ADA-deficient SCID is associated with a specific microenvironment and bone phenotype characterized by RANKL/OPG imbalance and osteoblast insufficiency

Adenosine deaminase (ADA) deficiency is a disorder of the purine metabolism leading to combined immunodeficiency and systemic alterations, including skeletal abnormalities. We report that ADA deficiency in mice causes a specific bone phenotype characterized by alterations of structural properties and impaired mechanical competence. These alterations are the combined result of an imbalanced receptor activator of nuclear factor-kappa B ligand (RANKL)/osteoprotegerin axis, causing decreased osteoclastogenesis and an intrinsic defect of osteoblast function with subsequent low bone formation. In vitro, osteoblasts lacking ADA displayed an altered transcriptional profile and growth reduction. Furthermore, the bone marrow microenvironment of ADA-deficient mice showed a reduced capacity to support in vitro and in vivo hematopoiesis. Treatment of ADA-deficient neonatal mice with enzyme replacement therapy, bone marrow transplantation, or gene therapy resulted in full recovery of the altered bone parameters. Remarkably, untreated ADA-severe combined immunodeficiency patients showed a similar imbalance in RANKL/osteoprotegerin levels alongside severe growth retardation. Gene therapy with ADA-transduced hematopoietic stem cells increased serum RANKL levels and children`s growth. Our results indicate that the ADA metabolism represents a crucial modulatory factor of bone cell activities and remodeling. The trials were registered at www.clinicaltrials.gov as #NCT00598481 and #NCT00599781. (Blood. 2009; 114: 3216-3226)

Progression of Lipid Peroxidation Measured as Thiobarbituric Acid Reactive Substances, Damage to DNA and Histopathological Changes in the Liver of Rats Subjected to a Methionine-Choline-Deficient Diet

Methionine-choline-deficient diet represents a model for the study of the pathogenesis of steatohepatitis. Male rats were divided into three groups, the first group receiving a control diet and the other two groups receiving a methionine-choline-deficient diet for 1 month (MCD1) and for 2 months (MCD2), respectively. The livers of the animals were collected for the determination of vitamin E, thiobarbituric acid reactive substances (TBARS), GSH concentration, DNA damages, and for histopathological evaluation. The hepatic TBARS and GSH content was higher (P < 0.05) in the groups receiving the experimental diet (MCD1 and MCD2) compared to control diet, and hepatic vitamin E concentration differed (P < 0.05) between the MCD1 and MCD2 groups, with the MCD2 group presenting a lower concentration. Damage to hepatocyte DNA was greater (P < 0.05) in the MCD2 group (262.80 DNA injuries/100 hepatocytes) compared to MCD1 (136.4 DNA injuries/100 hepatocytes) and control diet (115.83 DNA injuries/100 hepatocytes). Liver histopathological evaluation showed that steatosis, present in experimental groups was micro- and macro-vesicular and concentrated around the centrolobular vein, zone 3, with preservation of the portal space. The inflammatory infiltrate was predominantly periductal and the steatosis and inflammatory infiltrate was similar in the MCD1 and MCD2 groups, although the presence of Mallory bodies was greater in the MCD2 group. The study describes the contribution of a methionine-choline-deficient diet to the progression of steatosis, lipid peroxidation and hepatic DNA damage in rats, serving as a point of reflection about the role of these nutrients in the western diet and the elevated non-alcoholic steatohepatitis rates in humans.

Effects of the utilization of homeopathic elements in commercial diluent on swine sperm viability

It has been speculated that the homeopathic treatment of sperm cells in order to improve semen quality could be promising. However, few data is available and its use in spermatozoa requires investigation. It is well established that mitochondrial membrane potential is an important viability parameter of spermatozoa and it is intimately related to reproductive efficiency. In this manner, new technologies in order to improve the activity of sperm cells and, finally, the fecundity of swine herds are of extremely importance. Due to the lack of knowledge of homeopathic treatment effect on spermatozoa, the aim of the present study was to verify the effect of three different homeopathic treatments on viability of boar sperm cells. Three homeopathic treatments composed by Pulsatila CH6, Pulsatila and Avena CH6, Avena CH6 and one control treatment (sucrose) were added to diluted boar semen, which were cooled for 24 or 48 h. Interestingly, no positive effect of homeopathic treatments was observed over semen viability. However, it was demonstrated that the 24 h of cooling storage provided more viable sperm cells when compared to the 48-h period. This effect of storage period on sperm viability was assessed by intact plasmatic membrane, intact acrosome and mitochondrial membrane potential evaluation.

Bovine oocyte vitrification: Effect of ethylene glycol concentrations and meiotic stages

Success in oocyte cryopreservation is limited and several factors as cryoprotectant type or concentration and stage of oocyte meiotic maturation are involved. The aim of the present study was to evaluate the effect of maturation stage and ethylene glycol (EG) concentration on survival of bovine oocytes after vitrification. In experiment 1, kinetics of oocyte in vitro maturation (IVM) was evaluated. Germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I (MI), and metaphase II (MII) oocytes were found predominantly at 0, 0-10, 10-14, and 18-24 h of INK respectively. In experiment 2, in vitro embryo development after in vitro fertilization (IVF) of oocytes exposed to equilibrium (ES) and vitrification solution VS-1 (EG 30%), or VS-2 (EG 40%) at 0, 12 or 18 It of IVM was evaluated. Only blastocyst rate from oocytes vitrified in SV-2 after 18 h of IVM was different from control oocytes. Hatched blastocyst rates from oocytes vitrified in VS-1 after 12 and 18 h, and SV-2 after 18 h of IVM were different from unvitrified oocytes. In experiment 3, embryo development was examined after IVF of oocytes vitrified using VS-I or VS-2 at 0, 12 or 18 h of IVM. Rates of blastocyst development after vitrification of oocytes in VS-1 at each time interval were similar. However, after vitrification in VS-2, blastocyst rates were less at 18 h than 0 h. Both cleavage rates and blastocyst rates were significantly less in all vitrification groups when compared to control group and only control oocytes hatched. In conclusion, both EG concentration and stage of meiotic maturation affect the developmental potential of oocytes after vitrification. (C) 2007 Elsevier B.V. All rights reserved.

Effect of culture media on porcine embryos produced by in vitro fertilization or parthenogenetic activation after oocyte maturation with cycloheximide

This study evaluated the effects of reversible meiotic inhibition and different culture media (PZM3 or NCSU23) on production of porcine embryos by either in vitro fertilization (IVF) or parthenogenetic activation (PA). Oocytes from abattoir-derived ovaries were allocated into two groups for maturation: CHX (5 mu g/ml cycloheximide for 10 h) or Control (no CHX). The percentage of metaphase II (MII) oocytes was determined at 36, 40 or 44 h of in vitro maturation. For IVF and PA, denuded oocytes were fertilized with purified sperm for 6 h or activated by electric stimuli. Zygotes were then subdivided into two culture groups: NCSU23 or PZM3. No effect of treatment with CHX and culture media was observed on cleavage (D3) and blastocyst (D7) rates in IVF and PA groups. There are no differences of quality or development rates between IVF-derived embryos cultured in NCSU23 or PZM3. However, we observed high quality PA embryos in PZM3 compared with NCSU23. Maturation arrest with CHX decreased the average blastocyst cell number in IVF while it was increased in PA embryos. As older oocytes are more effectively activated, CHX-blocked oocytes reached the mature stage faster than the control group. In conclusion, the CHX treatment for 10 h, followed by oocyte maturation for 40 h, is an efficient protocol to produce high quality parthenote embryos, especially when they are cultured in PZM3. However, this protocol is not satisfactory for IVF embryos production. In this case, a shorter maturation period could provide better embryo quality.

Bovine sperm cells viability during incubation with or without exogenous DNA

The aim of this study was to assess the effect of exogenous DNA and incubation time on the viability of bovine sperm. Sperm were incubated at a concentration of 5 x 10(6)/ml with or without plasmid pEYFP-NUC. Fluorescent probes, propidium iodide/Hoechst 33342, FITC-PSA and JC-1, were used to assess plasma membrane integrity (PMI), acrosome membrane integrity (AMI) and mitochondrial membrane potential (MMP) respectively at 0, 1, 2, 3 and 4 h of incubation. Exogenous DNA addition did not affect sperm viability; however, incubation time was related to sperm deterioration. Simultaneous assessment of PMI, AMI and MMP showed a reduction in the number of sperm with higher viability (integrity of plasma and acrosome membranes and high mitochondrial membrane potential) from 58.7% at 0 h to 7.5% after 4 h of incubation. Lower viability sperm (damaged plasma and acrosome membranes and low mitochondrial membrane potential) increased from 4.6% at 0 h to 25.99% after 4 h of incubation. When PMI, AMI and MMP were assessed separately we noticed a reduction in plasma and acrosome membrane integrity and mitochondrial membrane potential throughout the incubation period. Therefore, exogenous DNA addition does not affect sperm viability, but the viability is reduced by incubation time.

Effects of gonadotropin-exposed medium with high concentrations of progesterone and estradiol-17 beta on in vitro maturation of canine oocytes

During the process of maturation in the oviduct, canine oocytes in the germinal vesicle stage are exposed to decreasing levels of estradiol-17 beta and increasing levels of progesterone. However, hormone concentrations in the microenvironments in which they act are higher than serum concentrations. Therefore, the aim of the present study was to compare the meiotic competence of canine oocytes harvested from anestrous bitches in culture medium containing high concentrations (20 mu g ml(-1)) of estradiol-17 beta and/or progesterone in association to gonadotropins (luteinizing hormone and follicle-stimulating hormone) using three different maturation periods (48, 72, and 96 h). Oocytes were cultured in tissue culture medium (TCM-199) and arranged in four experimental groups: group control, group E2 (estradiol-17 beta), group P4 (progesterone), and group E2 + P4. Regardless of the maturation period, groups P4 and E2 + P4 presented statistically higher rate of germinal vesicle breakdown oocytes compared to the group control and group E2. There were no significant differences among groups on germinal vesicle, metaphase I, metaphase II, and degenerated or unidentifiable oocytes rates. The mean percentage of metaphase II oocytes was higher at 96 h when compared to 72 h. Results of the present research indicate no influence of estradiol-17 beta supplementation, unless in association with progesterone. There is an evidence of the positive effect of progesterone on germinal vesicle breakdown. Results also showed that extended periods of in vitro maturation affect positively maturation rates to metaphase II of low competent oocytes harvested from anestrous bitches, independent of the maturation media. In conclusion, high concentrations of steroids, especially progesterone, have positive effect on in vitro oocyte maturation when the oocytes are derived from the anestrous status.

Remifentanil versus dexmedetomidina como coadjuvantes de técnica anestésica padronizada em pacientes com obesidade mórbida

Justificativa e objetivos: comparou-se a ação de duas drogas coadjuvantes da anestesia, remifentanil e dexmedetomidina, na recuperação anestésica e na evolução do pH e da PaCO2, em pacientes com obesidade mórbida que foram submetidos à cirurgia de Capella. Método: o estudo foi aleatório, prospectivo e duplamente encoberto. Noventa e dois pacientes foram designados a um de dois grupos e submetidos à técnica anestésica (geral/peridural) padronizada. O grupo Remifentanil (Grupo R) e o da Dexmedetomidina (Grupo D) receberam infusão contínua por via venosa destas drogas (0,1 µg.kg-1.min-1 e 0,5 µg.kg-1.h-1 peso ideal mais 30% para ambas) logo após a intubação traqueal. Os pacientes foram monitorizados com pressão arterial média invasiva, oximetria de pulso, EEG bispectral (BIS), capnografia, estimulador de nervo periférico e ECG. Foram avaliados: 1) diferentes tempos de recuperação anestésica (abertura dos olhos, reinicio da respiração espontânea, tempo de extubação traqueal, tempo para de alta da sala de recuperação pós-anestésica e hospitalar), 2) a evolução da gasometria arterial, e 3) analgesia pós-operatória. Resultados: oitenta e oito pacientes foram avaliados. Os pacientes do grupo R apresentaram abertura ocular precoce (9,49 ± 5,61 min versus 18,25 ± 10,24 min, p < 0,0001), menor tempo para reiniciar a ventilação espontânea (9,78 ± 5,80 min versus 16,58 ± 6,07 min, p < 0,0001), e menor tempo para a extubação traqueal (17,93 ± 10,39 min versus 27,53 ± 13,39 min, p < 0,0001). Não houve diferença quanto ao tempo para alta anestésica (105,18 ± 50,82 min versus 118,69 ± 56,18 min) e para alta hospitalar (51,13 ± 6,37 horas versus 52,50 ± 7,09 horas). Os dois grupos apresentaram diminuição dos valores de pH e da PaO2 imediatamente após a extubação traqueal comparados com valores pré-operatórios, e que se manteve até a alta da SRPA. O grupo D apresentou valores maiores de PaCO2 após a extubação traqueal, comparados com valores pré-operatórios no mesmo grupo (p < 0,05), divergente do Grupo R; 41% dos pacientes do Grupo R e 60% do Grupo D (p < 0,02) requisitaram medicação analgésica de resgate no primeiro dia de pós-operatório. Conclusões: na população avaliada, a associação de remifentanil em técnica anestésica padronizada resultou em recuperação anestésica mais rápida, manutenção dos valores de PaCO2 durante o período pós-operatório imediato e menor consumo de analgésicos de resgate no período pós-operatório, quando comparada à dexmedetomidina.

Postharvest biological control of brown rot in peaches after cold storage preceded by preharvest chemical control 1

ABSTRACT Pathogenic fungi cause skin darkening and peach quality depreciation in post harvest. Therefore, alternative techniques to chemical treatment are necessary in order to reduce risks to human health. The aim of this study was to evaluate the effect of the application of Trichoderma harzianum in association with different fungicides applied before harvest to 'Eldorado' peaches for brown rot control and other quality parameters during storage. The treatments consisted of five preharvest fungicide applications (control, captan, iprodione, iminoctadine and tebuconazole) associated with postharvest application of T. harzianum, after cold storage (with and without application), in three evaluation times (zero, two and four days at 20 °C), resulting in a 5x2x3 factorial design. The application of T. harzianum only brought benefits to the control of brown rot when combined with the fungicide captan, at zero day shelf life. After two days, there was a greater skin darkening in peaches treated with T. harzianum compared with peaches without the treatment, except for peaches treated with the fungicide iprodione and T. harzianum The application of T. harzianum during postharvest showed no benefits for the control of brown rot, however, the association with fungicides reduced the incidence of Rhizopus stolonifer during the shelf life.

Coalizões em educação no Brasil: relação com o governo e influência sobre o Plano de Desenvolvimento da Educação (PDE)

Atualmente, duas coalizões lutam pela melhoria da qualidade da educação no Brasil: a Campanha Nacional pelo Direito à Educação e o Todos pela Educação. Essas duas coalizões diferenciam-se dos movimentos históricos por sua capacidade de agregar atores provenientes de instituições governamentais e não governamentais, de diversos níveis, que possuem valores e crenças comuns em uma aliança única, aproximando-se do conceito de coalizões advocatórias proposto por Sabatier e Jenkins-Smith (1993). Apesar da bandeira comum, elas possuem origens, composições, metas e formas de atuação completamente diferentes - a relação com o governo é um dos pontos que mais as diferencia. O lançamento do Plano de Desenvolvimento da Educação (PDE), com uma das medidas intitulada "Plano de Metas Compromisso Todos pela Educação", gerou questionamentos com relação à similaridade dos nomes entre um plano de governo e uma das coalizões. Neste artigo, apresentaremos as duas coalizões e a relação de ambas com o governo, utilizando o PDE como estudo de caso.

La Iglesia católica en Iberoamérica: las instituciones locales en una época de cambios (siglo XVIII)

El presente artículo considera de modo general el conjunto de las reconfiguraciones en el ámbito religioso que tuvieron lugar en Iberoamérica durante el siglo XVIII. De modo específico busca mostrar a partir de la bibliografía disponible un orden de procesos no demasiado visible en torno al papel de algunas instituciones eclesiásticas y religiosas - como las parroquias y las cofradías - en los procesos de estructuración social y política, así como las consecuencias de un tipo de intervención política como el que se verifica durante las reformas borbónicas y pombalinas.

Presencia de Triatoma sordida Stäl, 1859 en ecotopos urbanos de la ciudad de Corrientes, Argentina

Con el objeto de verificar la presencia de T. sordida Stäl 1859, en ecotopos urbanos de la ciudad de Corrientes, Argentina se tomaron como base un habitat extradomiciliario y la vivienda humana. En el palomar situado en el Parque Mitre fue realizado un censo completo de población de T. sordida para lo cual fueron disecados 400 nidos de paloma (Columbia livia). Los ejemplares de triatominos detectados en las viviendas fueron enviados por los pobladores al Laboratorio de Artrópodos. Los triatominos fueron determinados sistemáticamente y por clase de edad. La materia fecal de los mismos fue examinada al microscopio para la identificación del Trypanosoma cruzi. En el palomar se colectaron: 6 huevos, 45 ninfas de primer estadío, 110 ninfas de segundo estadío, 226 ninfas de tercer estadío, 303 ninfas de cuarto estadío, 1.103 ninfas de quinto estadío, 12 hembras y 11 machos. Se constató la presencia de esta especie en 27 viviendas localizadas en 8 barrios de la Ciudad. Se capturaron 13 huevos, 33 ninfas de diferentes estadíos, 27 hembras y 9 machos, totalizando 82 individuos. Ningún triatomino resultó infectado por T. cruzi. Los adultos de T. sordida se desplazaron hacia el domicilio en primavera y verano. Es de destacar que el 33% del material hallado en las viviendas correspondió a hembras. Se ha comprobado la adaptación de T. sordida a un habitat extradomiciliario y su tendencia a invadir la vivienda humana, hecho que se debería ser tenido en cuenta en los programas de control de vectores.

Interacción entre poblaciones de Triatoma infestans y Triatoma sordida

Para conocer si se producía algún tipo de interacción entre poblaciones de Triatoma infestans y Triatoma sordida que convivían en una unidad experimental y explotaban el mismo recurso alimentario (ave) se las estudió desde setiembre/1988 a abril/1989. La composición etaria inicial para cada especie fue: 27 N1, 7 N2, 11 N3, 3 N4, 8 N5, 4 machos y 10 hembras. La dinámica de población, el estado nutricional, la predación y la conducta gregaria, utilizados como parámetros de comparación, fueron estimados mediante censos mensuales. Triatoma infestans logró mayor éxito colonizador que T. sordida dado los valores alcanzados en: fecundidad (146 huevos/hembra), longevidad (157,8 días) y mortalidad (39,4%), en comparación a los obtenidos por T. sordida: 118 huevos/hembra, 81,1 días y 54,0% respectivamente. La población de T. infestans tuvo un crecimiento de tipo exponencial, con una alta tasa de renovación ninfal, en contraposición, la población de T. sordida se extinguió tempranamente. En general, el peso promedio de los ejemplares de T. infestans se mantuvo en valores próximos al inicial, mientras que en T. sordida se redujo. Los grupos gregarios se conformaron mayoritariamente en el sector inferior de la pared 1 (próximo al hospedador), observándose mayor nivel de contagio en T. infestans. A partir de los resultados obtenidos se formula la hipótesis de la superioridad competitiva en T. infestans.

Estadísticos poblacionales de Triatoma sordida Stäl 1859 (Hemiptera : Reduviidae) en condiciones experimentales

Se realizó el seguimiento longitudinal de cuatro cohortes de 100 huevos cada una. Los insectos fueron criados a 28°C ± 3°C y 63 % ± 10% de humedad relativa, y alimentados cada siete días sobre gallina (Gallus domesticus), durante 40 min. Las cohortes fueron controladas una vez por semana. Se observó un valor constante de supervivencia en las cuatro cohortes, sin picos importantes. La expectativa de vida de los machos, en promedio, fue de 37,4 semanas y la de las hembras 36,8. El tiempo generacional demandó 61,7 semanas. La tasa reproductiva neta indica que esta población se incrementará 143,3 veces con cada generación. La tasa intrínseca de crecimiento natural fue de 0,082 por semana. El máximo valor reproductivo se verificó, en promedio, a la tercer semana del ingreso al estado adulto. La distribución estable de edades de T. sordida sería de 32% huevos, 26% ninfas de primer estadio, 19% ninfas de segundo estadio, 13% ninfas de tercer estadio, 6% ninfas de cuarto estadio, 3% ninfas de quinto estadio y 1% de adultos. Es la primera cita para Argentina sobre la tabla de vida de T. sordida. El análisis de estos parámetros permite considerar que esta especie se comporta como "K estratega", en condiciones experimentales.