The ubiquitin editing enzyme A20 maintains immune homeostasis and prevents autoimmunity

Dissertation presented to obtain the Doctorate degree (Ph.D.) in Biology at Instituto de Tecnologia Química e Biológica da Universidade Nova de LisboaThe immune system is vital to ensure the surveillance of organisms against pathogens or malignant cells. However, the negative regulation of the immune system is equally essential, and defects in the termination of immune signals can result in autoimmunity and other pathologies. The functioning of the immune system results from the integration of signals between and within cells. For some time, studies in immune signaling have focused on the molecular events playing a role in activating such cascades. However, little was known about how to turn off those signals. Just recently, new research started shedding light into mechanisms that ensure the negative regulation of immune signaling.(...

Cuidados paliativos em doentes com cancro

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências FarmacêuticasNos últimos anos as doenças oncológicas têm vindo a sofrer um grande aumento, passando por muitos a ser vistas como uma nova epidemia. A busca incessante da cura para estas doenças promove a constante negação da morte, deixando para segundo plano as intervenções na saúde dos doentes terminais. As intervenções que têm como objetivo proporcionar ao doente um fim de vida digno e com o mínimo de sofrimento, bem como paz e estabilidade à família que o acompanha em doenças carregadas de sofrimento não só físico como psicológico denominam-se cuidados paliativos. Neste âmbito surge esta dissertação de mestrado, introduzindo os cuidados paliativos com o devido enquadramento teórico. Relata-se a sua história, desde o aparecimento deste tipo de cuidados. É feita uma abordagem aos cuidados paliativos onde se descreve os seus princípios e filosofias, as unidades onde se efetuam e prestam este tipo de cuidados bem como o funcionamento das suas equipas e quais os seus modelos de atuação. Procura-se através da análise de casos de doentes com cancro avaliar a evolução de sintomas associados a este tipo de doenças. Por fim enfatiza-se o funcionamento e desenvolvimentos dos Cuidados Paliativos em Portugal nos últimos anos, presta-se ainda uma cuidada atenção aos cuidados paliativos pediátricos relatando o seu desenvolvimento e as suas diferenças em relação aos mesmos quando prestados em adultos. In the last years the incidence of oncological diseases has increased being seen by many as a new epidemic. The relentless pursuit of a cure promotes a constant denial of death, leaving aside active and effective interventions on the healthcare of these terminal patients that would dignify his life. These interventions are named as hospice care. Those cares provide the patient a worthy end of life with a minimal suffering, as well peace and stability to the family with all the physical and psychological suffering. In this context this dissertation arises, introducing palliative care with proper theoretical framework. Relates the history of hospice cares as since its foundation, describing its principles and philosophies as units in which they take place this type of care as well as the functioning of their work teams and their role models. Through the analysis of cancer patients, to evaluate the evolution of the symptoms associated to such diseases. Finally describe the operation and development of hospice care in Portugal in recent years and take a careful attention to paediatrics hospice care and his development and which differences they have relative to the cares of the adults

Óxido nítrico exalado em doentes com síndrome de apneia/hipopneia obstrutiva do sono

Tese de mestrado, Ciências do Sono, Faculdade de Medicina, Universidade de Lisboa, 2010Introdução: A fracção exalada do óxido nítrico ou óxido nítrico exalado (FENO) é um marcador de inflamação da via aérea. Tendo em consideração os eventos respiratórios recorrentes que ocorrem durante o sono nos doentes com Síndrome de Apneia/Hipopneia Obstrutiva do Sono (SAHOS), é esperada a existência de níveis elevados de FENO. Objectivos: Verificar se os níveis de FENO numa amostra de doentes com SAHOS se encontram elevados e identificar possíveis variações dos níveis de FENO ao longo do dia. Métodos: Os níveis de FENO foram determinados antes (22:30 h) e depois (7:30h) da polissonografia em 89 indivíduos com SAHOS e 30 controlos, todos do sexo masculino e não fumadores. Num subgrupo de doentes e indivíduos saudáveis (n=62) os valores de FENO foram também determinados às 12:30 h e às 17:30 h. Todos os indivíduos preencheram um questionário clínico, com preenchimento da Escala de sonolência de Epworth, e um questionário de cefaleias. O grupo de doentes com SAHOS apresentou uma média etária de 45,1 ± 7,7 anos, um índice de distúrbio respiratório (RDI) médio de 34,5 ± 26,9 eventos/h e um índice de massa corporal (IMC) médio de 30,9 ± 4,6 Kg/m2. No grupo de controlo, a média etária foi de 33,1 ± 6,4 anos, o RDI médio de 2,5 ± 1,5 eventos/h e o IMC médio de 25,5 ± 4,2kg/m2. Resultados: Nos doentes com SAHOS os níveis de FENO foram superiores aos registados nos indivíduos saudáveis: 7:30 h (SAHOS: 25.8 ± 15.8 ppb / Controlos: 18.1 ± 7.3 ppb; p=0.005); 12:30 h (SAHOS: 25,4 ± 17.2 ppb / Controlos: 16,0 ± 5.2 ppb; p=0.013) e 22:30 h (SAHOS: 23.6 ± 13.9 ppb / Controlos: 16.3 ± 7.4 ppb; p=0.005). Os níveis de FENO diminuíram durante o dia em ambos os grupos, com um decréscimo médio entre as determinações da manhã e da noite de -2,4 ± 6,9 ppb (p<0,005) nos doentes com SAHOS e -1,8 ± 4,4 ppb (p<0,039) no grupo de indivíduos saudáveis. A descida dos valores de FENO foi também significativa nos doentes com SAHOS entre as determinações das 7:30h-17:30h (-2,5 ± 6,1ppb; p=0,009), das 12:30 h - 17:30 h (-2,6 ± 5,8 ppb; p=0,004) e das 12:30 h-10:30 h (-2,6 ± 5,4 ppb; p=0,004), bem como no grupo de controlo entre as 12:30 h-22:30 h (-1,9 ± 2,9 ppb; p=0,022). Não se constataram diferenças significativas nos valores de FENO tendo em conta a presença ou não de cefaleias, de sonolência diurna ou de acordo com a gravidade da SAHOS. A presença de eventos respiratórios (RDI e IAH) e o IMC nos doentes com SAHOS não se correlacionou com os níveis de FENO. No grupo de doentes verificou-se uma correlação negativa entre a saturação de O2 mínima e os níveis de FENO medidos pela manhã (p=0,027; rho=-0,238) e à noite (p=0,021; rho=-0,245) e uma correlação positiva com a percentagem de saturação de O2 inferior a 90% (manhã: p=0,041; rho=0,221 e noite: p=0,028; rho=0,235). Numa análise de regressão linear multivariada, a idade foi a única variável que explicou as diferenças nos níveis de FENO entre os grupos de doentes e de controlo, com os níveis mais elevados verificados nos indivíduos de faixa etária superior ( r2=0,21; p=0,013). Conclusão: Neste estudo detectou-se um padrão de variação circadiária nos níveis de FENO, quer em indivíduos saudáveis, quer em doentes com SAHOS, com um decréscimo dos níveis de FENO ao longo do dia. Os níveis superiores de FENO em doentes com SAHOS foram influenciados pela idade e não pelos eventos respiratórios que ocorrem durante o sono. Atendendo a que as variações de FENO não parecem ser explicadas nem pela SAHOS, nem pela sua gravidade, a determinação de FENO não pode ser utilizada como marcador biológico de inflamação na VAS, neste grupo de doentes.Background: Fractional exhaled nitric oxide (FENO) is a marker of airway inflammation. Due to sleep repetitive apneas, Obstructive Sleep Apnea (OSA) patients are expected to have morning elevated levels of FENO. Objectives: To evaluate if FENO levels are elevated in a group of OSA patients and study diurnal variation in FENO in these patients and in healthy subjects. Methods: FENO levels were determined before (10:30 pm) and after overnight polysomnography (07:30 am) in 89, non smoking OSA male patients with a mean age: 45.1 ± 7.7 years, a mean body mass index (BMI): 30.9 ± 4,6 Kg/m2 and a mean respiratory disturbance index (RDI): 34.5 ± 26.9/h, and also in 30 healthy male subjects (mean age 33.1 ± 6.4 years; mean BMI 25,5 ± 4,2 kg/m2 and mean RDI: 2.5 ± 1.5/h). In a subgroup of OSA and healthy subjects (n=62) FENO was also determined at 12:30 pm and 05:30 pm. All subjects filled in a clinical form with the Epworth Sleepiness Scale (ESS) and a headache questionnaire. Results: OSA patients had higher FENO compared to healthy subjects: 7.30 am (OSA: 25.8 ± 15.8 / Healthy: 18.1 ± 7.3 ppb; p=0.005); 12:30 pm (OSA: 25,4 ± 17.2 / Healthy: 16,0 ± 5.2 ppb; p=0.013) and 10:30pm (OSA: 23.6 ± 13.9 / Healthy: 16.3 ± 7.4 ppb; p=0.005). FENO levels decreased during the day with a mean decrease between morning and evening of -2,4 ± 6,9ppb (p<0,005) in OSA patients and -1,8 ± 4,4ppb (p<0,039) in healthy subjects. Decrease of FENO was also statistically significant in OSA between 7:30 am - 05:30 pm (-2,5 ± 6,1 ppb; p=0,009), 12:30 pm - 05:30 pm (-2,6 ± 5,8 ppb; p=0,004) and 12:30 pm - 10:30 pm (-2,6 ± 5,4 ppb; p=0,004) and in controls between 12:30 pm - 10:30 pm (-1,9 ± 2,9 ppb; p=0,022). In OSA group there was a significant negative correlation with the minimal O2 saturation and FENO (morning: p=0,027; rho=-0,238; evening: p=0,021; rho=-0,245) and positive for the % O2 saturation <90% (morning: p=0,041; rho=0,221; evening: p=0,028; rho=0,235). There were no differences in FENO values concerning presence of headache, sleepiness by Epworth and OSA severity. RDI, IAH and BMI were not correlated with FENO on patients with OSA. In a regression analysis model, age was the only variable that explained the differences between OSA and healthy subjects (r2=0,21; p=0,013). Conclusion: OSA patients and healthy subjects showed a circadian pattern of FENO with a decrease of values throughout the day. Higher FENO values in OSA patients seem to be explained by age and not by respiratory events. No other significant differences were detected, therefore the role of FENO as a inflammatory marker of OSA was not confirmed by our data

Abordagem clínica da leucoplasia verrucosa proliferativa

O cancro oral é um dos 10 cancros mais comuns no mundo, apresentando taxas de mortalidade elevadas. É na maioria das vezes diagnosticado tardiamente. Contudo sabe-se que existe um grupo de lesões, designadas, de potencialmente malignas, que têm capacidade de evoluir e progredir até cancro oral. Entre este encontra-se a leucoplasia oral, caracterizada como uma mancha ou placa branca que não pode ser identificada clínica ou patologicamente como nenhuma outra lesão branca e sem etiologia conhecida para além do tabaco (OMS, 1978). Existem vários tipos de leucoplasias, contudo há uma rara, sem etiologia conhecida e com caracter bastante agressivo, designada, leucoplasia verrucosa proliferativa. É caracterizada como uma leucoplasia de progressão lenta, com características multifocais, que se apresenta clinicamente com aspeto verrucoso e com prolongamentos exofíticos e proliferativos. Apresenta elevadas taxas de transformação maligna e de recidiva após tratamento, não havendo ainda um tratamento totalmente eficaz. Devido a sua elevada taxa de transformação maligna e a evolução para carcinoma verrucoso e carcinoma de células escamosas, é importante que seja feito um diagnóstico precoce, para que se tente travar a sua evolução, o que melhora o prognóstico e a morbilidade associada à lesão. Diminuindo, assim, também, a incidência de cancro oral, ou pelo menos a incidência de cancro oral extensivo.Oral Cancer is one of the 10 most common cancers in the world, with high mortality rates. Is most often diagnosed late. However, it is known that there is a group of potentially malignant lesions that have the ability to evolve into oral cancer. Spot or white plaque that can not be identified clinically or pathologically as no other white lesion and no known etiology other than tobacco (WHO, 1978). There are several types of leukoplakias, however there is a leukoplakia without kown etiology, rare and with a very aggressive character, called proliferative verrucous leukoplakia. Is characterized as a slow progression leukoplakia, with multifocal features, that presents clinically verrucous appearance and with exophytic and proliferative projections. Presents with high rates of malignant transformation and relapse after treatment, with no totally effective treatment. Because of its high rate of malignant transformation and evolution in to verrucous carcinoma and squamous cell carcinoma, it is important that exists early diagnosis to try to halt its evolution and reducing the incidence of oral cancer or extensive oral cancer

Rare autosomal dominant hereditary hemochromatosis associated with SLC40A1 gene: ferroportin disease or type 4 hereditary hemochromatosis?

Ferroportin (FPN1), encoded by the SLC40A1 gene, is the unique cellular iron exporter identified in mammals. FPN1 transfers iron from the intestine and macrophages into the bloodstream. This function is negatively regulated by hepcidin. Mutations in SLC40A1 may affect FPN1 function, originating distinct autosomal dominant diseases: (i) the Ferroportin Disease (FD), due to loss-of-function mutations, is characterized by decreased iron export from enterocytes and severely affected iron transfer in macrophages, giving rise to a marked iron accumulation in spleen and liver; and (ii) the Type 4 Hereditary Hemochromatosis (HH), resulting from gain-of-function mutations conferring resistance to hepcidin-mediated FPN1 degradation and consequently high cellular iron export. In this study, 335 individuals suspected of having non-classic HH were enrolled. Six genes related with iron metabolism were analysed by SSCP, dHPLC or NGS. The latter used TruSeq or Nextera XT libraries and a MiSeq platform (Illumina). Genetic variants found were validated by Sanger sequencing. Predictive consequences at protein level were evaluated using Polyphen-2 and SIFT softwares. From all patients analysed, three SLC40A1 pathogenic variants were detected in heterozygosity in three women: two missense, c.238G>A, p.Gly80Ser and c.610G>A, p.Gly204Ser; and one deletion, c.485_487delTTG; p.Val162del. These variants had been reported in public databases, but they were not known to be present in the Portuguese population. The p.Gly80Ser and the p.Val162del are FPN1 loss-of-function mutations and were found associated with hyperferritinemia and low transferrin saturation (FD). In contrast, the p.Gly204Ser induced a gain of FPN1 function with a full iron export capacity giving the patient a type 4-HH phenotype, which includes iron overload, hyperferritinemia and high transferrin saturation. Detailed clinical evaluation of the suspected patients are useful to unravel the effect of different mutations in FPN1 function, expression and regulation.This work was partially supported by INSA_2013DGH910 and GenomePT (POCI-01-0145-FEDER-022184).info:eu-repo/semantics/publishedVersio

Caracterização da estrutura populacional de isolados de Escherichia coli provenientes de suiniculturas de produção intensiva e extensiva de Portugal

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências FarmacêuticasEscherichia coli é uma bactéria pertencente à numerosa família Enterobacteriaceae. Está normalmente associada a infeções do trato urinário, meningites do recém-nascido, infeções intestinais e septicemia. E. coli constitui também uma das bactérias mais prevalentes na flora intestinal dos animais e do Homem, sendo por isso frequentemente usada como indicador da pressão seletiva causada pelo uso de antibióticos no Homem e nos animais, com especial ênfase em animais para consumo humano. Dados recolhidos na Europa demonstram que há uma elevada ocorrência de estirpes de E.coli consideradas comensais provenientes tanto do Homem como de animais resistentes a vários antibióticos. Os isolados de E.coli pertencentes aos grupos filogenéticos A e B1 são tradicionalmente considerados comensais. Contudo, cada vez mais têm sido associados a infeções extraintestinais tal como acontece nos grupos de maior virulência B2 e D. O presente trabalho tem como objetivo principal caracterizar a estrutura populacional de isolados de E.coli provenientes de suiniculturas portuguesas, avaliar a sua resistência a antibióticos frequentemente usados clinicamente e relacionar essas resistências com a virulência tipicamente associada aos grupos filogenéticos, pois ainda são escassos os estudos neste nicho. É também objetivo deste estudo contribuir para a melhoria de estratégias de prevenção e controlo da transmissão da resistência aos antibióticos ao Homem, quer pela cadeia alimentar, quer através do ambiente, diminuindo desta forma os riscos para a Saúde Pública. Para isso, foram analisados 155 isolados bacterianos, provenientes de amostras de animais e do ambiente (águas, ar, rações, entre outras) de uma suinicultura Portuguesa de produção intensiva, e outra de produção extensiva. Depois de realizada a extração do DNA bacteriano nos isolados analisados, identificou-se a presença de E. coli por PCR. Para os isolados identificados E. coli, procedeu-se a um PCR multiplex para determinação do grupo filogenético, tendo os produtos de PCR sido analisados por electroforese em gel de agarose. Efetuou-se ainda a análise da suscetibilidade aos antibióticos dos isolados de E. coli através do método de difusão por discos, tendo sido testados antibióticos pertencentes a diferentes classes. Dos 155 isolados bacterianos analisados foram identificados como sendo E. coli 103 isolados (66%). Foi ainda observado que os isolados provenientes das duas suiniculturas estudadas foram mais frequentemente resistentes aos antibióticos espectinomicina (100%), neomicina (95%) e estreptomicina (85%). De uma forma geral, foram registadas maiores percentagens de resistência na suinicultura de produção intensiva. O grupo filogenético A é predominante no total de isolados considerados resistentes. Embora em menos quantidade, o filogrupo D aparece associado a isolados de E. coli resistentes a quase todos os antibióticos analisados. As suiniculturas portuguesas parecem assim ser potenciais reservatórios de bactérias virulentas e com resistências a antibióticos de uso clinico Humano e possivelmente de genes de resistência a antibióticos com elevada capacidade de dispersão. O tema da resistência aos antimicrobianos é um assunto de elevada importância, contudo é uma área que ainda tem uma grande lacuna, sobretudo no papel dos animais na epidemiologia da resistência a antibióticos, sendo necessário realizar mais estudos para que seja possível implementar medidas eficazes no controlo de resistências a antibióticos. Escherichia coli is a bacteria part of the numerous family known as Enterobacteriaceae. It is normally associated with urinary tract infections, newborn meningitis intestinal infections and septicemia. E. coli is also one of the most prolific bacteria both in men’s and animal’s intestinal flora, which is why it is frequently used as an indicator of the selective pressure caused by the use of antibiotics in men and animals, especially when it comes to the production of animals for human consume. Data gathered around Europe shows a high occurrence of E.coli commensal strains present in men and animals which are resistant to various antibiotics. E.coli isolated belonging to phylogenetic groups A and B1 are traditionally considered to be commensal. Nevertheless, they have increasingly been associated with extra-intestinal infections, something common among the more virulent groups B2 and D. The main objective of this study is to characterize the population structure of E.coli isolated obtained from Portuguese pig farms, evaluate their resistance to some of the most clinically used antibiotics and to relate those resistances with the virulence showed by the phylogenic groups, as there are very few studies on the matter. This study also intends to define strategies or enhance those already in place in order to help prevent the transmission of resistances to men, may it be through the food chain or the environment, ultimately decreasing public health’s risks. With that aim in mind, 155 bacteria isolated were analyzed using samples of animals and the environment (water, air, rations, among others) belonging to an intensive production Portuguese pig farm and another one of extensive production. After extracting bacterial DNA from the isolated, it was possible to identify the presence of E. coli by PCR. For the identified E. coli isolated it was done a PCR multiplex to determine the phylogenic group, including the analysis of every single PCR product by the way of agar gel electrophoresis. It was also conducted the susceptibility analysis of the E. coli isolated through the disc diffusion method (Kirby – Bauer method), with different classes antibiotics being tested. Of the 155 bacterial isolated analyzed, 103 isolated (66%) were identified as being E. coli. Additionally, it was also observed that the isolated originating from the two studied pig farms were frequently more resistant to the spectinomycin antibiotic (100%), neomycin (95%) and streptomycin (85%). In general, higher percentages of resistance were recorded in the intensive production pig farm. The phylogenetic group A is predominant in the total number of isolated considered to be resistant. Although in a lesser quantity, phylogenetic group B appears associated to E. coli. isolated resistant to almost all antibiotics tested. Portuguese pig farms seem to be potential virulent bacteria repositories with resistance to human clinic antibiotics and possibly also repositories of antibiotic resistant genes with high capacity of dispersion. The theme of antimicrobial resistance is a matter of huge importance but still an area with a lot to discover, mainly in the role played by animals in the epidemiology of antibiotic resistance. It is necessary to conduct more studies in order to make it possible to implement efficient measures in the control of antibiotic resistances

Ovarian Response, Follicular Function and Oocyte Developmental Competence in Gonadotropin Treated Prepubertal Calves

Sexual immaturity during the prepubertal period in cattle is characterized by low pulse frequency of LH, anovulatory waves of follicle development, absence of corpora lutea (i.e., progesterone) and oocytes of reduced developmental capacity in vitro when compared to oocytes of sexually mature animals. However, calf ovaries are responsive to exogenous gonadotropin treatment early in life, and the use of prepubertal animals as a source of oocytes for in vitro embryo production may have significant potential to decrease generation interval and accelerate the rate of genetic gain. The overall objective of this thesis was to investigate the effect of exogenous FSH treatment on the ovarian response, hormonal profiles, ovulation and oocyte developmental competence in prepubertal calves. In the first study (Chapter 3) the effect of cumulative dose (200 mg vs. 350 mg) and duration (4 vs. 7 days) of FSH treatment on the ovarian response and the number of spontaneous and induced ovulations in 5-month-old calves were compared. Calves (n=24) were selected for gonadotropin treatment from a group of spring-born calves (n=51) based on the antral follicle counts (AFC) at the time of wave emergence. Calves were classified in low, medium and high AFC, and the ones in the medium classification (250.8, range of 20 to 32 follicles) were used in this study. At the end of the FSH treatment and 24 hours after treatment with pLH, the number of follicles 9 mm was greater in the 7-day than in the 4-day treatment group and in calves given a cumulative dose of 350 mg of FSH compared to those given 200 mg. Spontaneous ovulations were observed in 14 calves between Day 4 of FSH treatment and 12-hours post-LH treatment. The number of total and spontaneous ovulations was higher in the 7-day treatment groups than in the 4-day groups, and the number of spontaneous ovulations was higher in calves given a cumulative dose of 200 mg FSH than 350 mg. Numbers of ovulations in response to exogenous LH did not differ among groups. In the second study (Chapter 4), data showed in prepubertal calves (n=46) that the number of follicles at the beginning of a wave was predictive of the number recruited into subsequent waves and that after FSH treatment, the number of medium and large sized follicles available for follicular aspiration was positively associated with the number of follicles 1 mm at the time of wave emergence. In calves with low (n=12) and high (n=10) AFC at wave emergence, 7 days of FSH treatment resulted in a higher number of large than small size follicles than the 4 days of FSH treatment. High AFC at wave emergence resulted in a greater number of follicles 6 mm available for aspiration and a greater number of cumulus oocyte complexes (COC) collected than low AFC. The third study (Chapter 5) was designed to investigate the relationship between the antral follicular counts and plasma concentrations of AMH and FSH at the time of wave emergence in prepubertal calves and to compare the effects of age and duration of gonadotropin treatment on ovarian response of prepubertal and pubertal cattle. The AFC and the plasma concentrations of AMH at the time of wave emergence were positively correlated and both were positively correlated with the number of follicles 6 mm at the time of oocyte collection. Ovarian response was greater in calves at 4 months of age than at 7 months and was characterized by a higher number of medium to large (i.e., 6mm) sized follicles and higher number of recovered oocytes following transvaginal ultrasound-guided aspiration. Prolonged follicular growth by the 7-day treatment resulted in a greater number of large follicles at the end of the treatment and a greater degree of follicular maturation, characterized by lower intrafollicular E2:P4 ratio and higher proportion of fully expanded COC after LH treatment. In the fourth study (Chapter 6), different methods to control endogenous LH release and prevent ovulations during 7 days of exogenous FSH treatment were investigated. A long-acting progesterone (Long-acting P4) treatment at the time of follicular ablation and the GnRH antagonist Cetrorelix given at 48 hours intervals during FSH treatment were effective in preventing endogenous LH release and spontaneous ovulations during 7 days of exogenous FSH treatment. Ovulations were observed after FSH treatment in the Long-acting P4 treatment groups, while ovulations were prevented in calves of the Cetrorelix group. Luteal structures formed due to spontaneous ovulations were functional and a positive correlation was observed between the number of structures and the plasma concentrations of progesterone. In Chapter 7, oocyte developmental competence following in vitro vs. in vivo maturation of oocytes from prepubertal calves and in vivo maturation of oocytes from calves and pubertal heifers given 4 or 7 days of exogenous FSH treatment were evaluated in Study I. Spontaneous ovulations before oocyte collection were observed in calves and prevented meaningful comparisons among groups, resulting in lower rates of cleavage and blastocyst in the 7-day in vivo group. Blastocyst rates did not differ between 4-day in vivo vs. 4- and 7-day in vitro. In Study II, 4, 6 and 7 days of exogenous FSH treatment under controlled endogenous LH release were compared and six days of exogenous FSH support was associated with the greatest developmental competence of oocytes collected from 5-month-old calves

Users' Perceptions of Signage Systems at Three Portuguese Hospitals

This article is a report on the quantitative data collected from patients, family members, and visitors using the outpatient areas of three hospitals in Portugal. It details the users' views regarding the existing signage and presents suggestions to improve the design and implementation of the signage systems. A questionnaire was used with 1,287 respondents. The results showed that almost all users had a positive opinion regarding the current signage. However, some of the users' answers and observed behaviors indicated that the majority tended to ignore the signs and preferred to ask staff for help. Additionally, when asked for suggestions, many of the respondents were able to point out existing problems that affected their wayfinding. Although the signage was generally evaluated as good, many of the users perceived a variety of problems and, as already mentioned, asked the staff for directions, which results in lost time and hidden costs for the institutions