Update of the cooling protocol for antibiotic-free storage of boar semen at 5°C improves sperm quality and maintains low bacterial counts.

Preserving boar semen at 5°C instead of the conventional storage temperature of 17°C would enable a reduction of antibiotic use in pig insemination. To protect the chilling-sensitive boar spermatozoa, holding the extended semen at a higher temperature before cooling could be beneficial and facilitate the implementation of the innovative preservation concept in practice, provided that bacterial growth is kept at a low level. The aim of this study was to introduce a holding time (HT) at 17°C before cooling and to examine the effect on sperm quality and bacterial growth compared to the original cooling protocol for antibiotic-free 5°C semen storage. A series of experiments with semen doses from eight boars extended in Androstar® Premium without conventional antibiotics revealed that sperm kinematics and the integrity of sperm plasma membranes and acrosomes were improved with HT between 16 and 24 h followed by delayed cooling with 0.04°C/min when compared to the original protocol for semen preservation at 5°C (p < 0.05). Both a shorter HT of 6 h and a faster cooling rate of 0.07°C/min reduced sperm quality (p < 0.05). The HT for 24 h did not compromise the inhibitory effect on bacterial growth during long-term semen storage at 5°C, not even in semen doses spiked with Serratia marcescens. In conclusion, semen storage at 5°C with the modified cooling protocol improved sperm quality and is antimicrobially efficient. It thus presents a ready-to-use tool for a reduction or replacement of antibiotics in pig insemination.

Storage of boar semen at 17°C without conventional antibiotics in an extender containing an organic bactericidal substance.

Introduction: Facing the global threat of antimicrobial resistance, the reduction of antibiotic use in semen extenders is a main goal in artificial insemination (AI) of pigs. The aim of this study was to investigate the potential of a commercial extender containing an organic bactericidal supplement in the absence of conventional antibiotics to control bacterial growth and to maintain the quality of boar spermatozoa during long-term semen storage for up to 144 h at 17°C. Methods: Semen from 233 boars housed at 16 European AI centers was split and diluted in the long-term extender "Androstar Plus without antibiotics + organic bactericidal supplement" (APlus) and in the control extender Beltsville Thawing Solution (BTS) with gentamicin, which is routinely used in many AI centers. Sperm motility was assessed with computer-assisted semen analysis (CASA) and membrane integrity was evaluated with flow cytometry. The number of bacteria was determined by counting colonies on agar plates. Results: At the end of storage, bacterial counts were ≥ 106 CFU/mL in 10.7% of the APlus and in 0.4% of the BTS samples. At the same time, bacterial counts were only weakly correlated with sperm motility (r = -0.23, p < 0.05), and there was no correlation with sperm membrane integrity (p > 0.05). Among the 12 identified bacterial species in APlus samples, loss of sperm quality was exclusively observed in the presence of >106 CFU/mL Serratia marcescens and Klebsiella oxytoca. Both these bacterial species, despite their known multi-drug resistance and the continuous use of gentamicin in Europe, proved sensitive to this antibiotic, thus indicating an efficient quality assurance program and responsible antibiotic use. Conclusion: Long-term storage of boar semen at 17°C without conventional antibiotics in an extender containing an organic bactericidal supplement is an option if semen samples are regularly tested for the presence of S. marcescens and K. oxytoca, and the source of contamination is eliminated.

Growth Dynamic and Threshold Values for Spermicidal Effects of Multidrug-Resistant Bacteria in Extended Boar Semen.

The aim of this study was first to examine the prevalence of bacteria-associated loss of sperm quality in samples from insemination centers during a seven-year semen monitoring program and, second, to investigate the growth dynamic of four different multidrug-resistant bacterial species and their impact on sperm quality during semen storage. A reduced sperm quality associated with bacterial contamination was found in 0.5% of 3219 of the samples from insemination centers. In samples spiked with Serratia marcescens and Klebsiella oxytoca, bacterial growth by six log levels was seen during storage at 17 °C, causing loss of sperm motility, membrane integrity, membrane fluidity, and mitochondrial membrane potential at >107 CFU/mL (p < 0.05). Storage at 5 °C in the Androstar Premium extender efficiently inhibited their growth. Achromobacter xylosoxidans and Burkholderia cepacia showed limited growth up to two log levels at 17 °C and did not impair sperm quality. In conclusion, spermatozoa tolerate moderate loads of multidrug-resistant bacteria, and hypothermic, antibiotic-free semen storage effectively limits bacterial growth. The constant use of antibiotics in semen extenders should be reconsidered.

Hypermobile Patients With Femoroacetabular Impingement Syndrome Can Be Effectively Treated Utilizing Hip Arthroscopy With Periportal Capsulotomy Closure: A Matched Cohort Analysis Compared to Patients Without Joint Hypermobility.

PURPOSE: To assess the 2-year outcomes of arthroscopic treatment with periportal capsulotomy closure for femoroacetabular impingement syndrome (FAIS) in patients with generalized ligamentous laxity (GLL). METHODS: A retrospective analysis was performed from a prospectively collected database of FAIS patients undergoing hip arthroscopy. FAIS patients with GLL were identified as having Beighton score ≥4. FAIS patients with GLL were treated with arthroscopic labral repair, osteochondroplasty, via periportal capsulotomy with subsequent capsular closure. These patients were matched by age, sex, and body mass index (BMI) with a cohort of FAIS patients without GLL who underwent the same procedure via periportal capsulotomy without capsular closure. Preoperatively, and 2 years postoperatively, patients completed patient-reported outcomes (PRO) scores, including the Hip Disability and Osteoarthritis Outcome Score (HOOS), 12-item Short-Form survey (SF-12) and the visual analog scale (VAS). RESULTS: Forty patients (5 male, 35 female) with FAIS and GLL were included (age: 29.7 ± 9.0; BMI: 23.3 ± 4.1). FAIS patients with GLL demonstrated similar significant PRO score improvements compared to a matched cohort of FAIS patients without GLL at 2 years after surgery (VAS Pain: (-)2.5 ± 3.0, (-)2.7 ± 2.7; SF-12 PCS: 17.7 ± 14.2, 16.7 ± 15.0; HOOS-Symptoms: 26.3 ± 24.0, 20.6 ± 18.1; HOOS-Pain: 29.8 ± 20.4, 24.4 ± 9.0; HOOS-ADL: 24.9 ± 18.4, 22.0 ± 19.9; HOOS-Sports: 43.6 ± 26.1, 33.1 ± 29.8; and HOOS-QOL: 44.2 ± 27.6, 41.7 ± 27.1, respectively). Both cohorts achieved minimal clinically important differences (MCID) for each HOOS subscore at equivalent high rates (70-88%). CONCLUSIONS: Patients with GLL in the setting of FAIS can be effectively treated with arthroscopy via periportal capsulotomy and capsular closure. These patients demonstrate significant improvements in PRO scores at 2 years, similar to normal laxity FAIS patients undergoing arthroscopic treatment via periportal capsulotomy without capsular closure. LEVEL OF EVIDENCE: Level III, retrospective comparative therapeutic trial.

Patient-Reported Outcome Surveys for Femoroacetabular Impingement Syndrome Demonstrate Strong Correlations, High Minimum Clinically Important Difference Agreement and Large Ceiling Effects.

PURPOSE: To determine the correlation between different patient-reported outcome (PRO) measurements used to assess outcomes after arthroscopic treatment of femoroacetabular impingement syndrome (FAIS) in a single cohort of patients. METHODS: Patients undergoing primary hip arthroscopy for FAIS (without dysplasia, arthritis, or joint hypermobility) were retrospectively analyzed from a prospectively collected cohort. PROs collected before surgery and at 2-year follow-up included the Visual Analog Scale (VAS) for pain, modified Harris Hip Score (mHHS), Hip Disability and Osteoarthritis Outcome Scores (HOOS) with subscales for symptoms, pain, activities of daily living (ADL), sport, and quality of life (QoL), and the physical and mental components of the Short Form-12 (SF-12 PCS and SF-12 MCS). RESULTS: Three hundred patients with 2 years' follow-up (mean age 35.1 ± 11.3, BMI 24.7 ± 3.8, 52.7% female, median Tönnis grade 1) were identified. All patients underwent femoroplasty and labral repair. There was a strong correlation among nearly all the PRO surveys at 2-year follow-up, with the highest correlations identified between mHHS and HOOS-Pain (r = .86, P < .001) and mHHS and HOOS-ADL (r = .85, P < .001). Preoperative scores and the change from preoperative to postoperative scores demonstrated an overall moderate correlation between surveys. There was a consistently weak correlation between the SF-12 MCS and all other PROs. There were strong agreements (67%-77%) in the patients achieving minimal clinically important differences (MCID) for each PRO survey. All surveys except the SF-12 demonstrated a ceiling effect after surgery, with 13% to 43% of patients achieving the maximum score. CONCLUSIONS: PRO surveys used for FAIS demonstrate strong correlations, especially in the evaluation of patients during the postoperative period. MCID for VAS, mHHS, and HOOS demonstrate strong agreement, whereas large ceiling effects were seen with the mHHS and HOOS. The results support a more efficient use of PRO scores while being able to accurately capture patient outcomes. LEVEL OF EVIDENCE: IV, retrospective case series.

Antimicrobially Active Semen Extenders Allow the Reduction of Antibiotic Use in Pig Insemination.

Antibiotic use in semen extenders for livestock may contribute to the development and spreading of multi-drug resistance. Antimicrobial control in semen doses for artificial insemination of pigs is indispensable due to the relatively high storage temperature (17 °C). The objectives of this study were first, to examine whether the antimicrobial capacity differs between antibiotic-free extenders and second, to determine whether an antimicrobial active extender provides the possibility to reduce antibiotics. Antibiotic-free semen extenders Beltsville Thawing Solution (BTS) and Androstar Premium were inoculated at 103 to 104 CFU/mL with four pure bacterial strains isolated from boar ejaculates or a mixture thereof, and then stored for 144 h at 17 °C. Bacterial counts after aerobic culture decreased in BTS up to one log level and decreased in Androstar Premium by 2 to 3.5 log levels (p < 0.05). In semen samples from nine boars stored in the inoculated Androstar Premium extender containing half of the standard concentration of gentamicin, bacteria counts were below 101 CFU/mL. Likewise, half of the standard dose of apramycin and ampicillin was fully antimicrobially active and sperm quality was maintained. In conclusion, semen extenders with intrinsic antimicrobial activity allow a reduction in antibiotic use in pig insemination.

The Incidence of Noninvasive Follicular Thyroid Neoplasm with Papillary-Like Nuclear Features: A Meta-Analysis Assessing Worldwide Impact of the Reclassification.

Background: Noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) reclassification has significantly influenced the field of thyroidology. However, the extent of this impact depends upon the incidence of NIFTP in a given population. In this meta-analysis, we aimed to obtain robust information about the actual incidence of NIFTP worldwide by reviewing the published data. Methods: Comprehensive literature search was performed using electronic databases of PubMed and Web of Science over a five-year period (January 1, 2016, to January 30, 2021). The incidence of NIFTP was calculated by dividing the number of NIFTPs by the number of papillary thyroid carcinomas (PTCs). Meta-analysis of proportion and their 95% confidence interval [CI] were pooled using the random-effect model. Heterogeneity across the included studies was assessed using I2 statistics. Egger's regression test and funnel plot of estimates were used to evaluate the publication bias. p-Value <0.05 was considered significant. Results: From 505 publications, we included 50 studies, all retrospective, with 100,780 PTCs and 3990 NIFTP from 92 institutions worldwide. The overall incidence of NIFTP was 6.0% [CI 4.4-8.2] among PTCs or thyroid malignancies with a high level of heterogeneity among the included studies (I2 = 98.6%). NIFTP incidence was largely similar in North America and Europe (9.3% vs. 9.6%), with a significantly lower overall rate in Asia (2.1%). There was a significant decline in the reported incidence of NIFTP in non-Asian studies published after 2017 (p = 0.002). On applying our data on global thyroid cancer statistics, this reclassification would affect ∼30,881 patients annually, with a lower impact in Asia compared with North America and Europe. Conclusions: This comprehensive meta-analysis confirms that the worldwide NIFTP incidence is much lower than estimated initially. The NIFTP rates are significantly lower in Asian compared with North American and European countries. Apart from geography, NIFTP rates are significantly influenced by the nature of study, type of database used for sample collection, and the diagnostic criteria used. Introduction of NIFTP may potentially spare over 30,000 patients worldwide annually from clinical and psychological consequences of a thyroid cancer diagnosis.

Treatment of Labral Calcification in the Setting of Femoroacetabular Impingement Syndrome With Arthroscopic Calcification Excision, Labral Repair, and Osteoplasty Improves Outcomes.

PURPOSE: To describe the diagnosis and 2-year outcomes of arthroscopic treatment for labral calcification in the setting of femoroacetabular impingement syndrome (FAIS). METHODS: A retrospective analysis was performed from a prospectively collected database of patients with FAIS undergoing hip arthroscopy. Patients with FAIS with labral calcification were differentiated radiographically from patients with other paralabral radiopaque densities such as os acetabuli, acetabular rim fractures, and labral ossification. Patients with FAIS with labral calcification were treated with arthroscopic calcification excision, labral repair, and osteoplasty and matched by age, sex, and body mass index with a cohort of patients with FAIS without labral calcifications who underwent labral repair and osteoplasty. Pre- and 2 years postoperatively, patients completed patient-reported outcome (PRO) scores including the modified Harris Hip Score, Hip disability and Osteoarthritis Outcome Score (HOOS), 12-item Short-Form survey, and visual analog scale. RESULTS: In total, 40 hips (21 male, 19 female) with FAIS and labral calcification were included (age 36.8 ± 8.1, body mass index, 25.9 ± 4.5). Patients with FAIS with labral calcification demonstrated similar significant PRO score improvements compared with a matched cohort of patients with FAIS without labral calcification at 2 years after surgery (visual analog scale: (-)2.3 ± 0.4, (-)2.7 ± 0.5, modified Harris Hip Score: 16.1 ± 2.6, 17.1 ± 3.2; HOOS symptoms: 21.9 ± 3.7, 18.6 ± 3.6; HOOS pain: 22.1 ± 3.0, 25.0 ± 3.5; HOOS activities of daily living: 20.2 ± 2.8, 23.8 ± 3.3; HOOS sport: 35.6 ± 5.0, 35.6 ± 4.1; HOOS quality of life: 36.9 ± 4.5, 37.5 ± 4.4; 12-item Short-Form survey physical component summary: 15.5 ± 2.3, 20.1 ± 2.1, respectively). Both cohorts achieved minimal clinically important differences at equivalent rates (60%-82.5%) for all PRO scores. CONCLUSIONS: Patients with labral calcification in the setting of FAIS can be effectively treated with arthroscopic calcification excision, labral repair, and osteoplasty. These patients demonstrate significant improvements in patient-reported outcomes and achievement of minimal clinically important differences at 2 years similar to patients undergoing arthroscopic treatment for FAIS without labral calcification. LEVEL OF EVIDENCE: Level III, matched cohort study.

The interaction between TERT promoter mutation and MGMT promoter methylation on overall survival of glioma patients: a meta-analysis.

BACKGROUND: There are controversial results concerning the prognostic implication of TERT promoter mutation in glioma patients concerning MGMT status. In this meta-analysis, we investigated whether there are any interactions of these two genetic markers on the overall survival (OS) of glioma patients. METHODS: Electronic databases including PubMed and Web of Science were searched for relevant studies. Hazard ratio (HR) and its 95% confidence interval (CI) for OS adjusted for selected covariates were calculated from the individual patient data (IPD), Kaplan-Meier curve (KMC), or directly obtained from the included studies. RESULTS: A total of nine studies comprising 2819 glioma patients were included for meta-analysis. Our results showed that TERT promoter mutation was associated with a superior outcome in MGMT-methylated gliomas (HR = 0.73; 95% CI = 0.55-0.98; p-value = 0.04), whereas this mutation was associated with poorer survival in gliomas without MGMT methylation (HR = 1.86; 95% CI = 1.54-2.26; p-value < 0.001). TERT-mutated glioblastoma (GBM) patients with MGMT methylation benefited from temozolomide (TMZ) treatment (HR = 0.33; 95% CI = 0.23-0.47; p-value < 0.001). MGMT methylation was not related with any improvement in OS in TERT-wild type GBMs (HR = 0.80; 95% CI = 0.56-1.15; p-value = 0.23). CONCLUSIONS: The prognostic value of TERT promoter mutation may be modulated by MGMT methylation status. Not all MGMT-methylated GBM patients may benefit from TMZ; it is possible that only TERT-mutated GBM with MGMT methylation, in particular, may respond.

Fast Starters and Slow Starters After Hip Arthroscopy for Femoroacetabular Impingement: Correlation of Early Postoperative Pain and 2-Year Outcomes.

BACKGROUND: Patients experience varying degrees of pain and symptoms during the early recovery period after hip arthroscopy for femoroacetabular impingement (FAI). Some "fast starters" report minimal discomfort and are eager to advance activities, while "slow starters" describe severe pain and limitations. The relationship between these early postoperative symptoms and 2-year outcomes after hip arthroscopy is unknown. PURPOSE: To analyze the relationship between early postoperative pain and 2-year patient-reported outcomes (PROs) after hip arthroscopy for FAI. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Patients without arthritis or dysplasia who were undergoing primary hip arthroscopy for FAI were prospectively enrolled and completed validated PROs. Scores for visual analog scale (VAS) for pain were collected preoperatively and at 1 week, 6 weeks, and 2 years postoperatively. Scores for the modified Harris Hip Score (mHHS), Hip disability and Osteoarthritis Outcome Score (HOOS), and 12-Item Short Form Health Survey (SF-12) were collected preoperatively and 2 years postoperatively. Paired t tests were used to evaluate PRO score changes, and correlation analyses were used to assess relationships between early postoperative pain and 2-year postoperative outcomes. RESULTS: A total of 166 patients were included (55% female; mean ± SD age, 35.29 ± 9.6 years; mean body mass index, 25.07 ± 3.98 kg/m2). Patients demonstrated significant improvements in PRO scores (VAS, SF-12 Physical Component Score, mHHS, and all HOOS subscales) at 2 years after hip arthroscopy for FAI (P < .001). There was a significant correlation between lower 1-week VAS pain level (fast starters) and lower 2-year VAS pain level (R = 0.31; P < .001) as well as higher 2-year PRO scores (SF-12 Physical Component Score, mHHS, and all HOOS subscales: R = -0.21 to -0.3; P < .001). There was no correlation between 1-week VAS pain and 2-year SF-12 Mental Component Score (P = .17). Preoperative VAS pain levels showed positive correlations with 1-week postoperative pain scores (R = 0.39; P < .001) and negative correlations with 2-year patient outcomes (R = -0.15 to -0.33, P < .01). There was no correlation between 6-week postoperative pain scores and 2-year PRO scores. CONCLUSION: Fast starters after hip arthroscopy for FAI experience sustained improvements in outcomes at 2 years after surgery. Patient pain levels before surgery may delineate potential fast starters and slow starters.

Efficacy and Safety of Crizotinib in the Treatment of Advanced Non-Small-Cell Lung Cancer with ROS1 Rearrangement or MET Alteration: A Systematic Review and Meta-Analysis.

BACKGROUND: Crizotinib has been approved for the treatment of non-small-cell lung cancer (NSCLC) with ROS proto-oncogene 1 (ROS1) gene fusion. This drug has also been granted breakthrough designation for NSCLCs with MET exon 14 alterations. OBJECTIVE: This systematic review and meta-analysis aimed to investigate the efficacy and safety of crizotinib in patients with these diseases. METHODS: We searched PubMed and Web of Science for relevant studies. Meta-analysis of proportions was conducted to calculate the pooled rate of complete response, partial response, stable disease, progressive disease, disease control rate (DCR), objective response rate (ORR), and drug adverse effects (AEs) of crizotinib in NSCLCs with ROS1 rearrangement or MET alterations. RESULTS: A total of 20 studies were included for meta-analysis. Among patients with ROS1-positive NSCLC, crizotinib exhibited a pooled DCR of 93.2% (95% confidence interval [CI] 90.8-95.5) and a pooled ORR of 77.4% (95% CI 72.8-82.1). The median progression-free survival (PFS) and overall survival (OS) of patients in this group was 14.5 and 32.6 months, respectively. For NSCLC with MET alterations, crizotinib was associated with a lower efficacy (DCR 78.9% [95% CI 70.3-87.4] and ORR 40.6% [95% CI 28.3-53.0]). The median PFS was 5.2 months, and median OS was 12.7 months. The most common drug AEs were vision impairment (43.7%), edema (42.9%), and fatigue (40.1%). CONCLUSION: Our study highlighted and confirmed the efficacy of crizotinib in patients with NSCLC with ROS1 or MET genetic alterations. Crizotinib had remarkable effects on advanced NSCLC with ROS1 fusion, as previously reported. However, the role of this targeted therapy in MET-altered NSCLC remains investigational.