Evaluation of hormones as candidate early biomarkers of Holstein bull performance around the first semen collection at artificial insemination centres.

Artificial insemination (AI) centres select bulls as calves according to their genetic breeding values and raise them until the first semen collection; yet, a high dropout rate of reared bulls is a problem for AI centres. Potential hormonal indicators of bull sexual maturation (cortisol, dehydroepiandrosterone (DHEA), testosterone, oestradiol, insulin-like growth factor 1 (IGF-1)) were observed and evaluated in relation to the performance parameters to perhaps identify candidate biomarkers allowing an early selection of bulls as suitable sires. Blood samples from 102 German Holstein calves at 4 ± 1, 8 ± 1 and 12 ± 2 months of age from six AI centres were analysed using validated immunoassays for cortisol, DHEA, testosterone, oestradiol and IGF-1. Semen analyses included native and thawed diluted semen. Bulls were classified at the first semen collection into groups with good versus poor performance (GP vs. LP). After 2 years, the subsequent differentiation was done in high (HPP), medium (MPP) and low performance persistency (LPP). Age at first semen collection was an important factor for sperm quality. Cortisol concentrations decreased with age, but the cortisol/DHEA ratio decreased with age only in GP bulls (p < .05). Oestradiol and testosterone concentrations both correlated with libido behaviour (p < .05). Testosterone and IGF-1 concentrations were higher at the time of first semen collection in GP bulls and increased with age (p < .05). In conclusion, testosterone and IGF-1 concentrations at first semen collection are associated with performance at first semen collection and future performance persistency, and might be useful early biomarkers for consistent sperm producing bulls on AI centres.

Acinetobacter in fresh faecal samples is related to sperm motility: A case report in Piétrain boars housed on different deep-litter bedding materials.

The swine gastrointestinal tract contains a great variety of microbes, forming a complex and dynamic ecosystem. Various internal and external factors (e.g. age, breed and diet) may influence its composition. This study aimed to investigate the gut microbial diversity of German Piétrain boars housed on different deep-litter bedding materials (regional wood shavings, linen, hemp, spelt husks, and wood shavings) via 16S-rDNA sequencing. Additionally, short-chain fatty acids were analysed using gas chromatography. Fresh faecal samples (n = 80) from 40 Piétrain boars were collected twice during the trial. Although it can be assumed that boars ingest bedding orally, no differences in the microbiome composition could be found. The main phyla were Firmicutes and Bacteroides. Acinetobacter was identified as a biomarker for sperm quality differences (total sperm motility) in breeding boars.

Surgical Management of Periprosthetic Joint Infections in Hip and Knee Megaprostheses.

Periprosthetic joint infection is a feared complication after the megaprosthetic reconstruction of oncologic and non-oncologic bone defects of including the knee or hip joint. Due to the relative rarity of these procedures, however, optimal management is debatable. Considering the expanding use of megaprostheses in revision arthroplasty and the high revision burden in orthopedic oncology, the risk of PJI is likely to increase over the coming years. In this non-systematic review article, we present and discuss current management options and the associated results focusing on studies from the last 15 years and studies from dedicated centers or study groups. The indication, surgical details and results in controlling infection are presented for debridement, antibiotics, irrigation and retention (DAIR) procedure with an exchange of the modular components, single-stage implant exchange, two-stage exchanges and ablative procedures.

Evaluation of time to reimplantation as a risk factor in two-stage revision with static spacers for periprosthetic knee joint infection.

INTRODUCTION: We investigated the time to reimplantation (TTR) during two-stage revision using static spacers with regard to treatment success and function in patients with chronic periprosthetic joint infection (PJI) of the knee. METHODS: 163 patients (median age 72 years, 72 women) who underwent two-stage exchange for chronic knee PJI between 2012 and 2020 were retrospectively analyzed (based on the 2011 Musculoskeletal Infection Society criteria). A cutoff TTR for increased risk of reinfection was identified using the maximally selected log-rank statistic. Infection control, aseptic revisions and overall survival were analyzed using Kaplan-Meier survival estimates. Adjustment for confounding factors-the Charlson Comorbidity Index (CCI) and C-reactive protein (CRP)-was done with a Cox proportional hazards model. RESULTS: When TTR exceeded 94 days, the adjusted hazard of reinfection was increased 2.8-fold (95% CI 1.4-5.7; p = 0.0036). The reinfection-free rate was 67% (95% CI 52-79%) after 2 years and 33% (95% CI 11-57%) after 5 years for a longer TTR compared to 89% (95% CI 81-94%) and 80% (95% CI 69-87%) at 2 and 5 years, respectively, for a shorter TTR. Adjusted overall survival and number of aseptic revisions did not differ between the longer TTR and shorter TTR groups. Maximum knee flexion was 90° (IQR 84-100) for a longer TTR and 95° (IQR 90-100) for a shorter TTR (p = 0.0431), with no difference between the groups in Oxford Knee Score. Baseline characteristics were similar (body mass index, age, previous surgeries, microorganisms) for the two groups, except that there was a higher CCI (median 4 vs. 3) and higher CRP (median 3.7 vs 2.6 mg/dl) in the longer TTR group. CONCLUSION: A long TTR is sometimes unavoidable in clinical practice, but surgeons should be aware of a potentially higher risk of reinfection. LEVEL OF EVIDENCE: III, retrospective comparative study.

Moderate Continuous and Modified High-Intensity Interval Training in Patients With Left Ventricular Assist Devices: The Prospective Train-the-LVAD Trial.

BACKGROUND: High-intensity interval training (HIIT) of 4 minutes at 80%-90% of peak oxygen consumption (VO2peak) has been shown to be feasible in patients with left ventricular assist devices (LVADs). The effect of shorter bouts of HIIT, which reduce the anaerobic burden, has not been investigated compared to moderate continuous training (MCT). METHODS AND RESULTS: We conducted a prospective, monocentric study (NCT05121077) randomizing patients with LVADs into 20 minutes of MCT (n = 10) or short bouts (≤ 90 seconds) of HIIT (n = 10) following cardiopulmonary exercise testing at 50%-60% and 80%-90% of VO2peak. Each of the 18 supervised sessions (3×/week, t0-t1) included 10 minutes of strengthening training. The primary outcome was the increase of VO2peak in the 2 groups between t0 and t1. Secondary outcomes were changes in the 12-item Kansas City Cardiomyopathy Questionnaire, the 6-minute walk distance and the percentage of VO2peak at the first ventilatory threshold. VO2peak significantly increased with HIIT (13.0 ± 4.6mL/kg/min vs 14.6 ± 4.3mL/kg/min; P = 0.037), but not with MCT (11.8 ± 3.3mL/kg/min vs 13.1 ± 3.3mL/kg/min; P = 0.322), without between-group differences (P = 0.853). Secondary outcomes improved from t0-t1 in MCT and HIIT, without differences between the groups. CONCLUSIONS: Short bouts of HIIT are feasible, and they improved VO2peak and functional parameters in patients in this pilot prospective study.

Science-based quality control in boar semen production.

The ever-increasing understanding of sperm physiology, combined with innovative technical advances, continuously furthers the development of boar semen production management. These improvements pave the way for the future implementation of modified quality assurance concepts. This review provides an overview of current trends and new approaches in boar semen production, focusing on: the improvement of hygienic standards, alternatives to the use of antibiotics including the application of cold temperature storage and the utilization of antimicrobial additives, as well as the implementation of new quality control tools. Furthermore, the influence of dilution and temperature management, as well as new possibilities for an improvement of boar semen shipping and storage conditions are reviewed.

Genetic and phenotypic spectrum in the NONO-associated syndromic disorder.

The non-POU domain-containing octamer-binding (NONO) protein is involved in multiple steps of gene regulation such as RNA metabolism and DNA repair. Hemizygous pathogenic variants in the NONO gene were confirmed to cause a rare X-linked syndromic disorder. Through our in-house diagnostics and subsequent matchmaking, we identified six unrelated male individuals with pathogenic or likely pathogenic NONO variants. For a detailed comparison, we reviewed all published characterizations of the NONO-associated disorder. The combined cohort consists of 16 live-born males showing developmental delay, corpus callosum anomalies, non-compaction cardiomyopathy and relative macrocephaly as leading symptoms. Seven prenatal literature cases were characterized by cardiac malformations. In this study, we extend the phenotypic spectrum through two more cases with epilepsy as well as two more cases with hematologic anomalies. By RNA expression analysis and structural modeling of a new in-frame splice deletion, we reinforce loss-of-function as the pathomechanism for the NONO-associated syndromic disorder.

Variability in Cochlear Implantation Outcomes in a Large German Cohort With a Genetic Etiology of Hearing Loss.

OBJECTIVES: The variability in outcomes of cochlear implantation is largely unexplained, and clinical factors are not sufficient for predicting performance. Genetic factors have been suggested to impact outcomes, but the clinical and genetic heterogeneity of hereditary hearing loss makes it difficult to determine and interpret postoperative performance. It is hypothesized that genetic mutations that affect the neuronal components of the cochlea and auditory pathway, targeted by the cochlear implant (CI), may lead to poor performance. A large cohort of CI recipients was studied to verify this hypothesis. DESIGN: This study included a large German cohort of CI recipients (n = 123 implanted ears; n = 76 probands) with a definitive genetic etiology of hearing loss according to the American College of Medical Genetics (ACMG)/Association for Molecular Pathology (AMP) guidelines and documented postoperative audiological outcomes. All patients underwent preoperative clinical and audiological examinations. Postoperative CI outcome measures were based on at least 1 year of postoperative audiological follow-up for patients with postlingual hearing loss onset (>6 years) and 5 years for children with congenital or pre/perilingual hearing loss onset (≤6 years). Genetic analysis was performed based on three different methods that included single-gene screening, custom-designed hearing loss gene panel sequencing, targeting known syndromic and nonsyndromic hearing loss genes, and whole-genome sequencing. RESULTS: The genetic diagnosis of the 76 probands in the genetic cohort involved 35 genes and 61 different clinically relevant (pathogenic, likely pathogenic) variants. With regard to implanted ears (n = 123), the six most frequently affected genes affecting nearly one-half of implanted ears were GJB2 (21%; n = 26), TMPRSS3 (7%; n = 9), MYO15A (7%; n = 8), SLC26A4 (5%; n = 6), and LOXHD1 and USH2A (each 4%; n = 5). CI recipients with pathogenic variants that influence the sensory nonneural structures performed at or above the median level of speech performance of all ears at 70% [monosyllable word recognition score in quiet at 65 decibels sound pressure level (SPL)]. When gene expression categories were compared to demographic and clinical categories (total number of compared categories: n = 30), mutations in genes expressed in the spiral ganglion emerged as a significant factor more negatively affecting cochlear implantation outcomes than all clinical parameters. An ANOVA of a reduced set of genetic and clinical categories (n = 10) identified five detrimental factors leading to poorer performance with highly significant effects ( p < 0.001), accounting for a total of 11.8% of the observed variance. The single strongest category was neural gene expression accounting for 3.1% of the variance. CONCLUSIONS: The analysis of the relationship between the molecular genetic diagnoses of a hereditary etiology of hearing loss and cochlear implantation outcomes in a large German cohort of CI recipients revealed significant variabilities. Poor performance was observed with genetic mutations that affected the neural components of the cochlea, supporting the "spiral ganglion hypothesis."

What Is the Risk of Dislocation and Revision in Proximal Femoral Replacement with Dual-mobility Articulation After Two-stage Revision for Periprosthetic Hip Infection?

BACKGROUND: Dislocation is a major complication of revision THA after two-stage exchange for periprosthetic joint infection (PJI). The likelihood of dislocation can be particularly high if megaprosthetic proximal femoral replacement (PFR) has been performed during a second-stage reimplantation. Dual-mobility acetabular components are an established way of reducing the instability risk in revision THA; however, the likelihood of dislocation for dual-mobility reconstructions in the setting of a two-stage PFR has not been studied systematically, although patients with these reconstructions might be at an increased risk. QUESTIONS/PURPOSES: (1) What is the risk of dislocation and revision for dislocation in patients who underwent PFR with a dual-mobility acetabular component as part of two-stage exchange for hip PJI? (2) What is the risk of all-cause implant revision and what other procedures were performed (apart from revision for a dislocation) in these patients? (3) What potential patient-related and procedure-related factors are associated with dislocation? METHODS: This was a retrospective study from a single academic center including procedures performed between 2010 and 2017. During the study period, 220 patients underwent two-stage revision for chronic hip PJI. Two-stage revision was the approach of choice for chronic infections, and we did not perform single-stage revisions for this indication during the study period. Thirty-three percent (73 of 220) of patients underwent second-stage reconstruction with a single-design, modular, megaprosthetic PFR because of femoral bone loss, using a cemented stem. A cemented dual-mobility cup was the approach of choice for acetabular reconstruction in the presence of a PFR; however, 4% (three of 73) were reconstructed with a bipolar hemiarthroplasty to salvage an infected saddle prosthesis, leaving 70 patients with a dual-mobility acetabular component and a PFR (84% [59 of 70]) or total femoral replacement (16% [11 of 70]). We used two similar designs of an unconstrained cemented dual-mobility cup during the study period. The median (interquartile range) patient age was 73 years (63 to 79 years), and 60% (42 of 70) of patients were women. The mean follow-up period was 50 ± 25 months with a minimum follow-up of 24 months for patients who did not undergo revision surgery or died (during the study period, 10% [seven of 70] died before 2 years). We recorded patient-related and surgery-related details from the electronic patient records and investigated all revision procedures performed until December 2021. Patients who underwent closed reduction for dislocation were included. Radiographic measurements of cup positioning were performed using supine AP radiographs obtained within the first 2 weeks after surgery using an established digital method. We calculated the risk for revision and dislocation using a competing-risk analysis with death as a competing event, providing 95% confidence intervals. Differences in dislocation and revision risks were assessed with Fine and Gray models providing subhazard ratios. All p values were two sided and the p value for significance was set at 0.05. RESULTS: The risk of dislocation (using a competing-risks survivorship estimator) was 17% (95% CI 9% to 32%) at 5 years, and the risk of revision for dislocation was 12% (95% CI 5% to 24%) at 5 years among patients treated with dual-mobility acetabular components as part of a two-stage revision for PJI of the hip. The risk of all-cause implant revision (using a competing-risk estimator, except for dislocation) was 20% (95% CI 12% to 33%) after 5 years. Twenty-three percent (16 of 70) of patients underwent revision surgery for reinfection and 3% (two of 70) of patients underwent stem exchange for a traumatic periprosthetic fracture. No patients underwent revision for aseptic loosening. We found no differences in patient-related and procedure-related factors or acetabular component positioning for patients with dislocation with the numbers available; however, patients with total femoral replacements had a higher likelihood of dislocation (subhazard ratio 3.9 [95% CI 1.1 to 13.3]; p = 0.03) and revision for a dislocation (subhazard ratio 4.4 [95% CI 1 to 18.5]; p = 0.04) than those who received PFR. CONCLUSION: Although dual-mobility bearings might be an intuitive potential choice to reduce the dislocation risk in revision THA, there is a considerable dislocation risk for PFR after two-stage surgery for PJI, particularly in patients with total femoral replacements. Although the use of an additional constraint might appear tempting, published results vary tremendously, and future studies should compare the performance of tripolar constrained implants to that of unconstrained dual-mobility cups in patients with PFR to reduce the risk of instability. LEVEL OF EVIDENCE: Level III, therapeutic study.

Dual use of breeding stallions is possible without affecting the sperm quality.

Artificial insemination (AI) is commonly used in the equine industry to enhance the genetic value in breeding programs and to effectively utilize ejaculates. Many stallions are used as breeding stallions as well as in high-level sports competitions to improve their market value. The goal of the present study was to investigate whether this dual use of stallions influences the animals´ stress levels and/or the quality of their ejaculates. For this purpose, 18 stallions were grouped into two categories: breeding stallions with (BSC = breeding stallion competition), and breeding stallions without secondary use in competitions (BS = breeding stallion). Two ejaculates were collected at a one-week interval and analysed with an extended spectrum of spermatological methods. Furthermore, saliva, as well as seminal plasma samples were taken, and the concentration of cortisol therein was determined. Additionally, dehydroepiandrosterone (DHEA) and the cortisol/DHEA ratio were analysed and calculated for seminal plasma. After statistical analysis of the correlations and interdependences between the two groups, the results showed that the BSC group had significantly higher saliva cortisol levels (p = .027) and tendentially higher DHEA concentrations in their seminal plasma (p = .056). No difference between BS and BSC could be found in regard to the sperm quality parameters and the cortisol concentration in seminal plasma samples. It can be concluded that while active participation in competitions represents a stress factor, the dual use of stallions in breeding programs and sports competitions is possible without negative effects on their sperm quality.

Bacterial killing activity and lysozymes: A stable defence mechanism in stallion seminal plasma?

During insemination, bacterial contamination of the ejaculate can lead to reduced sperm quality and transmission of pathogens to the female, thus should be avoided. The semen of a variety of animal taxa possess antimicrobial properties against a wide range of bacterial species through antimicrobial molecules, such as lysozyme, but their variance and the factors influencing it are unknown for most species. In this study, the antibacterial defence (bacterial killing activity (BKA) against Escherichia (E.) coli and Staphylococcus (S.) aureus as well as lysozyme concentration) was studied in seminal fluid from two consecutive ejaculates of 18 stallions. All ejaculates showed BKA against the tested bacteria, which correlated between the two consecutive ejaculates (rS  = 0.526, p = .025 for E. coli and rS  = 0.656, p = .003 for S. aureus) and appeared to be stable over the tested period. The lysozyme concentration (LC) showed no significant correlation between the consecutive ejaculates (rS  = 0.161, p = .681). However, LC had a positive correlation to the ratio of apoptotic spermatozoa within the ejaculates (rS  = 0.426, p = .019). In contrast to other livestock (e.g., boar, bull), the BKA in stallion semen did not correlate significantly with the age of the animal nor sperm quality characteristics.

Estimation of sperm concentration limits to produce intrauterine insemination doses in swine.

This study evaluated the effect of sperm concentration of boar semen doses, for intrauterine artificial insemination (IUAI), on semen quality and established concentration limits for their production. Twenty ejaculates from four crossbred mature PIC® boars were collected to produce 50 mL semen doses in a split sample, reaching the following sperm concentrations: ~20, 30, 60, and 100 × 106 cells/mL. Doses were produced using Androstar® Plus, stored at 17°C, and evaluated until 120 h of storage. There was a linear decrease in sperm motility as the sperm concentration increased (p linear < .01). The concentration which no longer affected the total and progressive motility was 59 and 55 × 106 cells/mL, respectively (corresponding to 71% and 62%, respectively). The pH linearly decreased as the sperm concentration increased (p < .01); yet, at 72 and 120 h, the parameter dramatically reduced in boar semen doses with 60 and 100 × 106 cells/mL. The percentage of cells with intact plasma and acrosomal membranes or with high mitochondrial membrane potential was not influenced by the sperm concentration (p ≥ .15). In conclusion, sperm motility was negatively affected in highly (60 and 100 × 106 cells/mL) concentrated doses. To achieve suitable sperm motility, boar semen doses may not surpass the sperm concentration of 55 × 106 cells/mL. The effect of low-concentrated boar semen doses on sperm quality still needs to be better evaluated, mainly considering the influence of extender type and thermo-resistance conditions.

Blood-saving dissection with monopolar tungsten needle electrodes and Teflon-coated spatula electrodes in tumor orthopedics.

INTRODUCTION: Resection of musculoskeletal tumors and reconstruction with tumor endoprostheses often results in blood loss requiring transfusion of blood products. We assessed the blood-saving potential of using monopolar tungsten needle electrodes and polytetrafluoroethylene (PTFE)-coated spatula electrodes (intervention) compared with conventional dissection with sharp instruments and coagulation with uncoated steel electrodes (control). METHODS: We retrospectively analyzed data of 132 patients (79 interventions, 53 controls) undergoing surgery by one single experienced surgeon in our tertiary referral center between 2012 and 2021. RESULTS: Intraoperative blood loss in the intervention group was reduced by 29% [median (IQR): 700 (400-1200) vs 500 (200-700) ml; p = 0.0043]. Postoperative wound drainage decreased by 41% [median (IQR): 1230 (668-2041) vs 730 (450-1354) ml; p = 0.0080]. Additionally, patients in need of PRBCs during surgery declined from 43% to 15% (23/53 vs 12/79; p = 0.0005), while the transfusion rate after surgery did not change notably. The number of patients in need of revision surgery due to wound healing disorders was low in both groups (control group: 4/53 vs intervention group: 4/79). Only one patient in the control group and two patients in the intervention group underwent revision surgery due to hemorrhage. Baseline characteristics were similar between groups (sex, Charlson Comorbidity score, tumor entity). CONCLUSION: Dissection with tungsten needle electrodes and PTFE-coated spatula electrodes appears an effective surgical blood-saving measure without increased risk of wound healing disorders. LEVEL OF EVIDENCE: III, retrospective comparative study. CLINICAL TRIAL REGISTRATION: The study was registered at ClinicalTrials.gov. Identifier: NCT05164809.

Diagnostic Yield of Targeted Hearing Loss Gene Panel Sequencing in a Large German Cohort With a Balanced Age Distribution from a Single Diagnostic Center: An Eight-year Study.

OBJECTIVES: Hereditary hearing loss exhibits high degrees of genetic and clinical heterogeneity. To elucidate the population-specific and age-related genetic and clinical spectra of hereditary hearing loss, we investigated the sequencing data of causally associated hearing loss genes in a large cohort of hearing-impaired probands with a balanced age distribution from a single center in Southwest Germany. DESIGN: Genetic testing was applied to 305 hearing-impaired probands/families with a suspected genetic hearing loss etiology and a balanced age distribution over a period of 8 years (2011-2018). These individuals were representative of the regional population according to age and sex distributions. The genetic testing workflow consisted of single-gene screening (n = 21) and custom-designed hearing loss gene panel sequencing (n = 284) targeting known nonsyndromic and syndromic hearing loss genes in a diagnostic setup. Retrospective reanalysis of sequencing data was conducted by applying the current American College of Medical Genetics and Genomics/Association for Molecular Pathology guidelines. RESULTS: A genetic diagnosis was established for 75 (25%) of the probands that involved 75 causal variants in 35 genes, including 16 novel causal variants and 9 medically significant variant reclassifications. Nearly half of the solved cases (47%; n = 35) were related to variants in the five most frequently affected genes: GJB2 (25%), MYO15A, WFS1, SLC26A4, and COL11A1 (all 5%). Nearly one-quarter of the cases (23%; n = 17) were associated with variants in seven additional genes (TMPRSS3, COL4A3, LOXHD1, EDNRB, MYO6, TECTA, and USH2A). The remaining one-third of single cases (33%; n = 25) were linked to variants in 25 distinct genes. Diagnostic rates and gene distribution were highly dependent on phenotypic characteristics. A positive family history of autosomal-recessive inheritance in combination with early onset and higher grades of hearing loss significantly increased the solve rate up to 60%, while late onset and lower grades of hearing loss yielded significantly fewer diagnoses. Regarding genetic diagnoses, autosomal-dominant genes accounted for 37%, autosomal-recessive genes for 60%, and X-linked genes for 3% of the solved cases. Syndromic/nonsyndromic hearing loss mimic genes were affected in 27% of the genetic diagnoses. CONCLUSIONS: The genetic epidemiology of the largest German cohort subjected to comprehensive targeted sequencing for hereditary hearing loss to date revealed broad causal gene and variant spectra in this population. Targeted hearing loss gene panel analysis proved to be an effective tool for ensuring an appropriate diagnostic yield in a routine clinical setting including the identification of novel variants and medically significant reclassifications. Solve rates were highly sensitive to phenotypic characteristics. The unique population-adapted and balanced age distribution of the cohort favoring late hearing loss onset uncovered a markedly large contribution of autosomal-dominant genes to the diagnoses which may be a representative for other age balanced cohorts in other populations.

Reprotoxic effects of fenpropimorph on the fertilizing potential of AI boars: A case study.

This case study describes the effects of a contamination of boar bedding material with reprotoxic compounds in an AI centre in southern Germany. The origin of the investigations was an extreme decline in the production output of the boars. In July 2021, more than 54% of boars were not in production and over 45% of ejaculates had insufficient sperm quality and quantity, which is a significant drop in comparison with the other months. This drop was accompanied by oligozoospermia (azoospermia), asthenozoospermia and teratozoospermia. Through intensive troubleshooting, the changes could be attributed to fenpropimorph, an ergosterol biosynthesis-inhibiting fungicide with reprotoxic potential, which was found in the sawdust used as bedding as well as in liver samples of affected animals, reaching a concentration (mean ± SD) between 0.20 ± 0.36 mg/kg and 0.019 ± 0.001 mg/kg respectively. Furthermore, autopsy findings revealed hyperaemia of the testis, histologically focal degeneration of the germinal epithelium and signs of reduced spermatogenesis and spermiogenesis.

Live cells are not affected by dead sperm in liquid boar semen: New insights based on a thermo-resistance test.

This study evaluated the effect of different proportions of dead spermatozoa on the quality of liquid boar semen during a thermo-resistance test (TRT). After 3 days of storage (17°C), 54 conventional artificial insemination semen doses (~23 × 106 sperm/ml in ~88 ml of BTS) were split into three 15 ml-treatments (25%, 50%, and 75% dead sperm cells) by mixing two subsamples containing 75% (I) and 0% (II) of live cells. Spermatozoa were evaluated after TRT at 30 (on-test) and 300 min (off-test) incubation at 38°C. At the on-test, treatments of 25%, 50%, and 75% dead sperm cells showed medians for total sperm motility of 77.6%, 50.2%, and 25.6%, respectively. Considering the absolute variation of sperm motility during TRT, doses with 25% dead sperm lost more percentage points (pp) (-9.4 pp) compared to doses containing 50% (-8.2 pp) and 75% dead sperm (-4.5 pp). The lowest loss was observed for doses with 75% dead sperm (p < .01). However, data showed that treatments lost similar proportion of motile cells over the TRT: 25% dead sperm = -11.9%, 50% dead sperm = -16.0%, and 75% dead sperm = -17.5% (p = .31). Regarding the flow cytometry parameters (plasma and acrosomal membrane integrity, mitochondrial activity of cells with intact plasma membrane, high degree of lipid disorder, and apoptotic cells), the absolute variations did not surpass values of -1.8, 3.4, -5.4, and 4.7 pp, respectively. Furthermore, the relative variation suggested that dead sperm did not substantially change their values over the TRT. In conclusion, dead sperm cells did not influence the quality of contemporary live cells during the period and in conditions of a TRT.

α/ß-Hydrolase D16B Truncation Results in Premature Sperm Capacitation in Cattle.

Recently it was shown that a specific form of male infertility in Holstein cattle was caused by a nonsense variant in the α/ß-hydrolase domain-containing 16B (ABHD16B) gene resulting in a protein truncation at amino acid position 218 (p.218Q*) and loss of function. Lipidomics showed that the absence of ABHD16B influenced the content of phosphatidylcholine (PC), ceramide (Cer), diacylglycerol (DAG), and sphingomyelin (SM) in variant carrier sperm membranes. However, the exact cause of infertility in affected sires has remained unclear until now. To elucidate the cause of infertility, we analyzed (i) standard sperm parameters (i.e., total sperm number, morphological intact sperm, total sperm motility), (ii) in vitro fertilizability and effects on early embryonic development, and (iii) sperm survival rates (i.e., capacitation time). The affected spermatozoa showed no changes in the usual sperm parameters and were also capable of fertilization in vitro. Furthermore, the absence of ABHD16B did not affect early embryonic development. Based on these results, it was concluded that the affected spermatozoa appeared to be fertilizable per se. Consequently, the actual cause of the inability to fertilize could only be due to a time- and/or place-dependent process after artificial insemination and before fertilization. A process fundamental to the ability to fertilize after insemination is capacitation. Capacitation is a biochemical maturation process that spermatozoa undergo in the female genital tract and is inevitable for the successful fertilization of the oocyte. It is known that the presence and concentration of certain sperm membrane lipids are essential for the correct course of capacitation. However, precisely these lipids are absent in the membrane of spermatozoa affected by the ABHD16B truncation. Since all other causes of fertilization inability were excluded in the previous experiments, consequently, the only remaining hypothesis was that the loss of function of ABHD16B leads to a capacitation disruption. We were able to show that heterozygous and homozygous affected spermatozoa exhibit premature capacitation and therefore decay before fertilization. This effect of the loss of function of ABHD16B has not been described before and our studies now revealed why sires harboring the variant in the ABHD16B gene are infertile.

[Update on 3D printing in the surgery of musculoskeletal tumors]. / Update 3D-Druck in der Chirurgie muskuloskeletaler Tumoren.

The importance of 3D printing applications in the surgery of musculoskeletal tumors has increased in recent years. Even prior to the era of 3D printing, computer-assisted techniques, such as navigation, have proved their utility. Due to the variable appearance of bone tumors, there is a need for individual solutions. The 3D printing can be used for the development of anatomical demonstration models, the construction of patient-specific instruments and custom-made implants. For these three applications, different regulatory hurdles exist. Especially for the resection of pelvic tumors, 3D printing technologies seem to provide advantages due to the complicated anatomy and the proximity to relevant neurovascular structures. With the introduction of titanium printing, construction of individualized implants that fit exactly into the defect became feasible.

Synovial fluid pH is as specific as synovial leukocyte count but less sensitive for the diagnosis of chronic prosthetic joint infection.

BACKGROUND: Synovial fluid markers (synovial leukocyte count and differential) have been shown to be most accurate in diagnosing prosthetic joint infection (PJI). An inverse correlation for synovial leukocyte count and synovial pH is known assuming that leukocyte metabolism causes synovial fluid acidosis. This study's purpose is to analyze the use of synovial pH as a potential diagnostic marker for PJI. MATERIALS AND METHODS: 92 patients who presented with painful total joint arthroplasty (TJA) of the hip (THA; n = 25) or knee (TKA, n = 67) were prospectively investigated. In our cohort 33% (30/92) had PJI and 67% (62/92) were diagnosed non-infected based on the modified Musculoskeletal Infection Society (MSIS) criteria of 2018. Receiver operating curves and the Youden's index were used to define an ideal cut-off value for synovial pH and the sensitivity and specificity were calculated using cross-tables. Additionally, the sensitivity and specificity were calculated for synovial white blood cell (WBC) count (cut-off > 3000 leukocytes) and percentage of neutrophils (PMN%, cut-off > 80%). RESULTS: The median synovial pH level was significantly lower in the group with chronic PJI compared to implants with aseptic failure (7.09 vs. 7.27; p < 0.001). The calculated optimal cut-off value was 7.11 (AUC 0.771) with a sensitivity of 53% and specificity of 89%. However, the sensitivity and specificity of synovial WBC count were 90% and 88% and for synovial PMN% 73% and 98%, respectively. CONCLUSION: Synovial pH may be a useful adjunct parameter in the diagnosis of chronic PJI after hip or knee arthroplasty, but showed low sensitivity in this preliminary cohort. Future studies with larger numbers are needed. LEVEL OF EVIDENCE: 2a, diagnostic study. Trial registration German Clinical Trials Register (Registration number: DRKS00021038).

Material properties of human vertebral trabecular bone under compression can be predicted based on quantitative computed tomography.

BACKGROUND: The prediction of the stability of bones is becoming increasingly important. Especially osteoporotic vertebral body fractures are a growing problem and an increasing burden on the health system. Therefore, the aim of this study was to provide the best possible description of the relationship between the material properties of human vertebral trabecular bone measured under the most physiological conditions possible and the bone mineral density (BMD) determined by clinical quantitative computed tomography (QCT). METHODS: Forty eight cylindric cancellous bone samples with a diameter of 7.2 mm obtained from 13 human fresh-frozen lumbar vertebrae from 5 donors (3 men, 2 women) have been used for this study. After the specimens were temporarily reinserted into the vertebral body, the QCT was performed. For mechanical testing, the samples were embedded in a load-free manner using polymethylmetacrylate (PMMA). The surrounding test chamber was filled with phosphate buffered saline (PBS) and heated to 37 °C during the test. After 10 preconditioning load cycles, destructive testing was performed under axial compression. After determining the fracture site, BMD has been evaluated in this region only. Regression analyses have been performed. RESULTS: Fracture site had an average length of 2.4 (±1.4) mm and a position of 43.9 (±10.9) percent of the measurement length from the cranial end. No fracture reached the embedding. The average BMD at the fracture site was 80.2 (±28.7 | min. 14.5 | max. 137.8) mgCaHA/ml. In summary the results of the regression analyses showed for all three parameters a very good quality of fit by a power regression. CONCLUSION: The results of this study show that QCT-based bone density measurements have a good predictive power for the material properties of the vertebral cancellous bone measured under near to physiological conditions. The mechanical bone properties of vertebral cancellous bone could be modelled with high accuracy in the investigated bone density range.