Sire contribution to fertilization failure and early embryo survival in cattle.

Despite passing routine laboratory tests of semen quality, bulls used in artificial insemination (AI) exhibit a significant range in field fertility. The objective of this study was to determine whether subfertility in AI bulls is due to issues of sperm transport to the site of fertilization, fertilization failure, or failure of early embryo or conceptus development. In experiment 1, Holstein-Friesian bulls (3 high fertility, HF, and 3 low fertility, LF) were selected from the national population of AI bulls based on adjusted fertility scores from a minimum of 500 inseminations (HF: +4.37% and LF: -12.7%; mean = 0%). Superovulated beef heifers were blocked based on estimated number of follicles at the time of AI and inseminated with semen from HF or LF bulls (n = 3-4 heifers per bull; total 19 heifers). Following slaughter 7 d later, the number of corpora lutea was counted and the uteri were flushed. Recovered structures (oocytes/embryos) were classified according to developmental stage and stained with 4',6-diamidino-2-phenylindole to assess number of cells and accessory sperm. Overall recovery rate (total structures recovered/total corpora lutea) was 52.6% and was not different between groups. Mean (± standard error of the mean) number of embryos recovered per recipient was 8.7 ± 5.2 and 9.4 ± 5.5 for HF and LF, respectively. Overall fertilization rate of recovered structures was not different between groups. However, more embryos were at advanced stages of development (all blastocyst stages combined), reflected in a greater mean embryo cell number on d 7 for HF versus LF bulls. Number of accessory sperm was greater for embryos derived from HF than for LF bulls. The aim of experiment 2 was to evaluate the effect of sire fertility on survival of bovine embryos to d 15. Day 7 blastocysts were produced in vitro using semen from the same HF (n = 3) and LF (n = 3) bulls and transferred in groups of 5-10 to synchronized heifers (n = 7 heifers per bull; total 42 heifers). Conceptus recovery rate on d 15 was higher in HF (59.4%,) versus LF (45.0%). Mean length of recovered conceptuses for HF bulls was not affected by fertility status. In conclusion, while differences in field fertility among AI sires used in this study were not reflected in fertilization rate, differences in embryo quality were apparent as early as d 7. These differences likely contributed to the higher proportion of conceptuses surviving to d 15 in HF bulls.

Evaluation of delayed timing of artificial insemination with sex-sorted sperm on pregnancy per artificial insemination in seasonal-calving, pasture-based lactating dairy cows.

The objective was to use ovulation synchronization with timed artificial insemination (TAI) to evaluate the effect of timing of artificial insemination (AI) with frozen sex-sorted sperm on fertility performance in pasture-based compact calving herds. Ejaculates from 3 Holstein-Friesian bulls were split and processed to provide frozen sex-sorted sperm (SS) at 4 × 106 sperm per straw, and frozen conventional sperm at 15 × 106 sperm per straw (CONV). A modified Progesterone-Ovsynch protocol was used for estrous synchronization, with TAI occurring 16 h after the second GnRH injection for cows assigned to CONV, and either 16 h (SS-16) or 22 h (SS-22) for cows assigned to SS. Pregnancy diagnosis was conducted by transrectal ultrasound scanning of the uterus 35 to 40 d after TAI (n = 2,175 records available for analysis). Generalized linear mixed models were used to examine the effects of treatment on pregnancy per artificial insemination (P/AI). Fixed effects included treatment (n = 3), bull (n = 3), treatment by bull interaction, parity (n = 4), days-in-milk category (n = 3), and treatment by days-in-milk category, with herd (n = 24) included as a random effect. Pregnancy per AI was greater for CONV compared with both SS-16 and SS-22 (61.1%, 49.0%, and 51.3%, respectively), and the SS treatments did not differ from each other (relative P/AI for SS-16 and SS-22 vs. CONV were 80.2% and 84.0%, respectively). There were significant bull and treatment by bull interaction effects. Additional analysis was undertaken using a model that included herd as a fixed effect. This analysis identified marked herd-to-herd variation (within-herd relative P/AI for the combined SS treatments vs. CONV ranged from 48-121%). The tertile of herds with the best performance achieved a mean relative P/AI of 100% (range = 91-121%), indicating that P/AI equivalent to CONV is achievable with SS. Conversely, the tertile of herds with the poorest performance achieved a mean relative P/AI of 67% (range = 48-77%). We found that SS resulted in poorer overall P/AI compared with CONV sperm regardless of timing of AI. Marked variation existed between herds; however, one-third of herds achieved P/AI results equal to CONV. Identification of factors responsible for the large herd-to-herd variation in P/AI with SS, and development of strategies to reduce this variation, warrant further research.

Characteristics of offspring derived from conventional and X-sorted bovine sperm.

The objective of this retrospective study was to compare survival during the first year of life and adult performance of offspring derived from artificial insemination (AI) with X-sorted or conventional sperm processed from the same ejaculates. We analyzed a data set that included AI of dairy heifers and lactating cows with fresh conventional sperm (3 × 106 sperm per straw), fresh X-sorted sperm (1 or 2 × 106 sperm per straw), or frozen X-sorted sperm (2 × 106 sperm per straw). The data set contained records of 5,179 offspring born on 396 farms. Offspring were classified as born from conventional sperm (CONV) if they were the product of an insemination with fresh conventional sperm, or born from X-sorted sperm (SS) if they were product of any of the 3 X-sorted sperm treatments. Generalized linear mixed models were used to evaluate the effect of sperm treatment on (1) survival during the first year of life; (2) reproductive performance, lactation performance, and survival of female offspring; and (3) slaughter characteristics of male offspring. Stillbirth rates and mortality rates during the first 2 mo of life were greater for male calves (2.8 and 5.0%, respectively) than for female calves (1.6 and 2.0%, respectively). No differences between offspring derived from SS and CONV were detected for incidences of stillbirth or mortality during the first 12 mo of life within sex of calf. Reproductive performance, milk volume, milk fat, milk protein yields during first; second; and third lactations, and survival to third lactation did not differ between female offspring derived from CONV and SS. Across all age groups, CONV steers had heavier carcasses than SS steers (325.3 vs. 318.3 kg), but there were no differences in weight between CONV and SS steers within any of the age groups (≤24, 25-27, 28-30, and >30 mo of age). The distribution of slaughter age did not differ between CONV and SS steers when the analysis was restricted to herds that reared steers derived from both types of sperm. Carcass conformation and fat scores of steers were not affected by sperm treatment. There was no difference in carcass weight between young bulls (≤2 yr) derived from CONV or SS. In conclusion, the results provide no evidence of differences in survival during the first year of life between offspring derived from CONV or SS, or for any of the reproductive and lactation performance characteristics studied between female offspring derived from CONV or SS. Modest differences in carcass weight between CONV and SS steers were detected, but this may reflect differences in management and husbandry in the rearing herds rather than the sex-sorting process. A controlled study using steers derived from conventional or X-sorted sperm from split ejaculates and reared under the same husbandry conditions is needed to clarify whether there is a true difference in body weight gain due to the sex-sorting process.

Fertility of frozen sex-sorted sperm at 4 × 106 sperm per dose in lactating dairy cows in seasonal-calving pasture-based herds.

The objective was to evaluate the reproductive performance of frozen sex-sorted sperm at 4 × 106 sperm per dose (SexedULTRA 4M, Sexing Technologies, Navasota, TX) relative to frozen conventional sperm in seasonal-calving pasture-based dairy cows. Semen from Holstein-Friesian (n = 8) and Jersey (n = 2) bulls was used. Four of the Holstein bulls used were resident at or near a sex-sorting laboratory (Cogent, UK, or ST Benelux, the Netherlands). The remaining 6 bulls were located at studs in Ireland. For these 6 bulls, ejaculates were collected, diluted with transport medium, and couriered to Cogent in parcel shippers. Transit time from ejaculation to arrival at the sorting laboratory was 6 to 7 h. For all bulls, ejaculates were split and processed to provide frozen conventional sperm (CONV) at 15 × 106 sperm per straw and frozen sex-sorted (SS) sperm at 4 × 106 sperm per straw and used to inseminate lactating dairy cows after spontaneous estrus. Pregnancy diagnosis was performed by ultrasound scanning (n = 7,246 records available for analysis). Generalized linear mixed models were used to examine effects on pregnancy per AI (P/AI) at first artificial insemination, with sperm treatment (CONV vs. SS), bull (n = 10), and treatment × bull interaction as the fixed effects, and herd (n = 142) as a random effect. Overall, P/AI was greater for cows inseminated with CONV than for those inseminated with SS (59.9% vs. 45.5%; 76.0% relative to CONV). This study was not designed to compare resident bulls vs. shipped ejaculates, but the magnitude of the difference between P/AI achieved by CONV and SS was apparently less for resident bulls (60.3% vs. 50.2%) than for shipped ejaculates (58.6% vs. 40.7%). We discovered a treatment × bull interaction for shipped ejaculates (P/AI ranged from 45 to 86% relative to CONV) but not for the resident bulls (P/AI ranged from 81 to 87% relative to CONV). Relative P/AI of SS compared with CONV was greater in cows with high or average fertility potential (76.1% and 78.3%, respectively) than in cows with low fertility potential (58.1%). In 33.1% of the enrolled herds, the P/AI achieved with SS was 90% or more of the P/AI achieved with CONV; this was mainly explained by herds in which SS performed exceptionally well but CONV performed poorly. In conclusion, SS had lower overall P/AI compared with CONV; however, P/AI achieved with SS was dependent on the bull, fertility potential of the cow, and herd. Strategies to improve the P/AI with SS in seasonal-calving pasture-based lactating dairy cows require further research.

Fertility of fresh and frozen sex-sorted semen in dairy cows and heifers in seasonal-calving pasture-based herds.

Our objective in this study was to evaluate the reproductive performance of dairy heifers and cows inseminated with fresh or frozen sex-sorted semen (SS) in seasonal-calving pasture-based dairy herds. Ejaculates of 10 Holstein-Friesian bulls were split and processed to provide (1) fresh conventional semen at 3 × 106 sperm per straw (CONV); (2) fresh SS at 1 × 106 sperm per straw (SS-1M); (3) fresh SS semen at 2 × 106 sperm per straw (SS-2M); and (4) frozen SS at 2 × 106 sperm per straw (SS-FRZ). Generalized linear mixed models were used to evaluate the effect of semen treatment and other explanatory variables on pregnancy per artificial insemination (P/AI) in heifers (n = 3,214) and lactating cows (n = 5,457). In heifers, P/AI was greater for inseminations with CONV (60.9%) than with SS-FRZ (52.8%) but did not differ from SS-1M (54.2%) or SS-2M (53.5%). Cows inseminated with CONV had greater P/AI (48.0%) than cows inseminated with SS, irrespective of treatment (SS-1M, SS-2M, and S-FROZEN; 37.6, 38.9, and 40.6%, respectively). None of the SS treatments differed from each other with regard to P/AI in either heifers or cows. The relative performance of SS compared with CONV was also examined [i.e., relative P/AI = (SS P/AI)/(CONV P/AI) × 100]. Frozen SS achieved relative P/AI >84%. Bull affected P/AI in both heifers and cows, but no bull by semen treatment interaction was observed. In heifers, P/AI increased with increasing predicted transmitting ability for milk protein percentage. In cows, P/AI increased with increasing Economic Breeding Index (EBI) and with days in milk (DIM) at AI but decreased with increasing EBI milk subindex, parity and with DIM2. Cows in parity ≥5 had the lowest P/AI and differed from cows in parities 1, 2, or 3. Dispatch-to-AI interval of fresh semen did not affect P/AI in lactating cows, but a dispatch-to-AI interval by bull interaction was detected whereby P/AI was constant for most bulls but increased with greater dispatch-to-AI intervals for 2 bulls. In conclusion, frozen SS achieved greater P/AI relative to conventional semen than was previously reported in lactating cows. Fresh SS did not achieve greater P/AI than frozen SS, regardless of whether the sperm dose per straw was 1 × 106 or 2 × 106. A bull effect for all semen treatments, as well as a dispatch-to-AI interval by bull interaction for fresh semen, highlights the importance of using a large team of bulls for breeding management.

Global transcriptomic response of bovine endometrium to blastocyst-stage embryos.

The aims of this study were (i) to investigate changes in the global transcriptome of bovine endometrial explants induced by exposure to blastocysts, (ii) to investigate if male and female blastocysts elicit a differential response in the endometrial transcriptome in vitro and (iii) to determine whether bovine endometrium responds to the presence of murine embryos. In Experiment 1, endometrial explants from the same uterus were cultured for 6 h with or without 20 in vitro-produced bovine blastocysts. In Experiment 2, endometrial explants were cultured with male or female bovine blastocysts produced in vitro by IVF either using sex-sorted semen or conventional unsorted semen followed by embryo sexing based on a biopsy. In Experiment 3, endometrial explants were cultured alone or in the presence of bovine blastocysts (n = 25) or murine blastocysts (n = 25). Following culture, explants were snap frozen and stored at -80°C until RNA extraction, qPCR or RNA-Seq. Culture with bovine blastocysts increased endometrial expression of 40 transcripts, all of which were interferon-tau induced. Culture with male or female bovine blastocysts increased transcript abundance of five classic interferon-stimulated genes (MX1, MX2, ISG15, OASY1, RSAD2) in explants; however, there was no difference in abundance of transcripts previously reported to be related to embryonic sex (IFNAR1, IFNAR2, CTGF, ARTN, SLC2A1, SLC2A5). Exposure to murine blastocysts did not elicit any detectable change in transcript abundance. These findings, coupled with our previous data, indicate that very local, interferon-tau-induced changes in endometrial gene expression occur in response to blastocysts; whether such changes play any role in subsequent pregnancy recognition remains to be established.

Malignant carotid paraganglioma: a case report. / Paraganglioma carotídeo maligno. A propósito de un caso.

Tumors of the carotid body are uncommon neoplasms that originate in the neuroectoderm. These tumors are slow growing but well vascularized and very destructive. Between 5% and 13% of carotid body tumors are malignant, behaving aggressively at the local level, invading the adjacent lymph nodes, and resulting in remote metastases. We present the case of a 60-year-old man who was examined for pain and paresthesias in his arm. Magnetic resonance imaging showed a large tumor in the left carotid space. Histologic study of a biopsy specimen from the tumor revealed that it was a paraganglioma. Further studies to determine the extent of disease detected metastases in bone and lung, confirming the tumor's malignancy.

Prevalence of sarcopenia in a media and long stay Unit. / Prevalencia de sarcopenia en una unidad de media y larga estancia.

BACKGROUND: Sarcopenia is characterized by the loss of skeletal muscle mass and strength and represents a risk factor for frailty, loss of independence and physical disabilities. This study aimed to assess the prevalence of sarcopenia in a population of patients admitted to a media and long-stay unit. PATIENTS AND METHODS: Descriptive, cross-sectional observational study that included 166 patients over 18 years who were admitted consecutively for a year to palliative care, convalescence or rehabilitation. Administrative and anthropometric data were recorded. Sarcopenia detection was performed by assessing the strength in the scapular and pelvic girdle muscles, grip strength in the hands, muscle mass and physical functional capacity. RESULTS: The overall prevalence of sarcopenia was 77.6%. Most patients had severe sarcopenia (91.2%), with reduced levels of the 3 assessed parameters: muscle strength (88.2%), muscle mass (85.5%) and physical performance (94.70%). CONCLUSION: The prevalence of sarcopenia in patients admitted to our media and long-stay unit is higher than that reported in noninstitutionalized populations, and often has a significant degree of severity.

[Prevalence of diabetic retinopathy in the region of Girona. Study of related factors]. / Prevalencia de la retinopatía diabética en la población de diabéticos diagnosticados en las comarcas de Girona. Estudio de los factores asociados.

PURPOSE: To determine of the prevalence of diabetic retinopathy in the general diabetic population in the Girona region and epidemiologic study of the related factors. METHODS: A population-based cross-sectional study of a randomised sample taken of 401 diabetics in the Girona region. Descriptive statistical analysis of the ocular lesions in relation to diabetic retinopathy and analysis of logistic regression in order to determine the related factors. RESULTS: 30.6% (IC 95%: 26.5-35.5) of the sample presented some degree of diabetic retinopathy, 19.4% (IC 95%: 15.4-24.0) at the initial stage, 8.4% (IC 95%: 5.8-1.9) pre-proliferative and 2.6% (IC 95%: 1.3-5.1) at a proliferative stage. Regarding the type of diabetes, 53% of the patients with type 1 diabetes and 34.2% of those with type 2 diabetes had retinopathy. Retinopathy was observed in 15.7% of diabetics with less than 10 years of diabetes evolution, 51.3% in those between 10 and 20 years and 62.8% in those with more than 20 years. The concentration of HbA1C was significantly associated with the degree of retinopathy. Of those patients with retinopathy, 60.3% were insulin-treated, 30.6% with oral hypoglucemiants and 10.3% on diet only. Patients with family antecedents of retinopathy presented a prevalence three times greater than those without. CONCLUSIONS: Approximately one in three diabetic patients exhibited some degree of retinopathy in this study dealing with a population-based cohort of diabetic patients in Spain. The prevalence was greater among type I diabetic patients. As expected, years of evolution of the disease, type of treatment and diabetic metabolic control were the most significant factors influencing the presence of retinopathy.

[Upper limb deep venous thrombosis. Risk factors, outcome, and posthrombotic syndrome]. / Trombosis venosa profunda de miembros superiores. Factores de riesgo, evolución y síndrome postrombótico.

OBJECTIVE: Our objective was that of describing a series of episodes of upper limb deep venous thrombosis, with special emphasis on the incidence of posthrombotic syndrome and its impact in the quality of life of the patients. METHODS: 33 episodes (29 patients) of upper limb deep venous thrombosis were studied, between January 1995 and January 2002; episodes until September 1999 (18 episodes) were considered retrospectively while those which took place starting in this date (15 episodes) were considered prospectively. In all, 24 patients were cared in hospital clinic. It was possible to obtain complete data in 16 cases, in which evolution toward posthrombotic syndrome, venous revascularization, and impact of the pathological process in the global quality of life of patients were studied. RESULTS: Average age of patients was 58.9 years. In 25 episodes (75.8%) some risk factor for venous thrombosis existed, the presence of a venous catheter being the most frequent. Subclavian axillary affectation was predominant (18 episodes, 55%) and in the left side (25 episodes, 76%). Twenty-eight episodes were diagnosed with echography (84.8%). Mean D-dimers concentration (N = 16) was 1,046 ng/ml (standard deviation [SD]: 826.9). One of the episodes (3%) was associated to lung thromboembolism. Treatment consisted on the administration of low molecular weight heparin during the acute phase in 70% of the cases. In 13 episodes (39%) additional treatment of thrombosis (endovascular in most) was carried out. In 29 episodes (88%) chronic treatment was administered with acenocoumarol, while in 4 episodes (12%) the administration of low molecular weight heparin was kept. No patient died during hospitalization. In 15 patients (52%) control of revascularizacion was performed. We detected complete revascularization in 6 patients (40%), revascularization with collateral circulation in 4 patients (27%), partial revascularization in other 4 patients (27%) and presence of permeability but with expansion and tortuosity of deep venous system in 1 case (6%). Posthrombotic syndrome was considered mild or nonexistent in 10 patients (62.5%) and moderate in 6 patients (37.5%). Quality of life of patients with moderate posthrombotic syndrome was significantly worse than that of patients with mild posthrombotic syndrome (73 vs 90, p < 0.035). CONCLUSIONS: Most of upper limb deep venous thrombosis are associated to the use of intravenous catheters. Echography is the primary diagnostic technique. At the time of the diagnosis, patients tend to show elevation in D-dimers levels. Evolution toward posthrombotic syndrome is frequent; this syndrome does not use to be serious but indeed influences the quality of life of patients.

Efficacy of low-dose OKT3 as cytolytic induction therapy in heart transplantation.

BACKGROUND: The need for prophylactic cytolytic treatment in heart transplantation is a controversial issue. Its use, however, might prevent the onset of cellular rejection in the immediate postoperative period, facilitating patient management. It has recently been suggested that the administration of these products at low doses might have the same immunologic impact and would reduce secondary effects and the cost of treatment. METHODS: In a nonrandomized retrospective study, we assessed 45 consecutive patients who underwent orthotopic heart transplantation in 1992 and 1993. Six patients who died before receiving the complete OKT3 dose were excluded. Twenty-three patients were treated with 5mg/day doses of OKT3 for 7 consecutive days. Another 16 patients received 2.5 mg of OKT3 for 7 consecutive days. RESULTS: There were no significant differences between the two groups with respect to CD3 counts on days 2 (0.1% +/- 0.3% versus 0.04% +/- 0.25%; p > 0.05) and 6 (0.2% +/- 0.45% versus 0.1% +/- 0.3%; p > 0.05), number of rejection episodes (1.45% +/- 0.8% per year of follow-up versus 1.7% +/- 1.2%, p = 0.66), number of infectious complications (8 versus 3, p > 0.05), total methylprednisolone dose used to treat rejection crises (3900 +/- 2765 versus 3600 +/- 1963 mg; p = 0.71), adverse effects attributed to OKT3 (two versus none), or length of the postoperative hospital stay (36.8 +/- 19 versus 30.2 +/- 20.9 days). CONCLUSIONS: As cytolytic induction therapy in heart transplantation, a daily regimen of 2.5 mg of OKT3 for 7 days achieves the same clinical and immunologic effect as the conventional 5 mg/day dose. In addition, it results in a considerable reduction in the cost of treatment.

[Right ventricular arrhythmogenic dysplasia. The role of heart transplantation in its management]. / Displasia arritmogénica ventricular derecha. El papel del trasplante cardíaco en su manejo.

Arrhythmogenic right ventricular dysplasia is a rare disease that usually presents with ventricular arrhythmias and sometimes with heart failure. Rarely symptoms become severe and refractory to conventional therapy. We present the case of a 36-year-old man with this disease who had sustained ventricular arrhythmias and severe right heart failure. Because these symptoms were poorly controlled with medical therapy, cardiac transplantation was finally chosen as definitive treatment. The patient had a favorable course and now leads near-normal life.

[A heart transplant in a patient with doxorubicin-induced myocardiopathy as a complication of the treatment of a lymphoma]. / Trasplante cardíaco en un paciente con miocardiopatía por doxorrubicina como complicación del tratamiento de un linfoma.

We present the case of a young man who developed severe cardiomyopathy eight years after receiving high-dose doxorubicin therapy as treatment for diffuse lymphoblastic lymphoma. He underwent cardiac transplantation and eighteen months later the tumor has not recurred and the patient has no cardiac symptoms.