Sodium caseinate improves longevity and fertility of frozen bull semen.

This study aimed to evaluate the effect of sodium caseinate added into freezing extender on the sperm parameters of cryopreserved bull semen and in vitro and in vivo fertility. One ejaculate of 30 bulls was used and processed using Botu-Bov (Botupharma, Botucatu, Brazil) with the addition of 20% egg yolk (EY) or 15% egg yolk with 2% sodium caseinate (EY + SC), subsequently submitted to freezing. Semen from both groups were evaluated immediately after thawing (T0) and after thermic stress at 37 °C for 90 min (T90), for sperm kinetics, by CASA method, and plasma membrane integrity (PMI), superoxide (O2-) concentration and high mitochondrial potential (HMP) by flow cytometry. In vitro fertilization (IVF) was performed to assess embryo cleavage rate on day 3, and blastocyst rate on day 8. The in vivo fertility test was performed using fixed-time artificial insemination (FTAI). In sperm evaluation, trajectory velocity, linear velocity, curvilinear velocity, and lateral head movement were higher (P < 0.05) in EY + SC at T0. At T90, while rectilinearity and linearity did not differ between EY and EY + SC (P > 0.05), the other parameters evaluated were higher in EY + SC. Similarly, the integrity of the plasma and acrosomal membranes (iPAM) was higher (P < 0.05) at T90 in EY + SC, but did not differ (P > 0.05) between the groups at T0. For O2- and HMP, the values were lower (P < 0.05) in EY + SC group in both moments; furthermore, EY + SC showed higher cleavage and blastocyst rates in IVF. Likewise, pregnancy rates by FTAI were higher (P < 0.05) in the EY + SC group. In conclusion, the addition of sodium caseinate into freezing extender improves sperm parameters of frozen-thawed bull semen and fertility rates on during in vitro and in vivo tests.

Safe and successful protocol for desensitization to abiraterone

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Characterization of agar/soy protein biocomposite films: Effect of agar on the extruded pellets and compression moulded films.

Agar/soy protein biocomposite films were successfully processed by extrusion and compression moulding, obtaining transparent and homogeneous films. The conformational changes occurred during the extrusion process and the effect of agar on the final properties were analyzed. As shown by differential scanning calorimetry (DSC) and specific mechanical energy (SME) values, during the extrusion process protein denatured and unfolded protein chains could interact with agar. These interactions were analyzed by Fourier transform infrared spectroscopy (FTIR) and the secondary structure was determined from the amide I band. Those interactions were supported by the decrease of film solubility. Furthermore, the good compatibility between agar and soy protein was confirmed by the images from scanning electron microscopy (SEM).

Giant cell tumour of bone: new treatments in development

Giant cell tumour of bone (GCTB) is a benign osteolytic tumour with three main cellular components: multinucleated osteoclast-like giant cells, mononuclear spindle-like stromal cells (the main neoplastic components) and mononuclear cells of the monocyte/macrophage lineage. The giant cells overexpress a key mediator in osteoclastogenesis: the RANK receptor, which is stimulated in turn by the cytokine RANKL, which is secreted by the stromal cells. The RANK/RANKL interaction is predominantly responsible for the extensive bone resorption by the tumour. Historically, standard treatment was substantial surgical resection, with or without adjuvant therapy, with recurrence rates of 20–56 %. Studies with denosumab, a monoclonal antibody that specifically binds to RANKL, resulted in dramatic treatment responses, which led to its approval by the United States Food and Drugs Administration (US FDA). Recent advances in the understanding of GCTB pathogenesis are essential to develop new treatments for this locally destructive primary bone tumour (AU)

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Giant cell tumour of bone: new treatments in development.

Giant cell tumour of bone (GCTB) is a benign osteolytic tumour with three main cellular components: multinucleated osteoclast-like giant cells, mononuclear spindle-like stromal cells (the main neoplastic components) and mononuclear cells of the monocyte/macrophage lineage. The giant cells overexpress a key mediator in osteoclastogenesis: the RANK receptor, which is stimulated in turn by the cytokine RANKL, which is secreted by the stromal cells. The RANK/RANKL interaction is predominantly responsible for the extensive bone resorption by the tumour. Historically, standard treatment was substantial surgical resection, with or without adjuvant therapy, with recurrence rates of 20-56 %. Studies with denosumab, a monoclonal antibody that specifically binds to RANKL, resulted in dramatic treatment responses, which led to its approval by the United States Food and Drugs Administration (US FDA). Recent advances in the understanding of GCTB pathogenesis are essential to develop new treatments for this locally destructive primary bone tumour.

Seawater injection barrier recharge with advanced reclaimed water at Llobregat delta aquifer (Spain).

The main aquifer of the Llobregat delta (Barcelona, Spain) has been affected by seawater intrusion since the 1960s. The Catalan Water Agency (ACA) has sponsored the construction of a positive hydraulic barrier in order to stop the progress of seawater intrusion advance due to the intensive aquifer development. The hydraulic barrier consists of 15 wells into which highly treated reclaimed water from the waste water treatment plant of the Baix Llobregat is injected. Water is subjected, prior to the distribution to the injection wells, to secondary and tertiary treatments, and later to ultrafiltration, UV disinfection without chlorination, and salinity reduction through reverse osmosis. A preliminary pilot phase of the project was started in late 2007, with highly positive results, and the second phase started in mid 2010. Hydrogeological and hydrochemical monitoring data indicate an efficient performance and aquifer improvement. The evaluation of such efficiency and operational costs has been analyzed and discussed.

Implementación universal de un cribado de defectos congénitos de garantía en un área sanitaria: área hospitalaria de Valme, Sevilla, España / Implementation of a standardised and universal screening on congenital defect in a health area: Valme hospital area, Seville, Spain

Antecedentes: La posibilidad de que un recién nacido presente algún tipo de defecto congénito al nacimiento es de un 2-4 por ciento y la aplicación de métodos de cribado de cromosomopatías y de malformaciones estructurales puede reducir la prevalencia de estos defectos congénitos al nacimiento. Objetivos: Demostrar que es posible la implantación de un cribado de malformaciones congénitas de garantía (sensibilidad de diagnóstico para malformaciones estructurales mayores y para síndrome de Down del 80 por ciento) y universal (aplicado al 90 por ciento de gestantes). Proponemos que la implantación de este cribado supone una disminución de la tasa de los defectos congénitos no diagnosticados al nacimiento a menos del 0,5 por ciento de los recién nacidos. Método: Estudio prospectivo. Hemos valorado 12.478 gestantes (julio 2006-septiembre de 2009). Método de cribado de defecto congénitos: test combinado asociado a ecografía morfológica (18-22 semanas) Resultados: La prevalencia de defecto congénito fue de 2,26 por ciento [IC 95 por ciento: 1,9-2,5] (282/12478). Valoración ecográfica fue del 99,2 por ciento de las gestantes. Tasa de diagnóstico de malformaciones estructurales fue de 79,3 por ciento [IC 95 por ciento: 74,3-84,4] (196/247) y 95,6 por ciento [IC 95 por ciento: 91,8-99,3] (110/115) para las malformaciones mayores. Se ofertó un cribado de cromosomopatias al 95,1 por ciento de las gestantes con una tasa de diagnóstico del 88,5 por ciento [IC 95 por ciento: 79,9-99] (31/35). Conclusiones: Un cribado de defectos congénitos universal y de garantías logró disminuir la prevalencia de defectos congénitos al nacimiento sin diagnosticar a un 0,5 por ciento.

Background: The probability of a newborn presenting some kind of congenital defect at birth is 2-4 percent and the application of methods of screening for chromosomal and structural abnormalities can reduce the prevalence of these defects at birth. Objectives: The aim of this study is to prove that it is possible to implement a screening for congenital malformations that is standardised (diagnostic sensitivity [Sen]>80 percent for major structural deformations and Down's syndrome) and universal (90 percent of pregnant women). We also want to prove that this screening reduces the rate of undiagnosed congenital defects at birth. Methods: Prospective study. We assessed 12,478 pregnant women (July 2006- September 2009). A morphological ultrasound (18-22 weeks) and a combined test were carried out as the methods for screening for congenital defects. Results: The prevalence of congenital defects was 2.26 percent [95 percent CI: 1.9-2.5] (282/12478). The ultrasound scan was performed on 99.2 percent of the pregnant women. There was a Sen of 79.3 percent [95 percent CI: 74.3-84.4] (196/247) for structural malformation and 95.6 percent [95 percent CI: 91.8-99.3] (110/115) for major malformations). Screening for chromosomal anomalies was performed on 95.1 percent of pregnant women with a Sen of 88.5 percent [95 percent CI: 79.9-99] (31/35). Conclusions: A standardised and universal screening for congenital defects reduced the prevalence of undiagnosed congenital defects at birth to 0.5 percent.

Hearts with concordant ventriculoarterial connections but parallel arterial trunks.

OBJECTIVES: To determine the characteristic morphological features of hearts with concordant ventriculoarterial connections and parallel arterial trunks, and to provide unequivocally a method to describe their anatomy. DESIGN, METHODS AND PATIENTS: The entire cardiac database and cardiac pathological archive at the Hospital for Sick Children, Toronto, Ontario, Canada, was interrogated to identify all patients with concordant ventriculoarterial connections and parallel arterial trunks. The clinical records, autopsy reports and actual cardiac specimens of those who underwent autopsy, were reviewed. RESULTS: 8 cases meeting our criteria were identified. The infundibular anatomy was variable, including four hearts with bilateral infundibulums, three with subpulmonary infundibulums and one with bilaterally absent infundibulums. Considerable variability was also found in the type of atrial arrangement, along with the morphology of the atrioventricular junctions. The most common findings were the usual atrial arrangement (n = 5), left juxtaposition of the right atrial appendages (n = 3), an atrial septal defect (n = 6), univentricular atrioventricular connection (n = 5), ventricular septal defect (n = 8) and pulmonary obstruction (n = 4). In addition, five specimens had either a single coronary artery or two coronary arteries arising from the anticipated right coronary aortic sinus. CONCLUSIONS: Concordant ventriculoarterial connections with parallel arterial trunks can be found in a variety of segmental combinations. An accurate diagnosis of these rare hearts can be achieved by detailed analysis of not only the ventriculoarterial connections but also the infundibular anatomy and the spatial relationship of the arterial trunks. Particular attention to the coronary arteries is warranted.

Endoscopic retrograde cholangiopancreatography after pancreaticoduodenectomy for benign and malignant disease: indications and technical outcomes.

BACKGROUND AND STUDY AIMS: Patients undergoing pancreaticoduodenectomy develop postoperative complications related to surgery and their disease. Very little data are available on the role or success of endoscopic retrograde cholangiopancreatography (ERCP) in such patients. The aim of this study was to evaluate the indications and role of diagnostic and therapeutic ERCP after pancreaticoduodenectomy for both benign and malignant disease. PATIENTS AND METHODS: This study was a 10-year (1990 - 2000) single institution retrospective review of all ERCPs performed on patients who had undergone pancreaticoduodenectomy surgery. Indications for the ERCP and technical procedural success were studied. RESULTS: 29 patients with a pancreaticoduodenectomy underwent 56 ERCPs. Reasons for surgery were neoplasia and chronic pancreatitis. Indications for ERCP included evaluation of jaundice and pain. Technical success related to the clinical indication (jaundice 69 %, pain 54 %). CONCLUSION: ERCP plays an important role in the management of postpancreatic surgery problems including biliary and anastomotic strictures, and should be the modality of choice. However, surgical technique may make the afferent limb inaccessible, and the ductal anastomosis difficult to identify in patients with some types of pancreaticoduodenectomy. Closer collaboration between surgeon and endoscopist may allow alterations in surgical technique to improve postoperative ERCP success.

Radial endoscopic ultrasound and spiral computed tomography in the diagnosis and staging of periampullary tumors.

BACKGROUND AND STUDY AIMS: The early detection and accurate staging of pancreatic and ampullary cancer is of utmost importance for the achievement of surgical radical treatment. The aim of this study was to assess prospectively the role of endoscopic ultrasonography (EUS) in detection and staging of pancreatic and ampullary cancer, comparing its results to those obtained with spiral computed tomography (SCT). PATIENTS AND METHODS: Sixty-one patients with suspected pancreatic and ampullary tumors were included, 46 (75, 4%) of whom presented with obstructive jaundice. Patients underwent EUS and SCT within a 7-day period. Examiners were unaware of the previous imaging results, except conventional echography. Image interpretation was compared to surgical and histopathological findings. RESULTS: Fifty-six (91, 8%) patients were surgically explored. Clinical follow-up and/or tissue diagnosis determined the correct diagnosis in the remaining five patients. Pancreatic cancer and ampullary cancer were observed in 29 (47, 6%) and 10 (16, 4%) patients, respectively. Chronic pancreatitis and choledocholithiasis were the most common diagnosis in patients with non-neoplastic disease. EUS was more effective than SCT for the definition of the final diagnosis in patients with obstructive jaundice (87.0 vs. 67.4%, p = 0.04). Both exams were equally effective for detecting pancreatic cancer but EUS predicted more accurately the involvement of portal-mesenteric axis by the tumor (87.0 vs. 67.4%, p = 0.04). EUS was particularly useful in the diagnosis of cancer of papilla of Vater. CONCLUSION: In patients with pancreatic adenocarcinoma without unequivocal signs of distant metastatic disease, EUS is more accurate than SCT to predict venous involvement by the tumor. EUS is superior to SCT to detect ampullary adenocarcinoma. Both methods are equally ineffective to detect nodal involvement in pancreatic and ampullary cancer.

Citopatología de los carcinomas de mama / Cytopathology of the breast carcinomas

La principal indicación de la PAAF de mama es la existencia de una masa palpable y valorada conjuntamente con la exploración clínica y mamográfica (Triple test); su precisión diagnóstica es de un 99 por ciento. Los criterios citológicos que permiten establecer un diagnóstico de malignidad son: celularidad abundante (celularidad tumoral), grupos celulares tridimensionales y poco cohesivos, pleomorfismo celular, células aisladas con citoplasma, ausencia de núcleos bipolares desnudos y presencia de fondo sucio Aunque la mayoría de los carcinomas mamarios son de tipo ductal, existen parámetros citológicos que, aunque no siempre, permiten identificar subtipos específicos. El material obtenido con la PAAF permite la valoración de factores pronósticos, ya sea realizando técnicas inmunocitoquímicos para receptores hormonales o aplicando criterios que permiten valorar el grado nuclear. La PAAF de mama en las lesiones malignas aporta una sensibilidad del 93 por ciento y una especificidad del 94 por ciento (AU)

Synthesis of extracellular matrix and adhesion through beta(1) integrins are critical for fetal ventricular myocyte proliferation.

Extracellular matrix (ECM) regulates vascular smooth muscle cell proliferation. The role of ECM in myocardial growth is unexplored. We sought to determine whether human fetal ventricular myocytes (HFVMs) produce ECM and whether synthesis and attachment to ECM are necessary for their epidermal growth factor (EGF)-dependent and -independent proliferation. Cultured HFVMs proliferate in the presence but not absence of serum and EGF, as determined by increase in cell number and [(3)H]thymidine and [(14)C]leucine incorporation (measures of DNA and protein synthesis, respectively). Using a cyanogen bromide digestion technique to measure collagen and elastin and using affinity chromatography for fibronectin, we found that HFVMs synthesized collagen and fibronectin but not elastin. HFVMs grown on exogenous ECM (including fibronectin and type I collagen and laminin) demonstrated no change in proliferation or DNA and protein synthesis with or without EGF. However, inhibition of collagen synthesis using cis-4-hydroxyproline resulted in a decrease in EGF-related HFVM proliferation and DNA and protein synthesis, which was reversed by exposure to L-proline but not by growth on type I collagen. Use of beta(1) but not beta(3) integrin antibody to inhibit cell interaction with ECM resulted in a decrease in HFVM proliferation and DNA and protein synthesis in response to EGF. Furthermore, EGF-dependent proliferation was enhanced by alpha(1)beta(1) and alpha(5)beta(1) antibodies that act as functional ligands, but not alpha(3)beta(1), the only beta(1) subtype expressed in adult myocytes. In conclusion, proliferating HFVMs synthesize collagen and fibronectin. The proliferative response of HFVMs to EGF requires the synthesis of collagen as well as attachment to specific alpha/beta(1) integrin heterodimers.

Prenatal diagnosis of right ventricular outflow tract obstruction with intact ventricular septum, and detection of ventriculocoronary connections.

OBJECTIVES: To determine the accuracy of prenatal diagnosis of pulmonary atresia and intact ventricular septum (PAIVS), and pulmonary stenosis, including prenatal detection of ventriculocoronary connections, to evaluate heart size during the prenatal period, and to evaluate the outcome. DESIGN AND PATIENTS: Medical records of 20 cases with prenatally diagnosed PAIVS and pulmonary stenosis were reviewed retrospectively. Prenatal and postnatal echocardiography were also reviewed and dimensions of the ventricles and vessels were measured retrespectively. RESULTS: Of 20 prenatal diagnoses (15 PAIVS and five pulmonary stenosis), 16 were confirmed as correct. One critical pulmonary stenosis case had been diagnosed as PAIVS prenatally; three had no confirmation. Eight pregnancies were terminated, three had no active treatment, and nine were treated; all survived. Of 13 assessed with ventriculocoronary connections prenatally, seven were diagnosed correctly (four with, three without ventriculocoronary connections), but one was falsely positive; five had no confirmation. The more prominent hypoplasia of the main pulmonary artery and the tricuspid valve annulus, and the sigmoid shape of the ductus arteriosus, seemed to be associated with the presence of ventriculocoronary connections. CONCLUSIONS: Current prenatal echocardiography can accurately diagnose right ventricular outflow tract obstruction and ventriculocoronary connections. Prenatal detection of this constellation of abnormalities aids in family counselling and decisions on postnatal management.

Evolution of fetal ventricular aneurysms and diverticula of the heart: an echocardiographic study.

Due to the rarity of congenital ventricular diverticula and aneurysms, their natural history remains unclear. An excellent prognosis has been suggested for those cases diagnosed during fetal life: From October 1992 to January 1996 seven fetuses were diagnosed with ventricular diverticula or aneurysms. Gestational age ranged from 18 to 36 weeks. The indications for fetal echocardiogram were cardiomegaly, abnormal four-chamber view, a large pericardial effusion, and hydrothorax. Echocardiography revealed a moderate sized apical left ventricular aneurysm (2), a small subvalvular right ventricular diverticulum (1), small apical right ventricular diverticulum (2), a large submitral left ventricular aneurysm (1), and a large diverticulum arising from the lateral free wall of the left ventricle (1). Decreased left ventricular function was detected in three fetuses with left ventricular aneurysms. Two fetuses with large lesions, developed hydrops and died in utero. Postnatal echocardiograms confirmed prenatal findings in all survivors. All infants remained asymptomatic, with age on follow-up from 8 to 24 months. An accurate diagnosis of ventricular diverticula and aneurysms is feasible prenatally. Outcome depends on the size and progression of the lesion. The presence of a large pericardial effusion in three cases with a diverticula was noted.

Contribution of individual promoters in the ddlB-ftsZ region to the transcription of the essential cell-division gene ftsZ in Escherichia coli.

The essential cell-division gene ftsZ is transcribed in Escherichia coli from at least six promoters found within the coding regions of the upstream ddlB, ftsQ, and ftsA genes. The contribution of each one to the final yield of ftsZ transcription has been estimated using transcriptional lacZ fusions. The most proximal promoter, ftsZ2p, contributes less than 5% of the total transcription from the region that reaches ftsZ. The ftsZ4p and ftsZ3p promoters, both located inside ftsA, produce almost 37% of the transcription. An ftsAp promoter within the ftsQ gene yields nearly 12% of total transcription from the region. A large proportion of transcription (approximately 46%) derives from promoters ftsQ2p and ftsQ1p, which are located inside the upstream ddlB gene. Thus, the ftsQAZ genes are to a large extent transcribed as a polycistronic mRNA. However, we find that the ftsZ proximal region is necessary for full expression, which is in agreement with a recent report that mRNA cleavage by RNase E at the end of the ftsA cistron has a significant role in the contol of ftsZ expression.

Isolated ductal closure in utero diagnosed by fetal echocardiography.

In utero isolated ductal closure is uncommon and can lead to fetal hydrops and death if not recognized. Five fetuses were diagnosed prenatally by echocardiography. The reasons for referral were hydrops (2), cardiomegaly (1), polyhydramnios and choroid plexus cyst (1), and polyhydramnios and teratoma of the neck (1). Gestational age was median 32, range 18-41 weeks. Two mothers received indomethacin for polyhydramnios, with the echocardiogram performed 5 and 3 days after the last dose. Three had a negative history of drug ingestion. Fetal echocardiograms showed absent flow in the ductus arteriosus, dilated right ventricle with decreased function, and mild or moderate tricuspid and pulmonary insufficiency in all. The left ventricle was hypercontractile, with significantly increased left ventricular stroke output when compared to the right. Four fetuses were delivered by cesarean section and had an uneventful course. One fetus died shortly after birth due to airway obstruction from a large teratoma. Autopsy showed ductal constriction. Postnatal echocardiograms showed absent ductal flow and dilated right ventricle. On follow-up, survivors remained asymptomatic with cardiac size returning to normal. Premature closure of the ductus arteriosus should be considered in hydrops of unknown etiology, right ventricular dysfunction, and following indomethacin therapy. Urgent delivery results in an excellent prognosis.

Heart rate and cardiac output after atropine in anaesthetised infants and children.

PURPOSE: Heart rate is considered to be a major determinant of cardiac output in infants and small children but the relationships between age, heart rate and cardiac output in humans have never been clearly established. This study was designed to determine the change in cardiac output following atropine iv to anaesthetised infants and small children. METHODS: Following Institutional Ethics Committee approval and written-informed consent, 20 ASA I or II unpremedicated patients aged from 1 to 36 mo were studied. Anaesthesia was induced with 5 mg.kg-1 thiopentone, 2 micrograms.kg-1 fentanyl and maintained with halothane 0.5% in nitrous oxide 66% in oxygen. Vecuronium 0.1 mg.kg-1 was used to provide muscular relaxation. Cardiac output was measured by non-invasive transthoracic blind continuous-wave Doppler echocardiography before and after the administration of 0.02 mg.kg-1 atropine iv. RESULTS: Atropine increased both heart rate and cardiac index by 31.1 +/- 12.8% and 29.4 +/- 17.3% respectively (P < 0.05). The cardiac index before atropine was 5.1 +/- 1.2 L.min-1.m-2 and the increase after atropine varied widely from 1.4 to 52.1%. Although atropine did not alter the overall stroke index the recorded changes ranged from -20.8 to +18.0%. There was no association between age and either cardiac index or % change in cardiac index after atropine. However, there was a positive but weak correlation between percentage change in heart rate and cardiac output (r2 = 0.46). CONCLUSION: Atropine causes a variable increase in cardiac output in infants and children aged between 1 and 36 mo. The change in cardiac output, considering the limits of the transthoracic echocardiography methodology, suggests that this is related to the increase in heart rate but is not dependent of age.