Effect of Sperm Separation Methods on Morphology and Functions of Frozen Buffalo Spermatozoa

This work was planned to compare three methods for selection of active buffalo spermatozoa, examine the effects of these separation methods on morphology, viability and functions of spermatozoa used for IVF purposes. Ten frozen straws per trial (10 times) were pooled and divided into 4 aliquots:  A) First aliquot was considered as control without any separation method. B) Second aliquot was subjected to sperm selection by density gradient method (percoll:PureSperm) using 40-80% double density gradient. C) The third aliquot was subjected to swim-up in sp-TALP. D) The fourth aliquot was subjected to washing by centrifugation with sp-TALP. The percentage of motility increased for Percoll, swim up and washing than control (86.0, 73.0, and 66.5 vs. 56.5) respectively. Sperm abnormalities % was significantly decreased after Percoll, swim up and sperm wash separation methods. Spermatozoa obtained by Swim up and Percoll had the highest percentage of intact membrane. Different spermatozoa separation methods significantly increased the lytic activity of the recovered spermatozoa. Live spermatozoa percentage with reacted acrosome significantly increased after both swim up separation and washing. The percentage of dead spermatozoa with reacted acrosome significantly decreased after percoll separation but it did not change when the swim up method was used. Finally it is concluded that, density gradient centrifugation using PureSperm® could be considered as the method of choice for selection of frozen thawed buffalo spermatozoa and presumably with a high potential fertilizing ability. density gradient centrifugation using PureSperm® could be considered as the method of choice for selection of frozen thawed buffalo spermatozoa

Effect of Sperm Separation Methods on Morphology and Functions of Frozen Buffalo Spermatozoa

This work was planned to compare three methods for selection of active buffalo spermatozoa, examine the effects of these separation methods on morphology, viability and functions of spermatozoa used for IVF purposes. Ten frozen straws per trial (10 times) were pooled and divided into 4 aliquots:  A) First aliquot was considered as control without any separation method. B) Second aliquot was subjected to sperm selection by density gradient method (percoll:PureSperm) using 40-80% double density gradient. C) The third aliquot was subjected to swim-up in sp-TALP. D) The fourth aliquot was subjected to washing by centrifugation with sp-TALP. The percentage of motility increased for Percoll, swim up and washing than control (86.0, 73.0, and 66.5 vs. 56.5) respectively. Sperm abnormalities % was significantly decreased after Percoll, swim up and sperm wash separation methods. Spermatozoa obtained by Swim up and Percoll had the highest percentage of intact membrane. Different spermatozoa separation methods significantly increased the lytic activity of the recovered spermatozoa. Live spermatozoa percentage with reacted acrosome significantly increased after both swim up separation and washing. The percentage of dead spermatozoa with reacted acrosome significantly decreased after percoll separation but it did not change when the swim up method was used. Finally it is concluded that, density gradient centrifugation using PureSperm® could be considered as the method of choice for selection of frozen thawed buffalo spermatozoa and presumably with a high potential fertilizing ability. density gradient centrifugation using PureSperm® could be considered as the method of choice for selection of frozen thawed buffalo spermatozoa

Pooled analysis of WHO Surgical Safety Checklist use and mortality after emergency laparotomy

Background The World Health Organization (WHO) Surgical Safety Checklist has fostered safe practice for 10 years, yet its place in emergency surgery has not been assessed on a global scale. The aim of this study was to evaluate reported checklist use in emergency settings and examine the relationship with perioperative mortality in patients who had emergency laparotomy. Methods In two multinational cohort studies, adults undergoing emergency laparotomy were compared with those having elective gastrointestinal surgery. Relationships between reported checklist use and mortality were determined using multivariable logistic regression and bootstrapped simulation. Results Of 12 296 patients included from 76 countries, 4843 underwent emergency laparotomy. After adjusting for patient and disease factors, checklist use before emergency laparotomy was more common in countries with a high Human Development Index (HDI) (2455 of 2741, 89.6 per cent) compared with that in countries with a middle (753 of 1242, 60.6 per cent; odds ratio (OR) 0.17, 95 per cent c.i. 0.14 to 0.21, P <0001) or low (363 of 860, 422 per cent; OR 008, 007 to 010, P <0.001) HDI. Checklist use was less common in elective surgery than for emergency laparotomy in high-HDI countries (risk difference -94 (95 per cent c.i. -11.9 to -6.9) per cent; P <0001), but the relationship was reversed in low-HDI countries (+121 (+7.0 to +173) per cent; P <0001). In multivariable models, checklist use was associated with a lower 30-day perioperative mortality (OR 0.60, 0.50 to 073; P <0.001). The greatest absolute benefit was seen for emergency surgery in low- and middle-HDI countries. Conclusion Checklist use in emergency laparotomy was associated with a significantly lower perioperative mortality rate. Checklist use in low-HDI countries was half that in high-HDI countries.Peer reviewe

Global variation in anastomosis and end colostomy formation following left-sided colorectal resection

Background End colostomy rates following colorectal resection vary across institutions in high-income settings, being influenced by patient, disease, surgeon and system factors. This study aimed to assess global variation in end colostomy rates after left-sided colorectal resection. Methods This study comprised an analysis of GlobalSurg-1 and -2 international, prospective, observational cohort studies (2014, 2016), including consecutive adult patients undergoing elective or emergency left-sided colorectal resection within discrete 2-week windows. Countries were grouped into high-, middle- and low-income tertiles according to the United Nations Human Development Index (HDI). Factors associated with colostomy formation versus primary anastomosis were explored using a multilevel, multivariable logistic regression model. Results In total, 1635 patients from 242 hospitals in 57 countries undergoing left-sided colorectal resection were included: 113 (6·9 per cent) from low-HDI, 254 (15·5 per cent) from middle-HDI and 1268 (77·6 per cent) from high-HDI countries. There was a higher proportion of patients with perforated disease (57·5, 40·9 and 35·4 per cent; P < 0·001) and subsequent use of end colostomy (52·2, 24·8 and 18·9 per cent; P < 0·001) in low- compared with middle- and high-HDI settings. The association with colostomy use in low-HDI settings persisted (odds ratio (OR) 3·20, 95 per cent c.i. 1·35 to 7·57; P = 0·008) after risk adjustment for malignant disease (OR 2·34, 1·65 to 3·32; P < 0·001), emergency surgery (OR 4·08, 2·73 to 6·10; P < 0·001), time to operation at least 48 h (OR 1·99, 1·28 to 3·09; P = 0·002) and disease perforation (OR 4·00, 2·81 to 5·69; P < 0·001). Conclusion Global differences existed in the proportion of patients receiving end stomas after left-sided colorectal resection based on income, which went beyond case mix alone

Zearalenone (ZEN) disrupts the anti-inflammatory response of bovine oviductal epithelial cells to sperm in vitro

Dietary contamination by Zearalenone (ZEN) has a detrimental effect on bovine fertility. Recently, we showed a novel anti-inflammatory response of bovine oviductal epithelial cells (BOEC) to active sperm cells in vitro. The aim of the present study was to investigate the effect of ZEN exposure of BOEC on the immune-related cytokine expression in response to bovine sperm. At concentrations of 100 and 1000ng/mL, ZEN induced the expression of TNF and IL1B (pro-inflammatory cytokines) as well as IL8 (chemokine) in BOEC in a dose-dependent manner. Furthermore, ZEN induced PTGES expression and PGE2 secretion in BOEC. Sperm co-culture induced an anti-inflammatory response in BOEC with upregulation of TGFB, secretion of PGE2 and downregulation of TNF. Most importantly, ZEN at 1-1000ng/mL eliminated the response of BOEC to sperm. Estradiol-17β (5ng/mL) treatment did not produce the same effects as ZEN, suggesting that the response of BOEC to ZEN is, at least in part, not mediated by estrogen receptors. Taken together, ZEN can produce inflammatory effects on BOEC by stimulating the expressions of pro-inflammatory cytokines and disrupt the normal interaction between sperm and BOEC at the level of cytokine expressions and PGE2 production. Thus, exposure of the bovine oviduct to ZEN may negatively affect sperm survival and reduce fertility

Zearalenone (ZEN) disrupts the anti-inflammatory response of bovine oviductal epithelial cells to sperm in vitro

Dietary contamination by Zearalenone (ZEN) has a detrimental effect on bovine fertility. Recently, we showed a novel anti-inflammatory response of bovine oviductal epithelial cells (BOEC) to active sperm cells in vitro. The aim of the present study was to investigate the effect of ZEN exposure of BOEC on the immune-related cytokine expression in response to bovine sperm. At concentrations of 100 and 1000ng/mL, ZEN induced the expression of TNF and IL1B (pro-inflammatory cytokines) as well as IL8 (chemokine) in BOEC in a dose-dependent manner. Furthermore, ZEN induced PTGES expression and PGE2 secretion in BOEC. Sperm co-culture induced an anti-inflammatory response in BOEC with upregulation of TGFB, secretion of PGE2 and downregulation of TNF. Most importantly, ZEN at 1-1000ng/mL eliminated the response of BOEC to sperm. Estradiol-17β (5ng/mL) treatment did not produce the same effects as ZEN, suggesting that the response of BOEC to ZEN is, at least in part, not mediated by estrogen receptors. Taken together, ZEN can produce inflammatory effects on BOEC by stimulating the expressions of pro-inflammatory cytokines and disrupt the normal interaction between sperm and BOEC at the level of cytokine expressions and PGE2 production. Thus, exposure of the bovine oviduct to ZEN may negatively affect sperm survival and reduce fertility