Evaluation of the relationship between endometriosis and omega-3 and omega-6 polyunsaturated fatty acids

Endometriosis is a common chronic inflammation causing major problems including infertility. The role of omega-3 and omega-6 fatty acids as their potential anti-inflammatory effects in endometriosis needs to be further explored. The objective of this study was to compare serum phospholipid fatty acid profile in endometriosis patients with controls, and to explore the correlation of this profile with the severity of the disease. Sixty-four endometriosis patients and 74 control women, in reproductive age, participated in this study. Among the endometriosis patients, 19 cases were in stage I, 27 cases in stage II, 8 cases in stage III, and 10 cases in stage IV. Each patient underwent laparoscopy. Before surgery, 5 ml of blood was obtained. After extraction of the total lipids, serum total phospholipid fraction was isolated by thin layer chromatography. Fatty acid composition of the phospholipid fraction was determined by gas chromatography and the resulted profile was compared in endometriosis patients and controls. The profile was also compared in the endometriosis group based on the severity of disease. Stearic acid was significantly lower in the endometriosis group as compared to controls [P= 0.030]. No other fatty acid compositions were significantly different between patients and controls. Serum ratio of eicosapentaenoic acid [EPA] to arachidonic acid [AA] was in reasonable correlation with the severity of endometriosis [r = 0.34, P = 0.006]. According to these findings, levels of fatty acids in serum total phospholipids seem not to be a marker for endometriosis, but the EPA to AA ratio was a relevant factor indicating severity of illness

Non-invasive detection of esophageal cancer using genetic changes in circulating cell-free DNA

Cell free DNA [cfDNA] is a genetic biomarker that is present in serum or plasma in high concentration in many types of cancer. Identification of circu-lating cancer related DNA molecules in serum or plasma is a non-invasive tool for early diagnosis and prognosis in many cancer patients. For this review, study selection and data extraction were performed by the authors. Detection of point mutations, microsatellite alterations, DNA hypermethylations and losses of heterozygosity in circulating cell free DNA have been characterized in esophagus cancer. Application of circulating cell free DNA as a biomarker, provide the best opportunity for constructing non-invasive tests for early detection, prognosis and management of cancer patients, after therapy in many types of cancer

Genetically modified foods and social concerns

Biotechnology is providing us with a wide range of options for how we can use agricultural and commercial forestry lands. The cultivation of genetically modified [GM] crops on millions of hectares of lands and their injection into our food chain is a huge global genetic experiment involving all living beings. Considering the fast pace of new advances in production of genetically modified crops, consumers, farmers and policymakers worldwide are challenged to reach a consensus on a clear vision for the future of world food supply. The current food biotechnology debate illustrates the serious conflict between two groups: 1] Agri-biotech investors and their affiliated scientists who consider agricultural biotechnology as a solution to food shortage, the scarcity of environmental resources and weeds and pests infestations; and 2] independent scientists, environmentalists, farmers and consumers who warn that genetically modified food introduces new risks to food security, the environment and human health such as loss of biodiversity; the emergence of superweeds and superpests; the increase of antibiotic resistance, food allergies and other unintended effects. This article reviews major viewpoints which are currently debated in the food biotechnology sector in the world. It also lays the groundwork for deep debate on benefits and risks of Biotech-crops for human health, ecosystems and biodiversity. In this context, although some regulations exist, there is a need for continuous vigilance for all countries involved in producing genetically engineered food to follow the international scientific biosafety testing guidelines containing reliable pre-release experiments and postrelease track of transgenic plants to protect public health and avoid future environmental harm

Stress-induced proteomic changes in the hippocampus of pregnant wistar rats

Stress is a threatening factor that all living organisms encounter throughout life. Depending on the type of stress, there are several mechanisms for keeping body homeostasis to minimize stress effects. Brain is an organ which shows high sensitivity to stress conditions. Although many studies have shown induced-stress effects on rat embryos, little is known about the mechanisms involved in coping with stress by female rats during pregnancy. In the present study, restraint stress method was applied because this technique has been widely used in animal models to induce both psychological and physical stress. Restraint stress was applied in regular sessions [1 and 3 hrs] in two groups of 6 pregnant Wistar rats and similar number of animals was used as control group receiving no stress. ACTH and corticosterone levels in plasma samples were shown to increase in response to stress treatments. On the last day of pregnancy, rat hippocampus from the brain of each animal in all three groups was removed and analyzed using 2 Dimensional Gel Electrophoresis [2DE] technique. Using Image Master Software, approximately 2000 proteins were detected in the 2D gels analyzed, among which 34 proteins exhibited differential expression. These results indicate that the proteome patterns from the hippocampus of pregnant rats subjected to 1 and 3 hr of stress differs significantly from the control [unstressed] group. Future mass spectrometry identification of the 34 protein spots discovered in this study should allow a more precise understanding of molecules and cellular pathways involved in stress-induced responses during pregnancy

Bryostatin-1, fenretinide and 1alpha,25 [OH]2D3 induce growth inhibition, apoptosis and differentiation in T and B cell-derived acute lymphoblastic leukemia cell lines [CCRF-CEM and Nalm-6]

In many acute leukemias, normal differentiation does not occur. However, in many cell lines derived from hematologic malignancies, differentiation or apoptosis can be induced by variety of agents. Despite advances in the treatment of Acute Lymphoblastic Leukemia [ALL], in most patients long-term survival rates remain unsatisfactory, especially in T-cell derived ALL. Thus we studied the anti-cancer effects of fenretinide, 1alpha,25[OH]2D3, and bryostatin-1 in CCRF-CEM [T-cell derived] and Nalm-6 [B-cell derived] ALL cell lines. Using MTT assays, both cell lines were shown to exhibit increased inhibition of proliferation at micro [fenretinide] and nanomolar [1alpha,25[OH]2D3, bryostatin-1] concentrations. These anti-cancer agents were shown to induce apoptosis and activate caspase-3 pathway in both ALL cell lines. Furthermore, for the first time we are reporting consistent anti-proliferative and apoptotic effects of Bryostatin-1 in ALL T-cell derived cell line with the lowest ED50 [ranging 4.6 nM - 7.4 nM]. To evaluate the differentiation induction by fenretinide, 1alpha,25[OH]2D3, and bryostatin-1 in ALL cell lines, we assayed for the expressions of CD19, CD38 markers on Nalm-6 and CD7 marker on CCRF-CEM cell line. The flow cytometric analysis showed a significant increase in expression of CD markers in response to anticancer drug treatments. To assay the effects of anti-cancer drugs on cell cycle distribution, cell cycle analysis using flow cytometry was employed. These anti-cancer drugs appear to affect the CCRF-CEM and Nalm-6 cell cycles differently [G0/G1 and G2/M, respectively]. Overall results demonstrate that the anticancer agents used in this study are strong inhibitors of ALL cell proliferation and inducers of apoptosis and differentiation in vitro. These findings may be quite helpful if these drugs are to be used for differentiation therapy of ALL patients in clinics in the future. Further studies are warranted to establish the in vivo effect of these drugs particularly in patients with T-cell derived ALL

Flow cytometric analysis of 4-HPR-induced apoptosis and cell cycle arrest in acute myelocytic leukemia cell line [NB-4]

In many acute leukemias, normal differentiation does not occur. However, in many cell lines derived from hematologic malignancies, differentiation or programmed cell death [apoptosis] can be induced by variety of agents including: Vitamin analogs, demethylating agents, cyclic AMP analogs and anti-proliferative agents. To the best of our knowledge there had been not any study specifically to analyze apoptotic and anti-proliferative effects of 4-HPR [a vitamin analog] in NB-4 cell line. To test whether this drug has activity in acute myeloid leukemia [AML], we first analyzed the anti-proliferative effect of 4-HPR in on EAML cell line [NB-4] using MTT Assay. Next we tested whether this drug induced apoptotic cell death. The abilty of this compound to induce apoptosis of cancer cells was examined by Annexin V-FITC Assay using Flow cytometry. We also analyzed the cell cycle progression by PI staining using flow cytometry. Using MTT assay, NB-4 cells exhibited increased inhibition of proliferation at micromolar concentrations of 4-HPR at 24, 48 and 71 hrs post treatment. Flow cytometry analysis indicates that 4-HPR is a potent inducer of in vitro apoptotic cell death, and cell cycle analysis revealed an increase in 5 phase population. In total, the results indicate that 4-HPR is a strong inhibitor of AML cell proliferation and a potent inducer of in vitro apoptotic cell death. Further studies are required to evaluate the in vitro effects of 4-HDR in AML blasts derived from AML patients

Comparing seminal plasma biomarkers between normospermic and azoospermic men

Azoospermia affects more than 10% - 15% of infertile male subjects attending infertilty clinics. At present, testicular biopsy is the golden standard procedure for evaluating spermatogenesis status in men with azoospermia. Semen collection and analysis is a non-invasive method and has proven to be valuable in the evaluation of spermatogenesis. Identification of seminal plasma markers with testicular or extra-testicular origins have a great value in predicting the prescence of sperm in testicular tissue and presumptive cause of azoospermia. The aim of this study was to find such markers by comparing the content of seminal plasma using different methods in normospermic and azoospermic men. Semen samples were collected from 200 men attending Avicenna Infertility Clinic [AIC] in Tehran, Iran. Semen samples were analysed according to WHO guidlines. The subjects were divided into two groups: normospermic [n = 100; group one] and azoospermic men [n = 100; group two] according to semen analysis results. Seminal plasma was separated by high speed centrifuagation and stored in -20°C. Four markers including fructose, neutral alpha glucosidase [NaG], inhibin B and anti-Mullerian hormone [AMH] were measured in seminal plasma. Fructose and NaG were evaluated by spectrophotometry, while inhibin B and AMH were assessed by ELISA method. The spermatogenesis status in the azoospermic group was evaluated by histopathological method following testicular biopsy. Fructose concentration showed no difference between the two groups. However, it was significantly correlated with sperm count [p < 0.01, r = -0.408]. Seminal plasma inhibin B [OR: 1.01; 95%: CI: 1.005 - 1.016], AMH [OR: 1.63; 95% CI: 1.17 - 2.28] and N alpha G, [OR: 1.07; 95% CI: 1.04 - 1.1] levels were higher in normospermic subjects compared to azoospermic men. There were significant differences in inhibin B and AMH concentrations between the two groups based on the presence or absence of mature sperm in testicular biopsies [p < 0.01]. Inhibin B concentration was positively correlated with sperm count in the normospermic group, however, N alpha G concentration correlated with sperm count of normospermic men [p < 0.01, r = 0.345] and the subjects'age in both groups. Inhibin B and AMH were correlated with the presence of sperm in testicular tissue samples. According to non-specific changes in inhibin B and AMH concentrations, identification of more specific molecular markers in seminal plasma to definitely evaluate the status of spermatogenesis is recommended

role of microRNAs in human diseases

MicroRNAs [miRNAs] are short RNA molecules which bind to target mRNAs, resulting in translational repression and gene silencing and are found in all eukaryotic cells. Approximately 2200 miRNA genes have been reported to exist in the mammalian genome, from which over 1000 belong to the human genome. Many major cellular functions such as development, differentiation, growth, and metabolism are known to be regulated by miRNAs. Proximity to other genes in the genome and their locations in introns of coding genes, noncoding genes and exons have been reported to have a major influence on the level of gene expressions in eukaryotic cells. miRNAs are well conserved in eukaryotic system and are believed to be an essential and evolutionary ancient component of gene regulatory networks. Therefore, in recent years miRNAs have been studied as a likely candidate for involvement in most biologic processes and have been implicated in many human diseases

Tyrosine phosphorylation pattern in sperm proteins isolated from normospermic and teratospermic men

In mammalian system, spermatozoa are not able to fertilize the oocyte immediately upon ejaculation, thus they undergo a series of biochemical and molecular changes which is termed capacitation. During sperm capacitation, signal transduction pathways are activated which lead to protein tyrosine phosphorylation. Tyrosine phosphorylated proteins have an important role in sperm capacitation such as hyperactive motility, interaction with zona pellucida and acrosome reaction. Evaluation of tyrosine phosphorylation pattern is important for further understanding of molecular mechanisms of fertilization and the etiology of sperm dysfunctions and abnormalities such as teratospermia. The goal of this study is to characterize tyrosine phosphorylation pattern in sperm proteins isolated from normospermic and teratospermic infertile men attending Avicenna Infertility Clinic in Tehran. Semen samples were collected and the spermatozoa were isolated using Percoll gradient centrifugation. Then the C with 5% CO[2] in 3% Bovine Serum spermatozoa were incubated up to 6h at 37 Albumin-supplemented Ham's F-10 for capacitation to take place. The total proteins from spermatozoa were extracted and were subjected to SDS-PAGE before and after capacitation. To evaluate protein tyrosine phosphorylation pattern, western blotting with specific antibody against phosphorylated tyrosines was performed. The results upon western blotting showed: 1] at least six protein bands were detected before capacitation in the spermatozoa from normospermic samples. However, comparable levels of tyrosine phosphorylation was not observed in the spermatozoa from teratospermic samples. 2] The intensity of protein tyrosine phosphorylation appears to have been increased during capacitation in the normospermic relative to the teratospermic group. For the first time, these findings demonstrate and suggest that the differences in the types of proteins and diminished tyrosine phosphorylation efficiency in sperm from teratospermic men may be responsible for their compromised capacitation and low fertilization success rates