ABSTRACT
Background: After improvements in various cancer treatments, life expectancy has been raised, but success in treatment causes loss of fertility in many of the survived young men. Cryopreservation of immature testicular tissues or cells introduced as the only way to preserve fertility. However, freezing has some harmful effects. Melatonin, a pineal gland hormone, has receptors in reproductive systems of different species. It is assumed that melatonin has free radical scavenger properties
Objective:The aim of this study was to evaluate the effects of melatonin on the cryopreserved testicular cells in mouse
Materials and Methods: Cells from 7- 10 days old NMRI mice testes were isolated using two step enzymatic digestion. The testicular cells were divided into two groups randomly and cryopreserved in two different freezing media with and without the addition of 100 micro m melatonin. Finally, apoptosis of the cells was assayed by flow cytometry. Also, lactate dehydrogenase activity test was performed to assess the cytotoxicity
Results: The results of lactate dehydrogenase showed the nearly cytotoxic effect of melatonin. The results of flow cytometry showed increase in apoptosis in the cryopreserved cells in the media containing melatonin compared to the control group
Conclusion: The present study shows that melatonin has an apoptotic effect on cryopreserved mouse testicular cells
ABSTRACT
Objective: To evaluate the effect of Exendine-4 [EX-4], a Glucagon-like peptide 1 [GLP-1] receptor agonist, on the differentiation of insulin-secreting cells [IPCs] from rat adipose-derived mesenchymal stem cells[ADMSCs]
Materials and Methods: In this experimental study, ADMSCs were isolated from rat adipose tissue and exposed to induction media with or without EX-4. After induction, the existence of IPCs was confirmed by morphology analysis, expression pattern analysis of islet-specific genes [Pdx-1, Glut-2 and Insulin] and insulin synthesis and secretion
Results: IPCs induced in presence of EX-4 were morphologically similar to pancreatic islet-like cells. Expression of Pdx-1, Glut-2 and Insulin genes in EX-4 treated cells was significantly higher than the cells exposed to differentiation media without EX-4. Compared to EX-4 untreated ADMSCs, insulin release from EX-4 treated ADMSCs showed a nearly 2.5 fold [P<0.05] increase when exposed to a high glucose [25 mM] medium. The percentage of insulin positive cells in the EX-4 treated group was approximately 4-fold higher than in the EX-4 untreated ADMSCs
Conclusion: The present study has demonstrated that EX-4 enhances the differentiation of ADMSCs into IPCs. Improvement of this method may help the formation of an unlimited source of cells for transplantation
ABSTRACT
Objective: Zinc oxide nanoparticles [ZnO-NPs] are increasingly used in sunscreens, biosensors, food additives, pigments, manufacture of rubber products, and electronic materials. There are several studies about the effects of NPs on dermal fibroblast or keratinocytes, but very little attention has been directed towards adipose-derived mesenchymal stem cells [ASCs]. A previous study has revealed that ZnO-NPs restricted the migration capability of ASCs. However, the potential toxicity of these NPs on ASCs is not well understood. This study intends to evaluate the effects of ZnO-NPs on subcutaneous ASCs
Materials and Methods: In this experimental study, In order to assess toxicity, we exposed rat ASCs to ZnO-NPs at concentrations of 10, 50, and 100 Mug/ml for 48 hours. Toxicity was evaluated by cell morphology changes, cell viability assay, as well as apoptosis and necrosis detection
Results: ZnO-NPs concentration dependently reduced the survival rates of ASCs as revealed by the trypan blue exclusion and 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium-bromide [MTT] tests. ZnO-NPs, at concentrations of 10 and 50 Mug/ml, induced a significant increase in apoptotic indices as shown by the annexin V test. The concentration of 10 Mug/ml of ZnO-NPs was more toxic
Conclusion: Lower concentrations of ZnO-NPs have toxic and apoptotic effects on subcutaneous ASCs. We recommend that ZnO-NPs be used with caution if there is a dermatological problem
ABSTRACT
Leukemia inhibitory factor [LIF] is a glycoprotein, categorized as a subfamily of interleukin 6 cytokines which is known in many mammals. A pluripotent cytokine with a wide biological function range has numerous effects on target cells. The LIF regulates neuron survival, hematopoiesis and seen in LIF[-/-] knockout mice affects blastocyst implantation, also acts as pre-inflammatory cytokine, and regulates immune response. Further, it is able to maintain stem cells poly potency. The main object of present work was expression, optimizing, and purification of recombinant human leukemia inhibitory factor [rhLIF]. In this experimental study, Pet28 [+] carrying the LIF gene and kanamycin resistance marker was cloned in E. coli strain BL21. The induction was optimized by altering 3 factors including the temperature, the induction time, and the concentration of the Isopropyl beta-D-1-thiogalactopyranoside [IPTG] as inducer. The purification of the recombinant human LIF [rhLIF] was done by single step affinity chromatography. After the purification, method accuracy was proved by Sodium dodecyl sulfate [SDS] -PAGE electrophoresis and Western blotting. Optimizing of the expression was reached by changing various parameters, and purification has been done successful. rhLIF undergoes modification by glycosylation to get its full functionality. The produced rhLIF in prokaryotic host in this work is lacking of glycosylation. However, its proper function should be evaluated in further studies
ABSTRACT
Background: Interleukin [IL]-23 and IL-27 are two IL-12-related cytokines which their function may dramatically influence the inflammatory response to tumor development. IL-12 and IL-27 seem to have antagonistic roles with IL-23 in tumor site. In this study, IL-23 and IL-27 mRNA expressions were analyzed in peripheral blood of patients with breast cancer and healthy volunteers using quantitative real-time PCR
Methods: Peripheral blood samples were collected from 50 women with breast cancer and 50 healthy ones. The total RNA was extracted from peripheral blood after lysis with ammonium chloride and TRizol reagent and the cDNA was synthesized. The expression of IL-23 and IL-27 gene transcripts was determined with real-time polymerase chain reaction [qRT-PCR] using Syber Green PCR Master Mix
Results: It is found that IL-23 and IL-27 transcripts had significantly higher expression in peripheral blood of patients compared with the healthy controls. The ratio of IL-23 transcript expression to IL-27 was 3.4 fold lower in the studied patients compared with the normal individuals
Conclusion: It is concluded that the over expression of IL-23 and IL-27 gene transcript in peripheral blood of breast cancer patients may be an immune response against tumor development and the inflammatory response plays a critical role in tumor development via up regulating the corresponding cytokines. However, the IL-23/IL-27 ratio may play an important role in cytokine-based immunotherapy against cancer. Further research should be carried out to assess these cytokines in a larger sample size
ABSTRACT
Reactive oxygen species and oxidative stress impair beta-cell function and reduce insulin secretion. It has been shown that progesterone and cilostazol possess antioxidant properties. The present study was aimed to investigate in vitro pretreatment effect of progesterone and cilostazol on insulin secretion as well as their protective effects against hydrogen peroxide-induced oxidative stress in pancreatic isolated islets from mouse. Pancreatic islets were isolated from 84 male NMRI mice [25-30g] by collagenase digestion method and pretreated for 48h with cilostazol [10 microM], progesterone [0.5 microM] and glibenclamide [10 microM] in culture medium. Then islets were exposed to hydrogen peroxide [H2O2. 500 microM] for 2h. Next, culture mediums containing glucose concentration of 2.8 mM or 16.7 mM were added to them and incubated in this status for 1h. At the end, the rate of insulin output from islets, lipid peroxidation and antioxidant enzymes activities in islet tissues were assayed. Exposure of islets to H2O2, resulted in a significant decrease in insulin secretion, superoxide dismutase and catalase activities [P<0.001]. Also islets malondialdehyde levels were increased by H2O2, after addition of 2.8mM [P<0.05] and 16.7mM [P<0.001] glucose. 48h pretreatment of islets with cilostazol and progesterone, significantly reverted back this changes [P<0.05]. Results of present study showed that cilostazol and progesterone protect mice pancreatic islets against H2O2-induced oxidative stress. At the end, our results suggested that protective effects of progesterone and cilostazol are mediated by augmentation the antioxidant defence system of islets
ABSTRACT
Glycogen storage disease II [GSDII or Pompe disease, OMIM # 232300] is an autosomal recessive hereditary lysosomal disorder. Mutations in the GAA gene usually lead to reduced acid alpha-glucosidase [acid maltase, GAA, OMIM [asterisk] 606800, EC 3.1.26.2] activity, which results in impaired degradation and subsequent accumulation of glycogen within lysosomes. We present an Iranian boy, who was diagnosed with GSDII based upon clinical and biochemical findings. A single adenine insertion [insA] was detected at codon 693 that leads to a predicted premature stop codon at codon 736 in the GAA gene. The parents were heterozygous for the same change. According to the human genome mutation database [www.hgmd.org] and lecture reviews, the detected change is a novel mutation. We suppose that the discovered insertion in the GAA gene might lead to a reduced activity of the gene product. This assumption is in agreement with biochemical and clinical signs in the patient
Subject(s)
Humans , Male , Female , alpha-Glucosidases/genetics , Mutation , ChildABSTRACT
Previous studies have shown that some cytokines have protective effects on cartilage in joint diseases. In the current study, effects of IL-4 against morphological changes and tissue degradation induced by IL-1 Alpha on bovine nasal cartilage [BNC] explants were investigated. Fresh BNC samples were prepared from a slaughterhouse under sterile conditions. BNC explants culture was treated with both IL-l Alpha [10 ng/ml] and IL-4 [50 ng/ml] at the same time for 28 days. The morphological characteristics of explants were assessed by using histology techniques and invert microscopy. Matrix metalloproteinase-1 [MMP-1] production was assessed within different days by using Western blotting. IL-l Alpha induced prominent cartilage morphology degradation. The pro and active form of MMP-1 band substantially increased at day 21 of culture. In the presence of both IL-l Alpha and IL-4, chondrocytes preserved their ordinary normal phenotype with intact extracellular matrix. In addition, a significant reduction in pro-MMP-1and inhibition of active MMP-1 was seen. In conclusion, IL-4 could be regarded as a potential candidate in cartilage protecting against the degradation changes of IL-l Alpha. It seems that the preservation effect of IL-4 is associated with significant reduction of MMP-1
Subject(s)
Animals, Laboratory , Animals , Chondrocytes , Matrix Metalloproteinase 1 , Cattle , Nasal CartilagesABSTRACT
A study of the histological events under interleukin-1 alpha [IL-1alpha] induction of bovine nasal cartilage [BNC] could result in useful data to better understand the mechanisms involved in tissue breakdown in joint diseases. The aim of this study was to investigate the effects of IL-1alpha on chondrocyte phenotype and extracellular matrix [ECM] changes in BNC explants. In this experimental study, samples were divided into two groups. Group I [control group] BNC explants were cultured only in Dulbecco's modified Eagle's medium [DMEM]. In group II, BNC explants were treated with IL-1alpha [10 ng/ml] for 28 days. Then, samples were harvested on culture days 3, 7, 14, 21 and 28 and chondrocyte morphology and ECM alterations were assessed by invert microscopy and histology by hematoxylin and eosin [H and E] and Alcian blue. Cell viability was evaluated by the lactate dehydrogenase [LDH] assay test. Data were analyzed by the t test and p<0.05 was considered significant. IL-1alpha induced significant morphological changes in cartilage. In the presence of IL-1alpha, most chondrocytes transformed into a fibroblast-like morphology with a granular black point appearance. An increase in the cell: matrix ratio was observed and there were decreased numbers of chondrocytes.IL-1alpha induced breakdown of ECM. We observed partial degradation of ECM between days 7-14 and complete degradation occurred between days 21-28 of culture. The LDH levels increased. IL-1alpha induced morphological changes in chondrocytes and increased destruction of cartilage ECM. There was a parallel correlation between proteoglycan degradation and changes in chondrocyte morpholgy
Subject(s)
Animals , Chondrocytes/drug effects , Nasal Cartilages/drug effects , Extracellular Matrix , L-Lactate DehydrogenaseABSTRACT
Noise pollution has a high prevalence among the environmental pollutions and is considered as a teratogenic agent for reproductive system. This study was performed with the purpose of evaluation of the effect of honey and vitamin E on sex hormone levels in male rats exposed to noise pollution. Twenty-four adult male rats with the mean weight of 200 +/- 20g were randomly divided into 4 groups: 1 [honey+voice], 2 [vitamin E+voice], 3 [voice], and 4 [control]. Groups 1 and 2 received honey and vitamin E as gavage, in addition to voice; group 3 was only exposed to noise pollution. After 50 days, serum level of hormone in male rats were measured by ELISA technique after taking blood from heart. Then, the male rats were placed in a cage with female rats of the same breed with a 2:1 ratio. Weight and number of embryos from fertilization were assessed. Data were analyzed using ANOVA and Tukey statistical methods. In this study, it was found that the secretion of sex hormones [FSH, LH, and testosterone] impaired under the effect of noise pollution. Serum level of testosterone decreased in rats that were under noise stress [p<0.05], and use of honey and vitamin E as antioxidants were modulated the level of this hormone in male rats. Noise pollution in male rats increased in the serum level FSH, LH [p<0.05]. Weight and number of live embryos decreased because of this stress [p<0.05]. The use of Honey and vitamin E by male rats increased live embryos [p<0.05]. According to the results of this study, noise pollution has negative effects on the fertility of male rats. Also, use of honey and vitamin E increases the fertility in groups exposed to noise pollution
Subject(s)
Animals, Laboratory , Vitamin E , Gonadal Steroid Hormones , Honey , Rats, Wistar , Follicle Stimulating Hormone , Luteinizing Hormone , TestosteroneABSTRACT
Regulatory T cells [T-regs] have an important role in cancer by suppression of protective antitumor immune responses. Regulatory T cells express the forkhead/ winged helix transcription factor [FOXP3] and OX40 molecules which have important regulatory roles in the immune system. To evaluate FOXP3 and OX40 transcripts in the peripheral blood mononuclear cells of women with breast cancer. Blood samples from 40 women with histologically-confirmed infiltrating ductal carcinoma of the breast and 40 healthy volunteer women without a history of malignancy or autoimmune disorders were collected. The abundance of FOXP3 and OX40 gene transcripts were determined by quantitative real-time PCR [qRT-PCR]. There was a significant positive correlation between FOXP3 and OX40 gene expression in women with breast cancer in a stage dependent manner. This finding emphasizes the importance of T-regs as predominant targets for breast cancer immunotherapy
Subject(s)
Receptors, OX40 , Breast Neoplasms/immunology , Breast Neoplasms/genetics , T-Lymphocytes, Regulatory , Immunotherapy , Polymerase Chain Reaction , Gene Expression , Transcription Factors , ImmunotherapyABSTRACT
Cyclosporine A [CsA] is an important immunosuppressive agent; however, its clinical use is limited by several side effects such as hepatotoxicity. Vitamin C [ascorbic acid] is a very important and powerful antioxidant and protects membranes against oxidation. The aim of this study was to study protective role of vitamin C against CSA-induced hepatotoxicity. Thirty male Wister strain rats weighting 230-260g were randomly divided into 3 groups [n = 10]: group A was the control group and received placebo [Normal Saline], group B was the CSA-treated group and received 15mg/kg/day CsA for 21 days, group C was the CsA + vitamin C group and was received 200mg/kg/day vitamin C orally 3 hours before receiving 15mg/kg/day CsA. On 22[th] day rats serum obtained for measuring biochemical factors including bilirubin, alanine aminotransferase [ALT], aspartate aminotransferase [AST], triglyceride, alkaline phosphatase [ALP] and lactate dehydrogenase [LDH], total protein, and albumin. Bilirubin, ALT, AST, triglyceride, ALP, and LDH levels were lower in CsA + ascorbic acid group than that of CsA group [P < 0.05] while plasma total protein and albumin were significantly higher in CsA + ascorbic acid group than that of CsA group [P < 0.05]. In conclusion, we have shown that vitamin C administration provides protection against CSA-induced injury in rat liver function and may have hepatoprotective role in the patients experiencing CSA treatment
Subject(s)
Male , Animals, Laboratory , Cyclosporine/toxicity , Chemical and Drug Induced Liver Injury/drug therapy , Rats, WistarABSTRACT
The detection of nucleic acids using Real-Time CPR has many application. Quantitative analysis of mRNA using Real-Time PCR by Relative and Absolute methods. Widely used in biological studies. The purpose of this study was to compare the Relative and calculated PCR Efficiency by standard curve and LinRegPCR methods. After sampling and extraction of RNA and cDNA synthesis, the quantitative RT-PCR was performed, then the PCR Efficiency was calculated by Using the two methods of standard curve and LinRegPCR. At the end the results of the two methods were compared and analyzed. The efficiency of PCR for the GAPDH, TGF-beta and IL- 10 genes with the standard curve method were 1.99, 1.81 and 1.87, respectively. The PCR efficiency of these three genes were 1.98, 1.82 and 1.82 by using LinRegPCR software method, respectively. The analysis of the data obtained from PCR proliferation of cDNA of the three genes, GAPDH, TGF beta and IL-I0 showed no statistical difference between standard curve and LineRegPCR methods. Therefore, it is recommended to use the LinRegPCR software method instead of the expensive standard curve method
ABSTRACT
Lipoic acid [LA] is an effective anti-oxidant agent that can scavenge free radicals in biological systems. The aim of this research was to study the probable protective effect of LA in spinal ischemic/reperfusion [I/R] injury. Thirty male Wistar rats, weighing 230-285 g, were assigned randomly into 3 groups [10 animals in each group]: sham spinal I/R, and spinal I/R + LA. The spinal I/R + LA rats received LA 100 mg/kg subcutaneously 3 days prior to ischemia induction and 3 days after. The induction of ischemia lasted for 30 min. At 72 h postoperatively, the neurological status was worse in the I/R group than the sham group [p < 0.05]. The neurological status of animals in the LA-treated group appeared better than the I/R group [p < 0.05]. In the I/R group, tissue glutathione peroxidase [GPx] and super oxide dismutase [SOD] activity were significantly less compared to the control group [p < 0.05]. In the LA-treated group, tissue GPx and SOD levels were higher compared to the I/R group [p < 0.05]. LA pretreatment reduced neurologic injury in the rats, most probably by maintaining the oxidant/anti-oxidant ion balance during spinal cord ischemia. Reperfusion may have contributed to the protective effects seen in the LA pretreatment
Subject(s)
Male , Animals, Laboratory , Antioxidants , Spinal Cord Ischemia , Oxidative Stress , Free Radicals , Free Radical Scavengers , Rats, Wistar , Random AllocationABSTRACT
Current treatments for joint diseases are moderately successful, but unfortunately are associated with significant side effects. This study was undertaken to investigate the combination effects of IL-4 and prednisolone on tissue characteristics and production of matrix metalloproteinase-1[MMP-1] in IL-lalpha-treated bovine nasal cartilage [BNC] explants. BNC explants were cultured in DMEM with IL-lalpha [10 ng/ml], IL-4 [50 ng/ml] and prednisolone [1 or 1,000 nM] at the same time for 28 days. At days 3, 7, 14, 21and 28, the media were collected and replaced with fresh media, and the removed media were stored at -20[degree sign] C. The alterations of tissue characteristics were assessed by using histology techniques. Western-blot method was used to determine the effects of IL-4 and prednisolone combination on MMP-1 production. The cell viability was evaluated by using lactate dehydrogenase assay test. In the presence of IL-lalpha alone, most chondrocytes were transformed into fibroblast-like morphology with pyknotic nuclei at day 28. In addition, a clear band of MMP-1 and extracellular matrix [ECM] degradation were observed. In combination of IL-4 and prednisolone, chondrocytes preserved their ordinary normal features. MMP-1 band formation was completely inhibited and ECM absolutely showed normal characteristics. IL-4 and prednisolone did not show cytotoxicity effects on BNC explant culture. This combination can strongly preserve cartilage from degradation features and the data possibly suggest that the combination of IL-4 and prednisolone could be a candidate for alternative therapy in joint diseases
ABSTRACT
The aim of this study was to test the therapeutic efficacy of sodium alginate in a rat model of trinitrobenzene sulfonic acid [TNBS]-induced inflammatory bowel disease. This experiment was carried out using 77 Sprague-Dawley rats which were divided into six groups; normal, control, prophylactic, therapeutic and two experimental groups. Rats were sacrificed 1, 2, 3 and 6 weeks after colitis induction. Severity of colitis was graded macroscopically and assessed using serum and colonic mucosal cytokines and eicosanoids. Intrarectal TNBS [30 mg] produced a significant chronic ulcerative colitis. The lesions were most severe on day seven after TNBS instillation, and then declined, but lesions were still observed after six weeks. TNBS administration also significantly enhanced the serum and colonic mucosal cytokines [TNF-alpha and IL-6] and eicosanoids [LTB4 and PGE2] levels, which paralleled with the severity of colitis. Low viscosity sodium alginate [LVA] solution as therapeutic agent was administered orally as drinking water at concentration of 0.5% [W/V] for six weeks. Results showed that pre-treatment [in prophylactic group] and treatment with LVA were significantly able to reduce colonic damage score, serum level and colonic mucosal production of TNF-alpha, IL-6, LTB4 and PGE2 in pre-treated and treated animals compared with non-treated controls. LVA therapy is able to suppress chronic ulcerative colitis in experimental model