Iranian Journal of Basic Medical Sciences
Volume:17 Issue: 2, Feb 2014

Estrogen receptor-alpha (ERα) mediates estrogen action in regulation of different levels of the hypothalamic-pituitary-testis axis. It has a key role in spermatogenesis. Estrogen receptor alpha knock-out (ER koα) male mice were infertile and severe impairment in spermatogenesis and seminiferous tubules was observed. Recently, it has been reported that microRNA (miRNA) mir-100 and let-7b were predicted to target ERα gene. MiRNA are small, endogenous, single stranded RNA molecules that regulate gene expression and have been implicated in various disease states. It has been proved that some miRNAs expression is tissue- and disease-specific, giving potential for identifying miRNAs as a diagnostic tool. In this study, the change in the expression levels of mir-100, let-7b and ERα expression levels were evaluated in oligospermic infertile patients (n=43) compared to control fertile subjects (n=43). After washing and separating sperms, total RNA was isolated and then cDNA was synthesized. The expression levels of mir-100 and let-7b and ERα were evaluated by real time PCR. Mir-100, let-7b levels were significantly higher than those in control group (P=0.008 and P=0.009, respectively). We have found that, ERα level was significantly decreased in comparison with normal group (P< 0.0001). Changes in mir-100, let-7b and ERα expression levels in oligospermic patients may be associated with the susceptibility and progression of infertility. The results of this study indicate that miRNA can have a key role in spermatogenesis and might have a diagnostic and prognostic value in men infertility.

Gingivitis is an inflammatory disease that affects tooth-supporting tissues and is caused by a microbe-immune response. The aim of this study was to evaluate the effects of omega-3 fatty acids on immune system regulation and the prevention and treatment of gingivitis using an animal model. Gingival inflammation was induced by lipopolysaccharide (LPS) injection. Forty adult male rats were divided into four equal groups: 1. Negative control group (sterile saline was injected into gingival tissue followed by oral gavage with saline); 2. Positive control group (LPS injection was followed by oral gavage with saline); 3. Treatment group (LPS injection was followed by oral gavage with omega-3); 4. Prophylactic group (oral gavage with omega-3 was followed by LPS injection). After 24 days, the rats were sacrificed and histological tissue samples were randomly evaluated for the inflammatory tissue changes. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels were measured by ELISA. The levels of IL-1β and TNF-α in the prophylactic group and the level of TNF-α in the treatment group were significantly lower than those in the positive control group (P<0.05). The severity of inflammation was normal, slight, moderate, and severe in the negative control group, prophylactic group, treatment group, and positive control group, respectively. ANOVA was used for the statistical analyses, with P<0.05 regarded as significant. Prior consumption of omega-3 fatty acids is effective in reducing inflammation in induced rat gingivitis, resulting in a decreased level of biomarkers and fewer destructive effects.

Transplantation quality improvement and reduction of cellular damage are important goals that are now considered by researchers. Melatonin is secreted from the pineal gland and some organs such as testes. According to beneficial effects of melatonin (such as its antioxidant and antiapoptotic properties), researchers have proposed that the use of melatonin may improve transplantation quality. The aim of this study was to investigate the effects of melatonin on the spermatogonial stem cells transplantation in the azoospermic mice. The testes of the BALB/c mice pups (6-day-old) after vitrified-thawed, were digested with enzymes (collagenase, DNaseΙ, trypsin-EDTA) to disperse the cells. The SSCs, type A, were isolated from the rest of testicular cells by MACS. Spermatogonial stem cells were labeled with PKH26 fluorescent kit. Labeled spermatogonial stem cells were transplanted into the testes of infertile mice (busulfan 40 mg/kg). The mice died two months after transplantation and the efficiency of spermatogenesis was investigated. TNP2 and hematoxyline-eosin staining were used to detect the efficiency of cell transplantation. TNP2 were detected in the samples that received melatonin and spermatogonial stem cells transplantation, simultaneously. TNP2 was not detectable in the transplant recipient mice that received placebo for 10 weeks (control group). According to hematoxyline-eosin staining, melatonin improved structure of testes. Administration of melatonin (20 mg/kg) simultaneously with transplantation of spermatogonial stem cells in azoospermia mouse testis increases the efficiency of transplantation and improves structural properties of the testes tissue.

A growing interest has recently been attracted towards the identification of plant-based medications including those with protective effects against cognitive impairment. Sesamol has shown promising antioxidant and neuroprotective effects, therefore, we aimed to evaluate its therapeutic potential in epilepsy which is commonly associated with oxidative stress and cognitive impairment. Male Wistar rats received pentylenetetrazole (PTZ) (30 mg/kg, IP) once every other day until the development of kindling, i.e., the occurrence of stage 5 of seizures for three consecutive trials. After the completion of kindling procedure, behavioural tests including elevated plus maze and passive avoidance were performed in order to assess learning and memory. Oxidative stress was assessed by estimation of lipid peroxidation and reduced glutathione. The effects of pretreatment with sesamol (10, 20, and 30 mg/kg, IP) against PTZ-induced seizures, cognitive impairment and oxidative stress were investigated. 32.45 ± 1.86 days after treatment with PTZ, kindling was developed that was associated with myoclonic jerks and generalized tonic-clonic seizures. Moreover, PTZ kindling induced a remarkable cognitive impairment and oxidative stress. Sesamol (30 mg/kg) significantly delayed the development of kindling and prevented seizure-induced cognitive impairment and oxidative stress. Sesamol exerts ameliorative effects in the experimental model of epilepsy. This phytochemical may be considered as a beneficial adjuvant for antiepileptic drugs.

Breast cancer is the most common cancer in women. Every year, one million new cases are reported worldwide, representing 18% of the total number of cancer in women. Hereditary BRCA1 and BRCA2 mutations account for about 60% of inherited breast cancer and are the only known causes of hereditary breast cancer syndrome. The aim of this study was to determine the frequency of BRCA2 (exon2 and exon11) gene mutation in patients with early-onset breast cancer among Iranian Azeri-Turkish women. We obtained clinical information, family history and peripheral blood from 110 women under the age of 45 with early-onset breast cancer for scanning germline mutations in the exon2 and 11 of BRCA2 genes which comprises over 50% of the gene. Single-strand conformation polymorphism assay was used in order for screening potential mutations on amplified regions followed by direct sequencing analysis to determine the genotypes. Overall, 11 sequence variants were identified in this study group, including four homozygotes and seven heterozygotes silent substitution of c.3807T to C, p.Val1269Val (rs543304). Mutations in BRCA2 were surprisingly infrequent in the early onset breast cancer patients among Iranian Azeri-Turkish women.

Drug delivery through the skin can transfer therapeutic levels of drugs for pharmacological effects. Analgesics such as NSAIDs have gastrointestinal side effects and topical dosage forms of these drugs are mainly preferred, especially for local pains. Meloxicam is one of NSAIDs with no topical form in the market. In this research, we attempted to quantify the skin permeation of a meloxicam topical preparation and to show how permeation would be increased by using thymol as an enhancer. The effect of eutectic point of drug and thymol mixture on rate and extent of skin permeation was also studied. Different mixtures of thymol and meloxicam (2:8, 4:6, 5:5, 6:4, 8:2) were prepared and their melting point were obtained by differential scanning calorimetry. Then drug permeation was measured using diffusion cells and the Guinea pig skin. Mixtures in ratios 5:5 and 4:6 of meloxicam / thymol showed a new endotherm at 149 and 140°C in DSC thermograms. The permeability of meloxicam from the creams containing 6:4, 5:5 and 4:6 ratios of meloxicam to thymol were 4.71, 15.2, 22.06 μg/cm2 respectively. This was significantly different from the cream of pure meloxicam (3.76 μg/cm2). This study set out to determine that thymol plays as a skin permeation enhancer and increases the meloxicam skin absorption and this enhancement is significant at the eutectic point of drug-enhancer mixture.

Hyperglycemia worsens the neuronal death induced by cerebral ischemia. Previous studies demonstrated that diabetic hyperglycemia suppressed the expression of heat shock protein 70 and 60 (HSP70 and 60) in the liver. IgG antibody titres against heat shock protein 27 (anti HSP27) were measured to determine whether hyperglycemia exacerbates ischemic brain damage by suppressing the expression of heat shock protein 27 (HSP27) in the brain. A randomized controlled study of traumatic brain injury ICU patients treated either by intensive insulin treatment (IIT) or by conventional glucose control (CGC) was conducted. Patients received at least 50% of their estimated daily energy requirements parenterally. Serum anti HSP27 antibody concentration was assessed at baseline, day 7 and day 14. Twenty-six out of 29 patients (n=13 in each group) completed the study. At baseline, there were no differences between the two groups. On day 14, there was a significant reduction in anti HSP27 titre concentration in the IIT compared to the GCG group (0.47±0.27 mg/dl vs 0.60±0.15 mg/dl, P =0.03). In this study, intensive control of traumatic brain injury patients on parenteral nutrition reduced anti HSP27 titre, possibly suggesting a reduction in stress.

PCOS (polycystic ovary syndrome) produces symptoms in approximately 5% to 10% of women of reproductive age (12–45 years old). It is thought to be one of the leading causes of female subfertility. This study aimed to confirm the role of nutrition containing omega-3 (polyunsaturated fatty acid) on control of experimental PCO induced by estradiol-valerat in rats. Wistar female rats (n=40) were allocated into control (n=10) and test groups (n= 30), test group was subdivided into 3 groups: G1, received omega-3 (240 mg/kg/orally/daily); G2 and G3 groups were induced PCO by single injection of estradiol-valerate (16 mg/kg/IM). Group 3 received omega-3 (240 mg/kg/orally/daily) and low carbohydrate feeding for 60 subsequent days; on sixtieth day 5 ml blood samples and ovarian tissues of all rats in the group were removed and prepared for biochemical and hormonal analysis. Catalase, GPX (Glutathione peroxidase), SOD (Superoxide dismutase) in groups that received omega-3 showed higher levels, but MDA (malondialdehyde) level was significantly decreased (P<0.05) in comparison with other experimental groups. Ovarian weights in both experimental and control groups were similar (P<0.05). Level of serum FSH (follicle stimulating hormone) was decreased, but level of testosterone was significantly increased (P<0.05) in PCO group in comparison with control and omega-3 groups. Results revealed that administration of omega-3 plus lower carbohydrate food significantly controlled PCO syndrome and balanced FSH and testosterone.

Accumulating evidence has demonstrated that miRNAs contribute to various genetic and epigenetic modifications in the pathogenesis of gastric cancer (GC). Recent studies focused on the four single nucleotide polymorphisms (SNPs) of pre-miRNAs including rs11614913, rs3746444, rs2910164, and rs2292832. It was suggested that these four SNPs were significantly associated with the risk of GC and were described as candidate susceptibility factors. However, the previous results show conflicting findings. The aim of this study was to investigate whether these four SNPs are associated with GC in Chinese Han population. Genotype frequencies of these four SNPs of pre-miRNAs in 220 GC patients and 530 control subjects were performed using a PCR-RFLP assay. No significant differences in genotype and allelic distribution were found in these four SNPs between GC and control subjects in the Chinese Han population. However, we found that the allelic frequency distributions of control subjects in these four SNPs were significantly different from those of the Japanese and the Koreans (All the P <0.05). The four SNPs did not show any significant correlation with the development of GC in the Chinese Han population, based on the current study.

To determine the fetal discernment in suspected cases of sex linked recessive disease in the first trimester of pregnancy. After collection of 100 Chorionic Villi samples, the DNAs were extracted and baby gender was determined. Meanwhile, after increasing the sensitivity, the system was able to detect the sex of each cell which was obtained by biopsy. Early fetal gender of 100 Chorionic Villi samples were assessed by PCR. After increasing sensitivity of the assay, the sexes in 13 fetuses that were in different cellular stages were detected. Morover, sexes were detected in two unfertilized and one fertilized ovum but without any division. Sex detection of fetus before delivery in the first trimester of pregnancy, will prevent babies with abnormalities being born. It can also be used in detection of recessive sex related diseases in In Vitro Fertilization cases for sex detection and to transfer female fetus to the mother. Our optimized molecular detection system was designed on the basis of amelogenin gene, which can determine the sex in blood, chorionic villi, and single cell in vitro fertilization with high sensitivity and specificity.

We aimed to characterize the phenotype and genotype of Bacillus spp isolated from diabetic patients’ eyes, by studying the drug sensitivity patterns with a disc-diffusion method. Fifty eyes of 25 patients with type II diabetes mellitus, with at least 10 years of diabetes history, were included in the study. We analyzed the eyes for the presence of Bacillus spp.; presumptive isolates were identified by morphological, and biochemical tests, and confirmed by the VITEK system. Automated EcoRI ribotyping was performed with a RiboPrinter® Microbial Characterization System. We determined the antibiotic resistance of the isolates by the Kirby–Bauer disc diffusion test. Seven out of 25 patients were on insulin treatment; 7 on oral anti-diabetic medication; and 11 on combination therapy of insulin and oral medications. Among the 28 Bacillus spp isolates, 14 were B. cereus, 11 were B. pumilus, 2 were B. mojavensis and 1 was B. subtilis. Almost all the strains were either resistant or multiresistant, particularly towards cefuroxime, methicillin, and ceftazidime. Diabetic patients seem to be more prone to B. cereus infections than healthy individuals. It would be greatly beneficial to understand and recognize the prevalence of microorganisms and their resistance patterns for better outcome in ocular surgeries.

effects on health have been demonstrated. Recent studies suggest that GT may contribute to reduction of cancer risk and progression. The aim of this study was to evaluate the effects of whole GT on urinary bladder chemical carcinogenesis in male and female ICR mice. The GT characterization was performed using spectrophotometric methods. Urinary bladder lesions were induced using N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) by gavage during 10 weeks and whole GT (0.5%) was provided ad libitum during 20 weeks. Animals from groups BBN+GT and BBN only developed preneoplastic lesions. We did not observe any effects by GT infusion administration on urinary bladder cancer development.