Effect of Maca aqueous extract addition to a freezing extender for canine semen.

This study investigated the effect of the addition of Lepidium meyenii (Maca) to the freezing extender on the post-thaw quality of dog semen. Ten canine ejaculates were frozen following a two-step protocol using a tris-glucose-citrate egg yolk extender with or without the addition of 10 µl/mL of aqueous extract of Maca (Maca and ctrl groups, respectively). Prior to (fresh semen) and after freezing (T0) sperm motility, kinetic parameters, viability and mitochondrial membrane potential (MMP), as well as the levels of malondialdehyde (MDA) were evaluated. In addition, sperm motility, kinetic parameters, viability and MMP were examined up to 2 h of incubation of 37 °C after thawing (T1 and T2) to evaluate thermo-resistance. The addition of Maca reduced MDA concentration at T0 (p < 0.05) and increased total motility, the percentage of sperm with medium velocity and WOB at T1. Progressive motility decreased (p < 0.05) at T1 in the ctrl group, whereas it was not affected in Maca group at any time point. In addition, the percentage of hyperactivated spermatozoa remained constant at T1 in the ctrl, while in the Maca group an increase (p < 0.05) of this parameter was recorded. Although no differences were found for MMP between groups at any time points, a decrease of viable sperm with low MMP was observed in ctrl group between T0 and T1 and in Maca group between T1 and T2. The addition of Maca prior freezing reduced the extent of lipid peroxidation and activated canine sperm motility and hyperactivation after thawing.

Effect of crocin supplementation in the extender on the quality of chilled canine semen.

The aim of the study was to evaluate the effects of crocin on canine sperm quality parameters during prolonged storage at 4 °C. Ejaculates from 10 dogs were diluted in a TRIS- egg yolk extender supplemented with 0 (control group), 0.5, 1, and 2 mM crocin and stored at 4 °C. Sperm membrane functional integrity, motility, and kinetics were assessed after 3 h, 24 h, 4 days and 7 days of storage. Based on the results, the more efficient concentration of crocin (0.5 mM) was chosen to evaluate sperm intracellular ROS levels, lipid peroxidation, and DNA fragmentation vs. the control. Semen with the addition of 0.5 mM crocin with respect to the control exhibited: i) increased (P < 0.05) sperm membrane functionality at 4 and 7 days of storage; ii) higher (P < 0.05) average path (VAP), straight-line velocities (VSL), and beat cross frequency (BCF) at 4 d of storage at 4 °C; iii) decreased (P < 0.05) intracellular ROS levels after 3 and 24 h storage. No differences in lipid peroxidation and DNA fragmentation were recorded between the control and C0.5 groups at any time point. Lipid peroxidation did not increase over time, while DNA fragmentation increased (P < 0.05) in both groups after 4 days of storage. The results demonstrated that the enrichment of extender with crocin improves to a certain extent canine semen quality, particularly after 4 days of storage at 4 °C.

Assessment of an open-access CASA software for bovine and buffalo sperm motility analysis.

The aim of this study was to verify the reliability of an open access CASA software (BGM) to evaluate the sperm motility of cattle and buffalo, comparing motility and kinematic parameters to those of a commercial one (HTM). Thirty frozen-thawed samples for each species were analyzed with both HTM and BGM, after 1 h of incubation at 37 °C. Sperm viability and mitochondrial membrane potential (MMP) were evaluated through flow cytometric analysis. Agreement of all motility variables between the two systems was assessed. Correlation analysis was performed to identify relationships between motion parameters and sperm viability and MMP. Bland Altman analysis showed good agreement between methods for all motility parameters except for curvilinear velocity (VCL) in cattle, and for average path (VAP), VCL and (amplitude of lateral head displacement) ALH in buffalo, that showed a proportional bias (P > 0.05). In both systems, positive correlation between both viability and high MMP and total and progressive motility of cattle spermatozoa were found; viability and the sperm with high MMP were positive correlated only with VAP, straight-line (VSL), VCL and ALH evaluated with HTM system. Different results were found for buffalo sperm motility parameters, since viability had positive correlations and mitochondrial activity negative ones. Results suggested that motility assessment performed by these two systems are comparable. The discrepancy of VCL, VAP, and ALH could be due to the difference in the algorithms between software. The open-access CASA plug-in is a reliable alternative to the expensive commercial CASA system for sperm motility assessment in cattle and buffalo.

The Use of Virtual-Problem-Based Learning during COVID-19 Pandemic Emergency: Veterinary Students' Perception.

The COVID-19 global pandemic emergency forced us to replace the "traditional and in presence" clinical, pre-graduating, veterinary medical training with clinical virtual-problem-based learning (v-PBL). This prospective cross-sectional case-control study aimed to evaluate the students' perception of the v-PBLs compared to the traditional veterinary clinical training (t-VCT). The t-VCT consisted of supervised management of clinical cases admitted at the Veterinary Teaching Hospital and in the field. The v-PBL consisted of genuine clinical cases shared by tutors throughout an online platform. A survey was delivered to all the fifth-year students who completed the t-VCT or the v-PBL. The survey was completed by 49% of the students. Overall student satisfaction regarding the training experiences was high in both groups, but it was less in the v-PBL than in the t-VCT group. The students of the v-PBL group perceived that they could not improve their practical clinical skills through online sessions, and they emphasized how it could be employed as support for traditional practical activities. All the students are satisfied with the supervision and considered the training correctly focused on relevant learning objectives and the task clearly explained. Stimulating the integration of knowledge and lifelong learning skills replicating life experiences the v-PBLs represented an attractive curricular alternative for veterinary education.

Variations of follicular fluid extracellular vesicles miRNAs content in relation to development stage and season in buffalo.

In buffalo (Bubalus bubalis) reproductive seasonality, causing cycles of milk production, is one of the major factors affecting farming profitability. Follicular fluid (FF) contains extracellular vesicles (EVs) playing an important role in modulating oocyte developmental competence and carrying microRNAs (miRNAs) essential for in vitro fertilization outcomes. The aim of this work was to characterize the FF-EVs-miRNA cargo of antral (An) and preovulatory (pO) follicles collected in the breeding (BS) and non-breeding (NBS) seasons, to unravel the molecular causes of the reduced oocyte competence recorded in buffalo during the NBS. In total, 1335 miRNAs (538 known Bos taurus miRNAs, 324 homologous to known miRNAs from other species and 473 new candidate miRNAs) were found. We identified 413 differentially expressed miRNAs (DE-miRNAs) (FDR < 0.05) between An and pO groups. A subset of the most significant DE-miRNAs between An and pO groups targets genes which function is related to the lipid and steroid metabolism, response to glucocorticoid and oestradiol stimulus. Comparison between BS and NBS showed 14 and 12 DE-miRNAs in An-FF-EVs and pO-FF-EVs, which regulate IL6 release and cellular adhesion, respectively. In conclusion, these results demonstrated that the miRNA cargo of buffalo FF-EVs varies in relation to both follicular development and season.

Overnight holding aids in selection of developmentally competent equine oocytes.

The demand for equine in vitro produced embryos has increased over the last decade. The aim of this study was to compare the effects of an extended IVM or a prolonged period before fertilization, including holding time, on equine immature oocyte developmental competence. Oocytes, collected from abattoir-derived ovaries, were divided into 4 groups: H0/24 (n = 165) 0 h holding + standard 24-26 h IVM; H8/36 (n = 160) 8 h holding + 36 h IVM; H20/24 (n = 187) 20 h holding + 24 h IVM; H0/44 (n = 164) 0 h holding + 44 h IVM. Oocytes matured to MII were fertilized by intracytoplasmic sperm injection (ICSI) and cultured for 10 days. The oocyte degeneration rate was higher (P < 0.05) for H20/24 than the other groups (H0/24 38.2 %, H8/36 43.1 %, H20/24 54.5 %, H0/44 32.9 %). Cleavage was higher (P < 0.05) in H20/24 (70 %) compared to H0/24 (45 %) and H8/36 (54 %) but not to H0/44 (63 %). No differences among groups were observed in the number of blastocysts per oocyte. Injected oocytes that reached the blastocysts stage were higher (P < 0.05) for H20/24 (20 %) than H0/24 (7 %) and H0/44 (7 %) but not H8/36 (12 %). For cleaved oocytes, a higher blastocyst rate (P < 0.05) was observed for H20/24 (28 %) than H0/44 (11 %), while H0/24 (15 %) and H8/36 (21 %) were not different from any group (P > 0.05). Timing of blastocyst development was not different among groups. Overnight holding of equine immature oocytes followed by a standard IVM interval may induce a pre-selection of the most competent oocytes thereby improving cleavage and embryo development rates after ICSI.

Effect of Aqueous Extract of Maca Addition to an Extender for Chilled Canine Semen.

Antioxidant supplementation has been proposed as a new strategy to improve the long-term preservation of semen. The aim of this study was to evaluate the effect of Maca supplementation of semen extender on quality-related canine semen parameters during cooling. Ejaculates from nine dogs were cooled for 7 days in the absence (control group) or in the presence of 10, 20 and 50 µL/mL of an aqueous extract of Maca. Sperm were evaluated for sperm viability, motility, DNA fragmentation and lipid peroxidation after 3 h, 24 h, 4 days and 7 days of storage. The addition of 10 µL/mL of Maca preserved sperm DNA and plasma membrane integrity at 3 h and increased sperm curvilinear velocity after 24 h. Treatment with 20 and 50 µL/mL of Maca increased the percentage of hyperactivated sperm after 3 h. Moreover, semen treated with 20 µL/mL of Maca decreased lipid peroxidation at 24 h. A significant reduction of sperm DNA and plasma membrane integrity as well as of kinetics parameters between 3 and 24 h of refrigerated storage with the higher concentration tested was observed. Although Maca was not able to protect canine semen with extended refrigeration storage time, it increased hyperactivation and preserved DNA integrity in short-term storage.

ERAS Is Constitutively Expressed in the Tissues of Adult Horses and May Be a Key Player in Basal Autophagy.

ERas is a new gene of the Ras family found in murine embryonic stem (ES) cells. Its human ortholog is not expressed in human ES cells. So far ERas gene has only been found to be expressed in the tissues of adult cynomolgus monkeys and cattle; however, information about ERAS expression or its potential functions in equine tissues is lacking. This study was performed to investigate whether Eras is an equine functional gene and whether ERAS is expressed in the tissues of adult horses and determine its potential physiological role. Expression of the ERas gene was detected in all examined adult tissues, and the RT-PCR assay revealed ERAS transcripts. Protein expression was also detected by Western blot analysis. Quantitative real time RT-qPCR analysis revealed that different expression levels of ERAS transcripts were most highly expressed in the testis. Immunohistochemically, ERAS was found to be localized prevalently in the plasmatic membrane as well as cytoplasm of the cells. ERAS was a physical partner of activated PDGFßR leading to the AKT signaling. ERAS was found to interact with a network of proteins (BAG3, CHIP, Hsc70/Hsp70, HspB8, Synpo2, and p62) known to play a role in the chaperone-assisted selective autophagy (CASA), which is also known as BAG3-mediated selective macroautophagy, an adaptive mechanism to maintain cellular homeostasis. Furthermore, ERAS was found to interact with parkin. PINK1, BNIP3, laforin. All these proteins are known to play a role in parkin-dependent and -independent mitophagy. This is the first study demonstrating that Eras is a functional gene, and that ERAS is constitutively expressed in the tissues of adult horses. ERAS appears to play a physiological role in cellular proteostasis maintenance, thus mitigating the proteotoxicity of accumulated misfolded proteins and contributing to protection against disease. Finally, it is conceivable that activation of AKT pathway by PDGFRs promotes actin reorganization, directed cell movements, stimulation of cell growth.

Effects of Oral Administration of Lepidium meyenii on Morphology of Mice Testis and Motility of Epididymal Sperm Cells After Tetrahydrocannabinol Exposure.

Background: Tetrahydrocannabinol (THC) administration is associated with testicular damage and reduced semen quality. Oral administration of Lepidium Meyenii (maca) improves spermatogenesis and sperm motility and count and reduces spermatogenic damage. Objectives: The aim of this study was to evaluate the effect of administration of THC, maca, and their combination on testicular tissue and semen parameters. Materials and Methods: Thirty-six-week-old male mice were classified into control, THC, Maca, and THC + Maca groups. The mice were subjected to Eco Color Doppler ultrasound examination of the testicles before and after treatment. After euthanasia, the epididymis, testes, liver, and kidney were collected for histological examination. For morphometry of the testis, tubular diameters and seminiferous epithelium height were measured. Sperm concentration and sperm motilities were assessed. Differences among the groups were assessed using the Kruskal-Wallis and Dunn's post-hoc test. Results: In all the groups, there were no significant changes in testicular morphology before and after treatment. Histological assessment of the testes showed no alterations in control, no significant alterations in Maca, mild to moderate alterations in THC, and mild alterations in THC + Maca groups. Histological examination of the other organs showed no significant differences among the groups. Tubular diameter showed significantly increased thickening for THC and THC + Maca compared with that for Maca and control. Moreover, seminiferous epithelium height decreased for THC compared with that in the control, Maca, and THC + Maca groups. No statistically significant reduction in the spermatogenic index was observed for THC compared with that for Maca and THC + Maca. Epididymal cross-sections of the groups showed no significant alterations. Sperm concentration and motility were higher for control and THC + Maca groups than in group THC and Maca. Conclusion: In vivo maca administration reduced the deleterious effect of THC on testicular parenchyma and semen production.

Transabdominal ultrasonographic measurement of caudal vena cava to aorta derived ratios in clinically healthy neonatal foals.

BACKGROUND: Ultrasonographic measurement of the vena cava and aorta (Ao) diameters and their ratios have been suggested to be a reliable way of quantifying hypovolemia. OBJECTIVE: To evaluate the feasibility and reliability of an ultrasonographic technique for measurement of Ao and caudal vena cava (CVC) and derived ratios using three different acoustic windows in a population of healthy neonatal foals. Correlation between Ao and CVC measurements and ratios and foals' age or bodyweight were also investigated. METHODS: In 14 healthy foals aged less than 7 days, the diameters of the Ao and of the CVC in long and short axis were measured by two observers from images obtained using three different ultrasonographic imaging planes (left dorsal, left ventral and right views). The Ao and CVC cross-sectional area and the CVC/Ao diameter and area ratios were calculated. Image quality was subjectively assessed. Intraobserver and interobserver reliabilities for image quality scores and measurements were evaluated between the two observers. Simple linear regression models were used to identify correlations between the CVC/Ao measurements and ratios and the age and bodyweight of the foals. RESULTS: The left ventral view showed the highest reliability. A correlation between bodyweight and the short axis measurement of the CVC was found (R2  = 0.385; p = 0.018). Age was positively correlated with the long axis of measurement of the CVC (R2  = 0.426; p = 0.011) and CVC/Ao diameter ratio (R2  = 0.625; p = 0.001). CONCLUSIONS: The left ventral view allows the Ao and CVC cross sections to be easily visualized and measured in neonatal foals in right lateral recumbency.

The Use of Platelet-Rich Plasma for Treatment of Tenodesmic Lesions in Horses: A Systematic Review and Meta-Analysis of Clinical and Experimental Data.

The use of platelet-rich plasma (PRP) to enhance tenodesmic lesion healing has been questioned over the years. The aim of this study was to evaluate current literature to establish the effectiveness of PRP for treating tenodesmic lesions through a systematic review, in accordance with the PRISMA guidelines, and a meta-analysis. Studies comparing PRP with placebo or other treatments for horses with tenodesmic injuries or evaluated PRP effect on tendon and ligament explants were included. Outcomes were clinical, ultrasound, histologic, molecular evaluation, and adverse effects. Two authors independently extracted data and assessed each study's risk of bias. Treatment effects were evaluated using risk ratios for dichotomous data, together with 95% CI. Data were pooled using the random-effects model. The quality of the evidence for each outcome was assessed using GRADE criteria. Twenty-four trials met inclusion criteria for systematic review, while fifteen studies were included in the meta-analysis. Results showed no significant differences in the outcomes between PRP and control groups. Finally, there is no definitive evidence that PRP enhances tendons and ligaments healing. Therefore, there is a need for more controlled trials to draw a firmer conclusion about the efficacy of PRP as a treatment for tenodesmic lesions in the horse.

DNA fragmentation and morphometric studies in sperm of stallions supplemented with maca (Lepidium meyenii).

The reproductive performances of livestock play an essential role in the economic management of the farm. The improvement of semen quantity and quality through the use of food supplements that lack substances which are forbidden in animal feeding, or that may have detrimental effects, is an important goal. Maca (Lepidium meyenii) is a plant that has been used for centuries in the Andes for nutrition and fertility enhancement in humans and animals. The aim of this study was to evaluate the effects of food supplementation of stallions with maca during the breeding season on spermatozoa parameters such as DNA fragmentation and shape, which are two predictive indexes of spermatozoa functionality. For this purpose, ejaculate volume, semen gel-free volume, sperm concentration and motility, total sperm count, sperm DNA fragmentation and sperm head parameters (length, width, perimeter, area, shape factor, roughness) were measured in four stallions. Maca food supplementation in stallions during breeding reduced the percentage of spermatozoa with fragmented DNA, increased significantly sperm concentration and exerted an elongation of the spermatozoa head, a condition that is believed to improve spermatozoa functionality, suggesting that food supplementation of maca could be useful in horse breeding during the breeding season.

Venous blood gas parameters, electrolytes, glucose and lactate concentration in sick neonatal foals: Direct venipuncture versus push-pull technique.

BACKGROUND: Blood collection by indwelling intravenous catheter (IVC) avoids repeated venipuncture, which could cause thrombophlebitis risk, anxiety and pain in patients. OBJECTIVES: To compare blood gas parameters, electrolytes, glucose, lactate and haematocrit concentration obtained from venous blood samples collected via a jugular IVC by push-pull (PP) technique to those obtained by venipuncture in hospitalised foals, at the time of catheter placement (T0) and 24 hours after the beginning of intravenous therapy (T24). STUDY DESIGN: Prospective observational study. METHODS: Paired blood samples were drawn from hospitalised foals at T0 and T24. In each foal, one venous blood sample was collected via IVC by the following PP technique: 2.4 mL of blood was aspirated and immediately reinfused through the catheter three times consecutively, then 1 mL of blood was collected using a 1 mL heparinised syringe. Thereafter, another sample was collected by direct venipuncture of the contralateral jugular vein, with an identical 1 mL heparinised syringe, with a 1-inch, 20-G needle. All samples were analysed with an automated blood gas analyser within 10 minutes of collection. The agreement between the two techniques was assessed by Bland-Altman analysis and intraclass correlation coefficient (ICC). RESULTS: The level of agreement of blood gas values obtained by the two different techniques was high with very small bias and clinically acceptable ICC (>0.907 at T0; >0.794 at T24) for all variables, except for haematocrit (bias -3.52 at T0; -2.44 at T24) and PvO2 at T0 and T24 (ICC 0.669 and 0.733, respectively). MAIN LIMITATIONS: Potential sub-clinical catheter-related complications were not investigated by ultrasound or bacterial culture of the catheter; short duration of the study. CONCLUSIONS: PP technique appears to be acceptable for collection of blood samples for venous blood gas parameters, as well as electrolytes, glucose and lactate in sick neonatal foals.

Correlation Between Serum Activity of Muscle Enzymes and Stage of the Estrous Cycle in Italian Standardbred Horses Susceptible to Exertional Rhabdomyolysis.

Equine exertional rhabdomyolysis (ER) is a well-recognized clinical syndrome affecting racehorses. Prevalence analysis of ER showed that female sex was a significant risk factor. The aim of this research was to evaluate the differences and correlations in the serum activity of muscle enzymes and the stage of the estrous cycle in ER-susceptible and control (C) mares. Serum muscle enzyme activity before and after exercise and sex hormones were analyzed in the two groups of mares. Ten cyclic ER and 10 cyclic C mares were examined weekly for 4 weeks. During diestrus, ER horses had significantly higher resting and postexercise aspartate aminotransferase (AST) activity, but not creatine kinase (CK) activity, compared with controls; only postexercise AST activity was significantly higher during estrus compared with activity levels in controls. During estrus, 17ß-estradiol and AST activity were significantly negatively correlated in the control but not ER mares. Based on our results, further studies should be performed to characterize the presumptive different roles played by sexual hormones in horses susceptible to ER compared with healthy mares.

"Real-time" Assessment of Surgical Margins During Radical Prostatectomy: State-of-the-Art.

Histopathologic examination of the pathologic specimens using hematoxylin & eosin stains represents the backbone of the modern pathology. It is time-consuming; thus, "real-time" assessment of prostatic and periprostatic tissue has gained special interest in the diagnosis and management of prostate cancer. The current study focuses on the review of the different available techniques for "real-time" evaluation of surgical margins during radical prostatectomy (RP). We performed a comprehensive search of the Medline database to identify all the articles discussing "real-time" or intraoperative assessment of surgical margins during RP. Several filters were applied to the search to include only English articles performed on human subjects and published between January 2000 and March 2019. The search revealed several options for pathologic assessment of surgical margins including intraoperative frozen sections, confocal laser endomicroscopy, optical spectroscopy, photodynamic diagnosis, optical coherence tomography, multiphoton microscopy, structured illumination microscopy, 3D augmented reality, and ex vivo fluorescence confocal microscope. Frozen section represents the gold standard technique for real-time pathologic examinations of surgical margins during RP; however, several other options showed promising results in the initial clinical trials, and considering the rapid development in the field of molecular and cellular imaging, some of these options may serve as an alternative to frozen section.

Ultrasonographic assessment of normal jugular veins in Standardbred horses.

BACKGROUND: Ultrasonography (US) is the recommended imaging technique to evaluate jugular veins. This prospective randomized clinical study was designed to collect a series of B-mode US measurements of manually distended jugular veins in healthy Italian Standardbreds and to find possible correlations between ultrasound measurements and animal morphometric characteristics. Forty-two horses, eight males and 34 females (range 3-22 years; bodyweight 494.4 ± 41.7 kg), were included in the study. The diameters and wall thicknesses of both jugular veins were measured at three different sites of the neck. The differences in ultrasound measurements based on scans, age, gender, side, and site of the neck were evaluated by ANOVA or by the Kruskal-Wallis test. The effects of the morphometric measures on each ultrasound parameter were evaluated by MANOVA (P < 0.05). RESULTS: The ultrasound measurements did not differ significantly between the three different sites or between genders; hence, they were pooled together in the results. On the transverse scan, the mean dorsoventral and lateromedial diameters were 1.58 ± 0.23 and 2.20 ± 0.25 cm, respectively; the mean superficial and deep wall thicknesses (SWT and DWT) were 0.07 ± 0.01 and 0.08 ± 0.01 cm, respectively. On the longitudinal scan, the mean dorsoventral diameter was 1.59 ± 0.26 cm, and the SWT and DWT were both 0.08 ± 0.01 cm. Neck length, from the caudal edge of the mandible to the thoracic inlet, was related to the dorsoventral diameter in both longitudinal and transverse scan and to the SWT and DWT in transverse scan, whereas height at the withers (measured with tape) and estimated weight were related to the wall thickness. Dividing the subjects into groups by age in years ("young" 3-7, "mature" 8-14, "old" > 14), differences were found for the lateromedial diameter in the transverse scan and the SWT on the longitudinal scan. The main limitation of this study was that only one operator performed the measurements. CONCLUSION: The US measurements of the jugular veins and their relationship with morphometric measures reported in this manuscript might be considered as guidelines both for early diagnosis and monitoring jugular vein abnormalities in healthy Italian Standardbred horses.

Combined addition of superoxide dismutase, catalase and glutathione peroxidase improves quality of cooled stored stallion semen.

During cold storage stallion spermatozoa experience undergo oxidative stress, which can impair sperm function and fertilizing capacity. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) are the main endogenous enzymatic antioxidants in stallion seminal plasma, and counteract reactive oxygen species. Semen dilution reduces the endogenous antioxidant concentrations. The aim of this study was to investigate whether addition of 15 IU/mL each of SOD, CAT, and GPX to diluted stallion semen would ameliorate a reactive oxygen-mediated decrease in semen quality during 72 h of storage at 5 °C. Ejaculates (n = 7) were divided in two aliquots and diluted in INRA 96 without (control) or with addition of antioxidants. Semen analysis was performed at the time of dilution and every 24 h during chilled storage. Antioxidant supplementation completely inhibited the storage-dependent increase in activated caspase 3 (P < 0.05). Concomitantly, the antioxidant-supplemented samples had a greater percentage of viable, motile and rapidly moving sperm than control samples after 72 h storage (P < 0.05). The DNA damage, as evaluated by TUNEL assay and SCSA, increased with storage time (P < 0.05). Antioxidant supplementation did not prevent, but did significantly reduce the increase in DNA strand breakage. The results indicate part of the intrinsic apoptotic pathway leading to effector caspase activation was inhibited, although an activation of molecules with endonuclease activity still occurred. In conclusion, adding equal concentrations of SOD, CAT and GPX to a semen extender suppressed caspase-3 activation and improved preservation of stallion sperm motility and viability during 72 h of storage at 5 °C.

Quick assessment of cell-free DNA in seminal fluid and fragment size for early non-invasive prostate cancer diagnosis.

BACKGROUND: Liquid biopsy consists in the quantification and qualification of circulating cell-free DNA (cfDNA) and tumor-derived DNA (ctDNA) for cancer recognition. Recently, the characterization of seminal cfDNA (scfDNA) has been reported as a possible biomarker for prostate cancer (PCa) diagnosis. METHODS: Thirty patients with histologically proven PCa, 33 with benign prostate hyperplasia (BPH) and 21 healthy controls were enrolled. cfDNA was extracted from seminal fluid samples. cfDNA quantification and analysis were performed using Qubit ssDNA Kit and Agilent 2100 Bioanalyzer. Statistical analysis included: Levene's test, Shapiro-Wilk, Kolmogorov-Smirnov and Kruskal Wallis tests. RESULTS: Median cfDNA was significantly higher in PCa patients 428.45 ng/mL (173.93-1159.62) compared to BPH patients 77.4 ng/mL (18.23-501) and healthy controls 25.4 ng/mL (15.37-76.62). scfDNA fragments longer than 1000 base-pairs were more common in patients with PCa compared to those with BPH and controls. CONCLUSIONS: scfDNA concentration and fragment size differed significantly in the three groups of PCa, BPH and healthy controls. Both parameters are potential clinical biomarkers for PCa and can be used in both early diagnosis and follow-up. Using automated systems for high-throughput cfDNA quantification could improve the reproducibility of the method and facilitate the implementation of liquid biopsies in the clinical setting.

Chemical Analysis of Lepidium meyenii (Maca) and Its Effects on Redox Status and on Reproductive Biology in Stallions †.

The present study was conducted to assess the chemical composition of Yellow Maca (Lepidium meyenii) and its biological activity on stallions following oral administration of hypocotyl powder. Maca was subjected to methanolic extraction and the chemical analysis was carried out by LC-MS-QTOF (liquid chromatography-mass spectrometry). Our results showed that Maca contains some effective antioxidants, a high percentage of glucosinolates, and other important components with a high antioxidant capacity. To evaluate the plant biological activity in stallions fed with Maca powder for 60 days, the redox status and some reproductive parameters were investigated. Blood and semen samples were collected at 0, 30, 60, and 90 days from the beginning of this study. Blood samples showed a decrease of the reactive oxygen metabolites, evaluated by d-ROMs test, and an increase of the antioxidant barrier in terms of biological antioxidant potential (BAP test), powerful oxidant capacity (OXY-Adsorbent test), and thiols evaluation (-SHp test). Furthermore, semen samples showed a positive trend during Maca administration in the following parameters: ejaculate volumes and sperm concentrations, total and progressive motility, and acrosome integrity.

Reliability of the different versions of Partin tables in predicting extraprostatic extension of prostate cancer: a systematic review and meta-analysis.

INTRODUCTION: Accurate prediction of extraprostatic extension (EPE) of prostate cancer (PCa) is the keystone for deciding whether to perform a neurovascular bundle sparing (NVBs) radical prostatectomy or not, which will subsequently affect the postoperative functional outcomes especially potency. Partin tables are the most commonly used predictive tools (PTs) for prediction of EPE. Moreover, they are the most commonly externally validated. In these settings, the aim of our work is to perform a systematic review of the literature and a meta-analysis for the discriminative performance of the different versions of Partin tables for EPE prediction. EVIDENCE ACQUISITION: A systematic search of Medline, Scopus and Cochrane library was performed to include all the external validation (EV) studies that reported the discriminative performance (area under the curve [AUC]) of the different versions of Partin tables as a PT for EPE. Different versions of Partin tables (1997, 2001, 2007, 2010, and 2013) were included in separate meta-analyses. The pooled AUC with 95% CI were calculated to determine the weighted summary AUC using the random effect model. EVIDENCE SYNTHESIS: Twenty-six studies carried out in different countries including the USA, Korea, Germany, Ireland, China, Austria, France, Italy, the UK, and India were included. Considering the small number and generally low quality of the EV studies in literature, most of the included studies showed some sort of bias especially in the sample size & missing data domain. The pooled EPE AUC were 0.642 (95% CI; 0.601-0.682), 0.672 (95% CI; 0.617-0.727), 0.659 (95% CI; 0.623-0.695), 0.669 (95% CI; 0.623-0.715) and 0.644 (95% CI; 0.545-0.742) for the 1997, 2001, 2007, 2010 and 2013 versions, respectively. CONCLUSIONS: Despite being the most commonly used predictive tool for prediction of EPE, the pooled EPE AUC for different versions of Partin tables showed poor discriminative performance. Thus, surgeons must be cautious when referring to Partin tables for prediction of EPE. Further EV studies are required to confirm these results.