Caffeine attenuates spermatogenic disorders in mice with induced chronic scrotal hyperthermia.

OBJECTIVE: Chronic scrotal hyperthermia (SHT) can lead to serious disorders of the male reproductive system, with oxidative stress playing a key role in the onset of these dysfunctions. Thus, we evaluated the impact of caffeine, a potent antioxidant, on cellular and tissue disorders in mice with chronic SHT. METHODS: In this experimental study, 56 adult male NMRI mice were allocated into seven equal groups. Apart from the non-treated control group, all were exposed to heat stress. Two groups, termed "preventive" and "curative," were orally administered caffeine. The preventive mice began receiving caffeine immediately prior to heat exposure, while for the curative group, a caffeine regimen was initiated 15 consecutive days following cessation of heat exposure. Each treated group was subdivided based on pairing with a positive control (Pre/curative [Cur]+PC) or a vehicle (Pre/Cur+vehicle). Upon conclusion of the study, we assessed sperm characteristics, testosterone levels, stereological parameters, apoptosis, antioxidant and oxidant levels, and molecular markers. RESULTS: Sperm parameters, testosterone levels, stereological parameters, biochemical factors (excluding malondialdehyde [MDA]), and c-kit gene expression were significantly elevated in the preventive and curative groups, especially the former, relative to the other groups. Conversely, expression levels of the heat shock protein 72 (HSP72) and nuclear factor kappa beta (NF-κß) genes, MDA levels, and apoptotic cell density were markedly lower in both caffeine-treated groups relative to the other groups, with more pronounced differences observed in the preventive group. CONCLUSION: Overall, caffeine attenuated cellular and molecular abnormalities induced by heat stress in the testis, particularly in the mice treated under the preventive condition.

Decellularized human amniotic membrane loaded with epigallocatechin-3-gallate accelerated diabetic wound healing.

Diabetic wounds, as one of the most important complications of diabetes, face many challenges in treatment. Herein we investigated whether decellularized human amniotic membrane (dAM) loaded with epigallocatechin-3-gallate (EGCG) could promote healing in diabetic rats. Sixty diabetic rats were randomly planned into the untreated group, dAM group, EGCG group, and dAM + EGCG group. On days 7, 14, and 21, five rats from each group were sampled for stereological, molecular, and tensiometrical assessments. Our finding revealed that the wound closure rate, the total volumes of new epidermis and dermis, the numerical densities of fibroblasts, blood vessels, collagen density as well as tensiometrical parameters of the healed wounds were considerably increased in the treated groups than in the untreated group, and these changes were more obvious in the dAM + EGCG ones. Furthermore, the expression of TGF-ß, bFGF, and VEGF genes were significantly upregulated in all treated groups compared to the untreated group and were greater in the dAM + EGCG group. This is while expression of TNF-α and IL-1ß, as well as cell numerical densities of neutrophils and macrophages decreased more considerably in the dAM + EGCG group in comparison to the other groups. In conclusion, it was found that using both dAM transplantation and EGCG has more effect on diabetic wound healing.

Coenzyme Q10 attenuates neurodegeneration in the cerebellum induced by chronic exposure to tramadol.

BACKGROUND: Chronic use of tramadol can cause neurotoxic effects and subsequently cause neurodegeneration in the cerebellum. The main damage mechanisms identified are oxidative stress and inflammation. Currently, we investigated the effects of coenzyme Q10 (CoQ10) in attenuates of neurodegeneration in the cerebellum induced by chronic exposure to tramadol. MATERIAL AND METHODS: Seventy-two male mature albino rats were allocated into four equal groups, including; non-treated group, CoQ10 group (which received CoQ10 at 200 mg/kg/day orally for three weeks), tramadol group (which received tramadol hydrochloride at 50 mg/kg/day orally for three weeks), and tramadol+CoQ10 group (which received tramadol and CoQ10 at the same doses as the previous groups). Tissue samples were obtained for stereological, immunohistochemical, biochemical, and molecular evaluations. Also, functional tests were performed to evaluate behavioral properties. RESULTS: We found a significant increase in stereological parameters, antioxidant factors (catalase, glutathione, and superoxide dismutase), and behavioral function scores in the tramadol+CoQ10 group compared to the tramadol group (p < 0.05). In addition, malondialdehyde levels, the density of apoptotic cells, as well as the expression of pro-inflammatory (tumor necrosis factor-alpha, interleukin 1 beta, and interleukin 6) and autophagy (lysosome-associated membrane protein 2, autophagy-related 5, beclin 1, and autophagy-related 12) genes were considerably reduced in the tramadol+CoQ10 group compared to the tramadol group (p < 0.05). CONCLUSION: We conclude that the administration of CoQ10 has neuroprotective effects in the cerebellum of rats that have chronic exposure to tramadol.

Human Placental Mesenchymal Stem Cell-derived Exosomes in Combination with Hyperbaric Oxygen Synergistically Promote Recovery after Spinal Cord Injury in Rats.

Spinal cord injury (SCI) is a critical medical condition during which sensorimotor function is lost. Current treatments are still unable to effectively improve these conditions, so it is important to pay attention to other effective approaches. Currently, we investigated the combined effects of human placenta mesenchymal stem cells (hPMSCs)-derived exosomes along with hyperbaric oxygen (HBO) in the recovery of SCI in rats. Ninety male mature Sprague-Dawley (SD) rats were allocated into five equal groups, including; sham group, SCI group, Exo group (underwent SCI and received hPMSCs-derived exosomes), HBO group (underwent SCI and received HBO), and Exo+HBO group (underwent SCI and received hPMSCs-derived exosomes plus HBO). Tissue samples at the lesion site were obtained for the evaluation of stereological, immunohistochemical, biochemical, molecular, and behavioral characteristics. Findings showed a significant increase in stereological parameters, biochemical factors (GSH, SOD, and CAT), IL-10 gene expression and behavioral functions (BBB and EMG Latency) in treatment groups, especially Exo+HBO group, compared to SCI group. In addition, MDA levels, the density of apoptotic cells and gliosis, as well as expression of inflammatory genes (TNF-α and IL-1ß) were considerably reduced in treatment groups, especially Exo+HBO group, compared to SCI group. We conclude that co-administration of hPMSCs-derived exosomes and HBO has synergistic neuroprotective effects in animals undergoing SCI.

The effect of encomir-93 mimic transfection on the expression of miR-93 and PSA and androgen receptor in prostate cancer LNcap cell line.

OBJECTIVES: Prostate cancer (PCa) is one of the most common cancers in men with high mortality rate which is a major concern for men's health. However, the molecular mechanisms remain poorly understood. miR-93 is an important oncogene which may have important function in prostate cancer.So, this study aimed to predict that encomir-93 mimic transfection on the expression of miR-93 and PSA and AR in prostate cancer LNcap cell line. METHODS: Lymph node carcinoma of the prostate (LNCaP) was cultured and then miR-93 mimics was designed, synthesized and the transfected to LNCaP. The expression level of prostate-specific antigen (PSA) and androgen receptor (AR) was determined via Real-time PCR after treated with 15 pmol of miR-93 mimics. RESULTS: miR-93 mimic transfection led to significant increase in PSA and AR expression in comparison with control group (p≤0.05). CONCLUSIONS: The miR-93 and its target genes has important role in PCa progression via enhancement in PSA and AR expression. Further research on the function of the miR-93 and its target genes in tumorgenesis and progression PCa could be helpful for the treatment of prostate cancer.

Increased expression of androgen receptor and PSA genes in LNCaP (prostate cancer) cell line due to high concentrations of EGCG, an active ingredient in green tea.

OBJECTIVES: Androgen receptor (AR) play a key role in the onset and progression of prostate cancer. Epigallocatechin-3-gallate (EGCG) is a polyphenolic compound and the active ingredient in green tea, which is involved in modulating gene expression through epigenetic alterations. Previous studies have shown that EGCG at low concentrations reduces the expression of AR and prostate-specific antigen (PSA) in the LNCaP cell line of prostate cancer. In this study, the effect of higher EGCG concentrations on AR and PSA expression in LNCaP prostate cancer cell line was investigated. METHODS: In this study, LNCaP prostate cancer cell line was used and after MTT test, concentrations of 40, 60 and 80 µg/mL EGCG were used for treatment. Then, the expression of AR and PSA genes was evaluated by RT-PCR. AR protein expression was also assessed by Western blotting. RESULTS: The present study showed that treatment of LNCaPs cells by EGCG reduces cell proliferation. The IC50 value was 42.7 µg/mL under experimental conditions. It was also observed that EGCG at concentrations of 40 and 80 µg/mL increased the expression of AR and PSA (p<0.05). CONCLUSIONS: The present study showed that the effect of EGCG on AR expression was different at different concentrations, so that unlike previous studies, higher concentrations of EGCG (80 and 40 µg/mL) increased AR and PSA expression. It seems that due to the toxic effects of EGCG in high concentrations on cancer cells and the possibility of its effect on normal cells, more caution should be exercised in its use.

Cancer Drug Resistance Reduction via Co-treatment with Oxaliplatin and Nitazoxanide: Targeting the ABC Transporters.

OBJECTS: Shortly after cancer is diagnosed, a phenomenon develops in cancer cells called multidrug resistance (MDR), in which cell sensitivity against anti-cancer drugs is significantly reduced. The present investigation aimed to assess the effects of nitazoxanide (NTZ), a safe drug, on LS174T/OXP-resistant cells. METHODS: In the current in vitro research, the effects of NTZ and oxaliplatin (OXP) on the viability of LS174T and LS174T/OXP cell lines were evaluated through MTT assay. Then, the changes in expression levels of MDR1, MRP1, BCRP, and LRP genes and proteins were measured by RT-qPCR and western blotting methods, respectively. Lastly, the apoptosis status was assessed by annexin V-FITC/PI staining flow cytometry assay. RESULTS: The IC50 values for cells resistant or sensitive to OXP were revealed (11567 nM vs. 1745 nM; p <0.05 for 24 h incubation, and 5161 nM vs. 882.2 nM; p <0.05 for 48 h incubation). Moreover, NTZ plus OXP led to a leftward shift in the cytotoxicity curve (2004 nM; p = 0.007). This co-treatment significantly decreased the expression of all genes and proteins (p <0.05). Finally, the combination of NTZ and OXP induced a significant increase in apoptosis (p <0.001). CONCLUSION: The data showed that NTZ treatment could increase the sensitivity of LS174T/OXP cell line to the OXP cytotoxic effects. Thus, NTZ may be efficient in reducing drug resistance in clinics by means of the negative regulation of ATP-binding cassette (ABC) transporters. However, further studies are necessary to explain the exact mechanisms of NTZ.

Stem Cell Therapy, the Market, the Opportunities and the Threat.

Stem cell therapy is going to become the most widely used type of therapy in regenerative medicine. The stem cell therapy market has grown at an exponential rate in recent years. The purpose of the present paper is to review the stem cell market and the factors affecting it. The methods used included a literature review across reputable databases, and identifying articles and trusted financial reports related to the stem cell therapy market. Results show that the stem cell market growth rate is increasing, so that, the global stem cell market size was valued at US$297 million in 2022 and is anticipated to grow at a compound annual growth rate of 16.8% from 2022 to 2027, driven by factors such as clinical trials with promising results, increasing funding for stem cell research, growing number of technologies and facilities for cell therapy, and rising demand for regenerative medicine. However, the market also faces some challenges such as ethical concerns, regulatory hurdles, and the high cost of stem cell therapies and products. To enhance the development of the market further, policymakers and regulatory bodies must simplify the complicated process of obtaining regulatory approvals for clinical use. However, there are growing concerns about the increasing number of unapproved treatments using stem cells.

Decellularized human amniotic membrane engraftment in combination with adipose-derived stem cells transplantation, synergistically improved diabetic wound healing.

BACKGROUND: One of the most important and common complications of diabetes is a disorder and defect in diabetic wound healing. AIMS: The aim of present study was to investigate the synergistic effects of decellularized human amniotic membrane (dHAM) engraftment and adipose-derived stem cells (ADSs) transplantation in the healing of delayed and ischemic diabetic wound. METHODS: Sixty diabetic male rats were randomly divided into four groups (n = 15), including untreated (Control) group, engraftment by dHAM (dHAM) group, transplanted by ADSs (ADS) group, and engraftment by dHAM plus transplanted by ADSs (dHAM + ADS) group. Sampling was performed on Days 7, 14, and 21 after surgery. Evaluation tests included stereology, immunohistochemistry, molecular, and biomechanical. RESULTS: Our results showed that the wound closure rate, volumes of newly formed epidermis and dermis, density of fibroblasts and blood vessels, collagen deposition, density of proliferation cells, expression levels of TGF-ß and VEGF genes, and biomechanical characteristics were significantly higher in all treated groups compared with control group; however, these changes were considerable in the combination group. This is while that the density of neutrophils and expression levels of TNF-α and IL-1ß genes in the treated groups, especially in the combination group, were significantly reduced compared with control group. CONCLUSION: Generally, the simultaneous use of dHAM and ADS accelerates healing and improves the quality of repaired diabetic wounds.

2D materials coated on etched optical fibers as humidity sensor.

In this investigation, etched-fibers are coated by 2D layers such as Molybdenum disulfide (MoS2), Molybdenum diselenide (MoSe2) and composition of graphene and graphene oxide (G/GO) to modify humidity sensing. The relative differentiation of attenuations (RDA) in presence of relative humidity (RH) is measured by Optical Loss Test Set at two standard-wavelengths-telecommunication (1310 nm and 1550 nm). Results show that the etched single-mode fiber (ESMF) coated with G/GO has relatively high and one by one function for RDA versus RH (more than 30%). Also, its sensitivity and variance are reasonable. The MoSe2 based sensor is applicable at humidity below 30% because of higher RDA. However, it is not useful at humidity more than 30% due to the absence of one by one function for RDA versus RH. Besides, ESMF coated with MoS2 has indistinctive behavior and is not useful as a humidity sensor.

Epigallocatechin-3-gallate Enhances the Efficacy of MicroRNA-34a Mimic and MicroRNA-93 Inhibitor Co-transfection in Prostate Cancer Cell Line.

The potential role of microRNAs (miRNA or MIR) as therapeutic molecules has moved them from basic research to the field of cancer therapy. High expression of miR-93 and low expression of miR-34a have previously been indicated in prostate cancer (PC), which is the second leading cause of cancer-related death in men. Androgen receptor (AR) and prostate-specific antigen (PSA) play key roles in the initiation and progression of this cancer. Therefore, this study aimed to investigate the effects of the transfection and co-transfection of miR-34a mimic and miR-93 inhibitor with or without epigallocatechin-3-gallate (EGCG) on prostate cancer cell line and also to evaluate their effects on the expression of AR, PSA. Human lymph node carcinoma of the prostate (LNCaP) cells were treated with miR-34a mimic or/and miR-93 inhibitor with or without EGCG. Gene or protein expressions were assessed by real-time PCR or western blotting of lysates. The transfection with miR-34a mimics significantly reduced the mRNA expression of AR (p=0.0016), and PSA (p=0.038) compared to the control. Also, the miR-93 inhibitor led to a decrease in the mRNA expression of AR (p=0.0057) and PSA (p>0.05) compared to the control group. Furthermore, the co-transfection, along with EGCG, caused more decrease in both the AR (p<0.001) and the PSA (p=0.003) expression compared with the co-transfection without EGCG. Our study indicates that the reduced expression of AR and PSA in PC cells followed by treatment with miR-34a mimic and miR-93 inhibitor and their combination with EGCG as a natural substance may be a promising therapeutic way for controlling the growth of these malignant cells.

Iron hypothesis and coronary artery disease in geriatric patients.

Context and aims: Iron is a pro-oxidant factor in the pathogenesis of CAD. The association of body iron status was investigated relative to the occurrence and severity of CAD.Design and methods: The subjects consisted of 110 males and 115 females who were classified as either a CAD case or a control according to the results of coronary angiography.Results: A new parameter, the "serum free iron index," was defined as the ratio of serum iron to UIBC. The level of ferritin showed significant increase [97.2 (67.0-171.2) µg/L vs. 85.6 (52.5-129.4), p = .034], whereas serum total iron, free iron index, transferrin, UIBC and iron saturation were unchanged in CAD patients relative to control group. Among the indices of body iron only serum ferritin had significant association with the likelihood (OR of 1.004 (1.000-1.007), p = .04) and severity of CAD [χ2(3)= 7.99, p = .01], but the correlation was lessened in the presence of classical risk factors. Serum ferritin had also the highest and significant efficiency to predict CAD (AUC = 0.61, p = .020).Conclusions: Serum ferritin as a marker of intracellular iron has significant association with CAD; nevertheless, the correlation is not independent. Since iron deficiency is prevalent in elderly patients, iron hypothesis needs to expand to the both sides of iron deficiency and toxicity.

The prevalence of post-traumatic stress disorder among emergency nurses: a cross sectional study in northern Iran.

BACKGROUND AND OBJECTIVE: Posttraumatic stress disorder (PTSD) is one of the anxiety disorders which occurs in response to facing events and accidents accompanied by fear, frustration, and terror. Nurses who worked in the emergency departments witness unpleasant accidents and are exposed to stress and violence more than others. The aim of this study was to investigate the prevalence of PTSD among Iranian nurses working in the emergency department. MATERIALS AND METHODS: In this cross-sectional study, all nurses working in the emergency department of educational hospitals affiliated with Mazandaran University of Medical Sciences were included through census method (n = 131). Each of the participants in this study responded to the questionnaires individually. The first part of the questionnaire captured demographics, while the second part was the Civilian Mississippi Scale for PTSD, which was used to investigate the extent of PTSD in this study. Data were analyzed using SPSS version 22. RESULTS: In this study, 131 respondents were included. In the classification of age, nurses with 20-30-year- old had the maximum frequency (41.2 %,). Most nurses in the study (93.1%, n = 122) had a bachelor's degree in nursing and 84 were married. The prevalence of PTSD in emergency nurses was 82.96%, which was higher in nurses with shorter working background and nurses with lower levels of education. Further, the average score of PTSD was higher in married nurses. CONCLUSION: The results of this study revealed the high prevalence of PTSD among nurses who worked in emergency department. Therefore, it seems that designing and providing supportive and educational services to emergency nurses can be effective for preventing and managing this disorder, which probably can improve their performance.

Esterification of HDL-cholesterol is Decreased in Diabetes Mellitus and CAD and Enhanced Following Treatment with Statins.

BACKGROUND: The main goal of using statins is to reduce the level of plasma cholesterol, meanwhile they have a wide spectrum of actions. OBJECTIVES: To identify the effect of statins on fractional cholesterol esterification (FCE) as well as the complete profile of lipids and (apo)lipoproteins. DESIGN AND METHODS: In an age and sex matched case-control study, 400 subjects who were referred for coronary angiography were divided into two groups according using statins. RESULTS: Total cholesterol was decreased significantly following treatment with statins (165.6 ± 38.0 mg/dL vs. 205.3 ± 48.4, p£0.001). About 90% of the reduction was occurred in nonHDL and 10% in HDL fraction. Reduction of nonHDL cholesterol (125.2 ± 35.2 mg/dL vs. 162.8 ± 45.2, p£0.001) occurred on both unesterified (52.4 ± 21.5 mg/dL vs. 65.2 ± 25.5, p£0.001) and esterified cholesterol (74.7 ± 27.3 mg/dL vs. 96.6 ± 34.1, p£0.001). But the decrease in HDL cholesterol (40.4 ± 10.0 mg/dL vs. 42.3 ± 9.9, p£0.079) happened exclusively in unesterified fraction (10.9 ± 3.4 vs. 15.2 ± 5.1, p£0.001) and was counterbalanced with a significant increase in esterified portion (29.5 ± 8.2 mg/dL vs. 27.2 ± 9.5, p£0.020). The ratio of esterified- per total- cholesterol in HDL was 67.5 ± 8.1% in the control group and was decreased to 58.0 ± 14.9% (p£0.01) in diabetes and CAD and increased to 73.5 ± 6.9 (p£0.01) after using statins. CONCLUSIONS: The results suggest that the percent of esterified cholesterol in HDL fraction is decreased in diabetes and CAD patients and increased by using statins.

The Ratio of Unesterified/esterified Cholesterol is the Major Determinant of Atherogenicity of Lipoprotein Fractions.

BACKGROUND: The hypothesis is proposed that the atherogenicity of lipoporotein fractions is correlated with the content of unesterified cholesterol. OBJECTIVES: To evaluate the role and prognostic values of unesterified and esterified cholesterol in lipoprotein fractions for coronary artery disease (CAD). DESIGN AND METHODS: The study population consisted of 400 patients who were divided to CAD controls and cases according to the data of coronary angiography. Fractional cholesterol esterification (FCE) as well as the complete profile of lipids and (apo)lipoproteins were determined. RESULTS: Total cholesterol was increased significantly in CAD patients (196.3 ± 52.3 mg/dL vs. 185.7 ± 48.0, p≤ 0.049) and the increment occurred totally in unesterified portion (77.2 ± 28.4 mg/dL vs. 71.1 ± 24.4, p≤ 0.031). HDL cholesterol showed a significant decrease in CAD group (39.9 ± 9.5 mg/dL vs. 44.6 ± 10.5, p≤ 0.001), but the decrement occurred wholly in the esterified portion (26.2 ± 9.2 mg/dL vs. 31.1 ± 8.1, p≤ 0.001). NonHDL cholesterol was increased significantly in CAD group (156.8 ± 48.3 mg/dL vs. 140.3 ± 43.6, p≤ 0.001), and the changes occurred in both un- and esterified portions. FCE in HDL was diminished significantly in CAD patients (64.8 ± 13.9% vs. 69.3 ± 7.9, p≤ 0.01). In multivariate logistic regression analysis, unesterified cholesterol in NonHDL (UeNonHDLc) and esterified cholesterol in HDL (EsHDLc) excluded total cholesterol and HDLc respectively from the regression equation. In ROC analysis, the ratio of UeNonHDLc/EsHDLc was the strongest predictor for CAD among cholesterol subfractions. CONCLUSIONS: The results confirm that UeNonHDLc is atherogenic and EsHDLc is antiatherogenic and are independent risk factors for CAD.

Neurological disorders caused by two cerebral alveolar hydatid cysts in an old woman: a rare case report.

Alveolar hydatid disease, caused by Echinococcus multilocularis, is a life-threatening infectious disease which primarily occurs in the liver. Intracranial hydatid disease is a rare presentation with reported incidence of ~1% of all cases. Here we reported a 60-year-old woman, with the past history of hydatid cysts in her liver, who was presented to us with progressive symptoms consist of headaches, diminished vision, cognitive disorders and delusion. She was disoriented in time, space and person. Bilateral mild papilledema and exaggerated reflexes were observed. Magnetic resonance imaging of the brain revealed two intra-axial multilucular cystic masses in the fronto-pareital and parieto-occipital regions. The patient underwent two operations and the lesions were removed without any rupture. Medical therapy with Albendazole was started. Neurological symptoms disappeared a few weeks after the surgeries. Although multiple alveolar hydatid cysts are extremely rare, they should be considered in the differential diagnosis of intracranial cystic lesions.

Calculation of LDL-Cholesterol vs. Direct Homogenous Assay.

BACKGROUND: Low-density lipoprotein cholesterol (LDLc) can be calculated or measured directly and their accordance is the subject of controversy. OBJECTIVES: The aim of this study was to identify the independent predictors of LDLc, to formulate the best equation for calculating LDLc and to evaluate the validity of it and the published formulas, including the new method with adjustable coefficient. METHODS: The profile of serum lipids and (apo)lipoproteins of 310 subjects was used to determine the most accurate formula for calculating serum LDLc. Serum lipids, lipoproteins and apolipoproteins were measured by enzymatic, new homogenous and immunoturbidometric methods, respectively. RESULTS: Multiple linear regression analysis indicates that total cholesterol, apoB, HDLc and triglyceride are independent predictors of LDLc. We proposed four new formulas to calculate LDLc. As total cholesterol (TC) is the major determinant of LDLc, it can be estimated simply as 0.545 of total cholesterol. Inclusion of HDLc, triglyceride, apoB and a constant value improved the equation slightly. The equation of: LDLc (mg/dl) = 0.75 TC - 0.5 HDLc - 0.1 TG had the lowest mean and SD of difference among all the methods examined here. LDLc was also calculated by the new modified Friedewald's equation using adjustable factor from Martin's table, but it did not improve the results significantly. LDLc gap was correlated significantly and positively with triglyceride and negatively with cholesterol or its subfractions. CONCLUSIONS: Our data suggest the simplest formula: LDLc = 0.545 TC or a more detailed: LDLc = 0.75 TC - 0.5 HDLc - 0.1 TG be used for calculating serum LDLc.