A second look at the embryotoxicity of hydrosalpingeal fluid: an in-vitro assessment in a murine model

The purpose of this study was to evaluate whether hydrosalpingealfluid (HSF) is toxic to the mouse embryo as assessed by theblastocyst development rate (BDR) and by cell counting in vitro.HSF was collected from nine patients undergoing salpingoneostomyto correct hydrosalpinx. Two-cell embryos were obtained fromsuperovulated ICR mice. T6 medium and T6 + 0.4% bovine serumalbumin (BSA) were used as control media. T6 medium containing10% or 50% HSF and 100% HSF from each patient were used as testmedia. Nine to 15 embryos were cultured in microdrops preparedfrom each of these media. The BDR was examined after 72 h ofculture in these media. To assess the total cell number withineach blastocyst, the blastocysts were fixed and stained withHoechst 33342 to facilitate cell counting. The BDR was affectedadversely only by 100% HSF and not in media containing 10% or50% HSF. The mean BDR using T6 medium and T6 + BSA were 88.7%and 85.3%, respectively. The mean BDR using media containing10% HSF or 50% HSF were 90.0% and 89.4%, respectively. MeanBDR using 100% HSF was 75.2% (P < 0.05). The overall meancell counts (± SEM) using T6 medium and T6 + BSA were86.9 ± 3.2 and 91.0 ± 3.8 respectively. Mean cellscounts were decreased significantly only in blastocysts culturedin 100% HSF (63.3 ± 4.6; P < 0.01) but not in blastocystscultured in 10% or 50% HSF (90.8 ± 4.2 and 81.9 ±6.1 respectively). Thus, it is concluded that HSF has no embryotoxiceffect but has a mildly negative effect on embryonic growthand development.     

Andrology: Analysis of sperm movement in relation to the oxidative stress created by leukocytes in washed sperm preparations and seminal plasma

The addition of luminol to unprocessed semen samples resultedin the generation of chemiluminescent signals, the intensityof which was highly correlated with the level of leukocyte contamination.Despite the spontaneous oxidant-generating capacity of seminalleukocytes, no correlations were observed between leukocytecontamination and the fertility status of the subjects or anyaspect of the semen profile, including the motility of the spermatozoaor their performance in a hyaluronate penetration assay. Luminol-dependentchemiluminescence and leukocyte contamination were also correlatedin washed sperm suspensions prepared either by repeated centrifugationor on discontinuous Percoll gradients. However, in such spermsuspensions, the spontaneous generation of oxidants by contaminatingleukocytes (>2x10⁴ leukocytes/ml) was invariably associatedwith a decreased capacity for movement. Moreover, causativeassociations between leukocyte contamination, reactive oxygenspecies generation, lipid peroxidation and impaired sperm motilitywere revealed by experiments involving the selective additionor removal of activated leukocytes. From these observationswe can conclude that low concentrations of leukocytes are acommon feature of the human ejaculate and can impair sperm function,particularly in the absence of seminal plasma. These findingshave implications for our understanding of the importance ofleukocytospermia in defining the fertility of human spermatozoain vivo and in vitro.     

Calcium antagonists modulate oxidative stress and acrosomal reaction in rat spermatozoa

Introduction

Calcium ions are vital in many biological processes and qualify as an almost ubiquitous intracellular second messenger. This indicates the multiplicity of the effects associated with drug actions aimed at interfering with calcium ions. To examine the cellular process involved in the induction of infertility in males by calcium antagonist (CA) even in the presence of normal semen parameters, we studied the effects of different CA namely; nifedipine, verapamil and diltiazem on oxidative balance and acrosome reaction in the sperm.

Material and methods

For this purpose, lipid peroxidation, antioxidants such as superoxide dismutase, catalase and reduced glutathione, and acrosomal reaction were determined in sperm samples of rats.

Results

Calcium antagonist causes significant oxidative stress in the epididymal sperm with increased malondialdehyde level and a concomitant decrease in antioxidant activities of catalase and superoxide dismutase. The percentage value of acrosomal-reacted sperm in the nifedipine, verapamil and diltiazem-treated rats were 41 ±2.45, 39 ±2.92 and 42 ±1.22 respectively, compared with the control group value of 86 ±2.92.

Conclusions

It appears CA oxidatively modify the sperm resulting in functional inhibition of acrosomal reaction. Suppression of the sperm acrosomal reaction is known to have serious adverse implications for fertilization.     

Induction of oxidative stress by bisphenol A and its pleiotropic effects

Bisphenol A (BPA) has become a target of intense public scrutiny since concerns about its association with human diseases such as obesity, diabetes, reproductive disorders, and cancer have emerged. BPA is a highly prevalent chemical in consumer products, and human exposure is thought to be ubiquitous. Numerous studies have demonstrated its endocrine disrupting properties and attributed exposure with cytotoxic, genotoxic, and carcinogenic effects; however, the results of these studies are still highly debated and a consensus about BPA's safety and its role in human disease has not been reached. One of the contributing factors is a lack of molecular mechanisms or modes of action that explain the diverse and pleiotropic effects observed after BPA exposure. The increase in BPA research seen over the last ten years has resulted in more studies that examine molecular mechanisms and revealed links between BPA‐induced oxidative stress and human disease. Here, a review of the current literature examining BPA exposure and the induction of reactive oxygen species (ROS) or oxidative stress will be provided to examine the landscape of the current BPA literature and provide a framework for understanding how induction of oxidative stress by BPA may contribute to the pleiotropic effects observed after exposure. Environ. Mol. Mutagen. 58:60–71, 2017. © 2017 Wiley Periodicals, Inc.     

Divergent behaviour of oxidative stress markers 8-hydroxydeoxyguanosine (8-OHdG) and 4-hydroxy-2-nonenal (HNE) in breast carcinogenesis

Karihtala P, Kauppila S, Puistola U & Jukkola‐Vuorinen A
(2011) Histopathology 58, 854–862
Divergent behaviour of oxidative stress markers 8‐hydroxydeoxyguanosine (8‐OHdG) and 4‐hydroxy‐2‐nonenal (HNE) in breast carcinogenesis Aims: To clarify the role of oxidative stress during breast carcinogenesis by studying the expression of 8‐hydroxydeoxyguanosine (8‐OHdG) (a marker of oxidative DNA damage) and 4‐hydroxy‐2‐nonenal (HNE) (a marker of lipid peroxidation) during the different phases of breast carcinogenesis. Methods and results: The study material consisted of a total of 219 patients: 31 with usual ductal hyperplasia (UDH), 25 with atypical ductal hyperplasia (ADH), 30 with ductal carcinoma in situ (DCIS) and 133 with invasive carcinoma. The expression of 8‐OHdG and HNE were evaluated immunohistochemically. Both 8‐OHdG (77.4%) and HNE (45.8%) expression was already seen in UDH lesions. Interestingly, the trend of these two immunostainings during breast carcinogenesis was diverse. 8‐OHdG expression diminished significantly in invasive breast carcinomas compared to non‐invasive lesions (P < 0.005 when set against non‐invasive cohorts). Also within the same lesions, 8‐OHdG expression was the most intensive in benign cells. Conversely, HNE immunostaining was strongest in invasive breast carcinomas (UDH versus invasive cohort, P = 0.015). Conclusions: 4‐hydroxy‐2‐nonenal as a marker of lipid peroxidation increases during breast carcinogenesis, reflecting the role of oxidative stress in the pathogenesis of breast cancer. However, 8‐OHdG shows diminished levels in carcinomas, possibly resulting from the induction of DNA repair in these invasive lesions.     

Reactive oxygen species level in follicular fluid--embryo quality marker in IVF?

BACKGROUND: The impact of oxidative stress in female reproduction is not clear. Contradictory reports on the effect of various oxidative stress markers on follicular fluid, oocytes and embryo quality and fertilization potential exist. The objectives of this study were to examine reactive oxygen species (ROS) levels in follicular fluid of women undergoing IVF and to relate these levels to embryo formation and quality. METHODS AND RESULTS: A total of 208 follicular fluid samples were obtained from 78 women undergoing controlled ovarian stimulation and analysed for ROS and lipid peroxidation (LPO). These samples were divided into groups I and II which represented follicular fluid containing grade III and grade II oocytes, respectively. These groups were further subdivided into groups IA, IB, IIA and IIB according to embryo quality. Subgroups IA and IIA consisted of follicular fluid samples corresponding to grade I/II embryo formation. Subgroups IB and IIB represented fertilization failure/pro-nucleolus (PN) arrest/grade III embryos. No significant correlation was observed in ROS levels on comparing groups I and II (P > 0.05). However, ROS levels were observed to be significantly different on comparing groups IA and IB (P < or = 0.01) and groups IIA and IIB (P < or = 0.05). LPO levels further supported our results. CONCLUSION: ROS levels in follicular fluid appear to play a significant role in embryo formation and quality.     

Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa

The objectives of this cross-sectional observational study were: (i) to detect DNA damage and plasma membrane translocation of phosphatidylserine in purified sperm populations of high and low motility, and (ii) to analyse their relationship with the endogenous generation of reactive oxygen species. Ejaculates from infertile men were examined following gradient centrifugation. The main outcome measures were: sperm motion parameters (assessed with a computer analyser), generation of reactive oxygen species (measured by chemiluminescence), DNA damage (detected by terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling and monoclonal antibody labelling of single-stranded DNA) and translocation of membrane phosphatidylserine (examined with annexin V staining). DNA fragmentation and membrane translocation of phosphatidyl-serine were observed in the fractions with low and high sperm motility in all patients. The fractions with low sperm motility had significantly higher proportion of cells with DNA damage and production of reactive oxygen species than the fractions with high sperm motility (P < 0.005). DNA fragmentation was significantly and positively correlated with the generation of reactive oxygen species (r = 0.42; P = 0.02). In conclusion: (i) spermatozoa from infertile men display translocation of membrane phosphatidylserine as diagnosed by annexin V positive staining; (ii) DNA damage (fragmentation and presence of single-stranded DNA) can be detected in ejaculated spermatozoa from infertile men in fractions with low and high sperm motility, and (iii) there is a relationship between DNA damage and oxidative stress.     

Prooxidant-antioxidant state of the organism during oxidative stress and correction of the L-arginine-NO system

The prooxidant-antioxidant balance in rats with oxidative stress was studied during correction of the L-arginine-NO system. Oxidative stress was induced by intravenous injection of E. coli lipopolysaccharide. Under conditions of oxidative stress the prooxidant-antioxidant imbalance was least pronounced during selective correction of the L-arginine-NO system. L-Arginine and nonselective NO synthase inhibitor had little protective effect. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 141, No. 4, pp. 368–370, April, 2006     

糖尿病肾病与线粒体氧化应激

近年来的研究发现,氧化应激与糖尿病肾病的发生、发展有着密切联系.线粒体电子传递呼吸链氧自由基产生过多是高糖介导组织损伤主要原因之一,改善线粒体氧化应激状态成为防治糖尿病肾病新的研究方向.     

Relationship between ROS production, apoptosis and DNA denaturation in spermatozoa from patients examined for infertility

BACKGROUND: The aim of this study was to examine the role of apoptosis and reactive oxygen species (ROS) in inducing DNA damage in ejaculated spermatozoa. METHODS: We examined ejaculated spermatozoa from 31 patients examined for infertility and 19 healthy donors for apoptosis, production of ROS and DNA damage using annexin V binding, chemiluminescence assay and sperm chromatin structure assay. RESULTS: The percentage of spermatozoa that underwent apoptosis in the whole ejaculate and mature fraction was higher in the patients than in the donors (P<0.001 and P=0.009, respectively). Levels of ROS in the whole ejaculate and immature fraction were higher in the patients than in the donors (P=0.002 and P=0.009). Apoptosis was significantly correlated with ROS within patients in the whole ejaculate [r (95% confidence interval)=0.53 (0.19-0.86)] and in the mature [0.71 (0.39-1.00)] and immature spermatozoa [0.75 (0.45-1.00)]. Only apoptosis and the DNA fragmentation index (DFI) were significantly correlated within patients in the whole ejaculate [0.57 (0.18-0.97)]. CONCLUSIONS: DNA damage may be induced by oxidative assault. Apoptosis may not contribute significantly to the DNA damage.     

Andrology: Total antioxidant capacity of human seminal plasma

Although the importance of seminal plasma in the protectionof spermatozoa against reactive oxygen species is well known,only a few studies have investigated its antioxidative propertiesand the possible relationship between infertility and plasmaticantioxidant defences. The aim of the present study was to assessthe status of the total non-enzymatic antioxidant defences ofhuman seminal plasma. Semen samples were obtained from 101 patientsconsulting for infertility and 15 fertile donors. A total reactiveantioxidant potential (TRAP) in seminal plasma was determinedby luminol-chemiluminescence. The relationship of seminal TRAPvalues to lipid peroxidation was also evaluated. Our resultsshow that semen samples from fertile controls show total antioxidantcapacity at higher frequency and value than equivalent samplesfrom suspected subfertile men. This fact as well as the inverserelationship observed between antioxidant capacity and lipidperoxidation potential strongly suggest that impaired antioxidantdefences may play a role in Infertile disorders.     

Pathologic mechanism of hidden blood loss after total knee arthroplasty: oxidative stress induced by free fatty acids

Background: Hidden blood loss (HBL) often occurs after joint replacement; however, the mechanism of HBL is not clear. We conducted a prospective study to analyze the correlation between high-level free fatty acids (FFA) and erythrocyte injury, and explore the pathologic mechanism of hidden blood loss (HBL) after total knee arthroplasty (TKA). Methods: Perioperative blood indexes were tested in 120 patients who underwent unilateral total knee replacement for end-stage knee osteoarthritis. The changes in FFA, reactive oxygen species (ROS), hemoglobin (Hb), and red blood cells (RBC) in the blood samples were detected. The activity of glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD) and hydrogen peroxide (H₂O₂) levels were measured. Morphologic changes of blood cells were analyzed under a microscope. Results: HBL occurred in all patients after TKA. The Hb and RBC decreased significantly 24 h after surgery (P <0.05), while FFA and ROS concentration were substantially elevated, and heteromorphic red blood cells appeared under the microscope. The hemoglobin content decreased to its lowest level at 48 h after the operation (P<0.01). With the increase of FFA and ROS levels, HBL appeared more obvious (P<0.01). GSH-PX activity, T-SOD activity, and H₂O₂ levels significantly decreased compared to preoperative tested samples (P<0.01). Microscopically, atypical erythrocytes increased significantly with cellular rupture and lysis identified. Conclusions: High levels of FFA in blood can induce oxidative stress and damage red blood cells, leading to the occurrence of HBL after surgery. Trial Registration: Chinese Clinical Trials Registry (the trial number: ChiCTR17010681, URL: http://www.chictr.org.cn).     

Prediction of endometriosis with serum and peritoneal fluid markers: a prospective controlled trial

BACKGROUND: The objective of this prospective controlled trial was to investigate the ability of a group of serum and peritoneal fluid (PF) markers to predict, non-surgically, endometriosis. METHODS AND RESULTS: Serum and PF samples were obtained from 130 women while undergoing laparoscopy for pain, infertility, tubal ligation or sterilization reversal. Concentrations of six cytokines [interleukin (IL)-1beta, IL-6, IL-8, IL-12, IL-13 and tumour necrosis factor (TNF)-alpha] were measured in serum and PF, and reactive oxygen species (ROS) in PF, and levels were compared among women who were allocated to groups according to their post-surgical diagnosis. Fifty-six patients were diagnosed with endometriosis, eight with idiopathic infertility, 27 underwent tubal ligation or reanastomosis (control group) and 39 were excluded due to bloody PF. Only serum IL-6 and PF TNF-alpha could be used to discriminate between patients with and without endometriosis with a high degree of sensitivity and specificity (P < 0.001). A threshold of 15 pg/ml PF TNF-alpha provided 100% sensitivity and 89% specificity (positive likelihood ratio of 9.1 and negative likelihood ratio of 0). A threshold of 2 pg/ml for serum IL-6 provided a sensitivity of 90% and specificity of 67% (positive likelihood ratio of 2.7 and negative likelihood ratio of 0.14). CONCLUSIONS: By measuring serum IL-6 and PF TNF-alpha, it was possible to discriminate between patients with endometriosis and those without. Before these markers can be used as a non-surgical diagnostic tool, these data should be verified in a larger study.     

L-arginine reverses indomethacin-induced inhibition of inflammatory fluid secretion of the feline gall bladder

Impaired antioxidant defences may predispose to the increased resting and exercise-induced oxidative stress found in patients with insulin-dependent diabetes mellitus (IDDM). We investigated major erythrocyte antioxidant enzyme activities at rest and in response to sustained, moderate intensity physical exercise in young diabetic men (n = 9) previously reported to have markedly elevated plasma lipid peroxidation and blood glutathione levels compared with control men (n = 13) (Laaksonen et al. 1996). At rest, erythrocyte glutathione reductase activity was 15% higher in the diabetic group (P = 0.049). Se-glutathione peroxidase and glutathione-S-transferase activities were similar in both groups. Red cell Cu, Zn-superoxide dismutase and catalase activities were lower in the IDDM group (P = 0.033 and P = 0.023, respectively). After 40 min of exercise at 60% of the subjects' peak oxygen consumption, Se-glutathione peroxidase activity rose by about 14% in the control group (P = 0.003), but not in the IDDM group (P = 0.47). Exercise did not cause significant changes in other enzyme activities in either group. To conclude, lower erythrocyte Cu, Zn-superoxide dismutase and catalase activity in young men with IDDM at rest may contribute to increased oxidative stress. On the other hand, increased glutathione reductase activity may represent a compensatory upregulation of glutathione homeostasis in response to increased oxidative stress. Upregulation of Se-glutathione peroxidase activity in response to physical activity appeared to be impaired in men with IDDM.     

Increased oxidative stress indices in the blood of child swimmers

The blood redox status of child athletes is compared with that of age-matched untrained individuals. In the present study, 17 swimmers (10.1 ± 1.6 years) and 12 non-athletes (9.9 ± 1.1 years) participated. Reduced glutathione (GSH) was lower by 37% in swimmers compared to non-athletes (P < 0.01), oxidized glutathione (GSSG) was not different and their ratio (GSH/GSSG) was lower by 43% in swimmers compared to non-athletes (P < 0.01). Thiobarbituric acid-reactive substances concentration was higher by 25% in swimmers compared to controls. Catalase exhibited a strong trend toward lower levels in swimmers (P = 0.08). Finally, total antioxidant capacity was found lower by 28% in swimmers compared to controls (P < 0.05). In conclusion, we report that children participating in swimming training exhibit increased oxidative stress and less antioxidant capacity compared to untrained counterparts and suggest that children may be more susceptible to oxidative stress induced by chronic exercise.     

Relationships between adult asthma and oxidative stress markers and pH in exhaled breath condensate: a systematic review

Oxidative stress has a recognized role in the pathophysiology of asthma. Recently, interest has increased in the assessment of pH and airway oxidative stress markers. Collection of exhaled breath condensate (EBC) and quantification of biomarkers in breath samples can potentially indicate lung disease activity and help in the study of airway inflammation, and asthma severity. Levels of oxidative stress markers in the EBC have been systematically evaluated in children with asthma; however, there is no such systematic review conducted for adult asthma. A systematic review of oxidative stress markers measured in EBC of adult asthma was conducted, and studies were identified by searching MEDLINE and SCOPUS databases. Sixteen papers met the inclusion criteria. Concentrations of exhaled hydrogen ions, nitric oxide products, hydrogen peroxide and 8‐isoprostanes were generally elevated and related to lower lung function tests in adults with asthma compared to healthy subjects. Assessment of EBC markers may be a noninvasive approach to evaluate airway inflammation, exacerbations, and disease severity of asthma, and to monitor the effectiveness of anti‐inflammatory treatment regimens. Longitudinal studies, using standardized analytical techniques for EBC collection, are required to establish reference values for the interpretation of EBC markers in the context of asthma.     

The reactive oxygen species-total antioxidant capacity score is a new measure of oxidative stress to predict male infertility.

The imbalance between reactive oxygen species (ROS) production and total antioxidant capacity (TAC) in seminal fluid indicates oxidative stress and is correlated with male infertility. A composite ROS-TAC score may be more strongly correlated with infertility than ROS or TAC alone. We measured ROS, TAC, and ROS-TAC scores in semen from 127 patients and 24 healthy controls. Of the patients, 56 had varicocele, eight had varicocele with prostatitis, 35 had vasectomy reversals, and 28 had idiopathic infertility. ROS levels were higher among infertile men, especially those with varicocele with prostatitis (mean +/- SE, 3.25 +/- 0.89) and vasectomy reversals (2.65 +/- 1.01). All infertile groups had significantly lower ROS-TAC scores than control. ROS-TAC score identified 80% of patients and was significantly better than ROS at identifying varicocele and idiopathic infertility. The 13 patients whose partners later achieved pregnancies had a mean ROS-TAC score of 47.7 +/- 13.2, similar to controls but significantly higher than the 39 patients who remained infertile (35.8 +/- 15.0; P < 0.01). ROS-TAC score is a novel measure of oxidative stress and is superior to ROS or TAC alone in discriminating between fertile and infertile men. Infertile men with male factor or idiopathic diagnoses had significantly lower ROS-TAC scores than controls, and men with male factor diagnoses that eventually were able to initiate a successful pregnancy had significantly higher ROS-TAC scores than those who failed.     

氧化应激促发颅内动脉瘤的研究进展

氧化应激与炎性反应是颅内动脉瘤(CA)形成和破裂的核心因素。作用机制为:1)直接损伤脑动脉内膜;2)使血管平滑肌细胞(VSMC)由可收缩型向炎性反应型转化并凋亡;3)募集炎性细胞、分泌炎性因子,侵袭管壁;4)激活基质金属蛋白酶(MMP),重构和解体血管壁;5)介导脂质过氧化,引起动脉粥样硬化和高血压。初步研究显示阻断氧化应激可预防CA发展,但详细机制尚需进一步探究。