Counteracting oxidative stress improves quality of chilled stallion semen

Stallion semen experiences oxidative stress during cooling and transport, and is consequently damaged by reactive oxygen species. Two different approaches have been investigated to enhance the intrinsic antioxidant defense mechanisms against oxidative stress in liquid-preserved semen. First, a dietary antioxidant supplementation to improve antioxidant status in tissue, seminal plasma and spermatozoa was tested. Second, the combined addition of three enzymatic antioxidants to semen extender in order to increase the antioxidant status of seminal plasma was evaluated. In the first experiment, the effect of dietary supplementation with Lepidium meyenii (Maca) on fresh and chilled stallion semen characteristics were evaluated. Maca is a traditional Andean crop used as a nutraceutical for its fertility-enhancing properties which is linked with its antioxidant activity. The diet of five stallions was supplemented daily with 20 mg of Maca powder for a total of 60 days. A control group of another five stallions received the same diet without Maca. Semen was collected once before the administration of Maca (D0), twice during the administration at 30 and 60 days (D30-D60), and finally twice at 30 and 60 days after the end of the administration (D90-D120). Ejaculates were processed for cooled shipping at 5°C and evaluated in the laboratory for total and progressive motility, acrosome integrity and lipid peroxidation after collection and after 24 hours, 48 hours, and 72 hours storage, respectively. Dietary supplementation with Maca significantly improved sperm concentration and total sperm (p<0.05). The beneficial effects of Maca supplementation on motility and acrosome integrity in the raw semen appeared from the end of treatment with Maca (D60) until the end of the study (D120). Furthermore, total motility, progressive motility, and acrosome integrity decline slower during storage in the Maca-treated group than in the control group. The lipid peroxidation did not change during cooling storage in each group and did not show a significant difference between two groups. The results from this study indicated that the dietary supplementation with Maca was able to increase sperm production and in stabilizing semen quality during storage of chilled semen. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxide (GPX) constitute the principal enzymatic components of the endogenous antioxidant system of equine spermatozoa and seminal plasma. The objective of the second experiment was to evaluate the effect of adding a combination of SOD, CAT and GPX to a semen extender on the quality of stallion semen stored at 5°Cfor 72 hours. Ejaculates from seven stallions were split in two aliquots and diluted with semen extender without (control) or with the addition of 15 IU/mL SOD, CAT, and GPX. Semen analysis was performed within 3 hours after semen collection (T0) and every 24 hours during storage of chilled semen (T24, T48, T72). At each time point, total and progressive motility, kinetic parameters of sperm movement, DNA fragmentation and the levels of activated caspase-3 were evaluated. In the first 24 h of storage, almost no difference between control samples and treated samples was evident. After 48 and 72 hours, beneficial effects of a combination of antioxidants became evident. The antioxidant supplementation significantly inhibited the activation of activated caspase 3 and concomitantly maintained total motility and the percentage of rapid moving sperm cells at a higher level (p<0.05). A storage-dependent increase in DNA damage was alleviated only to a minor extend after prolonged storage time, i.e. 72 hours. The results suggest that the tested combination of SOD, CAT and GPX added to a cooling extender improves viability, motility and kinetic features and reduce DNA fragmentation in semen stored for more than 24 hours. In conclusion, a dietary supplementation of stallions with an antioxidant or measures to supplement the semen extender with a combination antioxidants are both valid tools to counteract oxidative stress and maintain the quality of chilled equine semen at a high level for a prolonged time

Endometritis and Infertility in the Mare – The Challenge in Equine Breeding Industry–A Review

Most major infertility problems are complex and several factors can cause failure to produce offspring. In the last few years, much of the efforts of practitioners and researchers working in equine breeding industry have been directed to individuate the pathophysiological mechanisms underlying poor reproductive performances in mares. Endometritis is on the talk in much of the recent research as the most frequent cause of subfertility in mares that cycle normally but do not conceive and in mares that cycle normally and conceive but then suffer early embryonic death. Post-breeding persistent endometritis, bacterial and other infective endometritis and poor uterine clearance have all been discussed in an attempt to define risk factors and a diagnostic algorithm. The aim of this chapter is to perform a thorough review of recent literature about endometritis. The diagnostic algorithms are carefully examined, highlighting pros as well as pitfalls of each diagnostic aid. Suggested therapeutic protocols are examined in the effort to detect what is actually recommended and what would better benefit from further corroboration. The idea that a better etiopathogenetical understanding of the endometritis remains the key to access to a correct diagnostic protocol and to a successful therapeutic plan will inspire this chapter

The atypical receptor CCRL2 (C-C Chemokine Receptor-Like 2) does not act as a decoy receptor in endothelial cells

C-C chemokine receptor-like 2 (CCRL2) is a non-signaling seven-transmembrane domain (7-TMD) receptor related to the atypical chemokine receptor (ACKR) family. ACKRs bind chemokines but do not activate G protein-dependent signaling or cell functions. ACKRs were shown to regulate immune functions in vivo by their ability to scavenge chemokines from the local environment. This study was performed to investigate whether CCRL2 shares two of the main characteristics of ACKRs, namely the ability to internalize and scavenge the ligands. Cell membrane analysis of CCRL2-transfected cells revealed a weak, constitutive, ligand-independent internalization, and recycling of CCRL2, with a kinetics that was slower than those observed with ACKR3, a prototypic ACKR, or other chemotactic signaling receptors [i.e., chemokine-like receptor 1 and C-X-C motif chemokine receptor 2]. Intracellularly, CCRL2 colocalized with early endosome antigen 1-positive and Rab5-positive vesicles and with recycling compartments mainly characterized by Rab11-positive vesicles. CCRL2-transfected cells and activated mouse blood endothelial cells, that endogenously express CCRL2, were used to investigate the scavenging ability of CCRL2. These experiments confirmed the ability of CCRL2 to bind chemerin, the only recognized ligand, but excluded the ability of CCRL2 to perform scavenging. Collectively, these results identify unique functional properties for this member of the non-signaling 7-TMD receptor family

Endometritis associated with Enterococcus casseliflavus in a mare: A case report

Abstract Infectious endometritis is one of the main causes of subfertility/infertility in the mare. In this report, we present the first case of endometritis in mare associated with a strain of Enterococcus casseliflavus, an unusual gram-positive bacterium which can also be a zoonotic agent. Furthermore, the isolated strain showed a worrying multidrug-resistant profile. The accurate finding of a successful antimicrobial treatment and consequently, the pregnancy diagnosis indicate the importance to isolate, identify and define the antibiotic resistance profile of bacteria associated with endometritis

Effectiveness of D-mannose, Hibiscus sabdariffa and Lactobacillus plantarum therapy in prevention of infectious events following urodynamic study

The urodynamic study is an invasive examination that allows a thorough evaluation of the functional activity of the lower urinary tract (bladder, urethra). The execution of urodynamic study exposes the patient to the risk of contracting infections of the lower urinary tract. Prevention of urinary tract infections consists in the avoidance of risk factors and prophylaxis with antimicrobial and non-antimicrobial measures. In this article, we aimed to evaluate the effectiveness of a phytotherapeutic product composed of D-mannose, Hibiscus sabdariffa, and Lactobacillus plantarum in the prevention of infectious events following invasive urodynamic examination.Introduction and aim: The urodynamic study is an invasive examination that allows a thorough evaluation of the functional activity of the lower urinary tract (bladder, urethra). The execution of urodynamic study exposes the patient to the risk of contracting infections of the lower urinary tract. Prevention of urinary tract infections consists in the avoidance of risk factors and prophylaxis with antimicrobial and non-antimicrobial measures. In this article, we aimed to evaluate the effectiveness of a phytotherapeutic product composed of D-mannose, Hibiscus sabdariffa, and Lactobacillus plantarum in the prevention of infectious events following invasive urodynamic examination. Materials and methods: We selected 100 female consecutive patients (age 19-87 years) that underwent urodynamic invasive procedure. We prescribed 14-day therapy with D-mannose, H. sabdariffa, and L. plantarum to these 100 patients after urodynamic invasive test. After that, we have examined urine cultures and urinary symptoms through telephone interviews and hospital outpatient visits to evaluate the possible presence of urinary tract infections. Results: Urinary tract infections were observed through urine culture in 13% of the selected patients; of the 13 urinary tract infections, 9 were observed in patients with urinary tract recurrent infections in the last year and the other 4 cases were associated with patients with no urinary infection in the last year. Three women reported some urinary symptoms and underwent antibiotic therapy. The other 10 cases were classified as asymptomatic bacteriuria. Three cases were related to patients presenting with pelvic organ prolapse. Eight cases were instead related to significant post-voiding residue (&gt;100). Six patients with urinary tract infection reported constipation in the last year. All patients completed the prescribed therapy due to the affordable price of the product and no side effects have been reported. Conclusion: Our study about the administration of D-mannose, H. sabdariffa, and L. plantarum after invasive urodynamic examination underlined how this phytotherapeutic product can reduce the risk of bacteriuria and urinary tract infection in women

Monitoring PD-L1 positive circulating tumor cells in non-small cell lung cancer patients treated with the PD-1 inhibitor Nivolumab

Controversial results on the predictive value of programmed death ligand 1 (PD-L1) status in lung tumor tissue for response to immune checkpoint inhibitors do not allow for any conclusive consideration. Liquid biopsy might allow real-time sampling of patients for PD-L1 through the course of the disease. Twenty-four stage IV NSCLC patients included in the Expanded Access Program with Nivolumab were enrolled. Circulating tumor cells (CTCs) were analyzed by CellSearch with anti-human B7-H1/PD-L1 PE-conjugated antibody. PD-L1 expressing CTCs were assessed at baseline, at 3 and 6 months after starting therapy, and correlated with outcome. At baseline and at 3 months of treatment, the presence of CTCs and the expression of PD-L1 on their surface were found associated to poor patients outcome. Nevertheless, the high frequency of PD-L1 expressing CTCs hampered to discriminate the role of PD-L1 in defining prognosis. Conversely although CTCs were found in all patients 6 months after treatment, at this time patients could be dichotomized into two groups based PD-L1 expression on CTCs. Patients with PD-L1 negative CTCs all obtained a clinical benefit, while patients with PD-L1 (+) CTCs all experienced progressive disease. This suggests that the persistence of PD-L1(+) CTCs might mirror a mechanism of therapy escape

Parentification, distress, and relationship with parents as factors shaping the relationship between adult siblings and their brother/sister with disabilities

According to parentification theory, when the siblings of a brother/sister with disabilities assume parent-like duties, this role reversal is known as sibling-focused parentification. It has a significant impact on these siblings’ distress and the quality of their family relationships; 605 Italian adult siblings (19–26 years) of people with disabilities completed the online survey. Measures of siblings’ parentification, distress, quality of family relationships, social support, and perceived benefits of parentification were used. The hypothesized model aims to test, on the target sample, the distress and the quality of the relationship with parents as mediators on the interplay between the siblings’ parentification and their sibling relationship. Additionally, social support and perceived benefits of parentification as protective factors were considered. Results showed that the distress and the low quality of the relationship with parents negatively affected the interplay between the siblings’ parentification and the relationship with their own brother/sister with disabilities. Social support and the perceived benefits of parentification decreased the siblings’ distress levels; the perceived benefits of parentification served as a protective factor for the quality of the relationship with parents. Current findings extend the knowledge regarding the risk and protective factors of the siblings’ mental health when disability occurs in the family. Additionally, they inform family-based intervention programs, which should involve the whole family system for reducing distress and improving the wellbeing of siblings without disabilities

The atypical receptor CCRL2 (C-C Chemokine Receptor-Like 2) does not act as a decoy receptor in endothelial cells

C-C chemokine receptor-like 2 (CCRL2) is a non-signaling seven-transmembrane domain (7-TMD) receptor related to the atypical chemokine receptor (ACKR) family. ACKRs bind chemokines but do not activate G protein-dependent signaling or cell functions. ACKRs were shown to regulate immune functions in vivo by their ability to scavenge chemokines from the local environment. This study was performed to investigate whether CCRL2 shares two of the main characteristics of ACKRs, namely the ability to internalize and scavenge the ligands. Cell membrane analysis of CCRL2-transfected cells revealed a weak, constitutive, ligand-independent internalization, and recycling of CCRL2, with a kinetics that was slower than those observed with ACKR3, a prototypic ACKR, or other chemotactic signaling receptors [i.e., chemokine-like receptor 1 and C-X-C motif chemokine receptor 2]. Intracellularly, CCRL2 colocalized with early endosome antigen 1-positive and Rab5-positive vesicles and with recycling compartments mainly characterized by Rab11-positive vesicles. CCRL2-transfected cells and activated mouse blood endothelial cells, that endogenously express CCRL2, were used to investigate the scavenging ability of CCRL2. These experiments confirmed the ability of CCRL2 to bind chemerin, the only recognized ligand, but excluded the ability of CCRL2 to perform scavenging. Collectively, these results identify unique functional properties for this member of the non-signaling 7- TMD receptor family

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