Optimizing ReBreed21 I: Evaluation of endocrine and ovarian dynamics in non-bred Bos indicus heifers.

The present study evaluated follicular and endocrine dynamics during ReBreed21, a reproductive strategy that allows resynchronization of ovulation every 21 days in Bos indicus (Nelore) heifers. A synchronized estrous cycle was induced using a standard timed ovulation protocol (d -10: P4 implant inserted + 2 mg estradiol benzoate; d -2: P4 removed+ 0.5 mg cloprostenol + 0.6 mg estradiol cypionate + 200 IU equine chorionic gonadotropin (eCG); d0: 8.4 µg buserelin) without AI to ensure nonpregnancy in heifers. Day of GnRH was designated d0 of estrous cycle. On d12, heifers (n = 80) were randomized into three experimental groups: (1) ReBreed21 (n = 28) d12 P4 device inserted, d19 P4 device withdrawal plus 200 IU eCG, and d21 8.4 µg buserelin (GnRH); (2) ReBreed21+G (n = 26) same as ReBreed21 plus GnRH (16.8 µg) treatment on d12; and (3) Control (n = 26) no treatment. ReBreed21+G increased two-fold (62.9%; 18/26) percentage of heifers with synchronized follicular wave emergence compared to Control (34.6%; 9/26) whereas ReBreed21 (53.6%; 15/28) was intermediate. The ReBreeed21 groups (eCG on d19) increased (P < 0.01) follicular growth between d19 and d21 in ReBreed21 (2.3 ± 0.2 mm) and ReBreed21+G (3.4 ± 0.2 mm) compared with Control (1.2 ± 0.3 mm), resulting in greater (P < 0.01) follicle diameter on d21 for ReBreed21 (10.7 ± 0.4 mm) and ReBreed21+G (10.8 ± 0.4 mm) compared with Control (9.1 ± 0.5 mm). Structural luteolysis was similar among groups (P = 0.51), although the average day when P4 was <1 ng/mL was later (P < 0.01) for ReBreed21 (20.5 ± 0.2) and ReBreed21+G (20.7 ± 0.2) compared to Control (19.2 ± 0.4). Overall ovulation at the end of the estrous cycle was increased (P = 0.03) for ReBreed21 groups (83.3%; 45/54) compared with Control (57.7%; 15/26). Synchronized ovulation on day 22-23 was greater (P < 0.01) for ReBreed21 (78.6%; 22/28) and ReBreed21+G (76.9%; 20/26) compared with Control (30.8%; 8/26). Thus, the ReBreed21 resynchronization program produced acceptable endocrine and follicular dynamics, including synchronized ovulation at the end of the protocol in nonpregnant heifers providing good rationale for testing the fertility and practical implementation of this protocol under field conditions.

Profiles of interferon-stimulated genes in multiple tissues and circulating pregnancy-associated glycoproteins and their association with pregnancy loss in dairy cows†.

Pregnancy loss (PL) in lactating dairy cows disrupts reproductive and productive efficiency. We evaluated the expression of interferon-stimulated genes (ISG) in blood leukocytes, vaginal and cervical epithelial cells, luteolysis-related genes, progesterone, and pregnancy-associated glycoprotein (PAG) profiles in lactating dairy cows (n = 86) to gain insight about PL. Expression of ISG on d17, d19, and d21 was greater in cows that maintained the pregnancy (P33) compared to nonpregnant with no PL (NP). Greater ISG differences between groups were observed in the cervix (96.7-fold) than vagina (31.0-fold), and least in blood leukocytes (5.6-fold). Based on individual profiles of ISG and PAG, PL was determined to occur either before (~13%) or after (~25%) d22. For cows with PL before d22, ISG expression was similar on d17 but by d21 was lower and OXTR was greater than P33 cows and similar to NP; timing of luteolysis was similar compared to NP cows suggesting embryonic failure to promote luteal maintenance and to attach to the endometrium (no increase in PAG). For cows with PL after d22, ISG expression was similar to P33 cows on d17, d19, and d21 and luteolysis, when it occurred, was later than NP cows; delayed increase in PAG suggested later or inadequate embryonic attachment. In conclusion, PL before d22 occurred due to embryonic demise/failure to signal for luteal maintenance, as reflected in reduced ISG expression by d21. Alternatively, embryos with PL between d22 and 33 adequately signaled for luteal maintenance (ISG) but had delayed/inadequate embryonic attachment and/or inappropriate luteolysis causing PL.

Extranodal NK/T-Cell Lymphoma, Nasal Type, Extranasal and Ulcerative Blister Variant, Case Report.

The extranodal natural killer (NK) T-cell lymphoma of nasal type is a form of lymphoma that falls within the WHO/EORTC 2018 classification of cutaneous T-cell lymphomas. It is characterized for being aggressive, infrequent, and destroying midline facial structures; however, it can also be in primary or secondary form at extranasal sites, such as the skin or the gastrointestinal tract, among others. We report the case of an 18-year-old patient with an extranodal NK/T-cell lymphoma located in an extranasal site. The clinical presentation is characterized for being multifocal and with erythematous-violaceous plaques that progress to hemorrhagic blisters and necrotic ulcers. Although this type of lymphoma has been reported previously by others, the presence of blisters is an atypical finding, which we believe has been described only in one case in the medical literature.

The impact of chemical pollution on the European eel (Anguilla anguilla) from a Mediterranean hypersaline coastal lagoon.

The European eel (Anguilla anguilla) is a critically endangered species. The impact of environmental contamination on this species has been highlighted as contributing to the decline in recruitment. The Mar Menor hypersaline coastal lagoon (SE Spain) is one of the most productive fisheries of European eel in Europe, making it a critical habitat for species conservation. The present study aimed to provide an initial overview of the impact of organic chemical contaminants on the European eel and the potential sublethal effects of chemical pollution on pre-migrating eels in this hypersaline habitat. We investigated muscle bioaccumulation of main persistent and hazardous organic contaminants (including some current-use pesticides) and genotoxicity, neurotoxicity, and xenobiotic detoxification system responses. The findings show that lagoon eels were exposed to high levels of legacy organochlorine contaminants, recently banned pesticides (chlorpyrifos), and some emerging chemicals. Some individuals surpassed the maximum levels of CBs authorized by the European Commission for human consumption. In this species, residuals of chlorpyrifos, pendimethalin, and chlorthal dimethyl have been reported for the first time. This field study provides relevant data to stock management and human health consumption and provides the first biomarker responses in European eel under permanent hypersaline conditions. Furthermore, the high frequency of micronuclei in peripheral erythrocytes of lagoon eels indicates sublethal genotoxic effects on the organism. Overall, the European eels growing and maturing in the Mar Menor lagoon are exposed to toxic and carcinogenic chemicals. The lack of seafood safety regulations for human consumption for some legacy chemicals that were measured in high concentrations in our study requires special action. Further biomonitoring and research are recommended to protect the animal, public, and environmental health.

Successful treatment of mucosal neuromas by radiofrequency ablation in a patient with multiple endocrine neoplasia type 2B.

This is the first report of mucosal neuromas being treated successfully with radiofrequency ablation in a patient with multiple endocrine neoplasia type 2B.

Temporality of ovarian steroids and LH/FSH pulse profiles encompassing selection of the dominant follicle in heifers†.

The tested hypotheses were (1) LH/FSH pulses and F2 diameter are diminished by P4 and, (2) E2 increases during the transition to deviation and alters LH/FSH pulses. On Day 5 (Day 0 = ovulation), heifers were randomized into an untreated group (HiP4, n = 11), and a prostaglandin analog treated group (NoP4, n = 10). On Day 6, a follicular wave was induced by follicle ablation. Ultrasound and blood collections were performed every 12 h from Days 7 to 11. Blood was collected every 15 min for 10 h on Day 9 (largest follicle expected to be ~7.5 mm). Estradiol was ~75% greater (0.36 ± 0.14 vs 0.63 ± 0.19 pg/mL) in heifers with F1 ≥ 7.2 mm than in heifers with F1 < 7.2 mm. The HiP4 had smaller second largest follicle (F2) diameter, lower estradiol (P = 0.06), LH pulse baseline and peak concentrations (P < 0.007), in addition to half the frequency of LH/FSH pulses (4.1 ± 0.3 vs 9.6 ± 0.7 in 10 h) than the NoP4. Within HiP4, heifers with F1 ≥ 7.2 mm had ~25% fewer (P = 0.03) LH pulses compared to heifers with F1 < 7.2 mm. In contrast, within the NoP4, heifers with F1 ≥ 7.2 mm had ~75% greater LH (P = 0.05) and FSH (P = 0.08) pulse amplitude. We propose that greater F2 diameter at deviation in low P4 is related to greater LH baseline and peak concentrations, and greater frequency of LH/FSH pulses. A greater increase in E2 after F1 reaches ~7.2 mm results in further stimulation of LH/FSH pulse amplitude. Elevated P4 not only diminished frequency of LH/FSH pulses but also converted an E2 increase into a negative feedback effect on LH/FSH pulse frequency leading to smaller F2 at deviation.

Processes involved in prostaglandin F2alpha autoamplification in heifers.

In brief: Endometrial and luteal synthesis of prostaglandin F2alpha (PGF2A) occurs before and during luteolysis and is critical for luteal regression. This study demonstrates that PGF2A stimulates further PGF2A synthesis (autoamplification) apparently from the corpus luteum. Abstract: Understanding the endocrine profile of prostaglandin F2alpha (PGF2A) autoamplification is fundamental to comprehend luteal and endometrial responses to PGF2A. On day 10 of postovulation (preluteolysis), heifers (n = 6/group) were treated intrauterine with saline or PGF2A (0.5 mg; hour 0). A third group received flunixin meglumine + PGF (FM+PGF) to prevent endogenous synthesis of PGF2A. Exogenous PGF2A was metabolized at hour 2 as measured by PGF2A metabolite (PGFM). From hours 5 to 48, concentrations of PGFM were greatest in the PGF group, smallest in the FM+PGF, and intermediate in the control suggesting endogenous synthesis of PGF2A only in PGF group. Progesterone (P4) concentrations decreased transiently between hours 0 and 1 in PGF and FM+PGF groups but rebounded to pretreatment concentrations by hours 6 and 4, respectively. No control or FM+PGF heifers underwent luteolysis during the experimental period. Conversely, in the PGF group, one heifer had complete luteolysis (P4 < 1 ng/mL), two heifers had partial luteolysis followed by P4 and CL resurgence by hour 48, and three heifers did not undergo luteolysis. Endogenous PGF2A appears to be of luteal origin due to the lack of pulsatile pattern of PGFM and lack of endometrial upregulation of oxytocin receptor (typical of endometrial synthesis of PGF2A), whereas luteal downregulation of PGF receptor and HPGD indicates a classic luteal response to PGF2A signaling although other specific mechanisms were not investigated. The hypothesis was supported that a single PGF2A treatment simulating the peak of a natural luteolytic pulse and the uteroovarian transport of PGF2A stimulates measurable endogenous PGF2A production.

The cholesteryl-ester transfer protein isoform (CETPI) and derived peptides: new targets in the study of Gram-negative sepsis.

BACKGROUND: Sepsis is a syndrome where the dysregulated host response to infection threatens the life of the patient. The isoform of the cholesteryl-ester transfer protein (CETPI) is synthesized in the small intestine, and it is present in human plasma. CETPI and peptides derived from its C-terminal sequence present the ability to bind and deactivate bacterial lipopolysaccharides (LPS). The present study establishes the relationship between the plasma levels of CETPI and disease severity of sepsis due to Gram-negative bacteria. METHODS: Plasma samples from healthy subjects and patients with positive blood culture for Gram-negative bacteria were collected at the Intensive Care Unit (ICU) of INCMNSZ (Mexico City). 47 healthy subjects, 50 patients with infection, and 55 patients with sepsis and septic shock, were enrolled in this study. CETPI plasma levels were measured by an enzyme-linked immunosorbent assay and its expression confirmed by Western Blot analysis. Plasma cytokines (IL-1ß, TNFα, IL-6, IL-8, IL-12p70, IFNγ, and IL-10) were measured in both, healthy subjects, and patients, and directly correlated with their CETPI plasma levels and severity of clinical parameters. Sequential Organ Failure Assessment (SOFA) scores were evaluated at ICU admission and within 24 h of admission. Plasma LPS and CETPI levels were also measured and studied in patients  with liver dysfunction. RESULTS: The level of CETPI in plasma was found to be higher in patients with positive blood culture for Gram-negative bacteria that in control subjects, showing a direct correlation with their SOFA values. Accordingly, septic shock patients showing a high CETPI plasma concentration, presented a negative correlation with cytokines IL-8, IL-1ß, and IL-10. Also, in patients  with liver dysfunction, since higher CETPI levels correlated with a high plasma LPS concentration, LPS neutralization carried out by CETPI might be considered a physiological response that will have to be studied in detail. CONCLUSIONS: Elevated levels of plasma CETPI were associated with disease severity and organ failure in patients  with Gram-negative bacteraemia, defining CETPI as a protein implicated in the systemic response to LPS.

Transcatheter tricuspid valve interventions: Current status and future perspectives.

The development of transcatheter therapies to treat valvular heart diseases has changed the rules of the game, particularly in the case of aortic stenosis and mitral regurgitation. In the last years, the tricuspid valve has also been the focus of these percutaneous techniques for several reasons: (1) tricuspid regurgitation is a frequent disease associated with poor clinical outcomes in spite of medical treatment, leading to end-stage right ventricular heart failure, (2) surgical tricuspid valve repair or replacement has been the treatment of choice for patients with tricuspid valve disease, but high mortality rates for isolated surgical tricuspid valve intervention have been reported in the literature, and (3) most patients with tricuspid pathology are ultimately denied cardiac surgery because of their comorbidity burden. Thus, in this context the development of less invasive catheter-based therapies would be of high clinical relevance. The present review provides an overview regarding the framework of chronic tricuspid regurgitation transcatheter therapeutic options, summarizes the transcatheter systems under clinical use or clinical evaluation with their technical features, and describes the updated current evidence in this challenging and evolving field.

Effect of elevating luteinizing hormone action using low doses of human chorionic gonadotropin on double ovulation, follicle dynamics, and circulating follicle-stimulating hormone in lactating dairy cows.

Double ovulation and twin pregnancy are undesirable traits in dairy cattle. Based on previous physiological observations, we tested the hypothesis that increased LH action [low-dose human chorionic gonadotropin (hCG)] before the expected time of diameter deviation would change circulating FSH concentrations, maximum size of the second largest (F2) and third largest (F3) follicles, and frequency of multiple ovulations in lactating dairy cows with minimal progesterone (P4) concentrations. In replicate 1, multiparous, nonbred lactating Holstein dairy cows (n = 18) had ovulation synchronized. On d 5 after ovulation, all cows had their corpus luteum regressed and were submitted to follicle (≥3 mm) aspiration 24 h later to induce emergence of a new follicular wave. Cows were then randomized to NoP4 (untreated) and NoP4+hCG (100 IU of hCG every 24 h for 4 d after follicle aspiration). Ultrasound evaluations and blood sample collections were performed every 12 h for 7 d after follicle aspiration. All cows were then treated with 200 µg of GnRH to induce ovulation. In replicate 2, cows (n = 16) were resubmitted to similar procedures (i.e., corpus luteum regression, follicle aspiration, randomization, ultrasound evaluations every 12 h, GnRH 7 d after aspiration). However, cows in replicate 2 received an intravaginal P4 device that had been previously used (∼18 d). Only cows with single (n = 15) and double (n = 16) ovulations were used in the analysis. No significant differences were detected for frequency of double ovulation, follicle sizes, and FSH concentrations across replicates (NoP4 vs. LowP4 and NoP4+hCG vs. LowP4+hCG), so data were combined. Double ovulation was 40% for control cows with no hCG (CONT) and 62.5% with hCG (hCG). Double ovulation increased as the maximum size of F2 increased: <9.5 mm and 9.5-11.5 mm (7.7%) and ≥11.5 mm (94.1%). The hCG group had more cows with F2 > 11.5 (69%) than with 9.5 ≥ F2 ≤ 11.5 (25%) and F2 < 9.5 (6%). In agreement, F2 and F3 maximum size were larger in the hCG group, but FSH concentrations were lower after F1 > 8.5 mm compared with CONT. In contrast, FSH concentrations were greater before deviation (F1 closest value to 8.5 mm) in cows with double ovulations than in those with single ovulations, regardless of hCG treatment. In addition, time from aspiration to deviation was shorter in cows with double rather than single ovulation and in cows treated with hCG as a result of faster F1, F2, and F3 growth rates before diameter deviation. In conclusion, greater FSH and follicle growth before deviation seems to be a primary driver of greater frequency of double ovulation in lactating cows with low circulating P4. Moreover, the increase in follicle growth before deviation and in the maximum size of F2 during hCG treatment suggests that increased LH may also have a role in stimulating double ovulation.

A Score to Assess Mortality After Percutaneous Mitral Valve Repair.

BACKGROUND: Risk stratification for transcatheter edge-to-edge mitral valve repair (TEER) is paramount in the decision-making process for treating severe mitral regurgitation (MR). OBJECTIVES: This study sought to create and validate a user-friendly score (MitraScore) to predict the risk of mortality in patients undergoing TEER. METHODS: The derivation cohort was based on a multicentric international registry that included 1,119 patients referred for TEER between 2012 and 2020. Score discrimination was assessed using Harrell's c-statistic, and the calibration was evaluated with the Gronnesby and Borgan goodness-of-fit test. An external validation was carried out in 725 patients from the GIOTTO registry. RESULTS: After multivariate analysis, we identified 8 independent predictors of mortality during the follow-up (2.1 ± 1.8 years): age ≥75 years, anemia, glomerular filtrate rate <60 mL/min/1.73 m2, left ventricular ejection fraction <40%, peripheral artery disease, chronic obstructive pulmonary disease, high diuretic dose, and no therapy with renin-angiotensin system inhibitors. The MitraScore was derived by assigning 1 point to each independent predictor. The c-statistic was 0.70. Per each point of the MitraScore, the relative risk of mortality increased by 55% (HR: 1.55; 95% CI: 1.44-1.67; P < 0.001). The discrimination and calibration for mortality prediction was better than those of EuroSCORE II (c-statistic 0.61) or Society of Thoracic Surgeons score (c-statistic 0.57). The MitraScore maintained adequate performance in the validation cohort (c-statistic 0.66). The score was also predictive for heart failure rehospitalization and was correlated with the probability of clinical improvement. CONCLUSIONS: The MitraScore is a simple prediction algorithm for the prediction of follow-up mortality in patients treated with TEER.

Endometrial and luteal responses to a prostaglandin F2alpha pulse: a comparison between heifers and mares†.

In heifers and mares, multiple pulses of prostaglandin F2alpha (PGF) are generally associated with complete luteal regression. Although PGF pulses occur before and during luteolysis, little is known about the role of minor PGF pulses during preluteolysis on subsequent luteal and endometrial PGF production that may initiate luteolysis. Heifers (n = 7/group) and mares (n = 6/group) were treated with a single minor dose of PGF (3.0 and 0.5 mg, respectively) during mid-luteal phase (12 and 10 days postovulation respectively). After treatment, a transient decrease in progesterone (P4) concentrations occurred in heifers between Hours 0 and 2 but at Hour 4 P4 was not different from pretreatment. In mares, P4 was unaltered between Hours 0 and 4. Concentrations of P4 decreased in both species by Hour 24 and complete luteolysis occurred in mares by Hour 48. Luteal and endometrial gene expression were evaluated 4 h posttreatment. In heifers, luteal mRNA abundance of PGF receptor and PGF dehydrogenase was decreased, while PTGS2, PGF transporter, and oxytocin receptor were increased. In the heifer endometrium, receptors for oxytocin, P4, and estradiol were upregulated. In mares, luteal expression of PGF receptor was decreased, while PGF transporter and oxytocin receptor were increased. The decrease in P4 between Hours 4 and 24 and changes in gene expression were consistent with upregulation of endogenous synthesis of PGF. The hypotheses were supported that a single minor PGF treatment upregulates endogenous machinery for PGF synthesis in heifers and mares stimulating endogenous PGF synthesis through distinct regulatory mechanisms in heifers and mares.

Células madre mesenquimales como tratamiento para la regeneración de patologías articulares degenerativas. Revisión Narrativa / Mesenchymal stem cells as a treatment for the regeneration of degenerative articular diseases. A narrative review

RESUMEN Introducción: La reconstrucción de las estructuras de la articulación temporomandibular (ATM) degenerada permanecen como un desafío clínico hasta el día de hoy, y su tratamiento se ha enfocado principalmente en el alivio del dolor y la eventual restauración de la función articular. Objetivo: Describir, mediante una revisión narrativa, las propiedades y características de diferentes células mesenquimales para el tratamiento regenerativo de patologías articulares. Material y método: Se realizó una búsqueda electrónica de la literatura en las bases de datos de PubMed, EBSCO, Web of Science, Scopus y The Cochrane Library a partir del año 2005, en idioma ingles y español, con énfasis en la regeneración articular de la rodilla y la ATM. Se excluyeron aquellos estudios donde la degeneración articular fuese producto de un trauma. Fueron evaluados dieciocho artículos. Resultados y discusión: Resaltan los diferentes protocolos utilizados con células madre mesenquimales derivadas de médula ósea, tejido adiposo y cordón umbilical humano, con resultados de regeneración articular en todos los casos a nivel radiográfico e histológico cuando fue posible. El tratamiento de las patologías articulares en la ATM ha sido poco descrito, con buenos resultados en reportes de caso y estudios observacionales.

ABSTRACT: Introduction: To date, temporomandibular joint (TMJ) reconstruction remains a clinical challenge, and its treatment has focused mainly on pain relief and the eventual restoration of joint function. Aim: To describe, through a narrative review, the properties and characteristics of different mesenchymal cells for the regenerative treatment of joint diseases. Material and method: An electronic search of the literature was carried out in the databases of PubMed, EBSCO, Web of Science, Scopus and The Cochrane Library since the year 2005, in English and Spanish, emphasizing joint regeneration of the knee and the TMJ. Studies where joint degeneration was the result of trauma were excluded. Eighteen articles were evaluated. Results and discussion: The reported literature highlights the different protocols used with mesenchymal stem cells derived from bone marrow, adipose tissue and human umbilical cord, with results of joint regeneration in all cases at radiographic and histological level when possible. The treatment of joint diseases in the TMJ has been poorly described, with good results in case reports and observational studies. Because of this, it is important to continue investigating this matter to establish clear treatment protocols in the temporomandibular joint.

Self-Propulsion of Boiling Droplets on Thin Heated Oil Films.

We report on the self-propulsion of boiling droplets which, despite their contact with viscous, immiscible oil films, attain high velocities comparable to those of levitating Leidenfrost droplets. Experiments and model reveal that droplet propulsion originates from a coupling between seemingly disparate short and long timescale phenomena due to microsecond fluctuations induced by boiling events at the droplet-oil interface. This interplay of phenomena leads to continuous asymmetric vapor release and momentum transfer for high droplet velocities.

Knowledge, Attitude, and Practice (KAP) Survey toward Skin Cancer among Ecuadorian Population.

BACKGROUND: Skin cancer is one of the most common cancers, and melanoma is a highly preventable cancer. In Ecuador, few studies have evaluated the awareness levels of the population about the disease. For this reason, the objective of this study was to measure the level of knowledge, attitudes, and practices regarding skin cancer and its determining factors. METHODS: A cross-sectional analysis using an online self-assessment questionnaire containing 40 questions was delivered. A total of 537 participants were included in this study. Knowledge, attitude, and practice scores were assigned to each participant based on the number of correct or appropriate responses. Logistic regression analysis was used to calculate crude and adjusted odds ratios. RESULTS: In total, 75% of participants referenced knowledge of the harmful effects related to noncontrolled solar exposure. Concerning sunscreen, 76.7% knew the reason for using it. The female group was 1.68 times more likely to get a higher score than the male group, and the groups between 61-70 and 71-80 years were 0.30 and 0.17 times less likely to get a higher score compared with the less than 20-years-old group, respectively. CONCLUSIONS: The findings of this study indicate the requirement to increase the population's knowledge about skin cancer and possible protection measures. For this reason, the prevention and health promotion programs at a national level from primary healthcare centers are recommended. Due to the limitation of the representativeness of the sample, the use of more studies among Ecuadorian residents of the low socioeconomic level and replication in different provinces of Ecuador is justified.

Practical application of an impractical bovine genotype: creating bilateral twin pregnancies in Trio allele carriers.

Bovine twin birth is associated with detriments, including increased embryo/fetal losses, malpresentation, and dystocia. Incidence of these is lessened in bilateral compared with unilateral twin pregnancy. This study was undertaken to assess the use of follicular ablation by aspiration to create bilateral twin pregnancies in females with genetic potential for ~3.5 ovulations per cycle (Trio allele carriers). In experiment 1, carriers (n = 30) and noncarriers (n = 10) were synchronized for ovulation and timed artificial insemination (TAI). Follicles (>5 mm) in excess of one per ovary were aspirated ~16 h preceding TAI. Follicle count for females with follicles on only one ovary was reduced to two. Blood was sampled 2 wk post-TAI to assess progesterone (P4) concentrations; embryo count was determined by ultrasound 6 wk post-TAI. Circulating P4 concentration post-TAI was significantly (P < 0.001) associated with both genotype and subsequent pregnancy status (pregnant noncarriers: 7.06 ± 0.68 ng/mL; pregnant carriers: 5.54 ± 0.55 ng/mL; nonpregnant noncarriers: 5.22 ± 1.05 ng/mL; nonpregnant carriers: 3.13 ± 0.42 ng/mL). Experiment 2 was undertaken to offset the negative effects of follicular aspiration on subsequent P4 concentration observed in experiment 1. Carriers (n = 38) and noncarriers (n = 32) were submitted to TAI and follicle ablation as described for experiment 1. Additionally, accessory corpora lutea (CL) were induced in carriers by the administration of human chorionic gonadotropin (carriers) at day 6 post-TAI. Consequently, P4 concentration post-TAI was significantly (P < 0.05) associated with subsequent pregnancy status (pregnant: 8.48 ± 0.61 ng/mL; nonpregnant: 6.70 ± 0.63 ng/mL) but not with genotype (carrier: 8.01 ± 0.59 ng/mL; noncarrier: 7.17 ± 0.64 ng/mL). Embryo number was greater in carriers (exp. 1: 1.64 ± 0.81; exp 2: 1.45 ± 0.09) vs. noncarriers (1.00 ± 0.00, both experiments). Single, twin, and triplet pregnancies occurred in carriers in experiment 1, whereas multiples in experiment 2 were limited to twin pregnancies. Genotype effects on pregnancy rate were not significant (P > 0.10) in either experiment. Results suggest that follicular ablation to create bilateral twin pregnancies in Trio carriers is feasible but requires the induction of accessory CL to offset the negative effects of follicular aspiration on subsequent P4 concentration and associated fertility outcomes.

Selection of fewer dominant follicles in Trio carriers given GnRH antagonist and luteinizing hormone action replaced by nonpulsatile human chorionic gonadotropin†.

Studying selection of multiple dominant follicles (DFs) in monovulatory species can advance our understanding of mechanisms regulating selection of single or multiple DFs. Carriers of the bovine high fecundity Trio allele select multiple DFs, whereas half-sib noncarriers select a single DF. This study compared follicle selection during endogenous gonadotropin pulses versus during ablation of pulses with Acyline (GnRH antagonist) and luteinizing hormone (LH) action replaced with nonpulsatile human chorionic gonadotropin (hCG) treatment in Trio carriers (n = 28) versus noncarriers (n = 32). On Day 1.5 (Day 0 = ovulation), heifers were randomized: (1) Control, untreated; (2) Acyline, two i.m. doses (Days 1.5 and D3) of 3 µg/kg; (3) hCG, single i.m. dose of 50 IU hCG on Day 1.5 followed by daily doses of 100 IU; and (4) Acyline + hCG. Treatments with nonpulsatile hCG were designed to replace LH action in heifers treated with Acyline. Acyline treatment resulted in cessation of follicle growth on Day 3 with smaller (P < 0.0001) maximum follicle diameter in Trio carriers (6.6 ± 0.2 mm) than noncarriers (8.7 ± 0.4 mm). Replacement of LH action (hCG) reestablished follicle diameter deviation and maximum diameter of DFs in both genotypes (8.9 ± 0.3 mm and 13.1 ± 0.5 mm; P < 0.0001). Circulating follicle stimulating hormone (FSH) was greater in Acyline-treated than in controls. Finally, Acyline + hCG decreased (P < 0.0001) the number of DFs from 2.7 ± 0.2 to 1.3 ± 0.2 in Trio carriers, with most heifers having only one DF. This demonstrates the necessity for LH in acquisition of dominance in Trio carriers (~6.5 mm) and noncarriers (~8.5 mm) and provides evidence for a role of GnRH-induced FSH/LH pulses in selection of multiple DFs in Trio carriers and possibly other physiologic situations with increased ovulation rate.

Development of a novel 21-day reinsemination program, ReBreed21, in Bos indicus heifers.

The aim was to develop a program for resynchronization of ovulation (ReBreed21) that allowed reinsemination of non-pregnant Bos indicus heifers every 21 d using timed AI (TAI) without the need for detection of estrus. The Rebreed21 program begins 12 d after previous TAI (Day 0) by inserting an intravaginal P4 implant (Day 12) that is removed 7 d later (Day 19) combined with treatment with 300 IU of eCG. On Day 21, early pregnancy diagnosis (Doppler PD) is performed based on CL vascularity. Non-pregnant (NP) heifers immediately received AI combined with 100 µg of GnRH. The program is replicated 12 d after second TAI to produce a breeding season (BS) of 42 d with 3 potential TAIs. Two experiments were conducted as a proof of concept for this rapid rebreeding program. In Experiment 1, 76 heifers were enrolled in ReBreed21, as explained above. In Experiment 2, 300 Nellore heifers were synchronized for 1st TAI and randomly assigned to one of two groups: ReBreed21 (n = 147) or another early resynchronization procedure, Resynch14 (n = 153) with P4 implant inserted 14 d after previous TAI plus 50 mg of long-acting injectable P4; 8 d later P4 implant removed (Day 22) and early Doppler PD performed; NP heifers received 150 µg of cloprostenol, 0.5 mg of ECP, and 300 IU of eCG with TAI on Day 24. In both experiments, the largest follicle (LF) was measured at each Resynch TAI. Ultrasound was later used to confirm the early Doppler PD and to determine ovulation (OV) to Resynch at 12 d after TAI in ReBreed21 (Day 33 of pregnancy) and 14 d after TAI in Resynch14 (Day 38 of pregnancy). Final PD was performed 40 d after 3rd TAI. Results for Experiment 1 were: diameter of LF 11.8 ± 0.23 mm; 88.9% OV; 20.5% false positives; 38.1% P/AI at 1st TAI; 44.4% overall P/AI for ReBreed21 TAIs; 72.3% total pregnant at end of BS. In experiment 2, Rebreed21 vs. Resynch14 were different for: diameter of LF (10.9 ± 0.17 vs. 10.0 ± 0.17 mm, P = 0.0003), heifers with LF < 8.5 mm (10.2 vs. 26.4%, P = 0.04), or LF ≥ 11 mm (50.0 vs. 37.2%, P = 0.001), and P/AI at first TAI (29.3% [43/147] vs. 20.3% [31/153], P = 0.074) but similar for OV (overall 86.8% [239/275], P = 0.82), false positives (P = 0.52) overall P/AI for Resynch TAIs (33.6 vs. 28.8%, P = 0.4), and total pregnant at end of BS (58.5% [86/147] vs. 55.6% [85/153], P = 0.64). In addition, median time to pregnancy was 9 d earlier (P = 0.0007) for ReBreed21 than Resynch14. Thus, ReBreed21 is a novel protocol that allows earlier reinseminations than Resynch14 but with similar fertility.

Toxicity characterization of surface sediments from a Mediterranean coastal lagoon.

The occurrence of bioactive compounds and contaminant-associated effects was assessed by means of in vivo and in vitro assays using different extractable fractions of surface sediments from a contaminated coastal lagoon (Mar Menor, SE Spain). Sediment elutriates and clean seawater, previously exposed to whole sediment, were used for assessing the in vivo toxicity on embryo development of the sea urchin Paracentrotus lividus. Agonist and antagonist activities relating to estrogen and androgen receptors and agonist activities on aryl hydrocarbon receptor (expressed as ethoxyresorufin-O-deethylase (EROD) activities) were investigated in sediment extracts by using HER-Luc, AR-EcoScreenTM and fibroblast-like RTG-2 cell lines. Embryotoxicity effects were greater for sediment elutriates than those incubated in sediment-water interphase, implying that diffusion of bioactive chemicals can occur from sediments to sea water column, favoured by sediment disturbance events. In vitro results show the occurrence in extracts of compounds with estrogen antagonism, androgen antagonism and dioxin-like activities. Multidimensional scaling analysis classified the sampling sites into four sub-clusters according to their chemical-physical and biological similarities, relating in vitro bioactivity with the total organic carbon and known organic chemical load, with particular reference to total sum of PAHs, PCB 180, p,p-DDE and terbuthylazine. Overall, results pointed to the presence of unknown or unanalyzed biologically-active compounds in the sediments, mostly associated with the extracted polar fraction of the Mar Menor lagoon sediments. Our findings provide relevant information to be considered for the environmental management of contaminated coastal lagoons.

Necessity for LH in selection and continued growth of the bovine dominant follicle.

Previous research demonstrated that acute treatment with GnRH antagonist, Acyline, allowed follicle growth until ~8.5 mm and no dominant follicle was selected. This study evaluated whether deficient LH was the underlying mechanism for Acyline effects by replacing LH action, using human chorionic gonadotropin (hCG), during Acyline treatment. Holstein heifers (n = 24) during first follicular wave were evaluated by ultrasound and randomized into one of three treatments: Control (saline treatments), Acyline (5 µg/kg Acyline), or Acyline+hCG (Acyline plus 50 IU of hCG at start then 100 IU every 12 h). Pulses of LH were present in Control heifers (9 Pulses/10 h) but not during Acyline treatment. Data were normalized to the transition to diameter deviation (day 0; F1 ~7.5 mm). Diameter deviation of the largest (F1) and the second largest (F2) follicle was not observed in Acyline-treated heifers, whereas control heifers had decreased growth of F2 at F1 ~7.5 mm, indicating deviation. Selection of a single dominant follicle was restored by providing LH activity in Acyline+hCG heifers, as evidenced by F1 and F2 deviation, continued growth of F1, and elevated circulating estradiol. Separation of F1 and F2 occurred 12 h (~7.0 mm) earlier in Acyline+hCG heifers than Controls. Circulating FSH was greater in Acyline than Controls, but lower in Acyline+hCG than Controls after day 1.5. In conclusion, dominant follicle selection and growth after follicle deviation is due to LH action as shown by inhibition of this process during ablation of GnRH-stimulated LH pulses with Acyline and restoration of it after replacement of LH action by hCG treatment.