Association between seminal plasma neopterin and oxidative stress in male infertility: a case-control study

Background: Neopterin is a significant and sensitive marker in estimating the activity of cellular immune system. Oxidative stress plays a role in the etiology of male infertility. Increased reactive oxygen species is accompanied with increase in neopterin level. Hence neopterin may be involved in male infertility

Objective: The objective of this case-control study was to determine neopterin level in idiopathic infertile and normospermic men; furthermore, to identify its relationship with oxidative stress markers including total oxidant, malondialdehyde, sperm DNA fragmentation, and total antioxidant capacity of seminal plasma

Materials and Methods: Forty seven infertile and forty three normospermic males were selected according to WHO criteria. Their semen and blood samples were taken; subsequently, the levels of neopterin, total oxidant, total antioxidant, malondialdehyde, and sperm DNA fragmentation were measured

Results: The levels of neopterin, total oxidant, and malondialdehyde in seminal plasma of infertile males were significantly higher than those of normospermic group [p=0.038, 0.018, and 0.028, respectively]. Furthermore, sperm DNA fragmentation in infertile men was higher than that of control group [p<0.001]. Moreover, total antioxidant capacity of seminal plasma in infertile males was significantly lower than that of normospermic subjects [p=0.002]. No significant difference was observed in serum neopterin, total oxidant, and malondialdehyde between the infertile and normospermic groups

Conclusion: The significant inverse correlation between seminal plasma neopterin and total antioxidant in the infertile males supports a possible role of neopterin in male infertility. Neopterin can be suggested as a marker in monitoring and diagnosis of idiopathic male infertility

Moderate Exercise Enhances the Production of Interferon-γ and Interleukin-12 in Peripheral Blood Mononuclear Cells

The purpose of this study was to explore the effect of two months moderate exercise on levels of IFN-γ, IL-12, IL-6 and IL-4 in serum and supernatants of in vitro mitogen-activated (PHA for 48 h) whole blood (WB) and peripheral blood mononuclear cells (PBMCs). Sixteen healthy males participated in running program (30 min/day, 5 days/week). Blood samples were collected in three stages; 24 h before to start exercise, 48 h and two months after the last session of the exercise. The samples were analyzed for the cytokines by ELISA. The levels of IFN-γ and IL-12 were increased significantly in activated PBMCs culture after exercise and were back to normal level after two months rest. A significant elevation of IFN-γ/IL-4 ratio was observed in activated PBMCs culture by acting possibly on IFN-γ. The results suggest that short moderate intensity exercise enhances Th1 immune inflammatory and anti-allergic conditions in response to mitogen.

relationship between some endometrial secretion cytokines and in vitro fertilization

Endometrial secretion analysis is a non-invasive and promising method in evaluation of endometrial receptivity. The aim of the present study was to assess the relationship between the success rate of IVF procedures and some endometrial secretion cytokines, including interleukin-1beta [IL-1beta], tumor necrosis factor [TNF-alpha], interferon gamma-induced protein 10 [IP-10], and monocyte chemoattractant protein [MCP]. In a prospective cohort study, 50 women selected for IVF met the study inclusion criteria. All the patients underwent endometrial secretion aspiration prior to embryo transfer. The level of IL-1beta, TNF-alpha, IP-10 and MCP were analyzed by enzyme-linked immunosorbent assay method using special standard kits. To detect successful implantation and pregnancy patients underwent serum human chorionic gonadotropin measurements and ultrasound evaluation. Five samples were excluded. Nine women [20%] had successful clinical pregnancies, which resulted in live birth. Other 36 women [80%] were classified as failed pregnancy. Comparison of cytokine levels showed lower concentrations of TNF-alpha, IP-10, and MCP in the group with successful clinical pregnancy compared to the group with failed pregnancy [p=0.007, 0.005 and 0.001, respectively]. However, no significant difference was revealed in IL-1beta levels between two groups [p=0.614]. The current study suggested that lower concentrations of TNF-alpha, IP- 10, and MCP in endometrial secretions might be associated with improved endometrial receptivity and IVF outcome. Regarding IL-1beta, no statistically significant differences were seen between the groups with and without successful pregnancy

Effect of ibuprofen on IL-1 beta, TNF- alpha and PGE2 levels in periapical exudates: a double blinded clinical trial

Bone resorption is one of the main features of inflammatory periapical lesions and is mainly mediated by interleukin-1 beta [IL-1 beta], tumor necrosis factoralpha [TNF- alpha] and prostaglandin-E2 [PGE2]. Recent investigations of these lesions revealed that pharmacological modulation may be possible. The aim of this study was to evaluate the effect of Ibuprofen on IL-1 beta, TNF- alpha and PGE2 levels in periapical exudates and compare the results with a group of placebo control. Thirty patients with non vital teeth and radiographic lesions were divided into two groups of case and control according to their entrance to the study. Periapical exudates were taken from root canals using absorbent paper points and followed by 400 mg Ibuprofen and placebo prescribed one tablet every 6 hour for three days and in the fourth day second samples were taken, then final cleaning, shaping and obturation of the canals were completed. IL-1 beta, TNF- alpha and PGE2 levels were determined by enzymelinked immunosorbent assays [ELISA]. Data were analyzed using paired t-test and student's t-test. The results showed that PGE2 levels were decreased significantly in the case group to 86.92 +/- 72.42 Pg/ml following Ibuprofen treatment comparing with the pre-treatment [164.96 +/- 12.255 Pg/ml] [p=0.02] and placebo group [154.2 +/- 97.13 Pg/ml] [p=0.001]. But there were no significant differences in IL-1 beta and TNF- alpha level between the two groups and in each group before and after treatment. The data indicate that Ibuprofen, as a non-steroidal anti-inflammatory drug [NSAID], can be used to block PGE2 release, enhance healing of inflammatory periapical lesions and possibly to inhibit bone resorption

Diagnostic multiplex polymerase chain reaction assay for the identification of Pseudomonas aeruginosa from the skin biopsy specimens in burn wound infections and detection of antibiotic susceptibility

To identify Pseudomonas aeruginosa [P. aeruginosa] from the skin biopsy specimens in burn wound infections by multiplex polymerase chain reaction [M-PCR] and detection of antimicrobial susceptibility of isolates from culture. We conducted this cross-sectional study in 140 patients with wound infections who admitted to the referral burn center of Motahari, Tehran, Iran, during a 12-month period from 2005-2006. Skin biopsy specimens were aseptically taken from each patient, one for PCR and one for bacterial culture. A M-PCR test based on the simultaneous amplification of 2 lipoprotein genes: oprI and oprL, was used to directly detect fluorescent pseudomonades and P. aeruginosa in skin biopsy specimens. The susceptibility of P. aeruginosa isolates to 16 antibiotics was determined using the disc diffusion method. Out of 140 biopsy specimens, M-PCR detected 66 [47.2%] isolates, while culture detected 57 [40.7%] isolates as P. aeruginosa. Positive results for both genes which observed only for P. aeruginosa, while only one gene, oprI, was amplified from other fluorescent pseudomonades n=12 and all other bacterial tested n=62 were negative by the amplification test. The most effective antibiotics against isolate of P. aeruginosa were cefepime [79%], azetreonam [76%], ticarcillin-clavulanic acid [68%], tobramycin [62%], and amikacin [61%]. Multiplex PCR assay appears promising for the rapid and sensitive detection of P. aeruginosa from the burned skin biopsy specimens. Simultaneous amplification of 2 lipoprotein genes: oprI and oprL, could detect P. aeruginosa, and oprI gene only for other fluorescent pseudomonades

Designing E1 deleted adenoviral vector by homologous recombination

Adenoviruses are used extensively to deliver genes into mammalian cells, particularly where there is a requirement for high-level expression of transgene products in cultured cells, or for use as recombinant viral vaccines or in gene therapy. In spite of their usefulness, the construction of adenoviral vectors [AdV] is a cumbersome and lengthy process that is not readily amenable to the generation of large collection of clones. In this project, to delete E1 gene in adenovirus, an adenoviral plasmid containing lateral sites of E1 region of adenovirus was made and recombination in the 293A cells between the homologous region of this linearized plasmid and the adenovirus genome resulted in the formation of the complete adenoviral recombinant. This recombination resulted in loss of E1 region and we constructed a recombinant adenovirus type 5 vector that E1 gene was deleted by homologous recombination. Homologous recombination is more easy and fast technique in the production of AdV

Inducible expression of human gamma interferon

The premature termination of high producer clones, which will be killed due to cell proliferation and proteins production antagonism, is one of the basic drawback in recombinant proteins technology. Furtheremore, it is supposed some toxic proteins like interferon which we intended to clone and express, inhibit host cells' proliferation. So, it is necessary to tightly control IFN-gamma production during growing and selecting of highly producing clones. In the present study, we constructed an expression vector, pMPGB43P2[6]K containing the cumate-regulatable expression cassette to high production of human IFN-gamma in Chinese Hamster Ovary- Cumate Transactivator [CHO-CTA]. The clones were selected in the cumate and without cumate treated medium. Our results showed induced IFN-lamda expression level was about 5 of magnitude higher than the constitive transgenic system. Application of cumate-regulatable expression cassette, which can be switch off and on by cumate, is useful to production of high producer clones and express toxic proteins in animal cells

Molecular cloning and expression of human Gamma Interferon[IFN-Lambda] Full cDNA in Chinese Hamster Ovary [CHO] Cells

IFN-gamma is mostly secreted by activated CD4+, CD8+ T cells and NK cells. This cytokine has immunomodulatory, anti-cancer and anti-microbial effects and is important for prophylaxis, diagnosis and treatment of chronic infections and cancers. The purpose of this study was to clone the full cDNA of human IFN-gamma and express it on CHO cell line. Lymphocytes from a healthy individual were isolated and activated by phytohaemagglutinin [PHA] in vitro. After 4 hours, total RNA extracted and first cDNA str and was synthesized. cDNA was amplified with primers containing EcoRI and NotI sites. The amplified fragment and the PcDNA3.1 vector were cut by EcoRI and NotI and ligated. The construct [pcDNA3.1-IFN-gamma] was transferred into E.coli [strain: DH5 alpha] using CaCl2 method and selected by plating on a medium containing ampicillin. The construct sequence was confirmed by PCR and sequence analysis. Construct expression was achieved by performing a calcium phosphate-mediated transfection into CHO cells and followed by selection of stable drug [G418] resistant clones by limiting dilution assay [LDA]. The IFN-gamma production by transected CHO cells was measured using ELISA technique. Out of 33 grown transformed bacterial colonies, only 6 had the entire sequences of the insert and one of them was used for the transfection experiment. Out of 768 wells, 5 clones produced more than 100 ng/ml/10[6] cells of IFN-gamma. Among the 5 clones, one with the maximum production of INF-gamma [143 ng/ml/10[6] cells] was selected and used for propagation