An in-depth multiphasic analysis of the chocolate production chain, from bean to bar, demonstrates the superiority of Saccharomyces cerevisiae over Hanseniaspora opuntiae as functional starter culture during cocoa fermentation.

Hanseniaspora opuntiae is a commonly found yeast species in naturally fermenting cocoa pulp-bean mass, which needed in-depth investigation. The present study aimed at examining effects of the cocoa isolate H. opuntiae IMDO 040108 as part of three different starter culture mixtures compared with spontaneous fermentation, regarding microbial community, substrate consumption, and metabolite production dynamics, including volatile organic compound (VOC) and phytochemical compositions, as well as compositions of the cocoa beans after fermentation, cocoa liquors, and chocolates. The inoculated H. opuntiae strain was unable to prevail over background yeasts present in the fermenting cocoa pulp-bean mass. It led to under-fermented cocoa beans after four days of fermentation, which was however reflected in higher levels of polyphenols. Cocoa fermentation processes inoculated with a Saccharomyces cerevisiae strain enhanced flavour production during the fermentation and drying steps, which was reflected in richer and more reproducible aroma profiles of the cocoa liquors and chocolates. Sensory analysis of the cocoa liquors and chocolates further demonstrated that S. cerevisiae led to more acidic notes compared to spontaneous fermentation, as a result of an advanced fermentation degree. Finally, different VOC profiles were found in the cocoa beans throughout the whole chocolate production chain, depending on the fermentation process.

Importance of body condition score and ovarian activity on determining the fertility in beef cows supplemented with long-acting progesterone after timed-AI.

The working hypothesis was that supplementation with progesterone (iP4) at early diestrus increases pregnancy rates of suckled beef cows with poor body condition score (BCS) and/or in anestrus when cows are submitted to timed insemination (TAI). The time of ovulation among suckled multiparous (n = 1270) and primiparous (n = 303) Nelore cows was synchronized using an estradiol/P4-based protocol for TAI (D0). At Day ‒10, visual evaluation of BCS was performed (scale = 1-5) and animals were classified according to an ovarian activity (OA) score: (OA1) absence of CL and presence of follicles ≥ 8 mm, (OA2) absence of CL and presence of follicles < 8 mm and (OA3) presence of corpus luteum (CL). On Day 4, animals were assigned randomly to receive 150 mg of injectable, long-acting P4 (iP4, n = 786) or non-iP4 (n = 787). Further, ultrasonic evaluations were performed on D0 and D4 for measurements of the largest follicle diameter (DF) and CL area, respectively. The BCS affected positively DF, CL area and OA. Supplementation with iP4 did not affect (P = 0.49) pregnancy rates and there was no significant interaction (P > 0.1) of P4 treatment with BCS or OA was detected for conception and pregnancy rates. Regardless of iP4, pregnancy rates of cows with moderate (2.75-3.25) (59.1%) and high (≥ 3.5) (57.8%) BCS were greater than those of cows with low (2.0-2.5) BCS (41.5%). Cows in OA2 had a greater P/AI (51.3%) than cows in OA1 (41.7%) or OA3 (41.9%). In conclusion, P4 supplementation after TAI did not improve P/AI and did not enhance the response to treatment of cows with a low BCS, regardless of OA.

Generation and characterization of a human iPS cell line from a patient-related control to study disease mechanisms associated with DAND5 p.R152H alteration.

A DAND5-control human iPSC line was generated from the urinary cells of a phenotypically normal donor. Exfoliated renal epithelial (RE) cells were collected and reprogrammed into iPSCs using Sendai virus reprogramming system. The pluripotency, in vitro differentiation potential, karyotype stability, and the transgene-free status of generated iPSC line were analyzed and confirmed. This cell line can be exploited as a control iPSC line to better understand the mechanisms involved in DAND5-associated cardiac disease.

Generation of human iPSC line from a patient with laterality defects and associated congenital heart anomalies carrying a DAND5 missense alteration.

A human iPSC line was generated from exfoliated renal epithelial (ERE) cells of a patient affected with Congenital Heart Disease (CHD) and Laterality Defects carrying tshe variant p.R152H in the DAND5 gene. The transgene-free iPSCs were generated with the human OSKM transcription factor using the Sendai-virus reprogramming system. The established iPSC line had the specific heterozygous alteration, a stable karyotype, expressed pluripotency markers and generated embryoid bodies that can differentiate towards the three germ layers in vitro. This iPSC line offers a useful resource to study the molecular mechanisms of cardiomyocyte proliferation, as well as for drug testing.

Functional study of DAND5 variant in patients with Congenital Heart Disease and laterality defects.

BACKGROUND: Perturbations on the Left-Right axis establishment lead to laterality defects, with frequently associated Congenital Heart Diseases (CHDs). Indeed, in the last decade, it has been reported that the etiology of isolated cases of CHDs or cases of laterality defects with associated CHDs is linked with variants of genes involved in the Nodal signaling pathway. METHODS: With this in mind, we analyzed a cohort of 38 unrelated patients with Congenital Heart Defects that can arise from initial perturbations in the formation of the Left-Right axis and 40 unrelated ethnically matched healthy individuals as a control population. Genomic DNA was extracted from buccal epithelial cells, and variants screening was performed by PCR and direct sequencing. A Nodal-dependent luciferase assay was conducted in order to determine the functional effect of the variant found. RESULTS: In this work, we report two patients with a DAND5 heterozygous non-synonymous variant (c.455G > A) in the functional domain of the DAND5 protein (p.R152H), a master regulator of Nodal signaling. Patient 1 presents left isomerism, ventricular septal defect with overriding aorta and pulmonary atresia, while patient 2 presents ventricular septal defect with overriding aorta, right ventricular hypertrophy and pulmonary atresia (a case of extreme tetralogy of Fallot phenotype). The functional analysis assay showed a significant decrease in the activity of this variant protein when compared to its wild-type counterpart. CONCLUSION: Altogether, our results provide new insight into the molecular mechanism of the laterality defects and related CHDs, priming for the first time DAND5 as one of multiple candidate determinants for CHDs in humans.

Implication of the RD(Rio) Mycobacterium tuberculosis sublineage in multidrug resistant tuberculosis in Portugal.

Multidrug and extensively drug resistant Mycobacterium tuberculosis are a threat to tuberculosis control programs. Genotyping methods, such as spoligotyping and MIRU-VNTR typing (Mycobacterial Interspersed Repetitive Units), are useful in monitoring potentially epidemic strains and estimating strain phylogenetic lineages and/or genotypic families. M. tuberculosis Latin American Mediterranean (LAM) family is a major worldwide contributor to tuberculosis (TB). LAM specific molecular markers, Ag85C(103) single nucleotide polymorphism (SNP) and RD(Rio) long-sequence polymorphism (LSP), were used to characterize spoligotype signatures from 859 patient isolates from Portugal. LAM strains were found responsible for 57.7% of all tuberculosis cases. Strains with the RD(Rio) deletion (referred to as RD(Rio)) were estimated to represent 1/3 of all the strains and over 60% of the multidrug resistant (MDR) strains. The major spoligotype signature SIT20 belonging to the LAM1 RD(Rio) sublineage, represented close to 1/5th of all the strains, over 20% of which were MDR. Analysis of published datasets according to stipulated 12loci MIRU-VNTR RD(Rio) signatures revealed that 96.3% (129/134) of MDR and extensively drug resistant (XDR) clusters were RD(Rio). This is the first report associating the LAM RD(Rio) sublineage with MDR. These results are an important contribution to the monitoring of these strains with heightened transmission for future endeavors to arrest MDR-TB and XDR-TB.

[Socio-demographic features of puerperas and pregnancy follow-up: what has changed in 17 years?]. / Características sócio-demográficas das puérperas e seguimento da gravidez: o que mudou em 17 anos?

In recent years, the time for motherhood has been postponed, which is related to social aspects and can lead to health problems for the pregnant women and the child. An appropriate follow-up of the pregnancy is essential for identification and early treatment of possible complications. The purpose of this study was to evaluate some social and demographic characteristics and the pregnancy follow-up among puerperas, in two different times separated by 17 years (1991 e 2008). In this study, we found that the birth of a first child occurs now later, is more planned and happens more frequently outside of marriage. The postponement of motherhood can be associated with an increased rate of miscarriages. There has been a decrease in the rate of induced abortions, which is probably the result of the implementation of strategies for family planning. We noticed a decline in the number of pregnancies that had not a proper follow-up, demonstrating the improvement of health care. The immigrant population is responsible for about one quarter of the births in the Algarve, with a higher percentage of unplanned pregnancies, which eventually might be related to socio-economic vulnerabilities, but no difference was found in the surveillance of pregnancy, demonstrating a good accessibility of this population to health care. It is essential the existence of information about the risks of a late pregnancy and of social infrastructures to support motherhood at younger ages. Pregnancy surveillance programs have been successful, but there are still gaps in obstetric monitoring. Despite good results in terms of surveillance of pregnancy, the immigrant population may present some vulnerabilities, which means that their access to maternal and child health care should be monitored.

Redução da primeira dose de GnRH em vacas holandesas de alta produção sincronizadas com Ovsynch ou Heatsynch / Reduction of first GnRH dosage in high-yielding holstein cows synchronized with Ovsynch or Heatsynch

The objective of this study was to evaluate the effect of treatment with 10 or 20 μg of GnRH and type of FTAI protocols (Ovsynch and Heatsynch) on pregnancy rate of lactating dairy cows submitted to the synchronization at FTAI (n = 581). The animals were distributed among four treatments, allocated factorial design 2x2: Treat 1- Ovsynch (20 μg GnRH on D0; PGF on D7; 10 μg GnRH on D9 and FTAI on D10; n = 141), Treat 2- ½ Ovsynch (10 μg GnRH on D0; PGF on D7; 10 μg GnRH on D9 and FTAI on D10; n = 159), Treat 3- Heatsynch (20 μg GnRH on D0; PGF on D7; 1 mg EB on D8 and FTAI on D10; n = 147) and Treat 4- ½ Heatsynch (10 μg GnRH on D0; PGF on D7; 1 mg EB on D8 and FTAI on D10; n = 134). The Group Ovsynch (Treat 1 and 2) presented greater pregnancy rate than Group Heatsynch (Treat 3 and 4) at 28 days (45.7% (137/300) and 39.9% (112/281), respectively; P = 0.07) and at 56 days of gestation (41.0% (123/300) and 34.2% (96/281), respectively; P = 0.06). There were no statistical difference (P > 0.10) on pregnancy rate at 28 (44.4% (128/288) vs 41.3% (121/293); P = 0.50) and 56 days (38.2% (110/288) vs 37.2% (109/293); P = 0.68) between the dosages of the GnRH. The pregnancies losses (between 28 and 56 days) were not different between the FTAI protocols and GnRH dosage (P > 0.10). We concluded that the Ovsynch protocol presented greater pregnancy rate at FTAI than Heatsynch protocol and the administration of 10 μg of buserelin Acetate, at the beginning of these protocols did not decrease the efficiency of FTAI protocols in high-yielding Holstein cows.

O objetivo desse estudo foi avaliar o efeito da redução da primeira dose de GnRH (10 vs 20 μg) e do tipo de protocolo de sincronização para a IATF (Ovsynch e Heatsynch) na taxa de prenhez de vacas holandesas de alta produção sincronizadas para IATF (n = 581). Os animais foram distribuídos entre os tratamentos e alocados em arranjo fatorial 2x2, sendo: Trat 1- Ovsynch (20 μg GnRH no D0; PGF no D7; 10 μg GnRH no D9 e IATF no D10; n = 141), Trat 2- ½ Ovsynch (10 μg GnRH no D0; PGF no D7; 10 μg GnRH no D9 e IATF no D10; n = 159), Trat 3- Heatsynch (20 μg GnRH no D0; PGF no D7; 1 mg BE no D8 e IATF no D10; n = 147) e Trat 4- ½ Heatsynch (10 μg GnRH no D0; PGF no D7; 1 mg BE no D8 e IATF no D10; n = 134). O Grupo Ovsynch (Trat 1 e 2) apresentou maior taxa de prenhez que o grupo Heatsynch (Trat 3 e 4) aos 28 dias (45,7% (137/300) e 39,9% (112/281), respectivamente; P = 0,07) e aos 56 dias de gestação (41,0% (123/300) e 34,2% (96/281), respectivamente; P = 0,06). A dose de GnRH, não influenciou (P > 0.10) a taxa de prenhez aos 28 dias (44,4% (128/288) vs 41,3% (121/293); P = 0,50) e aos 56 dias (38,2% (110/288) vs 37,2% (109/293); P = 0,68) entre os grupos tratados com 10 μg ou 20 μg de acetato de buserelina. As perdas gestacionais (ente 28 e 56 dias) não diferiram entre os protocolos para IATF e dose de GnRH (P > 0,10). Conclui-se que, protocolo Ovsynch apresentou maior taxa de prenhez à IATF que o Heatsynch e a administração de 10 μg de Acetato de Buserelina no início dos tratamentos de sincronização não comprometeu a eficiência do protocolo de sincronização para IATF em vacas holandesas de alta produção.

What brings newborns to the emergency department?: a 1-year study.

AIMS: Characterization of newborn visits to the pediatric emergency department (PED). Analysis of the main illnesses and establishment of association between certain conditions and severity of diseases. METHODS: Retrospective study information concerning newborns (aged <28 days) who presented to the PED of Hospital Central de Faro during 2005. We studied the clinical and demographic data referring to the PED's episode and to perinatal occurrences. RESULTS: A total of 540 neonates visited the PED (1.5% of the total PED use), with an average age of 14.0 days and a slight prevalence of males (53.1%). Of the total visits, 17.2% were primary physician referrals. The chief complaints were jaundice, excessive crying, and rash. Diagnostic tests were requested in 27.2% cases. The major diagnoses were nonapparent disease, infant colics, and physiologic jaundice. In 13.0%, hospital admission was necessary. Newborns with referral, those with less than 37 weeks of gestation and those whose delivery weight was less than 2500 g, were more likely to be admitted. CONCLUSIONS: Most PED visits were because of nonserious diseases, mainly because of insufficient caretaker knowledge and information. This highlights the great need for caretakers' education by health staff. It is also important that physicians are aware of the main illnesses in the newborn period and know how to correctly identify the conditions associated to serious pathology.

Consolidaciones viciosas del radio distal / Distal radius malunion

Se analiza una serie de 20 pacientes con consolidaciones viciosas del radio distal tratados con osteotomias entre 1994 y 2000. Las deformidades fueron clasificadas, segun la topografia en: extraarticulares, intraarticulares y combinadas ; la etiologia mas frecuente es la fractura de Colles tratada con yeso simple. Las osteotomias extraarticulares se basaron en la tecnica de injerto iliaco tricortical descrita por Fernandez, y sus resultados funcionales y anatomicos fueron excelentes y buenos en el 80 por ciento de los casos. Las deformidades intraarticulares se trataron con osteotomias focales, tecnicamente mas complejas y menos estandarizadas y los resultados funcionales fueron menos predecibles. Concluimos que la evaluacion detallada de la deformidad, la planificacion preoperatoria minuciosa y la ejecucion de la tecnica quirurgica en forma precisa brindan resultados funcionales uniformemente buenos

Consolidaciones viciosas del radio distal / Distal radius malunion

Se analiza una serie de 20 pacientes con consolidaciones viciosas del radio distal tratados con osteotomias entre 1994 y 2000. Las deformidades fueron clasificadas, segun la topografia en: extraarticulares, intraarticulares y combinadas ; la etiologia mas frecuente es la fractura de Colles tratada con yeso simple. Las osteotomias extraarticulares se basaron en la tecnica de injerto iliaco tricortical descrita por Fernandez, y sus resultados funcionales y anatomicos fueron excelentes y buenos en el 80 por ciento de los casos. Las deformidades intraarticulares se trataron con osteotomias focales, tecnicamente mas complejas y menos estandarizadas y los resultados funcionales fueron menos predecibles. Concluimos que la evaluacion detallada de la deformidad, la planificacion preoperatoria minuciosa y la ejecucion de la tecnica quirurgica en forma precisa brindan resultados funcionales uniformemente buenos

Estudio de la asociacion de condromalacia patelar en las lesiones meniscales y del ligamento cruzado anterior / Patellar chondromalacia associated with meniscal and LCA injuries

Entre junio de 1997 y marzo de 2000 se estudiaron en conjunto con el servicio de diagnostico medico Rosario, 1084 pacientes que presentaban rodillas dolorosas agrupadas en patologias inflamatorias, degenerativas, traumaticas, tumorales, ligamentarias. Los pacientes fueron evaluados siempre por el mismo cuerpo medico de radiologos y protocolos de RMI. Se estudio prevalencia segun edad y sexo. Confeccionamos una planilla de estudio con 4 campos a evaluar (menisco interno, menisco externo, ligamento cruzado anterior y un ultimo campo de lo denominamos observaciones, incluyendo en este, diversos hallazgos patologicos. De los diversos hallazgos patologicos, comprobamos que existia un predominio de condromalacia patelar en pacientes mayores de 55 años, con sobrepeso corporal, alteracin del angulo Q y altura de la patela. Otras lesiones secundarias que encontramos de jerarquia fueron: contusiones femorotibiales, displasia femoropatelar, quistes de Baker, sinovitis de Hoffa, osteocondritis, fracturas ocultas de platillo tibial, plicas sinoviales, gangliones, osteonecrosis, encondromas, edema de partes blandas periarticulares

Estudio de la asociacion de condromalacia patelar en las lesiones meniscales y del ligamento cruzado anterior / Patellar chondromalacia associated with meniscal and LCA injuries

Entre junio de 1997 y marzo de 2000 se estudiaron en conjunto con el servicio de diagnostico medico Rosario, 1084 pacientes que presentaban rodillas dolorosas agrupadas en patologias inflamatorias, degenerativas, traumaticas, tumorales, ligamentarias. Los pacientes fueron evaluados siempre por el mismo cuerpo medico de radiologos y protocolos de RMI. Se estudio prevalencia segun edad y sexo. Confeccionamos una planilla de estudio con 4 campos a evaluar (menisco interno, menisco externo, ligamento cruzado anterior y un ultimo campo de lo denominamos observaciones, incluyendo en este, diversos hallazgos patologicos. De los diversos hallazgos patologicos, comprobamos que existia un predominio de condromalacia patelar en pacientes mayores de 55 años, con sobrepeso corporal, alteracin del angulo Q y altura de la patela. Otras lesiones secundarias que encontramos de jerarquia fueron: contusiones femorotibiales, displasia femoropatelar, quistes de Baker, sinovitis de Hoffa, osteocondritis, fracturas ocultas de platillo tibial, plicas sinoviales, gangliones, osteonecrosis, encondromas, edema de partes blandas periarticulares