Expression of mRNA Encoding the LH Receptor (LHR) and LHR Binding Protein in Granulosa Cells from Nelore (Bos indicus) Heifers Around Follicle Deviation.

The time at which follicles acquire LHR in bovine granulosa cells is the subject of some controversy among researchers. The main objective of the present study was to assess the mRNA expression of LHR and LRBP (mRNA protein binding), a post-transcriptional suppressor of LHR mRNA expression, in granulosa cells from the two largest follicles around the expected time of follicle deviation in Nelore heifers. First, the interval between ovulation and follicle deviation in 20 Nelore heifers was determined (2.3 ± 0.2 days after ovulation). Ovulation was hormonally synchronized, and then, heifers were slaughtered on days 2, 2.5 and 3 after ovulation (before, during and after, respectively, the expected time of follicle deviation), and granulosa cells from the two largest follicles were collected. The mRNA abundance of an LHR fragment common to all isoforms (total LHR) and LRBP was assessed by real-time RT-PCR, and LHR alternative transcripts were assessed by semiquantitative RT-PCR followed by electrophoresis. LHR mRNA expression was not detected before the expected time of deviation. Total LHR mRNA abundance was greater in the largest follicle and increased from day 2.5 to 3. In contrast, LRBP mRNA was detected starting on day 2 and was more expressed in the second largest follicle on days 2.5 and 3. The present data suggest that the expression of LHR mRNA in bovine granulosa cells is established after follicle deviation and that the lower abundance of LRBP mRNA after the expected time of deviation may contribute to greater expression of LHR in the bovine dominant follicle.

Differential expression of IGF family members in heat-stressed embryos produced in vitro from OPU-derived oocytes of Nelore (Bos indicus) and Holstein (Bos taurus) cows.

The IGF system is related to embryo quality. We aim to determine the effect of the heat stress on the mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4, and PAPPA in in vitro production (IVP) blastocysts from Nelore and Holstein after ovum pick up (OPU) to better understand the differences between these breeds. Oocytes from four Nelore and seven Holstein were collected in six OPU sessions. Following in vitro maturation and fertilization using six Nelore or Holstein sires, embryos were divided into control (cultured at 39°C) and heat stress (HS; exposed to 41°C for 9 h). Blastocysts were submitted to RNA extraction. The IGF1 expression was higher in blastocysts under HS in both breeds, and the expression of IGFBP2 and IGFBP4 was higher in Holstein blastocysts under HS. The high PAPPA expression and the low expression of IGFBP2 and IGFBP4 are associated with a more efficient degradation of IGFBPs, which results in greater IGF bioavailability in Nelore blastocysts and may contribute to the superior HS tolerance in Nelore, when compared to Holstein.

Influence of clinical presentation on the outcome of acute B aortic dissection: evidences from IRAD.

AIM: In-hospital outcome of acute type B dissection (ABAD) is strongly related to preoperative aortic conditions. In order to clarify the influence of the clinical presentation on the outcome, we analyzed the patients of the International Registry of Acute Aortic Dissection (IRAD). All patients affected by complicated ABAD, enrolled in the IRAD from 1996-2004, were included. Complications were defined as the presence of shock, periaortic hematoma, spinal cord ischemia, preoperative mesenteric ischemia/infarction, acute renal failure, limb ischemia, recurrent pain, refractory pain or refractory hypertension (group I). All other patients were categorized as uncomplicated (group II). A comprehensive analysis was performed of all clinical variables in relation to in-hospital outcome. RESULTS: The overall in-hospital mortality among 550 patients was 12.4%. Mortality in group I (250 patients) was 20.0 %, compared to 6.1% in group II (300 patients) (P<0.001). Univariate predictors of ABAD complications were Marfan syndrome, abrupt onset of pain, migrating pain, any focal neurological deficits, need for higher number of diagnostic examinations and use of magnetic resonance and/or aortogram, abdominal vessels involvement at aortogram, larger descending aortic diameter, especially >6 cm, pleural effusion, and widened mediastinum on chest X-ray. Univariate predictors of a non complicated status were normal chest X-ray and medical management. In group I, in-hospital mortality following surgical and endovascular intervention were 28.6% and 10.1% (P=0.006), respectively. Independent predictors of overall in-hospital mortality included age >70 years, female gender, ECG showing ischemia, preoperative acute renal failure, preoperative limb ischemia, periaortic hematoma, and surgical management. The only independent variable protective for mortality was magnetic resonance as diagnostic test. CONCLUSION: ABAD is a heterogeneous disease that produces dissimilar clinical subsets, each of which can have specific clinical signs, management and in-hospital results. In IRAD ABAD uncomplicated patients, medical therapy was associated with best hospital outcome, while endovascular interventions were associated with better results than surgery when invasive treatments were required. Although selection bias may be possible, and irrespective of treatments, knowledge of significant risk factors for mortality may contribute to a better management and a more defined risk-assessment in patients affected by ABAD.

Neither plasma progesterone concentrations nor exogenous eCG affects rates of ovulation or pregnancy in fixed-time artificial insemination (FTAI) protocols for puberal Nellore heifers.

The objective was to evaluate the effects of plasma progesterone (P4) concentrations and exogenous eCG on ovulation and pregnancy rates of pubertal Nellore heifers in fixed-time artificial insemination (FTAI) protocols. In Experiment 1 (Exp. 1), on Day 0 (7 d after ovulation), heifers (n = 15) were given 2 mg of estradiol benzoate (EB) im and randomly allocated to receive: an intravaginal progesterone-releasing device containing 0.558 g of P4 (group 0.5G, n = 4); an intravaginal device containing 1 g of P4 (group 1G, n = 4); 0.558 g of P4 and PGF(2α) (PGF; 150 µg d-cloprostenol, group 0.5G/PGF, n = 4); or 1 g of P4 and PGF (group 1G/PGF, n = 3). On Day 8, PGF was given to all heifers and intravaginal devices removed; 24 h later (Day 9), all heifers were given 1 mg EB im. In Exp. 2, pubertal Nellore heifers (n = 292) were treated as in Exp. 1, with FTAI on Day 10 (30 to 36 h after EB). In Exp. 3, pubertal heifers (n = 459) received the treatments described for groups 0.5G/PGF and 1G/PGF and were also given 300 IU of eCG im (groups 0.5G/PGF/eCG and 1G/PGF/eCG) at device removal (Day 8). In Exp. 1, plasma P4 concentrations were significantly higher in heifers that received 1.0 vs 0.588 g P4, and were significantly lower in heifers that received PGF on Day 0. In Exp. 2 and 3, there were no significant differences among groups in rates of ovulation (65-77%) or pregnancy (Exp. 2: 26-33%; Exp. 3: 39-43%). In Exp. 3, diameter of the dominant ovarian follicle on Day 9 was larger in heifers given 0.558 g vs 1.0 g P4 (10.3 ± 0.2 vs 9.3 ± 0.2 mm; P < 0.01). In conclusion, lesser amounts of P4 in the intravaginal device or PGF on Day 0 decreased plasma P4 from Days 1 to 8 and increased diameter of the dominant follicle on Day 9. However, neither of these nor 300 IU of eCG on Day 8 significantly increased rates of ovulation or pregnancy.

Effects of temporary calf removal and eCG on pregnancy rates to timed-insemination in progesterone-treated postpartum Nellore cows.

The objective was to evaluate the effects of temporary calf removal (TCR), eCG administration, or both, in a progesterone-based protocol. Suckled Nellore cows (40-80 d postpartum, n=443) with body condition scores from 2.0 to 3.5 (5-point scale) on three farms were all given a synchronizing protocol (PEPE). At the start (designated Day 0), cows were given an intravaginal device (1.0 g of progesterone) and 2.5mg of estradiol benzoate (EB) im. On Day 8, the device was removed and cows were given PGF(2 alpha) (150 microg of D-cloprostenol im), followed in 24h by 1.0mg EB im, and 30-36 h thereafter, fixed-time AI. The design was a 2 x 2 factorial; main effects were TCR (54-60 h; from device removal to FTAI) and eCG treatment (300 IU im, concurrent with PGF(2 alpha)). Transrectal ultrasonography was done on Days -10 and 0 to detect anestrus (absence of a CL at both examinations) and approximately 30 d after FTAI (pregnancy diagnosis). Data were analyzed by logistic regression. The following variables did not significantly affect pregnancy rates: farm, postpartum interval, cyclicity, inseminators, and semen (sire). Overall, 77% of the cows were deemed anestrus. Pregnancy rates were similar (P>0.05) among treatment groups: Control (54/108=50.0%), TCR (44/106=41.5%), eCG (63/116=54.3%), and TCR+eCG (49/113=43.4%). Pregnancy rate was higher in multiparous than primiparous cows (186/360, 51.7% vs. 24/83, 28.9%, P<0.01), but was not significantly affected by cyclicity status or body condition score. In conclusion, temporary calf removal, eCG, or both, did not significantly increase pregnancy rate to timed-insemination in a progesterone-based synchronization protocol in postpartum Nellore cows with acceptable body condition.

Serum antibody response to Cryptococcus neoformans in cats, dogs and koalas with and without active infection.

Anti-cryptococcal antibodies were measured in normal cats, dogs, horses and koalas, and cats, dogs and koalas with cryptococcosis using an enzyme immunoassay. Antibody levels were expressed as absorbance readings. Over 80% of cats and dogs with cryptococcal infection had elevated antibody levels at the time of diagnosis, during or after successful therapy. Antibody levels in these patients either remained elevated or declined slowly after treatment. For cats, anti-cryptococcal antibody levels were higher in C. neoformans var. gattii than var. neoformans infections, and lower in mild than in moderate or severe infections. The persistence of increased anti-cryptococcal antibody levels in over half of the feline and canine cases following active infection suggested the use of antibody determinations as a seroepidemiologic marker of previous infection. Consequently, antibody measurements from 'normal' animals indicated a prevalence of previous cryptococcal infection of 10% in cats and dogs, compared with 3% in horses and 5% in koalas. Preliminary studies of young animals suggested that anti-cryptococcal antibody levels were substantially lower in the young cats but not the young dogs surveyed, compared with their mature counterparts. The cut-offs used in the present work may thus be erroneously high, with a corresponding underestimation of the prevalence of inapparent cryptococcosis.

Serum antibody response to active infection with Cryptococcus neoformans and its varieties in immunocompetent subjects.

Using a class-specific enzyme immunoassay IgG and IgA anticryptococcal antibody was measured in 400 serum specimens at 1 week to 11.7 years after diagnosis from 43 immunocompetent subjects with confirmed active cryptococcosis. The prevalence of IgG was 86% at diagnosis, rose to 100% by 2 weeks and remained high thereafter. IgA prevalence was 71% at diagnosis, rose to 75% at 2 weeks and then fell over 2 years. IgG and IgA prevalence in paediatric controls was 5% and 0%, respectively. Mean antibody levels showed the same pattern and neither levels nor prevalence were influenced by age, sex or site of infection. Patients with the variety gattii infection had a greater antibody response than those with the variety neoformans which was significant for IgA. Specific anticryptococcal antibody was regularly present in conjunction with cryptococcal antigen at diagnosis. IgG persists but IgA falls over 1-2 years. The assay described may be a useful tool to study the antibody response and seroepidemiology of infection with C. neoformans.

Relaçäo entre o nível sérico de gamaglobulinas e as atividades de gamaglutamiltransferase, fosfatase alcalina e aspartato aminotransferase de bezerros recém-nascidos / Gammaglobulin, gammaglutamyl transferase, alkaline phosphatase, and aspartate aminotransferase relationship in newborn calves

Analisaram-se 140 amostras de soro sangüíneo de bezerros da raça Holandesa, recém nascidos, com o objetivo de verificar a relaçäo entre a concentraçäo de gamaglobulina e as atividades das enzimas gamaglutamiltransferase, fosfatase alcalina e aspartato aminotransferase nos primeiros seis dias de vida. Näo se observou correlaçäo significativa entre os níveis de gamaglobulina e atividade de aspartato aminotransferase. A correlaçäo positiva significativa verificada no primeiro dia de vida, entre teor de globulina e atividade da fosfatase alcalina foi de pouca intensidade, näo sendo considerado um teste seguro de identificaçäo de hipogamaglobulinemia. Observou-se correlaçäo positiva significativa, com variaçöes intensas, ao redor de 24 a 30 horas pós-nascimento, entre níveis de gamaglobulina e atividade da gamaglutamiltransferase, quando comparados aos valores obtidos nas primeiras seis horas pós-natal. Pode-se considerar tal enzima como teste de identificaçäo indireta de bezerros hipogamaglobulinêmicos, por falha na transferência de imunoglobulinas colostrais