Multimodal distribution and its impact on the accurate assessment of spermatozoa morphological data: Lessons from machine learning.

Objective assessment of sperm morphology is an essential component for assessing ejaculate quality. Due to economic limitations, investigators often divert to conducting observational studies instead of experimental ones, which provide the strongest statistical power, yielding more heterogeneous data regardless of the number of data sources (barns/farms). Using such data inevitably leads to higher variances of estimates, which negatively impacts the statistical power of a study. In this article, we describe a statistical methodology called finite mixture modeling (FMM), which, based on the supplied data and assumed number of sub-classes, classifies the data into two or more homogeneous types of distributions and determines their fractional size relative to the entire cohort. The goal is to use statistical methods that will confound the variance of the sample. A figure from a previous publication was used to generate simulated data (n=1559) on the cytoplasmic droplet rate. We identified that a bi-modal distribution with two latent classes best described the simulated data. Post-hoc estimation showed that about 80 % of observations belonged to latent class 1, with 20 % in latent class 2. The FMM methodology identified a cutoff point of 8.7 %. Finally, when estimating the standard error for the total cohort, the FMM methodology yielded a 40 % reduction in the standard error compared to standard methodologies. In conclusion, here we show that FMM successfully confounded the variance of the data and, as such, yielded lower estimates of the variance than standard methodologies, increasing the statistical power of the cohort.

Adherence to Healthy and Sustainable Dietary Patterns and Long-Term Chronic Inflammation: Data from the EPIC-Potsdam Cohort.

OBJECTIVES: We explored the prospective associations between adherence to a priori chosen dietary patterns, including EAT-Lancet (EAT-L) and Mediterranean (tMDS) diet with long-term inflammatory responses in a German population sample. DESIGN AND SETTING: Prospective cohort study. PARTICIPANTS: A subsample of 636 predominantly healthy participants from the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam study who were on average 51-years old at baseline. MEASUREMENTS: Data was collected repeatedly between 1994/1998 - 2013. At baseline (1994/1998) and 6.8-years later (2001/2005), EAT-L and tMDS scores were derived from available food frequency questionnaires. Stable high, stable low, increasing, and decreasing adherence to EAT-L and tMDS were defined as scoring above/below baseline median at baseline and 6.8-years later. Long-term chronic inflammation was assessed based on the average values of repeated measurements of two inflammatory biomarkers - chemerin and high-sensitivity C-reactive protein (hs-CRP) - in plasma samples collected between 2010/2012 and 2013. Multivariable linear regression analysis adjusted for socio-demographic and lifestyle factors at baseline and in 2010/2012 was used to assess the association between diet adherence and long-term hs-CRP and chemerin concentrations. RESULTS: Stable high or increasing adherence to EAT-L diet compared to stable low adherence was associated with slight reduction of long-term chemerin concentrations on the long run (stable high: -4.4%; increasing: -4.0%), not reaching statistical significance. Increasing adherence to tMDS compared to stable low adherence was also associated with a minor reduction in chemerin concentrations (-3.6%). Decreasing adherence to tMDS compared stable high adherence was associated with 2.7% higher chemerin. The associations were even less pronounced when hs-CRP was used as an outcome. CONCLUSIONS: Adherence to healthy and sustainable dietary patterns defined using existing definitions for EAT-L and tMDS were associated with minor and not statistically significant reduction in the concentrations of inflammatory biomarkers on the long run. More research is needed to explore whether following these diets may represent a suitable approach for targeted prevention in the general population.

Rotation of liquid-preserved artificial insemination doses on roller benches affects sperm quality during storage in stallions.

Appropriate stallion semen handling is of great importance in equine artificial insemination (AI) industry. Optimal treatment of AI doses is aiming for best sperm preservation by excluding strong environmental influences and adding media that favour sperm survival. One method widely used in stallion sperm handling is the rotation of liquid-preserved semen samples on roller benches during storage. As previous studies in boars give rise to the fact that rotation should not be considered beneficial for spermatozoa anymore, the present study investigated the influence of continuous rotation of diluted stallion AI portions on sperm quality. Ejaculates (n = 15) were collected at a German AI centre and diluted with the two extenders EquiPlus and Gent (Minitüb GmbH) to a final concentration of 50 × 106 sperm/mL. Afterwards, samples were placed separately on roller benches at 5°C in the dark, obtaining a rotation frequency of 5 revolutions per minute (rpm) and 36 rpm for four consecutive days following a split-sample design. Both groups were analysed daily in comparison to a control group (0 rpm) with an extended spectrum of spermatological methods including computer-assisted sperm analysis and flow cytometry. Statistical analyses were based on the calculation of generalized linear mixed models for each sperm parameter. The research revealed a decrease in sperm quality parameters of rotated samples compared to non-rotated control groups, visible in total sperm motility (p < .001), decreased thermo-resistance (p < .01) and a drop in pH (p < .001). Interestingly, no differences (p > .05) were detected between rotation frequencies of 5 and 36 rpm. We conclude that the fertilizing capacity of stallion semen was negatively affected by rotation during storage in vitro, irrespective of the rotation frequency. Further studies need to investigate whether field fertility in horses is similarly affected by semen rotation on roller benches in vivo.

Assessment of the stallion sperm acrosome in two different extenders: Spermac stain in comparison with PNA/PSA/PI triple-staining.

Since the stallion acrosome is very small compared to other species and cannot be properly assessed without additional staining, several labelling techniques were developed to facilitate its assessment. The aim of this study was to compare the Spermac stain (Minitüb GmbH) and a PNA/PSA/PI triple-staining detected by flow cytometry with regard to method agreement for detecting non-intact acrosomes within two different extenders. For this purpose, eighteen stallion ejaculates were split in half and diluted with the semen extenders EquiPlus or Gent (Minitüb GmbH) to a final concentration of 50 × 106 sperm/mL, respectively. Subsequently, 126 semen samples were stained with both methods between 4 and 240 h (mean: 63.8 ± 48.9 h) after semen collection. Calculated Intraclass correlation coefficients revealed excellent correlations between both methods for EquiPlus (r = .77, p < .001) and fair correlations for Gent (r = .49, p < .001). Interestingly, flow cytometry detected more non-intact acrosomes in EquiPlus than in Gent (p < .001), whereas the Spermac stain showed no differences (p = .902) between extenders. The poorer method agreement in Gent could be caused by egg yolk artefacts, which made interpretation difficult, so flow cytometry might be preferred. The differences in detected non-intact acrosomes between extenders highlighted the importance of establishing adapted laboratory protocols for different extender types in order to generate comparable results.

Metabolically defined body size and body shape phenotypes and risk of postmenopausal breast cancer in the European Prospective Investigation into Cancer and Nutrition.

BACKGROUND: Excess body fatness and hyperinsulinemia are both associated with an increased risk of postmenopausal breast cancer. However, whether women with high body fatness but normal insulin levels or those with normal body fatness and high levels of insulin are at elevated risk of breast cancer is not known. We investigated the associations of metabolically defined body size and shape phenotypes with the risk of postmenopausal breast cancer in a nested case-control study within the European Prospective Investigation into Cancer and Nutrition. METHODS: Concentrations of C-peptide-a marker for insulin secretion-were measured at inclusion prior to cancer diagnosis in serum from 610 incident postmenopausal breast cancer cases and 1130 matched controls. C-peptide concentrations among the control participants were used to define metabolically healthy (MH; in first tertile) and metabolically unhealthy (MU; >1st tertile) status. We created four metabolic health/body size phenotype categories by combining the metabolic health definitions with normal weight (NW; BMI < 25 kg/m2 , or WC < 80 cm, or WHR < 0.8) and overweight or obese (OW/OB; BMI ≥ 25 kg/m2 , or WC ≥ 80 cm, or WHR ≥ 0.8) status for each of the three anthropometric measures separately: (1) MHNW, (2) MHOW/OB, (3) MUNW, and (4) MUOW/OB. Conditional logistic regression was used to compute odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Women classified as MUOW/OB were at higher risk of postmenopausal breast cancer compared to MHNW women considering BMI (OR = 1.58, 95% CI = 1.14-2.19) and WC (OR = 1.51, 95% CI = 1.09-2.08) cut points and there was also a suggestive increased risk for the WHR (OR = 1.29, 95% CI = 0.94-1.77) definition. Conversely, women with the MHOW/OB and MUNW were not at statistically significant elevated risk of postmenopausal breast cancer risk compared to MHNW women. CONCLUSION: These findings suggest that being overweight or obese and metabolically unhealthy raises risk of postmenopausal breast cancer while overweight or obese women with normal insulin levels are not at higher risk. Additional research should consider the combined utility of anthropometric measures with metabolic parameters in predicting breast cancer risk.

Central venous catheter contamination rate in suspected sepsis patients: an observational clinical study.

BACKGROUND: More than 160,000 central-line-associated bloodstream infections (CLABSIs) are estimated for Europe each year, leading to about 25,000 deaths. AIM: To characterize the contamination of administration sets in suspected CLABSI cases in the intensive care unit (ICU). METHODS: In ICU patients (from February 2017 to February 2018) with suspected CLABSI, all sampled central venous catheters (CVCs) were examined in four segments (from CVC tip to connected tubing systems) for contamination. A risk factor analysis using binary logistic regression was performed. FINDINGS: Fifty-two consecutively sampled CVCs with 1004 elements were analysed with 45 elements being positive for at least one micro-organism (4.48%). There was a significant association with the duration of catheterization (P = 0.038, N = 50) with a daily increase of contamination risk by 11.5% (odds ratio: 1.115). The mean number of CVC manipulations was 40 within 72 h (standard deviation: 20.5), with no association with contamination risk (P = 0.381). The contamination risk of the CVC segments decreased from proximal to distal. Non-replaceable components of the CVC had a high risk (14 times higher; P = 0.01). A significant positive correlation was detected between positive tip cultures and microbial growth in the administration set (r(49) = 0.437; P = 0.001). CONCLUSION: Although only a minority of CLABSI-suspect patients had positive blood cultures, the contamination rate of CVCs and administration set was high, possibly indicating a relevant underreporting. The finding of identical species in adjacent segments underlines the role of upward or downward spread of micro-organisms within the tubes; therefore, aseptic tasks should be emphasized.

Dietary fatty acids and endometrial cancer risk within the European Prospective Investigation into Cancer and Nutrition.

BACKGROUND: Diet may impact important risk factors for endometrial cancer such as obesity and inflammation. However, evidence on the role of specific dietary factors is limited. We investigated associations between dietary fatty acids and endometrial cancer risk in the European Prospective Investigation into Cancer and Nutrition (EPIC). METHODS: This analysis includes 1,886 incident endometrial cancer cases and 297,432 non-cases. All participants were followed up for a mean of 8.8 years. Multivariable Cox proportional hazard models were used to estimate hazard ratios (HR) and 95% confidence intervals (CI) of endometrial cancer across quintiles of individual fatty acids estimated from various food sources quantified through food frequency questionnaires in the entire EPIC cohort. The false discovery rate (q-values) was computed to control for multiple comparisons. RESULTS: Consumption of n-6 γ-linolenic acid was inversely associated with endometrial cancer risk (HR comparing 5th with 1st quintileQ5-Q1=0.77, 95% CI = 0.64; 0.92, ptrend=0.01, q-value = 0.15). This association was mainly driven by γ-linolenic acid derived from plant sources (HRper unit increment=0.94, 95%CI= (0.90;0.98), p = 0.01) but not from animal sources (HRper unit increment= 1.00, 95%CI = (0.92; 1.07), p = 0.92). In addition, an inverse association was found between consumption of n-3 α-linolenic acid from vegetable sources and endometrial cancer risk (HRper unit increment= 0.93, 95%CI = (0.87; 0.99), p = 0.04). No significant association was found between any other fatty acids (individual or grouped) and endometrial cancer risk. CONCLUSION: Our results suggest that higher consumption of γ-linolenic acid and α-linoleic acid from plant sources may be associated with lower risk of endometrial cancer.

The use of machine learning methods to predict sperm quality in Holstein bulls.

The aim of this study was to develop prediction models for total sperm motility, morphological abnormalities and sperm output based on 1,551 ejaculate records of 58 Holstein bulls. The data was collected from September 2019 to November 2020 in a single artificial insemination (AI) center located in Eastern Germany. Factors considered for the prediction models include barn climate conditions, semen collector, number of false mounts, libido, semen collection frequency, breed and age (10-74 months). In this study, the prediction models Lasso, Group Lasso and Gradient Boosting were evaluated. The best model for each sperm quality parameter was chosen using cross validation. The models were estimated with five algorithms for sperm motility and sperm morphology and three algorithms for the number of total sperm per ejaculate (sperm output). For sperm motility and morphology a binary classification algorithm was applied, reaching an accuracy of over 80% for all models. For sperm output, no such classification was used and the only variable selected by all three algorithms was age. Furthermore, for sperm morphology, climate variables were frequently selected. Additionally, network diagrams from Group Lasso show the interdependencies between the major variable groups influencing sperm motility and morphology. In conclusion, the implementation of such prediction tools could help AI centers to optimize management factors and stabilize bull semen production in the future.

A Novel Approach for Non-Invasive Continuous In-Line Control of Perfusion Cell Cultivations by Raman Spectroscopy.

Continuous manufacturing is becoming more important in the biopharmaceutical industry. This processing strategy is favorable, as it is more efficient, flexible, and has the potential to produce higher and more consistent product quality. At the same time, it faces some challenges, especially in cell culture. As a steady state has to be maintained over a prolonged time, it is unavoidable to implement advanced process analytical technologies to control the relevant process parameters in a fast and precise manner. One such analytical technology is Raman spectroscopy, which has proven its advantages for process monitoring and control mostly in (fed-) batch cultivations. In this study, an in-line flow cell for Raman spectroscopy is included in the cell-free harvest stream of a perfusion process. Quantitative models for glucose and lactate were generated based on five cultivations originating from varying bioreactor scales. After successfully validating the glucose model (Root Mean Square Error of Prediction (RMSEP) of ∼0.2 g/L), it was employed for control of an external glucose feed in cultivation with a glucose-free perfusion medium. The generated model was successfully applied to perform process control at 4 g/L and 1.5 g/L glucose over several days, respectively, with variability of ±0.4 g/L. The results demonstrate the high potential of Raman spectroscopy for advanced process monitoring and control of a perfusion process with a bioreactor and scale-independent measurement method.

Dihydroceramide- and ceramide-profiling provides insights into human cardiometabolic disease etiology.

Metabolic alterations precede cardiometabolic disease onset. Here we present ceramide- and dihydroceramide-profiling data from a nested case-cohort (type 2 diabetes [T2D, n = 775]; cardiovascular disease [CVD, n = 551]; random subcohort [n = 1137]) in the prospective EPIC-Potsdam study. We apply the novel NetCoupler-algorithm to link a data-driven (dihydro)ceramide network to T2D and CVD risk. Controlling for confounding by other (dihydro)ceramides, ceramides C18:0 and C22:0 and dihydroceramides C20:0 and C22:2 are associated with higher and ceramide C20:0 and dihydroceramide C26:1 with lower T2D risk. Ceramide C16:0 and dihydroceramide C22:2 are associated with higher CVD risk. Genome-wide association studies and Mendelian randomization analyses support a role of ceramide C22:0 in T2D etiology. Our results also suggest that (dh)ceramides partly mediate the putative adverse effect of high red meat consumption and benefits of coffee consumption on T2D risk. Thus, (dihydro)ceramides may play a critical role in linking genetic predisposition and dietary habits to cardiometabolic disease risk.

Real-World Efficacy of Nintedanib Plus Docetaxel After Progression on Immune Checkpoint Inhibitors: Results From the Ongoing, Non-interventional VARGADO Study.

AIMS: To evaluate the efficacy and safety of nintedanib plus docetaxel in patients with advanced adenocarcinoma non-small cell lung cancer (NSCLC) who progressed after chemotherapy and immune checkpoint inhibitor (ICI) therapy. MATERIALS AND METHODS: VARGADO (NCT02392455) is an ongoing, prospective, non-interventional, real-world study of nintedanib plus docetaxel after first-line chemotherapy in the routine clinical treatment of patients with locally advanced, metastatic or locally recurrent adenocarcinoma NSCLC. Data were collected during routine visits. We report the results from cohort B (n = 80), who received third-line nintedanib plus docetaxel after first-line chemotherapy and second-line ICI therapy. RESULTS: The median duration of follow-up was 12.4 months. Median progression-free survival from initiation of third-line nintedanib plus docetaxel was 6.4 months (95% confidence interval 4.8, 7.3); median overall survival was 12.1 months (95% confidence interval 9.4, 13.5). The 1-year overall survival rate after initiation of third-line nintedanib plus docetaxel treatment (primary end point) was 52% (95% confidence interval 38.0%, 64.4%). Among 64 patients with a documented response, the objective response rate was 50% (n = 32; one complete response and 31 partial responses) and the disease control rate was 86% (n = 55). There were no new safety signals or unexpected toxicities. Among all treated patients, 74% (n = 59) experienced drug-related adverse events, most commonly (nintedanib-related/docetaxel-related) diarrhoea (34%/24%), a decreased white blood cell count (11%/19%) and nausea (13%/16%). CONCLUSIONS: Nintedanib plus docetaxel demonstrated a high response rate and disease stabilisation in the third-line setting after failure of prior chemotherapy and ICI treatment, with a manageable safety profile. These results suggest that nintedanib plus docetaxel represents an efficient treatment option after failure of prior ICIs. The ongoing VARGADO study provides valuable real-world data to inform clinical decision-making regarding treatment sequencing after chemotherapy and ICI failure in patients with adenocarcinoma NSCLC.

Cooled storage of semen from livestock animals (part I): boar, bull, and stallion.

This review is part of the Festschrift in honor of Dr. Duane Garner and provides an overview of current techniques for cooled storage of semen from livestock animals. The first part describes the current state of the art of liquid semen preservation in boars, bulls, and stallions, including the diluents, use of additives, processing, temperature, and cooling of semen. The species-specific physiology and varying extents of cold shock sensitivity are taken into consideration. In addition, factors influencing the quality of cooled-stored semen are discussed. Methods, trends, and the most recent advances for improving sperm quality during cold-temperature storage are highlighted and their respective advantages and disadvantages are contrasted. There has been much progress in recent years regarding cold-temperature storage of boar sperm and there is great potential for a large-scale use to replace the current 17 °C temperature storage regime and the associated use of antibiotics in the future. For stallion sperm, there is an opposite trend away from previous low-temperature storage towards storage at higher temperatures to increase sperm viability and longevity. In bulls, liquid storage of sperm is mostly used in the seasonal dairy production systems of New Zealand and Ireland, but with further research focusing on shelf-live elongation of liquid preserved sperm, there is potential for an application in breeding programs worldwide.

Post-operative hearing among patients with labyrinthine fistula as a complication of cholesteatoma using "under water technique".

INTRODUCTION: During surgery in patients with labyrinthine fistula the mandatory complete removal of the cholesteatoma while preserving inner ear and vestibular function is a challenge. Options so far have been either the complete removal of the cholesteatoma or leaving the matrix on the fistula. We evaluated an alternative "under water" surgical technique for complete cholesteatoma resection, in terms of preservation of postoperative inner ear and vestibular function. METHODS: From 2013 to 2019, 20 patients with labyrinthine fistula due to cholesteatoma were operated. We used the canal wall down approach and removal of matrix on the fistula was done as the last step during surgery using the "under water technique". The pre and postoperative hearing tests and the vestibular function were retrospectively examined. RESULTS: There was no significant difference between pre and post-operative bone conduction thresholds; 20% experienced an improvement of more than 10 dB, with none experiencing a postoperative worsening of sensorineural hearing loss. Among seven patients who presented with vertigo, two had transient vertigo postoperatively but eventually recovered. CONCLUSION: Our data show that the "under water technique" for cholesteatoma removal at the labyrinthine fistula is a viable option in the preservation of inner ear function and facilitating complete cholesteatoma removal.

Milk intake and incident stroke and CHD in populations of European descent: a Mendelian randomisation study.

Higher milk intake has been associated with a lower stroke risk, but not with risk of CHD. Residual confounding or reverse causation cannot be excluded. Therefore, we estimated the causal association of milk consumption with stroke and CHD risk through instrumental variable (IV) and gene-outcome analyses. IV analysis included 29 328 participants (4611 stroke; 9828 CHD) of the European Prospective Investigation into Cancer and Nutrition (EPIC)-CVD (eight European countries) and European Prospective Investigation into Cancer and Nutrition-Netherlands (EPIC-NL) case-cohort studies. rs4988235, a lactase persistence (LP) SNP which enables digestion of lactose in adulthood was used as genetic instrument. Intake of milk was first regressed on rs4988235 in a linear regression model. Next, associations of genetically predicted milk consumption with stroke and CHD were estimated using Prentice-weighted Cox regression. Gene-outcome analysis included 777 024 participants (50 804 cases) from MEGASTROKE (including EPIC-CVD), UK Biobank and EPIC-NL for stroke, and 483 966 participants (61 612 cases) from CARDIoGRAM, UK Biobank, EPIC-CVD and EPIC-NL for CHD. In IV analyses, each additional LP allele was associated with a higher intake of milk in EPIC-CVD (ß = 13·7 g/d; 95 % CI 8·4, 19·1) and EPIC-NL (36·8 g/d; 95 % CI 20·0, 53·5). Genetically predicted milk intake was not associated with stroke (HR per 25 g/d 1·05; 95 % CI 0·94, 1·16) or CHD (1·02; 95 % CI 0·96, 1·08). In gene-outcome analyses, there was no association of rs4988235 with risk of stroke (OR 1·02; 95 % CI 0·99, 1·05) or CHD (OR 0·99; 95 % CI 0·95, 1·03). Current Mendelian randomisation analysis does not provide evidence for a causal inverse relationship between milk consumption and stroke or CHD risk.

Cooled storage of semen from livestock animals (Part II): Camelids, goats, and sheep.

This review is part of the Festschrift in honor of Dr. Duane Garner and provides an overview of current techniques in cooled storage of semen from livestock animals such as camelids, goats, and sheep. Facing worldwide environmental changes and a trend towards more conscious and healthy eating behaviors, the development of a stable animal breeding industry is a significant challenge for the near future. In the present review, factors influencing semen handling in camelids, goats and sheep are described and relevant methods as well as current trends to improve liquid-storage of cooled semen are discussed, including extenders, additives, cooling rates, and storage temperatures. The species-specific physiology and resulting challenges are taken into consideration. While the main problem for camelid semen processing is the relatively greater viscosity as compared with that of some other animals, the deciding factor for successful artificial insemination (AI) in goats and sheep is the site (i.e., cervical or vaginal) of semen placement in the reproductive tract. Due to the type of cervical anatomy, the penetration of the cervix when using AI instruments is rather difficult. Furthermore, the seminal plasma of small ruminants affects the interaction with milk-based extenders and egg yolk which results in species-specific regimens for cooled liquid-preservation. Comparing all three species, the greatest pregnancy rates were obtained by AI with goat semen after cooled liquid-storage for several days.

Suitability of semen stress tests for predicting fertilizing capacity of boar ejaculates.

Besides classical semen parameters, semen stress tests (SSTs) are helpful tools to assess the fertilizing capacity of sperm. However, valid studies on SSTs in relation to fertility are rare because several corrections of common fertility parameters for female and male effects are required. Therefore, over a one-year period, we analyzed semen parameters of 260 ejaculates obtained from 130 Pietrain boars aged between 8 and 9 months in one AI center as well as 1521 corresponding insemination records for these ejaculates. Two consecutive ejaculates (4th and 5th) were collected from each boar and extended in DiluPorc™ BTS. In addition to routine semen evaluation, sperm motility was assessed after heat-resistance test (300 min incubation at 38 °C after seven days storage at 16 °C, HRT) and cold-resistance test (10 min incubation at 38 °C after three days storage at 6 °C, CRT). Generalized linear models (GLMs) were applied to analyze effects for the following predictors of farrowing rate (FR), number of total born (NTB) and live born (NLB) piglets: farm (P = 0.013 [FR], P = 0.001 [NTB], P = 0.023 [NLB]), parity (P = 0.679, P = 0.01, P < 0.001), weekday (P = 0.012, P = 0.08, P = 0.009) and year × season (P < 0.001, P = 0.688, P = 0.574). On boar level, GLMs revealed significant effects on FR, NTB and NLB for the predictors sow (all P < 0.001), total sperm number per dose (P = 0.007, P = 0.002, P < 0.001), total sperm motility (P = 0.002, P = 0.2, P = 0.003) and mitochondrial activity (P = 0.004, P < 0.001, P = 0.002). Moreover, FR and NTB were influenced by membrane integrity (both P < 0.001), FR and NLB by cold-resistance (P < 0.001, P = 0.043), and NTB and NLB by sperm morphology (P = 0.001, P < 0.001) and boar (both P < 0.001). NLB was additionally influenced by heat-resistance (P = 0.004) and farm (P = 0.018) and solely NTB was influenced by sperm output (P = 0.03). Boar and semen related factors explained 9% of the total variation in NTB and 7% of the total variation in NLB. Only 14.2% (n = 37) of the samples were both cold- and heat-resistant (≥65% of motile sperm). Cold- and heat-resistance were dependent factors (Chi-square, P = 0.001) and sperm motility after CRT and HRT showed a moderate positive correlation (rs = 0.40, P < 0.001, Spearman's rho). Finally, ROC curves demonstrated that neither SST can be used as a sole test for predicting the fertilizing capacity of boar ejaculates.

Development of a new antimicrobial concept for boar semen preservation based on bacteriocins.

The conventional storage temperature of 16-18 °C provides optimal conditions for the preservation of boar sperm quality, which are extremely cold sensitive cells. On the other hand, however, it requires the addition of antibiotics to inhibit bacterial growth. Rising numbers of antibiotic resistant bacteria call for alternatives to this conventional storing method. As potential alternative, three different bacteriocin candidates with known bacteriolytic activity against E. coli were examined on possible negative effects concerning the sperm quality and on their impact on bacterial growth of E. coli ILSH 02692 in BTS-extended semen w/o antibiotics. Although the lower concentrations (0.01 and 0.25%) of all bacteriocins did not show any impact on the quality of the semen, the higher concentrations (0.5 and 1.0%) of two bacteriocins led to a significant (P < 0.05) reduction in several sperm quality characteristics. The bacteriocin 860/1c after AMS/dialysis did not affect the sperm quality in any of the tested concentrations and in all tested extenders (BTS, MIII, Androstar Premium and Androhep all w/o antibiotics) at 16 °C as well as at 6 °C. This bacteriocin reduced growth of E. coli ILSH 02692 in BTS-extended semen by 50% compared to the control w/o bacteriocin. Furthermore, a preliminary insemination trial indicated no impact of the selected bacteriocin on fertility. These promising results show that the application of bacteriocins in liquid-preserved semen is a feasible possibility in the future.

Deep femoral artery: A new point of view based on cadaveric study.

INTRODUCTION: In diagnostic angiographic procedures, the knowledge of arterial variations in the femoral artery (FA), deep femoral artery (DFA) and lateral and medial circumflex femoral arteries (LCFA, MCFA) has a great impact. MATERIAL AND METHODS: The frequency of branching patterns of these arteries was investigated in 111 thighs of body donors. Gender and side differences were analyzed statistically. RESULTS: The median distance of separation of the DFA from the FA in relation to the inguinal ligament (IL) was 3.29 cm. High origins (1-2 cm below IL) and middle origins (3-5 cm below IL) of the DFA were found in an equal distribution of 39.3% and 41.1%, respectively. Low origins (6-10 cm below IL) were rare (19.6%) but showed a tendential significance toward expression in males (p = 0.096). The origin of the LCFA from the FA (19.8%) or DFA (70.2%) are in line with the findings of other groups. The origin of the MCFA from FA (14.4%) or DFA (74.7%) showed that circumflex femoral arteries arose mostly from DFA. A trifurcation of the FA into the DFA, LCFA and MCFA was only observed in 9.9% and, therefore, less frequently than reported by others. Branches of the femoral nerve (FN) passed mostly anterior (46.4%) or anterior and posterior (47.8%) to the LCFA. The rare constellation of branches of FN passing only posterior to the LCFA (5.8%) showed a tendential significance to left side expression (p = 0.084). CONCLUSIONS: Taken together, this is the first classification of the median distance of separation of the DFA from the FA in relation to the IL in three defined groups. The knowledge of DFA branching pattern is essential for recent therapy options of cardiac diseases using a femoral artery access: transcatheter aortic valve replacement, catheter-based miniaturized ventricular assist device and veno-arterial extracorporal membrane oxygenation. The variant topography of the branches of FN in relation to LCFA should be kept in mind when harvesting an anterolateral thigh flap.

Evaluation of a panel of spermatological methods for assessing reprotoxic compounds in multilayer semen plastic bags.

The increase of fertility performance in sows is one of the biggest achievements in pig production over the last 30 years. Nevertheless, pig farms using artificial insemination (AI) repeatedly experienced in recent year's fertility problems with dramatic consequences due to toxic compounds from plastic semen bags. In particular, bisphenol A diglycidyl-ether (BADGE) present in multilayer plastic bags can leach into the semen and could affect the functionality of the spermatozoa. Former studies could not find any alterations in spermatozoa based on the exposure to BADGE. The aim of the study was to evaluate effects of BADGE on boar spermatozoa using an extended panel of spermatological methods. In spring 2019, a large drop in farrowing rates from 92.6 ± 2.3% to 63.7 ± 11.1% in four sow farms in Croatia was detected. In migration studies, BADGE could be identified as a causal toxic compound and leached into the extended semen in concentration of 0.37 ± 0.05 mg/L. Detailed spermatological studies showed that significant predictors for effects on spermatozoa were different levels of motility and kinematic data after a prolonged storage time, thermo-resistance test (prolonged incubation time), mitochondrial activity, membrane integrity and fluidity. No serious effects were observed for sperm morphology and DNA fragmentation. These results provide new insights into the development of a new quality assurance concept for a detailed spermatological examination during testing of plastic materials for boar semen preservation. It could be shown that boar spermatozoa are an excellent biosensor to detect potential toxicity and fertility-relevant compounds.