Effects of Thymbra spicata extract and Thymol on morphine withdrawal syndrome in mice (insights to the liver function, antioxidant, and behavioral responses).

Safe chemicals for drug withdrawal can be extracted from natural sources. This study investigates the effects of clonidine and Thymbra spicata extract (TSE) on mice suffering from morphine withdrawal syndrome. Thymol, which is the active constituent in TSE, was also tested. A total of 90 mice were divided into nine groups. Group 1 was the control group, while Group 2 was given only morphine, and Group 3 received morphine and 0.2 mg/kg of clonidine. Groups 4-6 were given morphine along with 100, 200, and 300 mg/kg of TSE, respectively. Groups 7-9 received morphine plus 30, 60, and 90 mg/kg of Thymol, respectively, for 7 days. An oral naloxone challenge of 3 mg/kg was used to induce withdrawal syndrome in all groups. Improvement of liver enzyme levels (aspartate aminotransferase, alkaline phosphatase, and alanine transaminase) (p < .01) and behavioral responses (frequencies of jumping, frequencies of two-legged standing, Straub tail reaction) (p < .01) were significantly observed in the groups receiving TSE and Thymol (Groups 4-9) compared to Group 2. Additionally, antioxidant activity in these groups was improved compared to Group 2. Nitric oxide significantly decreased in Groups 4 and 6 compared to Groups 2 and 3 (p < .01). Superoxide dismutase increased dramatically in Groups 5, 8, and 9 compared to Groups 2 and 3 (p < .01). Groups 5-9 were significantly different from Group 2 in terms of malondialdehyde levels (p < .01). Certain doses of TSE and Thymol were found to alleviate the narcotics withdrawal symptoms. This similar effect to clonidine can pave the way for their administration in humans.

Are miR-26a and miR-26b microRNAs potent prognostic markers of gestational diabetes?

Background: Gestational diabetes mellitus is a common public health problem, accompanied by complications for the mother and fetus. So, introducing new biomarkers to identify early diabetes is essential. As serum miRNAs are potentially appropriate markers, we investigated miR-26a and miR-26b expression levels in pregnant women with and without gestational diabetes. Method: Demographic and clinical characteristics of 40 gestational diabetic patients and 40 healthy controls were assessed. The expression level of miR-26a and miR-26b microRNAs was measured by real-time PCR. Statistical analysis was done with GraphPad Prism software (version 8.4.3). Result: The findings of this study showed that the expression level of miR-26a and miR-26b increased in women with gestational diabetes compared with healthy pregnant women, but the increase in expression was only significant for miR-26a (p < 0.05). Conclusion: According to the statistical and ROC curves, we suggest miR-26a as a potential biomarker for the early diagnosis of gestational diabetes mellitus.

Attenuation of cannabis withdrawal symptoms by Prosopis farcta extract, its luteolin and melatonin in mice: Involvement of brain-derived neurotrophic factor and dopamine.

The aim of this study was the identification of luteolin in Prosopis farcta extract (PFE) and melatonin to evaluate its effect on THC withdrawal syndrome in mice. Luteolin was identified by high-performance liquid chromatography (HPCL). Signs of toxicity of mice in PFE and luteolin were monitored for LD50 calculation. The behavioral symptoms of THC withdrawal (stereotypies, ambulation, and inactivity time) induced by the rimonabant challenge were illustrated in THC-dependent mice receiving PFE, luteolin, and melatonin. The expression of mature BDNF (mBDNF) was evaluated by Western blot analysis. The dopamine concentrations were measured using HPLC. PFE and luteolin LD50 were 650 and 220 mg/kg, respectively. PFE (300 mg/kg), all doses of luteolin, and melatonin increased significantly the mBDNF expression and decreased the dopamine concentration. The findings suggest that PFE, luteolin, and melatonin are mighty in reducing the signs of THC withdrawal. It seems these effects were due to a decrease in dopamine concentration level and an increase in mBDNF protein expression in mice brains.

Chitosan/alginate scaffold enhanced with Berberis vulgaris extract for osteocyte differentiation of ovine fetal stem cells.

Designing biocompatible polymers using plant derivatives can be extremely useful in tissue engineering, nanomedicine, and many other fields of medicine. In this study, it was first looked into how chitosan/alginate scaffolds were made and characterized in the presence of berberine and barberry fruit extract. Second, the process of proliferation and differentiation of ovine fetal BM-MSCs (bone marrow-mesenchymal stem cells) was assessed on these scaffolds after BM-MSCs were extracted and confirmed by developing into osteocyte and adipose cells. To investigate the differentiation, treatment groups include (1) ovine fetal BM-MSCs were plated in Dulbecco's modified eagle medium culture medium with high glucose containing 10% fetal bovine serum and antibiotics (negative control), (2) ovine fetal BM-MSCs were plated in osteogenic differentiation medium (positive control group), (3) positive control group + barberry fruit extract, (4) positive control group + berberine, (5) ovine fetal BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate scaffold (hydrogel group), (6) ovine fetal BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/barberry fruit extract scaffold (hydrogel group containing barberry fruit extract), and (7) ovine fetal BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/berberine scaffold (hydrogel group containing berberine). Alkaline phosphatase (ALP) enzyme concentrations, mineralization rate using a calcium kit, and mineralization measurement by alizarin staining quantification were all found after 21 days of culture. In addition, real-time quantitative reverse transcription polymerase chain reaction was used to assess the expression of the ALP, COL1A2, and Runx2 genes. Days 5 and 7 had the lowest water absorption by the hydrogel scaffold containing barberry extract, which was significant in comparison to other groups (p < .05). Among the hydrogel scaffolds under study, the one containing barberry extract exhibited the lowest tensile strength, and this difference was statistically significant (p < .05). The chitosan/alginate hydrogel has the highest tensile strength of all of them. In comparison to the control and other treatment groups, the inclusion of berberine in the chitosan/alginate hydrogel significantly increased the expression of the ALP, Runx2, and COL1A2 genes (p < .05). The osteocyte differentiation of mesenchymal stem cells in in vitro settings appears to have been enhanced by the inclusion of berberine in the chitosan/alginate scaffold.

An overview of the role of metallic and nonmetallic nanoparticles and their salts during sperm cryopreservation and in vitro embryo manipulation.

The cryopreservation of spermatozoa and the in vitro embryo production are valuable tools used in a variety of species, including humans, livestock, fish, and aquatic invertebrates. Sperm cryopreservation has been used to maintain or increase the genetic diversity of threatened species. Reactive oxygen species (ROS) are molecules derived from oxygen, being formed as byproducts of cellular metabolism. During cryopreservation of sperm and other in vitro manipulations of oocytes and embryos, ROS production is dramatically increased. In cells, low, medium, and high levels of ROS lead to different outcomes, apoptosis, auto-phagocytosis, and necrosis, respectively. ROS produced by cells can be neutralized by intracellular antioxidant systems, including enzymes as well as non-enzymatic antioxidants. Free radicals and oxidative stress can be major factors influencing in vitro manipulations. In this review, we discuss the role that metallic and nonmetallic nanoparticles and their salts play in the modulation of oxidative stress during in vitro embryo production and cryopreservation of sperm.

Docosahexaenoic acid-loaded chitosan/alginate membrane reduces biofilm formation by P. aeruginosa and promotes MSC-mediated burn wound healing.

Aims: Chitosan, like docosahexaenoic acid (DHA) and mesenchymal stem cells (MSCs), is used in medicine as a wound healing accelerator. Thus, in this study, chitosan-alginate (CA) membranes containing DHA and MSCs were produced, and their antibacterial and antibiofilm activities against burn infections caused by Pseudomonas aeruginosa were investigated.Methods: Physicochemical properties were assessed by SEM, Fourier transform infrared (FTIR), and X-ray diffraction (XRD). Porosity, cytocompatibility, and antibacterial and antibiofilm activities were evaluated both in vitro and in vivo. The viability and apoptosis of MSCs were studied using flow cytometry. Wound healing effects were analyzed based on histopathological features, the wound contraction rate (WCR) ratio, and bacterial clearance.Results: The CA membranes showed antibiofilm activity both in vivo and in vitro, accompanied by reduced lasI and rhlI expressions and pyocyanin production. The membranes were highly porous and biocompatible and showed favorable physicochemical properties. Docosahexaenoic acid incorporation to CA membranes improved their antibacterial and antibiofilm activities, as well as MSCs' viability by reducing crystallinity and increasing porosity (p = .008). Treatment with CA-DHA-MSC accelerated burn wound healing (with complete healing being observed after 14 days, WCR = 85%) and augmented antibacterial and antibiofilm activities in vivo compared to CA-DHA and CA-MSC. The CA-DHA-MSC group delivered a significantly higher WCR and lower inflammation than the CA-MSC group (p = .0001).Conclusion: In combination with DHA-loaded CA membranes, MSCs reduced the healing time of burn wounds, offering a viable option for designing effective wound dressings.

Comparison of visfatin levels in patients with breast cancer and endometrial cancer with healthy individuals: A systematic review and meta-analysis.

Background and aims: Endometrial cancer (EC) and breast cancer (BC) are prevalent in women. Visfatin is an adipokine that, in addition to being involved in metabolism and inflammation, may also be interested in carcinogenesis. Visfatin measurement in cancer patients has shown that visfatin levels in cancer patients differed from those in healthy subjects. Various studies have shown that the level of visfatin is increased in people within EC and BC, and this difference has a significant relationship with prognosis. Methods: A comprehensive search of related articles from PubMed, Scopus, Web of Science, and the Google Scholar database was done by November 2021. Eligible articles measured visfatin levels in patients with breast cancer and EC. After selecting the eligible studies, the data were extracted and analyzed using the random effect method. Results: Given the effect size and the confidence interval obtained, the total level of visfatin in cancer patients was different from that in healthy individuals, and this difference was statistically significant. However, the difference in visfatin levels in patients with breast cancer was much more significant than in patients with EC compared to the control group. Conclusions: Due to the significant increase in visfatin levels in these patients, visfatin may be a potential prognostic factor in breast and ECs. Visfatin levels in cancer patients differed from those in healthy subjects, and this difference was also statistically significant (p-values = 0.00). Visfatin levels also differed between breast cancer patients and healthy individuals, which was statistically significant (p-values = 0.00). The difference in visfatin levels between patients with EC and healthy subjects was statistically significant (p-values = 0.047).

GFAP and Neuron Specific Enolase (NSE) in the Serum of Suicide Attempters.

Background: To determine whether neuronal damage and/or neuroinflammation exist in the brain of suicide attempters and to find a novel biological biomarker to help distinguishing high risk individuals with suicide behavior, we aimed to measure glial fibrillary acidic protein (GFAP), neuron specific enolase (NSE), and nerve growth factor (NGF) in suicide attempters. Methods: In the present case-control study, the serum level of NSE, GFAP, and NGF were measured quantitatively in 43 suicide attempters and 43 healthy control participants aged 18 to 35 years. Data were analyzed using the nonpaired t test followed by the Mann-Whitney posttest. Results: The mean serum level of NSE and GFAP were significantly higher in suicide attempters compared with healthy control individuals (p = 0.003, p = 0.001, respectively), while no significant difference was detected in NGF serum level between the 2 groups. Conclusion: Our findings of increased level of NSE along with the significant increase in GFAP would propose the presence of low grade neuroinflammation in the brain of these participants. NSE/GFAP might be good markers that is easily accessible and can be considered as prognostic markers in high-risk suicide attempters.

Investigation of biological effects of chitosan magnetic nano-composites hydrogel.

The growing concern about microorganism infections, especially hospital-acquired infections, has driven the demand for effective and safe agents in recent years. Herein, novel nanocomposites were prepared based on layered double hydroxides (LDH NPs), Fe2O3nanoparticles (Fe2O3NPs), and chitosan hydrogel beads in different concentrations. The characteristics and composition of the prepared materials were investigated by various techniques such as XRD, FESEM, and FTIR. The results indicate that the nanocomposites are synthesized successfully, and each component is present in hydrogel matrixes. Then, their biomedical properties, including antibacterial, antifungal, and antioxidant activity, were examined. Our findings demonstrate that the antimicrobial activity of nanocomposites significantly depends on the concentration of each component and their chemical groups. It shows itself in the result of the inhibitory zone of all bacteria or fungi samples. The obtained results indicate that the nanocomposite of Chitosan-hydrogel beads with 20% LDH and Fe2O3(CHB-LDH-Fe2O3%20) and Chitosan-hydrogel beads based on 20% LDH (CHB-LDH%20) showed excellent antibacterial and antifungal properties against all tested bacteria and fungi (P ≤ 0.01). In addition, the antioxidant effects of the synthesized materials (especially CHB-LDH Fe2O3%20 and CHB-LDH%20) were investigated, showing high antioxidant efficacy against DPPH free radicals (P ≤ 0.01). According to our findings, we can say that these materials are promising biomaterials for inhibiting some infectious bacteria and fungi.

Thymol Nanopolymer Synthesis and Its Effects on Morphine Withdrawal Syndrome in Comparison With Clonidine in Rats.

The drug delivery system is valuable in the treatment of the disease. A nanopolymer as a thymol and Thymbra spicata release system was synthesized and its effects on morphine withdrawal syndrome in comparison with clonidine in rats were studied. The nanopolymer was characterized by different methods, namely, IR, HNMR, CNMR, GPC, DLS, and AFM. Thymol in T. spicata extract was assessed. The loading and release rate of thymol and T. spicata extract on the nanopolymer were evaluated by HPLC. The median lethal dose (LD50) of the T. spicata extract, thymol, extract nanopolymer, and thymol nanopolymer was studied. The frequency of jumping, rearing, and teeth chattering in naloxone-induced morphine withdrawal syndrome was studied. Synthesized nanopolymer was desirable as a carrier for the drug. The loaded amount of extract and thymol on nanopolymer was estimated 55 ± 3.2% and 48 ± 2.6% and the drug released was 71 and 68%, respectively. LD50 of the T. spicata extract, thymol, extract nanopolymer, and thymol nanopolymer was 975, 580, 1,250, and 650 mg/kg, respectively. This study showed that thymol nanopolymer was more effective than clonidine to reduce the frequency of morphine withdrawal symptoms. Our results suggest that T. spicata extract, thymol, extract nanopolymer, and thymol nanopolymer are mighty in reducing the narcotic withdrawal signs. The mechanism of action and therapeutic potential is maybe similar to clonidine.

Changes in Physiological Levels of Cortisol and Adrenocorticotropic Hormone upon Hospitalization Can Predict SARS-CoV-2 Mortality: A Cohort Study.

There is some indication that coronavirus disease 2019 (COVID-19) causes hypothalamic-pituitary-adrenal axis insufficiency. However, being on glucocorticoids makes it difficult to fully investigate this axis, especially in patients with severe COVID-19. We aimed to discover if there was a connection between blood total cortisol and adrenocorticotropic hormone (ACTH) levels and mortality in patients with COVID-19. In Iran, 154 hospitalized patients with COVID-19 were studied in a prospective cohort study. ACTH and cortisol levels in the blood were measured on the first or second day of hospitalization. Most patients (52.6 vs. 47.4%) were men over 50 years old (55.8%), and 44.4% had an underlying illness. Serum cortisol and plasma ACTH medians were 15.6 (µg/dl) and 11.4 (pg/ml), respectively. 9.09% of the patients died. Cortisol levels were substantially lower in those who died (11.3 µg/dl) than in patients who were discharged (16.7 µg/dl, P < 0.01), while ACTH levels were unaffected. The most important factors determining mortality, according to the logistic model, were blood cortisol levels, the existence of an underlying disease, and the use of a mechanical ventilator. Cortisol levels that rose by one-unit correlated with a 26% lower risk of mortality. Comorbidities and mechanical ventilation increased the risk of death by 260 and 92 times, respectively. It can be concluded that in patients with COVID-19, a low cortisol level is linked to a high risk of mortality. Patients may sometimes have relative primary adrenal insufficiency. To judge and decide on therapeutic interventions, more reliable and long-term follow-up studies are required.

The Effect of Prosopis farcta and Its Bioactive Luteolin on the Hippocampus of Mice after Induced Ischemia Reperfusion.

BACKGROUND: Ischemia plays an important role in increasing damage to the nervous system. This study aimed to evaluate the effect of Prosopis farcta (PFE) and its bioactive luteolin (Lu) and forced swimming exercise on the hippocampus of mice after induced ischemia reperfusion. METHODS: The bioactive component of PFE (Lu) was identified by HPLC. Fifty-six male mice were divided into different groups. Ischemia was induced by ligation of the common carotid artery. After mice training (swimming exercise, 8 weeks) and consuming PFE and Lu, the mice's memory ability was evaluated in the shuttle box. Histological examination was performed by Nissel staining and immunohistochemistry. RESULTS: Results showed that the ischemic mice exercised and treated with PFE and Lu had higher step-through latency (STL) compared with the nonexercised mice, and this was confirmed with time spent in the dark compartment (TDC). The number of dark cells in the ischemic group exercising and receiving PFE and Lu decreased compared to that of the other groups in the hippocampus. DCX protein expression was increased in nonexercised groups compared to that of the exercised groups and those treated with PFE and Lu, while NeuN decreased. CONCLUSIONS: Forced swimming exercise following ischemia, as well as consumption of PFE and Lu, has reduced cell death and increased neurogenesis in the hippocampus and thus may help improve memory in ischemia.

The Role of Oxidant-Antioxidant Status in Suicide Behavior in Kurdish Ethnicity.

INTRODUCTION: Oxidative stress plays a critical role in the pathogenesis of neurodegenerative and neuropsychiatric disorders. However, its role in suicidal behavior has not been clarified yet. Consequently, we aimed to evaluate the oxidant-antioxidant status in the serum of suicide attempters in Ilam city. METHODS: Fifty suicide attempters and 40 control subjects (volunteers) aged 18-35 years were studied in the current experiment. To consider the oxidant-antioxidant status, serum levels of Malondialdehyde (MDA), Nitric Oxide (NO), Superoxide Dismutase (SOD), and the Total Antioxidant Capacity (TAC) were measured. RESULTS: Serum levels of SOD and TAC were significantly lower in the suicide attempters group compared to the controls. Furthermore, serum NO level was significantly higher in the suicide attempters compared to the control groups. Interestingly, the serum level of MDA was significantly lower in the suicide attempters compared to the control groups. CONCLUSION: The oxidative stress without MDA elevation, detected in suicide attempters, can be considered a biochemical hallmark in suicide behavior.

Immobilized Ag NPs on chitosan-biguanidine coated magnetic nanoparticles for synthesis of propargylamines and treatment of human lung cancer.

The magnetically isolable nanobiocomposites have significant impact as the modified new generation catalysts in recent days. This has persuaded us to design and synthesis of a novel Ag NPs decorated biguanidine-chitosan (Bigua-CS) dual biomolecular functionalized core-shell type magnetic nanocomposite (Ag/Bigua-CS@Fe3O4). Bigua-CS could be introducing polysaccharide materials as potential coating agent to immobilizing and stabilizing metal nanoparticles. The material was characterized using several advanced techniques like fourier transformed infrared spectroscopy (FT-IR), inductively coupled plasma (ICP), field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDX), atomic mapping, high resolution transmission electron microscopy (HR-TEM), vibrating sample magnetometer (VSM) and X-ray diffraction (XRD). Towards the chemical applications of the material, we headed the multicomponent synthesis of diverse propargylamines by A3 coupling in water, which ended up with excellent yields. Due to strong paramagnetism, the catalyst was easily isolable and reused in 9cycles without any leaching and considerable change in reactivity. In addition, the catalyst was engaged in biological assays like study of anti-oxidant properties by DPPH mediated free radical scavenging test using BHT as a reference molecule. Thereafter, on having a significant IC50 value in radical scavenging assay, we extended the bio-application of the catalyst in anticancer study of adenocarcinoma cells of human lungs. The three different cancer cell lines, PC-14, LC-2/ad and HLC-1 were used in this regard. The best result was achieved in the case of PC-14 cell line with strong IC50 values.

Effects of Different Concentrations of Reversine on Plasticity of Mesenchymal Stem Cells.

Dedifferentiation can be induced by small molecules. One of these small molecules used in this study in order to increase the plasticity of differentiation of stem cells was reversine. The objective of present study was to investigate the effect of different concentrations of reversine on the plasticity of ovine fetal bone-marrow mesenchymal stem cells (BM-MSCs). BM-MSCs were extracted from ovine fetal and cultured. Passaged-3 cells were evaluated for their differentiation potential into osteocytes and adipocytes cells. In the present study, BM-MSCs were culture plated in the presence of 0, 300, 600, 900 and 1200 nM of reversine. The number of viable cells was determined by MTT test after addition of different concentrations of reversine. Furthermore, expression of the nanog gene was evaluated. The culture without reversine was taken as the control group. Expression of nanog was analysed by immunocytochemistry. Multi-lineage differentiation showed that the BM-MSCs could be differentiated into adipose cells and osteocytes. Our results indicated that the addition of 1200 nM of reversine to medium significantly decreased overall proliferation compared to the other treatment groups (p > 0.05). Real-time PCR analysis showed that after addition of 600 nM of reversine significantly increased nanog expression compared to other treatments. All treatments received reversine were seen to be relative expression of nanog. Our findings confirm that low concentrations reversine increases the plasticity of ovine BM-MSCs.

Is there association between Trichomonas vaginalis infection and prostate cancer risk?: A systematic review and meta-analysis.

We performed a systematic review and meta-analysis to reveal the association between Trichomonas vaginalis (TV) infection and the risk of prostate cancer (PCa) development. Systematic searching (PubMed/Medline, Scopus, Embase, Web of Science, Cinhal, Wiley, Cochrane, Psychoinfo, ProQuest and Google Scholar) was done, up to March 2018 for case-control studies. Random effects model was applied to define odds ratios and their 95% confidence intervals. In total, 6 enteries were included in our meta-analysis, comprising 5590 individuals (2677 PCa cases and 2913 control individuals) examined for trichomoniasis, with a total positivity of 469 (17.51%) and 482 (16.54%) individuals, respectively. Totally, such association was documented in three countries, including USA (4 studies), Kuwait (one study) and Taiwan (one study). Based on pooled estimations, however a 1.17-time increase of PCa was observed among individuals with a previous exposure of TV, it was not statistically significant [OR = 1.17 (95% CI: 1.01 to 1.36)]. Egger's regression test demonstrated no publication bias among studies. Also, year of publication for included records was not significantly correlated to the relationship between trichomoniasis and PCa. Any further inferences should be based on future investigations for better understanding this relationship and shedding light on the cryptic pathogenesis of TV in PCa patients.

Novel synthesis of Falcaria vulgaris leaf extract conjugated copper nanoparticles with potent cytotoxicity, antioxidant, antifungal, antibacterial, and cutaneous wound healing activities under in vitro and in vivo condition.

Facile green synthesis of copper nanoparticles from different biological procedures has been indicated, but among all, biosynthesis of copper nanoparticles from medicinal plants is considered as the most suitable method. The use of medicinal plant material increases the therapeutical effects of copper nanoparticles. The aim of this study was green synthesis of copper nanoparticles from aqueous extract of Falcaria vulgaris leaf (CuNPs) and assessment of their cytotoxicity, antioxidant, antifungal, antibacterial, and cutaneous wound healing properties. These nanoparticles were characterized by X-ray diffraction (XRD), fourier-transform infrared spectroscopy (FT-IR), ultraviolet-visible spectroscopy (UV), transmission electron microscopy (TEM), and field emission scanning electron microscopy (FE-SEM) analysis. The synthesized CuNPs had great cell viability dose-dependently (Investigating the effect of the CuNPs on human umbilical vein endothelial cell (HUVEC) line) and indicated this method was nontoxic. Also, 2,2-diphenyl-1-picrylhydrazyl (DPPH) test was done to assess the antioxidant activities, which indicated similar antioxidant potentials for CuNPs and butylated hydroxytoluene. In part of cutaneous wound healing property of CuNPs, after creating the cutaneous wound, the rats were randomly divided into six groups: treatment with 0.2% CuNPs ointment, treatment with 0.2% CuSO4 ointment, treatment with 0.2% F. vulgaris ointment, treatment with 3% tetracycline ointment, treatment with Eucerin basal ointment, and untreated control. These groups were treated for 10 days. Treatment with CuNPs ointment remarkably increased (p ≤ .01) the wound contracture, vessel, hexosamine, hydroxyl proline, hexuronic acid, fibrocyte, and fibrocytes/fibroblast rate and substantially reduced (p ≤ .01) the wound area, total cells, neutrophil, and lymphocyte compared to other groups. In antibacterial and antifungal parts of this research, the concentration of CuNPs with minimum dilution and no turbidity was considered minimum inhibitory concentration (MIC). To determine minimum fungicidal concentration (MFC) and minimum bactericidal concentration (MBC), 60 µL MIC and three preceding chambers were cultured on Sabouraud Dextrose Agar and Muller Hinton Agar, respectively. The minimum concentration with no fungal and bacterial growth were considered MFC and MBC, respectively. CuNPs inhibited the growth of all fungi at 2-4 mg/mL concentrations and removed them at 4-8 mg/mL concentrations (p ≤ .01). In case of antibacterial effects of CuNPs, they inhibited the growth of all bacteria at 2-8 mg/mL concentrations and removed them at 4-16 mg/mL concentrations (p ≤ .01). The results of XRD, FT-IR, UV, TEM, and FE-SEM confirm that the aqueous extract of F. vulgaris leaf can be used to yield copper nanoparticles with notable amount of antioxidant, antifungal, antibacterial, and cutaneous wound healing potentials without any cytotoxicity. Further clinical trials are necessary for confirmation these therapeutical effects of CuNPs in human.

Characterization and Classification of Mesenchymal Stem Cells in Several Species Using Surface Markers for Cell Therapy Purposes.

Mesenchymal stem cells are multipotent cells capable of replicating as undifferentiated cells, and have the potential of differentiating into mesenchymal tissue lineages such as osteocytes, adipocytes and chondrocytes. Such lineages can then be used in cell therapy. The aim of present study was to characterize bone marrow derived mesenchymal stem cells in four different species, including: sheep, goat, human and mouse. Human bone-marrow mesenchymal stem cells were purchased, those of sheep and goat were isolated from fetal bone marrow, and those of mouse were collected by washing bone cavity of femur and tibia with DMEM/F12. Using flow-cytometry, they were characterized by CD surface antigens. Furthermore, cells of third passage were examined for their osteogenic and adipogenic differentiation potential by oil red and alizarin red staining respectively. According to the results, CD markers studied in the four groups of mesenchymal stem cells showed a different expression. Goat and sheep expressed CD44 and CD166, and weakly expressed CD34, CD45, CD105 and CD90. Similarly, human and mouse mesenchymal cells expressed CD44, CD166, CD105 and CD90 whereas the expression of CD34 and CD45 was negative. In conclusion, although all mesenchymal stem cells display plastic adherence and tri-lineage differentiation, not all express the same panel of surface antigens described for human mesenchymal stem cells. Additional panel of CD markers are necessary to characterize regenerative potential and possible application of these stem cells in regenerative medicine and implantology.

The combination of miR-122 overexpression and Let-7f silencing induces hepatic differentiation of adipose tissue-derived stem cells.

Human adipose tissue-derived stem cells (hADSCs) have been considered as a promising source for cell therapy of liver diseases due to their accessibility, abundance, and expression of hepatocyte markers. Currently, the important role of certain microRNAs (miRNAs) has been reported during hepatic differentiation of stem cells. However, the combination effect of miRNAs on hepatic differentiation of these cells needs to be more investigated. The present study seeks to determine whether the combination of miRNAs can enhance hepatic differentiation of hADSCs in the absence of any other stimulation. First, lentiviral transduction was used to overexpress miR-122 and silence d let-7f in hADSCs for up to 21 days. Then, hepatic functionality was evaluated by analyzing specific hepatocyte genes and biochemical markers at different time points of differentiation induction. Stable miR-122 overexpression and let-7f silencing together in hADSCs resulted in increased expression of hepatocyte markers including ALB, AFP, CK18, CK19, and HNF4a. In addition, urea and albumin production, immunocytochemistry, and glycogen staining confirmed that the treated cells differentiated toward hepatocyte-like cells. Therefore, our findings demonstrate the possibility of using microRNAs to induce hADSCs into functional hepatocyte-like cells.

Ovine fetal mesenchymal stem cell differentiation to cardiomyocytes, effects of co-culture, role of small molecules; reversine and 5-azacytidine.

The aim of the present study was to investigate the effect of small molecules: Reversine and 5-azacytidine (5-AC), in an indirect co-culture condition with the cardiac fibroblasts as well as non co-culture condition, in order to explore the effect of such molecules in the process of differentiation of the ovine bone-marrow mesenchymal stem cells (BM-MSCs) towards cardiomyocytes. Surface antigens of the isolated cells were analysed using flow-cytometry. In addition, following to three passages cells were examined for their differentiation capacity into osteocytes and adipose cells, in order to ensure the mesenchymal origin of the stem cells. Six types of treatments were carried out in the present investigation, such that, in the first treatment BM-MSCs were cultured for 28 days as control group; the second treatment was composed of culturing ovine fetal cardiac fibroblasts on inserts, aiming to use these inserts for culturing plates which were seeded with BM-MSCs (Chamber group). As the third treatment, BM-MSCs were supplemented with 10-µM 5-AC and incubated for 48 h. The fourth treatment was composed of supplementing BM-MSCs with the 600-nM reversine, incubated for 48 h, and subsequently the incubation was further extended for another 48 h in the presence of 5-AC. The fifth treatment was composed of supplementing the chamber group with 10-µM 5-AC and incubation for 48 h, and the last or the sixth treatment was such that chamber group was supplemented with 600-nM reversine and an incubation period of 48 h. Following to the incubation, medium was replaced with 10-µM 5-AC and further incubated for another round of 48 h. In all treatments, following to addition of the small molecules incubations were carried out for 28 days; same as controls. Expression of cardiac alpha-actinin was analysed by immunocytochemistry. BM-MSCs have shown to express CD44 and CD166 along with a weak expression of the CD90, CD34, in addition to CD45. Multilineage differentiation has indicated that BM-MSCs could differentiate into adipose and osteocytes cells as well. In the treatment 4 it was observed that FGF signalling involved genes and all cardiac-related genes (ANP, MYH6 and Troponin I) were significantly expressed, except connexin 43 compared to other treatments. All treatments received small molecules, either alone or as a co-culture were seen to express sarcomeric alpha-actinin. This finding was partially supported by immunocytochemistry. These results validate that reversine and 5-AC have an effect on ovine BM-MSC differentiation into cardiomyocytes. Copyright © 2016 John Wiley & Sons, Ltd.