. Previously, it has been reported that 5-Azacytidine (5-Aza), a DNA methyltransferase inhibitor, induces myogenesis in different types of stem cells. In the current study, we investigated the effect of 5-Aza incorporated muscle-derived hydrogel on the viability and proliferation of muscle-derived stem cells (MDSCs) in vitro and muscle regeneration in vivo. Wistar rat skeletal muscles were decellularized using a physico-chemical protocol. The decellularized tissue was analyzed using SEM, histological staining and evaluation of DNA content. Then, muscle-derived hydrogel was made from Pepsin-digested decellularized muscle tissues. 5-Aza was physically adsorbed in prepared hydrogels. Then, MDSCs were cultured on hydrogels with/without 5-Aza, and their proliferation and cell viability were determined using LIVE/DEAD and DAPI staining. Moreover, myectomy lesions were done in rat femoris muscles, muscle-derived hydroges with/without 5-Aza were injected to the myectomy sites, and histological evaluation was performed after three weeks. The analysis of decellularized muscle tissues showed that they maintained extracellular matrix components of native muscles, while they lacked DNA. LIVE/DEAD and DAPI staining showed that the hydrogel containing 5-Aza supported MDSCs viability. Histological analysis of myectomy sites showed an improvement in muscle regeneration after administration of 5-Aza incorporated hydrogel. These findings suggest that the combination of 5-Aza with skeletal muscle hydrogel may serve as an alternative treatment option to improve the regeneration of injured muscle tissue.

Opium may affect coronary artery disease by inducing inflammation but not through the expression of CD9, CD36, and CD68.

The molecular mechanisms of opium action with regard to coronary artery disease (CAD) have not yet been determined. The aim of this study was to evaluate the effect of opium on the expression of scavenger receptors including CD36, CD68, and CD9 tetraspanin in monocytes and the plasma levels of tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), malondialdehyde (MDA), and nitric oxide metabolites (NOx) in CAD patients with and without opium addiction. This case-control study was conducted on three groups: (1) opium-addicted CAD patients (CAD + OA, n = 30); (2) CAD patients with no opium addiction (CAD, n = 30); and (3) individuals without CAD and opium addiction as the control group (Ctrl, n = 17). The protein and mRNA levels of CD9, CD36, and CD68 were evaluated by the flow cytometry and quantitative polymerase chain reaction (RT-qPCR) methods, respectively. The consumption of atorvastatin, aspirin, and glyceryl trinitrate was found be higher in the CAD groups compared with the control group. The plasma level of TNF-α was significantly higher in the CAD + OA group than in the CAD and Ctrl groups (p = 0.001 and p = 0.005, respectively). MDA levels significantly increased in CAD and CAD + OA patients in comparison with the Ctrl group (p = 0.010 and p = 0.002, respectively). No significant differences were found in CD9, CD36, CD68, IFN-γ, and NOx between the three groups. The findings demonstrated that opium did not have a significant effect on the expression of CD36, CD68, and CD9 at gene and protein levels, but it might be involved in the development of CAD by inducing inflammation through other mechanisms.

Down-regulation of transforming growth factor-beta and interleukin-6 serum levels in the idiopathic chronic obstructive pulmonary disease.

BACKGROUND: Idiopathic chronic obstructive pulmonary disease (ICOPD) is a prevalent human disease. The etiology of the disease is yet to be clarified. The main aim of this project was to explore serum levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-ß) in the ICOPD patients in comparison to healthy controls. METHODS: In this cross-sectional study, serum levels of IL-6, TNF-α and TGF-ß were evaluated in the 70 non-smoker ICOPD patients and 70 sex and age matched controls, using ELISA technique by the commercial kits from Karmania Pars Gene Company. Analysis of data was performed by parametric independent and Pearson correlation test. RESULTS: Serum levels of IL-6 and TGF-ß, but not TNF-α, were significantly decreased in the ICOPD patients in comparison to controls. Serum levels of IL-6, TNF-α and TGF-ß were not altered in the ICOPD male in comparison to female and also in mild when compared to moderate ICOPD patients. CONCLUSIONS: Down-regulation of TGF-ß may be the main risk factor for deterioration of inflammation in the ICOPD patients. Decreased IL-6 may be related to the idiopathic type of COPD.

Transplantation of SDF-1α-loaded liver extracellular matrix repopulated with autologous cells attenuated liver fibrosis in a rat model.

Cell-based therapy and tissue engineering are promising substitutes for liver transplantation to cure end-stage liver disorders. However, the limited sources for healthy and functional cells and poor engraftment rate are main challenges to the cell-based therapy approach. On the other hand, feasibility of production and size of bioengineered tissues are primary bottlenecks in tissue engineering. Here, we induce regeneration in a rat fibrotic liver model by transplanting a natural bioengineered scaffold with a native microenvironment repopulated with autologous stem/progenitor cells. In the main experimental group, a 1 mm3 stromal derived factor-1α (SDF-1α; S) loaded scaffold from decellularized liver extracellular matrix (LEM) was transplanted (Tx) into a fibrotic liver and the endogenous stem/progenitor cells were mobilized via granulocyte colony stimulating factor (G-CSF; G) therapy. Four weeks after transplantation, changes in liver fibrosis and necrosis, efficacy of cell engraftment and differentiation, vasculogenesis, and liver function recovery were assessed in this (LEM-TxSG) group and compared to the other groups. We found significant reduction in liver fibrosis stage in the LEM-TxSG, LEM-TxS and LEM-TxG groups compared to the control (fibrotic) group. Liver necrosis grade, and alanine transaminase (ALT) and aspartate transaminase (AST) levels dramatically reduced in all experimental groups compared to the control group. However, the number of engrafted cells into the transplanted scaffold and ratio of albumin (Alb) positive cells per total incorporated cells were considerably higher in the LEM-TxSG group compared to the LEM-Tx, LEM-TxS and LEM-TxG groups. Serum Alb levels increased in the LEM-Tx, LEM-TxS, and LEM-TxG groups, and was highest in the LEM-TxSG group, which was significantly more than the fibrotic group. Small vessel formation in the LEM-TxSG group was significantly higher than the LEM-Tx and LEM-TxS groups. Totally, these findings support application of the in vivo tissue engineering approach as a possible novel therapeutic strategy for liver fibrosis.

Mn-incorporated nickel selenide: an ultra-active bifunctional electrocatalyst for hydrogen evolution and urea oxidation reactions.

We report Ni-Mn-Se supported on Ni foam as a highly active and stable bifunctional electrocatalyst that exhibits overpotentials of 28 and 122 mV to reach a current density of 10 mA cm-2 for the hydrogen evolution reaction (HER) and urea oxidation reaction (UOR), respectively, and maintains its stability for over 50 h in both reactions. In addition, an overall urea splitting cell voltage of 1.352 V is needed at the current density of 10 mA cm-2 for the optimized electrode.

Opium may affect coronary artery disease by inducing inflammation but not through the expression of CD9, CD36, and CD68.

The molecular mechanisms of opium with regard to coronary artery disease (CAD) have not yet been determined. The aim of the present study was to evaluate the effect of opium on the expression of scavenger receptors including CD36, CD68, and CD9 tetraspanin in monocytes and the plasma levels of tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), malondialdehyde (MDA), and nitric oxide metabolites (NOx) in patients with CAD with and without opium addiction. This case-control study was conducted in three groups: (1) opium-addicted patients with CAD (CAD+OA, n=30); (2) patients with CAD with no opium addiction (CAD, n=30); and (3) individuals without CAD and opium addiction as the control group (Ctrl, n=17). Protein and messenger RNA (mRNA) levels of CD9, CD36, and CD68 were evaluated by flow cytometry and reverse transcription-quantitative PCR methods, respectively. Consumption of atorvastatin, aspirin, and glyceryl trinitrate was found to be higher in the CAD groups compared with the control group. The plasma level of TNF-α was significantly higher in the CAD+OA group than in the CAD and Ctrl groups (p=0.001 and p=0.005, respectively). MDA levels significantly increased in the CAD and CAD+OA groups in comparison with the Ctrl group (p=0.010 and p=0.002, respectively). No significant differences were found in CD9, CD36, CD68, IFN-γ, and NOx between the three groups. The findings demonstrated that opium did not have a significant effect on the expression of CD36, CD68, and CD9 at the gene and protein levels, but it might be involved in the development of CAD by inducing inflammation through other mechanisms.

The effect of job stress management training on observance of professional ethics by nurses.

The present study was conducted to investigate the effect of job stress management training on observance of professional ethics by nurses. This quasi-experimental study was performed on 50 clinical nurses in Shiraz, Iran. Data collection tools included a questionnaire. The research intervention consisted of six two-hour sessions of stress management training. SPSS 21 were used to analyze the data. Data analysis showed that before intervention, the level of professional ethics of 64% of nurses was relatively good, but after intervention, the level of professional ethics of the majority of nurses (94%) was at the desired level. The results showed that the levels of professional ethics after job stress management training has increased significantly compared to before. The results showed that stress management training has led to improving the level of professional ethics of nurses.

Aqueous extract of Achillea millefolium significantly affects mesenchymal stem cell renewal and differentiation in a dose dependent manner.

Introduction: Achillea millefolium is an Iranian herbal medicine with various effects on the human cells. The aim of this study was to investigate the effects of the aqueous extract of Achillea millefolium (AEAM) on the proliferation and differentiation of mesenchymal stem cells (MSCs). Methods: In this study, bone marrow-MSCs (BM-MSCs) were obtained from Wister rat bone morrow and then cultured in Dulbecco's modified Eagle's medium /Nutrient Ham's Mixture F-12 (DMEM/F12) media. Then, the isolated MSCs were cultured in either osteocyte or adipocyte differentiation media containing 0.2 or 2 mg/mL AEAM and assessed using specific staining method. Results: The isolated BM-MSCs exhibited fibroblast-like morphology and were positive for CD73, and CD90, while negative for CD34 and CD45. AEAM significantly increased self-renewal of BM-MSCs at low dose (0.2 mg/ml, P= 0.001) and increased the pool stem cells in both osteocyte and adipocyte differentiation media. Conclusion: AEAM at low doses may be used in cases where there is a need for large number of stem cells, via increased numbers of MSCs, and help tissue repair and immunomodulation.

Brain magnetic resonance imaging without contrast significantly increased serum levels of IL-6, but not IL-10, IL-17A and TGF-ß.

Introduction: Magnetic Resonance Imaging (MRI) is a non-invasive imaging modality. However, the effects of MRI on the immune system in the in vivo conditions are yet to be clarified. In this study we explored the effects of routine brain MRI on the protein and mRNA peripheral blood levels of interleukin-6 (IL-6), IL-10, IL-17A and transforming growth factor-beta (TGF-ß).Material and methods: 40 subjects, who referred for brain MRI, were entered for evaluating effects of routine brain MRI on the protein and mRNA peripheral blood levels of IL-6, IL-10, IL-17A and TGF-ß. Accordingly, peripheral blood were collected before and 3 hours after MRI from the participants. Protein levels of the cytokines were evaluated using ELISA. Also, mRNA levels were analyzed using Real-Time PCR techniques.Results: Brain MRI without contrast led to an increase in protein levels of IL-6 in the peripheral serum, but did not change protein and mRNA levels of IL-10, IL-17A and TGF-ß. IL-6 mRNA levels after MRI were higher in the participants with mild anxiety compared to those without anxiety.Conclusion: brain MRI without contrast can induce secretion of IL-6 and may be associated with its functions, such as development of plasma cells or induction of inflammation.

Severe Acute Respiratory Syndrome Coronavirus 2: The Role of the Main Components of the Innate Immune System.

At the end of December 2019, the COVID-19 pandemic began in Wuhan of China. COVID-19 affects different people with a wide spectrum of clinical manifestations, ranging from asymptomatic with recovery without hospitalization up to a severe acute respiratory syndrome (SARS). The innate and adaptive immunity appears responsible for the defense against the virus and recovery from the disease. The innate immune system, as the first line of defense, is essential for the detection of virus and subsequent activation of acquired immunity. The innate immune response is carried out by sentinel cells such as monocytes/macrophages and dendritic cells and by receptors known as pattern recognition receptors (PRR). These receptors can recognize various components of the virus, which lead to intracellular signaling and subsequently the synthesis of various cytokines. These cytokines then recruit other immune cells, activate adaptive immune responses, and inhibit viral spreading. The most common receptors include Toll-like receptors, C-type lectin receptors, and RIG-I like receptors. This review describes the current knowledge about the interplay between innate immune responses and SARS-CoV-2 with a focus on the innate immune cells and the role of their receptors in viral RNA recognition, as well as their mechanisms for recognizing SARS-CoV-2.

Expression of toll like receptors before and after coronary angiography with isomolar contrast among those with and without coronary atherosclerosis.

Angiography is a safe technique for the detection of and treatment of cardiovascular diseases. However, the effects of the technique on the molecular response of the immune system are yet to be clarified. Toll like receptors (TLRs) are the important molecule participate in the innate immunity responses and induction of inflammation. This project was designed to explore the effects of angiography on the expression of TLR1, TLR2, TLR3 and TLR4. Fifty-five participants, including three separate groups (without artery stenosis, with one artery stenosis and more than one artery stenosis), were assessed in this project. TLR1, TLR2, TLR3 and TLR4 expression levels were evaluated in peripheral blood immune cells by measuring mRNA before and after angiography using Real-Time PCR techniques. mRNA levels of TLR1, TLR2 and TLR3 were significantly increased following angiography. Expression of TLR4 did not change after angiography. Other criteria also showed no correlation on TLR expression after angiography. TLR4 mRNA levels had a positive correlation with age in the participants without artery stenosis. Angiography may induce inflammation in subjects without artery stenosis via up-regulation of TLR1, 2 and 3 which may lead to cardiovascular diseases related complications.

The immune response against Toxoplasma gondii in BALB/c mice induced by mannose-modified nanoliposome of excreted/secreted antigens.

This study aimed to compare the immune response against Toxoplasma gondii (T. gondii) in BALB/c mice induced by excreted/secreted (E/S) antigens and mannose-modified nanoliposome of E/S antigens. Here, E/S antigens and mannose-modified nanoliposome of E/S antigens were firstly prepared, and then BALB/c female inbred mice were separately immunized. In the next step, anti-E/S antigen antibodies and the relative expression levels of IL-10 and IL-12 mRNA were detected by ELISA and real-time PCR, respectively. After immunization, mice were intraperitoneally challenged with 102 tachyzoites of T. gondii, and the survival rate was recorded. The ELISA analysis showed significant differences between the levels of anti-E/S antigen antibodies in the mice immunized by E/S antigens and those immunized by mannose-modified nanoliposome of E/S antigens at days 7, 10, 20, 25, and 30 (P < 0.05). Real-time PCR analysis showed that the relative expression of IL-10 was significantly decreased during 20 days. Yet, the relative expression of IL-12 was significantly increased during 20 days (P < 0.05). In T. gondii challenge test, significant differences were found between the survival rates of mice immunized by E/S antigens and mice immunized by mannose-modified nanoliposome with E/S antigens. This project evidenced that mannose-modified nanoliposome of E/S antigens induced a more powerful immune response against T. gondii in BALB/c mice when compared with excreted/secreted antigens alone.

Depression Is Associated with Downregulation of IPS-1 in Chronic HBV-Infected Patients in a Sex-Dependent Manner.

BACKGROUND: Depression and anxiety can modulate immune-related molecule expressions. The chronic HBV-infected (CHB) patients suffer from inappropriate immune responses. Additionally, psychological disorders are prevalent among the patients. Thus, depression and anxiety may alter immune-related molecule expression. This study aimed to examine IPS-1 and RIP1 mRNA levels in CHB patients suffering from various degrees of anxiety and depression. METHODS: Sixty patients with CHB participated in this research and completed standard questionnaires to evaluate depression and anxiety. The expression levels of IPS-1 and RIP1 were examined using real-time PCR techniques. RESULTS: The result revealed that although the expression of IPS-1 and RIP1 did not change in the CHB patients with various ranges of depression and anxiety, IPS-1 was significantly decreased in the male CHB patients who suffered from mild, moderate, and severe depression when compared to the patients with no depression. CONCLUSION: So, it was hypothesized that depression may be associated with alteration in the expression of IPS-1 in a sex-dependent manner. In other words, it appears that the male CHB patients are at risk of depression-related alteration in immune-related gene expression.

An automated deep learning convolutional neural network algorithm applied for soil salinity distribution mapping in Lake Urmia, Iran.

Traditional soil salinity studies are time-consuming and expensive, especially over large areas. This study proposed an innovative deep learning convolutional neural network (DL-CNN) data-driven approach for SSD mapping. Multi-spectral remote sensing data encompassing Landsat series images provide the possibility for frequent assessment of SSD in various regions of the world. Therefore, Landsat 7 ETM+ and 8 OLI images were acquired for years 2005, 2010, 2015 and 2019. Totally, 704 sample points collected from the top 20 cm of the soil surface, which 70% was used to train the network and the remains (30%) were utilized to validate the network. Accordingly, DL-CNN model trained using remote sensing (RS)-derived variables (land surface temperature (LST), Soil moisture (SM) and evapotranspiration) and geospatial data such as NDVI and landuse. To train the CNN, ReLu, Cross-entropy and ADAM were employed respectively as activation, loss/cost functions and optimizer. The results indicated the high confidence of OA 0.94.02, 0.93.99, 0.94.87 and 0.95.0 respectively for years 2005, 2010, 2015 and 2019. These accuracies demonstrated the best performance of automated DL-CNN for SSD mapping compared to RS soil salinity indexes. Furthermore, the FR and WOE models applied in order to generate a geospatial assessment of the DL-CNN classification results. According to the FR model, landuse, LST, LST and NDVI with the frequency ratio of 0.98.25, 0.94.03, 0.97.23 and 0.96.36 selected respectively as more effective factors for SSD in the study area for years 2005, 2010, 2015 and 2019. Also based on the WOE model, landuse, LST, landuse and NDVI with the WOE of 0.88.25, 0.91.88, 0.87.43 and 0.89.02 were ranked respectively for years 2005, 2010, 2015 and 2019 as efficient variables for SSD. In sum, our introduced method can be recommended for SDD spatial modelling in other favored areas with similar environmental conditions.

Mesenchymal Stem Cells: The Potential Therapeutic Cell Therapy to Reduce Brain Stroke Side Effects.

Tissue plasminogen activator (tPA) is the gold standard treatment for ischemic stroke in the time window of 3-4.5 hours after the onset of symptoms. However, tPA administration is associated with inflammation and neurotoxic effects. Mesenchymal stem cells (MSC)-based therapy is emerging as a promising therapeutic strategy to control different inflammatory conditions. This project was designed to examine the protective role of MSC administration alone or in combination with royal jelly (RJ) five hours after stroke onset. The mice model of middle cerebral artery occlusion (MCAO) was established and put to six groups, including intact (healthy mice without stroke), control (untreated stroke), treated with mouse MSC (mMSC), Sup (conditioned medium), RJ and combination of mMSC and RJ (mMSC/RJ). Thereafter, behavioral functions, serum and brain (in both infarcted and non-infarcted tissues) levels of interleukin (IL)-1ß, IL-4, IL-10, tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) the sizes of brain infarction have been determined in the groups. Administration of mMSC and mMSC/RJ significantly improved the behavioral functions when compared to the controls. mMSC, RJ and mMSC/RJ significantly decreased the infarcted volumes. RJ and mMSC/RJ, but not mMSC, significantly decreased the brain edema. The infarction increased the serum levels of the cytokines, except TNF-α, and treatment with mMSC, Sup and RJ reduced serum levels of the pro-inflammatory cytokines. mMSC reduced IL-1ß in the non-infarcted brain tissue. To conclude, data revealed that using mMSC/RJ combination significantly reduced stroke side effects, including brain edema and serum levels of pro-inflammatory cytokines, and suggested that combination therapy of MSCs with RJ may be considered as an effective stroke therapeutic strategy.