Heat stress affects the expression of key genes in the placenta, placental characteristics, and efficiency of Saanen goats and the survival and growth of their kids.

Heat stress is detrimental during gestation; however, the effects of heat stress on goat placental characteristics and kid survival remain unclear. The objective of this study was to evaluate the effects of heat stress at final gestation on cortisol concentration, placenta characteristics, and the expression of genes related to placenta. Forty-six primiparous and multiparous Saanen goats were subjected to control (CT; under a thermoneutral environment: air temperature between 12°C and 25°C and the relative humidity from 45 to 73%, n = 23) or heat stress (HS; under a climatic chamber: air temperature at 37°C and the relative humidity at 60 to 70% from 0800 to 1600 h, n = 23) from the last 60 d of pregnancy until the first colostrum suckling. The heat challenge imposed on HS goats during the prepartum period increased their rectal temperature, respiratory frequency, and cortisol levels in plasma and amniotic fluid versus CT goats. In the placenta, HS treatment also increased the expression of the HSPA1A gene. Heat-stressed goats also showed significantly lower expression of HSD11B2 and greater expression of MC2R and NR3C1 than CT goats, suggesting that heat stress decreased the effectiveness by which the HSD11B2 enzyme converts cortisol to cortisone and increased placental responsiveness to cortisol. The HS goats took longer to release the placenta with lighter placental cotyledons, and HS goats had a lower ratio between the kid's weight at birth and placenta weight than CT goats. There was no treatment effect on the kids' survival or weights at birth, but the kids from goats subjected to HS presented lesser cortisol concentration and greater mortality rates at weaning than kids from CT goats. Finally, the overexpression of HSPA1A by HS goats suggests a protective response of placenta. However, the heat stress negatively affected the placenta's expulsion length, placental cotyledons number, weight and area, the ratio between kid's weight and placenta weight, and cortisol signaling. Indeed, the upregulation of MC2R and NR3C1 and downregulation of HSD11B2 on placenta caused by heat stress were associated with greater cortisol concentrations in the amniotic fluid of HS goats. Although HS and CT kids had adequate weights and survival rate during the first weeks of life, the heat stress increased the mortality at weaning of HS kids versus CT kids, suggesting that the heat stress effect persists and can change the ability of kids to respond to weaning challenge.

Improved vitrification method allowing direct transfer of goat embryos.

The aim of this study was to design a vitrification method suited to field embryo transfer experiments in goat. In a first experiment, a standard vitrification protocol, previously designed for sheep embryos was compared to slow freezing of goat embryos. No significant difference was observed on kidding rate (48% versus 69%, respectively), nor on embryo survival rate (35% versus 45%). Second experiment: all embryos were vitrified. After warming, embryos were either transferred directly (direct transfer), or after in vitro dilution of the cryoprotectants (conventional transfer). The kidding rate was not affected by the transfer method (38% versus 23%, respectively). However, embryo survival rate tended to be higher after direct transfer (26% versus 14%). Third experiment: OPS vitrification was compared to standard vitrification. The kidding rate was not affected (22% versus 39%, respectively), but the embryo survival rate was lower after OPS (14% versus 28%). Fourth experiment: 0.4M sucrose was added with cryoprotectants in vitrification. The kidding rate after direct transfer was significantly enhanced after addition of sucrose (56% versus 27%, respectively), whereas embryo survival rate was not significantly affected (32% versus 18%). Fifth experiment: vitrification with sucrose supplementation was compared to slow freezing. No significant difference was observed after direct transfer on kidding rate (52% versus 31%, respectively), but embryo survival rate tended to be higher after vitrification (34% versus 21%). In conclusion, our results indicate that addition of 0.4M sucrose in association with direct transfer improves significantly the viability of goat vitrified embryos.

Efeito de condicionadores químicos na cama de frango sobre o desempenho de frangos de corte / Effect of chemical conditioners in the poultry litter on the broiler performance

Estudou-se o efeito de condicionadores químicos na cama de frango sobre o desempenho de frangos de corte criados em três lotes consecutivos. Foram utilizadas 1320 aves, 440 em cada lote, em delineamento experimental em blocos ao acaso, com cinco tratamentos (Trat. 1 - cama sem tratamento; Trat. 2 - cama tratada com sulfato de alumínio; Trat. 3 - cama tratada com gesso agrícola; Trat. 4 - cama tratada com superfosfato simples e Trat. 5 - cama tratada com cal hidratada) e quatro repetições. As aves e as rações foram pesadas no início e no final do período experimental para obtenção do peso final, do consumo de ração, da conversão alimentar e da viabilidade. O uso dos condicionadores não influenciou (P>0,05) o peso final, o consumo de ração e a conversão alimentar. A adição do sulfato de alumínio e do superfosfato simples reduziu (P<0,05) a viabilidade das aves. Não é necessário o uso dos condicionadores na cama de frango reutilizada até o terceiro lote.

Successful direct transfer of vitrified sheep embryos.

The use of a simple cryopreservation method, adapted to direct transfer of thawed embryos may help to reduce the costs of embryo transfer in sheep and increase the use of this technique genetic improvement of this species. Two experiments were made to test a vitrification method that is easy to apply in field conditions. All embryos were collected at Day 7 of the estrous cycle of FSH-stimulated donor ewes and were assessed morphologically, washed in modified PBS and incubated for 5 min in 10% glycerol, for 5 min in 10% glycerol and 20% ethylene glycol and were transferred into the vitrification solution (25% glycerol and 25% ethylene glycol). All solutions were based on mPBS. Embryos were loaded in straws (1 cm central part, the remaining parts being filled with 0.8 M galactose in mPBS) and plunged into liquid N2 within 30 sec of contact with the vitrification solution. The straws were thawed (10 sec at 20 degrees C) and the embryos were either transferred directly or after 5 min of incubation in the content of the straw (followed by washing in PBS) into the uterus of a recipient ewe. In Trial 1, the pregnancy rates at term (72 vs. 72%) as well as the embryo survival rates (60 vs 50% respectively) were not different between fresh (n = 48 embryos) and vitrified (n = 50) embryos. In a second trial no difference was observed between vitrified embryos transferred after in vitro removal of the cryoprotectant (n = 86 embryos) or directly after thawing (n = 72) both in terms of lambing rate (67 vs. 75%, respectively) and embryo survival rate (lambs born/embryos transferred; 49 vs. 53%). This method of sheep embryo cryopreservation provided high pregnancy and embryo survival, even after direct transfer of the embryos.

Recombinant FVIIa (NovoSeven) continuous infusion and total hip replacement in patients with haemophilia and high titre of inhibitors to FVIII: experience of two cases.

In this report we describe our experience of total hip replacement in two patients with severe haemophilia A and high titres of inhibitors to FVIII. We used rFVIIa replacement therapy by continuous infusion to perform the surgery. The total amount of rFVIIa used in these two patients was very similar but the manner of administration was quite different. In our experience, it is an advantage to use a higher dose for shorter periods than a lower dose for a longer treatment period. Tranexamic acid by continuous infusion, and parallel saline infusion were useful for good haemostasis and avoided local thrombophlebitis in the side of rFVIIa infusion.

Safety and efficacy of high-purity concentrates in haemophiliac patients undergoing surgery by continuous infusion.

In this study we explore the feasibility of high-purity double-inactivated concentrates by continuous infusion for the treatment of haemophiliacs in a group of patients undergoing different surgical procedures. The patients were enrolled in the study on the basis of their transfusion history, which was well known due to their long-term follow up at our Haemophilia Center. We did not perform a pre-operative pharmacokinetic study because one of the aims of this study was to demonstrate that continuous infusion can become a first choice standard treatment in patients with haemophilia. Fourteen haemophilia A and one haemophilia B patients who needed at least 5 days of replacement therapy were monitored for haemostatic efficacy, post-operative factor VIII and factor IX levels and evaluated for safety and flexibility of the products. The infusion rate of 3 IU kg-1 h-1 was demonstrated to be sufficient to ensure haemostasis and patients did not need additional bolus infusion during the post-operative period. Our study demonstrates the safety and feasibility of high-purity concentrates in patients undergoing surgery by continuous infusion, also in the absence of a previous pharmacokinetic study.

Predictor of the rate of CD4 lymphocyte loss in HIV-1-seropositive asymptomatic hemophiliacs by in vitro immunoglobulin synthesis.

The objective of the present study was to evaluate the prognostic utility in determining the risk of AIDS progression in HIV-1-infected asymptomatic hemophiliacs by in vitro immunoglobulin (Ig) synthesis. With this aim, a cohort of 28 HIV-1-seropositive hemophiliacs were studied. All showed the number of CD4 lymphocytes higher than 400 positive cells/mm3. In all cases the spontaneous and pokeweed mitogen-induced in vitro production of Ig by peripheral blood lymphocytes was evaluated at the beginning of the study and the ratio stimulated/spontaneous (Stim/Spon) synthesis was calculated. At the same time, the absolute CD8+ cell count, IgA serum immunoglobulin, p24 HIV-1 antigenemia, and beta2 microglobulin were calculated. These data were monitored during the 4-year follow-up of patients and compared with the stimulated/spontaneous Ig synthesis ratio to evaluate the predictive significance on the progression of HIV infection. According to the stimulated/spontaneous Ig synthesis ratio, hemophilic patients were separated into two categories. Group I included 12 subjects with a Stim/Spon ratio higher than 2 (the lowest value of normal controls) and group II included 16 cases with a ratio lower than 2. As control, in 36 HIV-1-negative hemophiliac individuals the stimulated/spontaneous Ig ratio ranged between 2 and 42; mean +/- SEM, 12.9 +/- 1.8. At the end of the 4-year follow-up, group I patients showed a CD4 count and clinical staging consistent with those of the first evaluation; in contrast group II demonstrated a significant decrease in CD4 lymphocytes and deterioration of clinical conditions. Our results show that a low Stim/Spon Ig ratio when the CD4 lymphocyte count was still normal appears to predict the depletion of this lymphoid subset and progression to AIDS before T CD8, IgA immunoglobulin, p24 HIV-1 antigenemia, and beta2 microglobulin abnormalities. In this setting, the stimulated/spontaneous Ig ratio may represent a useful tool for clinical decisions in HIV-1-infected hemophiliacs.

Hepatitis C virus genotypes and severity of chronic liver disease in haemophiliacs.

We studied the activity and stage of chronic liver disease in 45 HCV-seropositive/HIV-seronegative patients with severe haemophilia followed for at least 10 years. HCV-RNA was detected in serum in 36 patients (80%) Viraemic cases were further analysed for HCV genotypes: 10 (28%) were infected by type 1a, 10 (28%) by type 1b, seven (19%) by type 2, four (11%) by type 3, four (11%) had mixed infections (one by 1a + 1b, one by 1a + 2, one by type 2 + 3, and one by 1a + 2 + 3). ALT levels were within the normal range in 55% of the HCV-RNA negative patients but in only 11% of the viraemic cases. Results show a trend for higher levels of ALT in HCV-RNA-positive patients compared with those without viraemia (98 +/- 56 v 60 +/- 61), and particularly with patients with type 3 HCV infection (148 +/- 44). We suggest that a slow progression of chronic liver disease occurs in haemophilic HCV-positive/HIV-negative patients and conclude that presence of HCV-RNA in serum correlates well with cytolitic damage but, in the time-scale of our follow-up period, commonly used clinical-laboratory parameters cannot predict the chronic evolution of liver infection or identify differences in disease progression in relation to specific HCV subtypes.

Phenotypical and functional evaluation of CD8+/S6F1+ T lymphocytes in haemophiliac individuals with HIV-1 infection.

In this study we investigated the distribution of the S6F1 antigen, an epitope of the lymphocyte function-associated antigen, on CD8+ T lymphocytes in a series of 15 HIV-1+ and 20 HIV-1- haemophiliac patients. MoAbs recognizing the S6F1 antigen have been claimed to distinguish between killer effectors (brightly S6F1+ stained) and suppressor cells (dimly S6F1+ stained) within the CD8+ lymphoid population. In addition, we tried to find a correlation between the spontaneous in vitro immunoglobulin synthesis from patients' peripheral blood lymphocytes and the pattern of S6F1 expression. Although the total number of double-positive CD8+/S6F1+ cells was similar in both HIV-1+ and HIV-1- haemophiliac patients, a significant increase in the CD8+/S6F1+ population bright versus dim was documented in HIV-1-infected with respect to HIV-1- haemophiliacs (bright/dim ratio 3.97 +/- 0.61 versus 0.75 +/- 0.1, respectively, P < 0.005). This finding was correlated to a significant increase in spontaneous in vitro immunoglobulin production in HIV-1+ subjects compared with control haemophiliacs (P < 0.005). Purified CD8+ lymphocytes from HIV-1+ subjects showed a reduced suppressor activity on mitogen-induced immunoglobulin production. Taken together, these data suggest that HIV-1 infection favours the generation of CD8+/S6F1+ bright cells with putative cytotoxic-associated function, leading to a progressive reduction in the number of CD8+/S6F1+ dim suppressor lymphocytes. This phenomenon may contribute to the polyclonal hypergammaglobulinaemia present in HIV-1+ haemophiliac patients.

Orientación actual en el tratamiento quirúrgico de la sinovitis hemofilica con la sinovectomía / Recent surgical treatment of the synovitis haemophilic with the synovectomy

Han sido revisadas 350 sinovectomías (hombro, codo, cadera, rodilla, tobillo) con un seguimiento de 8 meses a 15 años. Del examen minucioso, se pudo observar que la sinovectomía da los mejores resultados cuando la artropatia presenta las siguientes características: HISTORIA DE HEMARTROSIS repetidas sin intervalos de completa remisión; signos de inicial flogosis articular; signos radiológicos de leve sufrimiento osteo-condoral. Se demostró no resolutiva cuando la actividad destructora de la sinovial ha llegado al ápice se ha extinguido, cuando los daños articulares son con deformación de los extremos óseos y desviaciones axiales y el síntoma más importante es el dolor a la carga (cadera, rodilla, tobillo). Los autores se refieren a una larga experiencia con la sinovectomía, recogida en 10 años, en el tratamiento de la artropatia hemofílica. Los resultados de una revisión a distancia de la casuística operatoria presentados en el Congreso de Dousseldorf (1985) fueron satisfactorios en la evolución de la artropatía y respecto al dolor y a la función articular

La seguridad transfusional de los hemofílicos: comparación entre el C.P. "bruto" de factor VIII y plasma de ánico donador y concentrados comerciales de "pools" / Transfusional security of the haemophilics: comparation between the C.P. of factor VIII and plasma from inique donor and commercial concentrate of \"pools\"

Se hace un estudio de la posibilidad de infección con el virus HIV en preparados de plasma, crios y comerciales

La osteotomía correctora de miembros inferiores en la artropatía hemofílica / The osteotomy corrector of lower extremite in the hemophilic arthropaty

En los pacientes afectados de Artropatía Hemofílica, es frecuente observar, especialmente en los miembros inferiores, la presencia de posiciones viciosas y de desviaciones axiales. Las alteraciones osteoarticulares prevalentes en los estados avanzados de la artropatía son las responsables del desequilibrio mecánico, de las recidivas de la hemartrosis y del dolor. Para el tratamiento de la artropatía con deformidad articular hemos recurrido alguna vez a intervenir realizando una osteotomía correctora asociada a sinovectomía o a "Rebaja Articular", y otras veces a intervenir realizando osteotomía solamente. Han estado seguidas 23 osteotomías (5 inter-trocantéricas para cadera; 6 supracondíleas y 12 osteotomías altas de tibia, para la rodilla) Tabla 1. Osteotomías 23, Cadera , Supracondilea fémur 6, Proximales de Tibia 12, Pacientes 20. Los controles a larga data (de 1 a 6 años), han constatado en la mayoría de los pacientes operados, además de la estabilidad de la corrección obtenida, el reinicio de una marcha más correcta y de la capacidad para el trabajo, la desaparición del dolor y de las recidivas de hemartrosis. Resultados que hemos obtenido también en aquellos pacientes en los cuales se les realizó osteotomía solamente. El juicio del paciente fue casi siempre muy satisfactorio e igualmente, si el cuadro radiológico de la articulación no ha mostrado variaciones relevantes, la mejor distribución de carga ha producido una función articular más correcta y con mejoramiento de la sintomatología

Repair of ventricular septal defect and aortic regurgitation associated with severe hemophilia B.

A child with severe factor IX deficiency who underwent an open-heart operation using extracorporeal circulation is described. The factor IX level was normalized immediately before operation and at the end of cardiopulmonary bypass by infusing prothrombin complex concentrate and fresh frozen plasma. Partial thromboplastin time and factor IX serum levels were monitored for 20 days postoperatively and showed factor IX activity higher than 50%.

Early synovectomy in haemophilia.

The Authors reviewed, after from 1 to 11 years, 28 joints of patients with A and B haemophilia operated on with synovectomy at an early stage. Investigation is made into the diagnostic elements for evaluating the earliness of arthropathy and indications for surgery. The results from early synovectomy are compared with the case histories reported in the literature when synovectomy was done at an advanced stage.