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Anethole, an antioxidant found in plants, appears to improve the survival of spermatozoa during semen cryopreservation. This study assessed the effects of commercial trans-anethole in ram semen cryopreservation. Thirty ejaculates from six rams were diluted in media containing anethole at the following concentrations: CONT (0 μM), AN10 (10 μM), AN50 (50 μM), and AN100 (100 μM). Semen was slow-frozen, preserved in liquid nitrogen, and thawed. Anethole at 10 μM or 50 μM did not compromise any studied sperm quality parameter but increased pre-freezing functionality of membrane and mitochondrial activity. At 10 μM, anethole reduced post-thawing spermatozoa lipoperoxidation. At 50 μM, anethole sustained higher mitochondrial activity after thawing, reduced minor defects in sperm, and increased the number of sperm binding to perivitelline membrane, while keeping lipoperoxidation levels as in control. Anethole at 100 μM promoted higher pre- and post-freezing mitochondrial activity and higher number of sperm binding to perivitelline membrane, in comparison to control. Additionally, some post-thawing kinematic parameters were enhanced by anethole at 100 μM. Of note, mitochondrial activity and lipoperoxidation were higher with anethole at 100 μM in comparison to 50 μM, not differing from control. At the hypoosmotic test, the highest concentration (100 μM) tested reduced sperm osmotic resistance. The results of this study indicate that using anethole in cryopreservation media promoted mostly positive effects on the fresh and post-thawed ram semen, and the advantages vary according to its concentration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The present study determined i) the presence of proteins (oviduct-specific glycoprotein, OVGP1; heat shock protein-70A, HSPA1A; heat shock protein-A8, HSPA8; annexin A1, ANXA1; annexin A5, ANXA5; and myosin-9, MYH9) known to be involved in early reproduction in the oviduct fluid (OF) of anestrous goats; and ii) the functional effect of during IVF on polyspermy modulation and embryonic development. In vitro-matured oocytes were co-cultured with spermatozoa (1.0, 2.0, or 4.0 x 10 6  cells/mL) for 18 h in SOF medium supplemented with 5 μg/mL of heparin, 4 μg/mL gentamicin, and 10% estrus sheep serum (CTRL1, CTRL2, and CTRL4 groups) or the same medium plus 10% OF (OF1, OF2, and OF4 groups) obtained from anestrus goats. The analysis of OF by western blotting confirmed the presence of the six proteins tested for. The increase in sperm concentration had no effect (P > 0.05) on the penetration rate in any group; however, monospermy rate decreased as sperm concentration was increased in both OF and CTRL. Regardless of the concentration used, when data were pooled, OF supplementation improved (P < 0.05) monospermy and tended (P = 0.057) to enhance IVF efficiency. Additionally, IVF efficiency was higher (P < 0.05) in OF1 than in OF4 [60 ± 13 vs 37 ± 5%). The development capacity was not affected (P > 0.05) by the sperm concentration and OF treatment, and the average values were cleavage (72 ± 2.6%), blastocyst (37 ± 3.0%), blastocyst in relation to the cleaved (51 ± 4.8%), hatched (62 ± 1.2%), and number of cells per blastocyst (174 ± 1.8%). In conclusion, the six proteins analyzed are present in the OF of anestrous goats, and the supplementation of this OF during IVF may modulate the polyspermy incidence and enhance IVF efficiency, especially when 1x10 6 sperm per mL is used., Competing Interests: Declaration of competing interest None of the authors have any conflict of interest to declare., (Copyright © 2021 Elsevier Inc. All rights reserved.)

This study evaluated the role of progesterone (P 4 ) and medroxyprogesterone acetate (MAP) on the molecular status of immature cumulus-oocyte complexes (COCs) and the implications for oocyte quality in sheep. The number of viable COCs per ewe and the rate of COCs screened for developmental competence by brilliant cresyl blue positive (BCB + ) were similar (P > 0.05), respectively, across treatments (P 4 : 7.7 ± 0.7 and 4.7 ± 1.2; MAP: 5.7 ± 1.0 and 3.5 ± 2.3; and control: 5.7 ± 1.1 and 3.6 ± 2.4). The COCs' gene expression was altered by exogenous progestogens compared with the control group: markers of steroidogenic pathway (FSH receptor [FSHr], LH receptor [LHr], and estradiol receptor α) and of quality (zygote arrest 1, growth differentiation factor 9, and B-cell lymphoma 2) were in abundance in P 4 (P < 0.05). In addition, reelin protein (RELN) was downregulated, and Bcl-2 was upregulated in MAP (P < 0.05). In the P 4 vs MAP comparison, FSHr, LHr, and RELN genes were upregulated (P < 0.05) in the P 4 group. In conclusion, P 4 and MAP promoted dissimilar effects on transcriptome profiling of immature BCB-selected COCs, possibly due to the differences in the chemical structure of progestogens and concentrations of serum P 4 . Exogenous P 4 impacted positively on the profile of genes related to oocyte quality., (Copyright © 2020. Published by Elsevier Inc.)

The effect of short-term administration of medroxyprogesterone acetate (MPA) or natural progesterone (P 4 ) during ovarian stimulation with FSH on oocyte recovery was investigated in Santa Inês ewes. Ewes were treated with an intravaginal sponge containing MPA for 6 d; GnRH was applied 36 h after sponge removal and FSH was given in 3 injections (40, 24, and 16 mg, respectively) every 12 h after (D0, approximate ovulation time). At the first FSH dose, the ewes received either a new MPA sponge (n = 10) or a controlled device for internal release impregnated with P 4 (n = 10) or did not receive any device (n = 10). Ovarian dynamics were assessed every 12 h by transrectal ultrasonography from D-3 to D2. Oocytes were recovered by laparoscopic ovum pick-up (LOPU) on D2 and graded by morphologic quality. The number of small, medium, and large follicles at D0 and D2 (ultrasound examinations), number of both follicles aspirated and oocytes recovered at LOPU, recovery rate, and oocyte grade did not differ (P > 0.05) among treatments. Thus, the short-term use of MPA or P 4 during ovarian stimulation did not affect the first-wave follicle population or morphologic quality of oocytes. We would suggest that, in this protocol, the use of exogenous progestin is unnecessary., (Copyright © 2019 Elsevier Inc. All rights reserved.)

This study examined the effects of bovine oviductal fluid (bOF) obtained during the follicular or luteal phase of the estrous cycle on ram sperm kinematics, capacitation status and plasma membrane (PM) integrity at various time points during the 24-h incubation period. Fresh ram spermatozoa were selected using the swim-up technique and then incubated separately with either follicular phase (FbOF) or luteal phase (LbOF) bovine oviductal fluid added to Fert-TALP medium (positive control - POSControl) or in Fert-TALP medium without capacitating agents (negative control - NEGControl) at 38 °C under 5% CO 2 . Incubation with FbOF or LbOF for 2 h and 4 h promoted an increase (P <  0.05) in most of the sperm motility parameters as compared with the NEGControl group, and bOF-induced changes in sperm kinematics were similar (P >  0.05) to those seen in the POSControl group. After 6 h of incubation, the stimulatory effect of FbOF or LbOF on ram sperm kinematics was no longer observed (P >  0.05). Sperm PM integrity was not affected (P >  0.05) by incubation in bOF-supplemented media or in absence of capacitating factors (NEGControl). Although neither FbOF nor LbOF had any effect on sperm capacitation rates, the proportion of acrosome-reacted spermatozoa was greater (P < 0.05) for bOF-containing media compared with the NEGControl group during the long incubation periods (18 h and 24 h). In conclusion, bOF from either follicular or luteal phase of the estrous cycle enhances ram sperm motility for up to 4 h and the rate of acrosome reaction after long (18-24 h) incubation periods without affecting sperm viability., (Copyright © 2019 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)

This study investigated the effect that bovine oviductal epithelial cell (BOEC) and ovine spermatozoa co-culture exposed to different hormonal environments had on ram sperm function over the course of a 24-h incubation period. Ram cooled-stored spermatozoa were selected by swim-up and then co-cultured separately for 24 h at 38.5 °C under 5% CO 2 with either: (1) Fert-TALP medium (positive control [POSControl]), (2) Fert-TALP medium supplemented with 17β-estradiol (E2) and progesterone (P4) at concentrations similar to follicular phase (Follicular NEGControl), (3) Fert-TALP medium supplemented with E2 and P4 concentrations similar to luteal phase (Luteal NEGControl), (4) BOEC cultured in the same medium as that of the Follicular NEGControl group (Follicular BOEC group), or (5) BOEC cultured in the same medium as that of the Luteal NEGControl group (Luteal BOEC group). The sperm kinematics, capacitation status, and plasma membrane (PM) integrity were evaluated at different intervals. Sperm PM integrity was not affected (P ˃ 0.05) by BOEC co-culture, regardless of the phase of the estrous cycle. After 4 h of incubation, the Luteal BOEC group presented lower (P <  0.05) progressive motility and total motility than the Luteal NEGControl group while the Follicular BOEC group showed lower (P <  0.05) velocimetric parameters and progressive motility than the Follicular NEGControl group. Throughout the incubation period, both BOEC co-culture groups showed a decrease (P <  0.05) in their capacitation rate in comparison to the POSControl group. Conversely, the Luteal BOEC group presented a higher (P <  0.05) non-capacitated rate than both the POSControl and Luteal NEGControl groups. In conclusion, BOEC co-culture with ovine spermatozoa at either the follicular or luteal phase decreases sperm kinematics and delays sperm capacitation., (Copyright © 2018 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)

The present study evaluated the effect of four ovarian stimulation protocols on the follicular population and molecular status of cumulus-oocyte complexes (COCs). Twelve Santa Inês ewes (in a cross-over design) received 80 or 120mg FSH alone in a multiple-dose (MD80 and MD120) regimen or in combination with 300IU equine chorionic gonadotrophin (eCG) in a one-shot (OS80 and OS120) protocol. The follicular population, COC recovery rate, mean COCs per ewe and the rate of brilliant Cresyl blue-positive (BCB+) COCs were similar among treatments (P>0.05). The expression of markers of oocyte competence (ZAR1, zygote arrest 1; MATER, maternal antigen that embryo requires; GDF9, growth differentiation factor 9; BMP15, bone morphogenetic protein 15; Bcl-2, B-cell lymphoma 2; BAX, Bcl-2 associated X protein) and the steroidogenic pathway (ERα, oestrogen receptor α; LHr, LH receptor; FSHr, FSH receptor; STAR, steroidogenic acute regulatory protein) was affected by stimulation. Specifically, the expression of markers of the steroidogenic pathway was reduced with increasing FSH dose in the OS protocol. FSH at a dose of 80mg reduced the expression of FSHr and ERα in the OS versus MD protocol. Conversely, in MD protocol, only LHr was affected by increasing FSH dose. In conclusion, 80mg FSH in the MD or OS protocol was sufficient to promote the development of multiple follicles and obtain fully grown (BCB+) oocytes. The MD protocol may be more appropriate for the production of better-quality oocytes.

This study aims to compare four different methods for selecting high responding sheep donors for in vivo embryo production. These methods include a pre-selection eCG test (eCG), antral follicle count (AFC), plasma anti-Müllerian hormone measurement (AMH) and genotyping for the presence of the FecG E mutation (a polymorphism in the GDF9 gene associated with increased ovulation rate). Santa Ines ewe lambs (n = 25) underwent superovulation (SOV) with 800 IU equine chorionic gonadotropin (eCG) and the corpus luteum (CL) count was recorded by laparoscopy after eight days. At the D0 eCG , blood samples for AMH and genotyping analysis were collected. Twenty-one days after the end of the eCG test, the same animals underwent SOV with 200 mg of FSH, administered in six decreasing doses, and then naturally mated. Immediately before the beginning of the FSH protocol (D0 FSH ), and at the moment of the first FSH dose (D9 FSH ), the AFC was assessed. Plasma AMH was again determined at the D9 FSH . After each screening process, animals were classified as having a high (HR), or low (LR), potential of response (using specific thresholds for each method). Then, the ewes' response to SOV and embryo yield for each screening method, classified as HR or LR, were compared. Animals classified as HR by AFC (HR AFC ) and by AMH concentration (HR AMH ) at the D9 FSH , produced more viable embryos than those classified as LR AFC and LR AMH (HR AFC 6.2 ± 3.2 vs LR AFC 2.8 ± 3.0 and HR AMH 6.6 ± 3.6 vs LR AMH 3.0 ± 2.9). Pre-selection tests with eCG and different FecG E genotypes, either heterozygous (+/E) or wild type (+/+), were unable to discriminate HR or LR animals. A tendency (P = 0.06) to have lower plasma AMH was observed in heterozygous FecG E (+/E) ewes. In conclusion, both AFC and plasma AMH can be used to select donor ewes with a higher potential of response for in vivo embryo production., (Copyright © 2018 Elsevier Inc. All rights reserved.)

This study evaluated the effect of two doses of prostaglandin at different intervals on reproductive parameters of crossbred ewes. In Experiment 1, 30 ewes received two doses of 120 μg cloprostenol at 7 (G 7 days ), 9 (G 9 days ), or 11.5 (G 11.5 days ) days apart. Ultrasound assessments were performed from the first and second cloprostenol administration for 5 days or ovulation detection. Estrus signs were checked by a teaser male. Plasma progesterone concentration was measured before each cloprostenol dose. In Experiment 2, 95 ewes were allocated into the same treatments and after the second dose, ewes in estrus were mated. At 30 days after breeding, pregnancy diagnosis was conducted and prolificacy was evaluated at lambing. In Experiment 1, at the first cloprostenol administration, 50% of ewes had an active CL and all showed estrus. At the second administration, 66.7% of ewes had an active CL and one did not present estrus. There was no difference (P > 0.05) after the second dose for as follows: overall estrous response (90%), interval from cloprostenol administration to estrous onset (42.0 ± 4.9 h), estrus duration (31.5 ± 2.1 h), ovulation rate (100.0%), and number of ovulations (1.5 ± 0.3). In Experiment 2, both pregnancy and prolificacy rates were similar (P > 0.05) for G 7 days (73.3; 145%), G 9 days (75.9; 125%), or G 11.5 days (75.9; 145%), leading to an overall pregnancy rate of 75.0% (66/88) and prolificacy rate of 137% . Therefore, the three treatments proposed were able to promote high pregnancy and prolificacy rates in crossbred ewes.

This study evaluated colour-Doppler ultrasound imaging (UI) as a substitute for laparoscopy to count the corpora lutea (CL) in superovulated sheep. Twenty-five Santa Ines ewes were superovulated three times at 21-day intervals. Corpora lutea were counted by colour-Doppler UI (CL DOPPLER ) 6 days after each superovulation and confirmed by laparoscopy (CL LAP ) 12 hr later. The mean number of CL was similar for both techniques (2.1 ± 2.5 vs. 2.1 ± 2.7 for CL DOPPLER and CL LAP , respectively) with a significant positive correlation (r = .94; r 2 =.89). Colour-Doppler UI effectively evaluated the ovarian response in superovulated ewes and efficiently identified animals that did not respond to superovulation., (© 2017 Blackwell Verlag GmbH.)

This study was designed to evaluate early pregnancy diagnosis using colour Doppler ultrasonography (US) for luteal vascularisation assessment. In Study 1, 28 ewes were artificially inseminated (Day 0), and luteal vascularisation was assessed from Day 12 to Day 19 by two evaluators using colour Doppler US, categorising the corpus luteum (CL) on a subjective scale ranging from 1 to 4. Females bearing a CL with score 2 or greater were presumably considered pregnant. Pregnancy was confirmed on Day 30 by B-Mode US. In Study 2, a predictive pregnancy diagnosis was performed on Day 17 in 197 ewes based on the criteria described in Study 1. Pregnancy was confirmed by B-mode US on Day 45. Agreement between evaluators was verified by an intraclass correlation coefficient (ICC) and Kappa index (κ). Performance of colour Doppler US for early pregnancy diagnosis was evaluated calculating sensitivity (Sens), specificity (Spec), negative predictive values (NPV), positive predictive values (PPV) and accuracy (Ac). In Study 1, luteal vascularisation assessment was unable to predict non-pregnant animals between 12 and 14 days after insemination, as all animals still had vascularised CL, and thus were considered pregnant. The colour Doppler US performance improved progressively until Day 17, when it reached maximum values (Sens = 100%, Spec = 76%, PPV = 73%, NPV = 100% and Ac = 86%). The subjective scale for luteal irrigation assessment showed medium to good agreement among evaluators on Day 12 and Day 13 (ICC = 0.66 and 0.68, respectively), and excellent agreement from Day 14 to Day 19 (ICC = 0.90, 0.80, 0.80, 0.84, 0.95 and 0.93, respectively). Agreement was almost perfect for score 1 CLs (κ = 0.87), and moderate for scores 2, 3 and 4 CLs (κ = 0.54, 0.48 and 0.41, respectively). In Study 2, performance of colour Doppler US as a tool to predict pregnancy status in ewes on Day 17 post-insemination was as follows: Sens = 93.5%, Spec = 80.8%, PPV = 85.6%, NPV = 91.1% and Ac = 87.8%. Subjective luteal vascularisation assessment using colour Doppler US to distinguish between pregnant and non-pregnant animals was considered a reliable tool which was highly efficient beginning 17 days after breeding., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: Primary hyperhidrosis (PH) can lead to mood changes due to the inconveniences it causes., Objective: This study aimed to examine the existence of anxiety and depression in patients with severe primary hyperhidrosis who sought treatment at a medical office., Methods: The questionnaire "Hospital Anxiety and Depression Scale" was used for 197 individuals, in addition to the chi square test and Fisher exact test, p <0.05., Results: There was an increased prevalence of anxiety (49.6%) but not of depression (11.2%) among patients with PH, with no link to gender, age or amount of affected areas. Palmar and plantar primary hyperhidrosis were the most frequent but when associated with the presence of anxiety, the most frequent were the axillary (p = 0.02) and craniofacial (p = 0.02) forms. There was an association between patients with depression and anxiety (p = 0.001)., Conclusions: the involvement of Primary hyperhidrosis was responsible for a higher prevalence of anxiety than that described among the general population and patients with other chronic diseases. Depression had a low prevalence rate, while mild and moderate forms were the most common and frequently associated with anxiety. The degree of anxiety was higher in mild and moderate types than in the severe form.