Estela Bastos

Effects of high-speed power training on functional capacity and muscle performance in older women.

The purpose of the study was to examine the effects of 12 weeks high-speed power training on isometric contraction (handgrip strength), maximal strength (1RM), muscle power (walking velocity, counter movement jump and ball throwing) and functional tasks of the arm and leg muscles (sit-to-stand and get-up and go). Fifty-six older women were divided into an experimental group and a control group [EG, n=28, 62.5 (5.4) years; CG: n=28, 62.5 (4.3) years]. The EG was submitted to a high-speed power training that consisted of 40% of one repetition maximum (1<RM) to 75% of 1RM); 3 sets 4-12 reps, countermovement jump and medicine ball (1.5 kg) throwing. Over the 12-week training period, the EG significantly increased dynamic and isometric strength performance (57% to 61%), muscle power (range from 14% to 40%) (P<0.05) and function (P<0.05). No significant magnitudes of increase were observed in the CG. These data indicate that high-speed power training is an effective exercise approach leading to large gains in upper and lower extremity muscle performance and function capacity.

An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues

Formalin-fixed paraffin-embedded tissues (FFPET) represent the largest source of archival biological material available for genomic studies. In this work we present an advanced protocol for extraction of high quality DNA from FFPET that can be applied in several molecular studies. Although cat mammary tumours (CMT) are the third most frequent tumour in cats the recovery of significant number of samples for molecular studies are in some way restricted to FFPET samples. We were able to obtain high quality DNA from FFPET of thirty six CMT that were subjected to pre-fixation and fixation processes routinely used in the veterinary hospitals. The quality of DNA obtained was tested by PCR amplification using six sets of primers that amplify single-copy fragments. The DNA fragments obtained were further sequenced. This protocol was able to provide FFPET gDNA that can be amplified and sequenced for larger fragments up to 1182bp.

Canine periodontitis: The dog as an important model for periodontal studies

Periodontal disease (PD) refers to a group of inflammatory diseases caused by bacterial plaque in the periodontium and ranges from an early stage (gingivitis) to an advanced stage (periodontitis). It is a multifactorial disease that results from the interaction of the host defence mechanisms with the plaque microorganisms. Early detection, diagnosis and treatment are essential in the control of this disease. PD has an enormous impact on human and veterinary medicine due to its high prevalence. The most common animal PD models use dogs and non-human primates, although other animals (rats, mice, hamsters, rabbits, miniature pigs, ferrets, and sheep) have also been employed. Dog models have contributed significantly to the current understanding of periodontology. The most important clinical aspects of canine PD are considered in this review and the various animal models are examined with an emphasis on the role of the dog as the most useful approach for understanding human PD and in the development of new therapeutic and preventive measures.

Molecular sexing of birds: A comparative review of polymerase chain reaction (PCR)-based methods.

Accurate identification of sex in birds is important for the management and conservation of avian wildlife in several ways, namely in the development of population, behavioral and ecological studies, as well as in the improvement of ex situ captive breeding programs. In general, nestlings, juveniles and adult birds of a wide number of sexually monomorphic species cannot be sexed based on phenotypic traits. The development of molecular methodologies for avian sexing overcame these difficulties, allowing a reliable gender differentiation for these species. The polymerase chain reaction (PCR)-based methods have been widely applied in molecular sexing of birds, using a large diversity of sex-linked markers. During the last 15 yrs, there was a continuous improvement in the PCR-based protocols for bird sexing, increasing the accuracy, speed and high-throughput applicability of these techniques. The recent advances in real-time PCR platforms and whole genome analysis methods provided new resources for the detection and analysis of novel specific markers and protocols. This review presents a comparative guide of classical and recent advances in PCR-based methods for avian molecular sexing, highlighting its strengths and limitations. Future research opportunities in this field are also addressed.

Polymerase chain reaction-single strand conformation polymorphism applied to sex identification of Accipiter cooperii

Determination of sex in birds is valuable for studying population dynamics and structure, habitat use, behavior and mating systems. The purpose of the present study was to optimize a DNA-based methodology to allow the sex identification in Accipiter cooperii nestlings. Chromo-helicase-DNA-binding (CHD1) gene was used in this work as a marker for sex identification. CHD-W and CHD-Z sequences should present length and/or sequence differences providing a way to identify gender. We used a non-invasive method for DNA extraction from feathers and performed polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method. The length difference between CHD-W and CHD-Z amplified fragments observed by electrophoresis in conventional agarose gel was not enough to provide a clear differentiation between males and females. However, patterns obtained by PCR-SSCP differentiated undoubtedly males and females in A. cooperii. This tool provides a precise gender identification assay and will be applied to confirm and refine morphometrically based sexing techniques used in the field.

An Efficient Method for Genomic DNA Extraction from Different Molluscs Species

The selection of a DNA extraction method is a critical step when subsequent analysis depends on the DNA quality and quantity. Unlike mammals, for which several capable DNA extraction methods have been developed, for molluscs the availability of optimized genomic DNA extraction protocols is clearly insufficient. Several aspects such as animal physiology, the type (e.g., adductor muscle or gills) or quantity of tissue, can explain the lack of efficiency (quality and yield) in molluscs genomic DNA extraction procedure. In an attempt to overcome these aspects, this work describes an efficient method for molluscs genomic DNA extraction that was tested in several species from different orders: Veneridae, Ostreidae, Anomiidae, Cardiidae (Bivalvia) and Muricidae (Gastropoda), with different weight sample tissues. The isolated DNA was of high molecular weight with high yield and purity, even with reduced quantities of tissue. Moreover, the genomic DNA isolated, demonstrated to be suitable for several downstream molecular techniques, such as PCR sequencing among others.

Interleukin-6 gene −174G>C and −636G>C promoter polymorphisms and prostate cancer risk

Prostate cancer (PCa) is one of the most commonly diagnosed internal malignancies affecting men. Due to the important roles of IL-6 in different physiological and pathophysiological processes, IL-6 polymorphisms may modulate PCa risk. IL-6 −174 G>C (rs 1800795, also designated −236 G>C) and −636 G>C (rs 1800796, also designated −572 G>C) promoter polymorphisms have been implicated in PCa susceptibility, albeit still controversial. A literature search using PubMed and Highwire databases was conducted, resulting in eight case–control studies concerning the IL-6 −174 G>C polymorphism (11,613 PCa cases and 13,992 controls) and four case–control publications regarding the IL-6 −636 G>C polymorphism (1,941 PCa cases and 3,357 controls). In order to derive a more precise estimation, a meta-analysis based upon these selected case–control studies was performed. There was no significant association between IL-6 −174 G>C polymorphism and PCa increased risk. Nevertheless, the presence of allele C and the CC genotype were statistically significantly associated with decreased PCa risk in the overall analysis for IL-6 −636 G>C polymorphism. Additional studies in larger samples and analyses of functional repercussions of these SNPs in prostate tumor cells are necessary to validate these findings.

Differential mortality of birds killed at wind farms in Northern Portugal

Capsule The Skylark Alauda arvensis had the highest overall mortality in ten Northern Portuguese wind farms surveyed between 2006 and 2011. Analysis from the integration of conventional and molecular techniques suggest a sex and age biased mortality affecting mainly adult males (90.9%), which may be related to their characteristic breeding male song-flights making them highly vulnerable to collision with wind turbines. The results highlight the added value of more complete population impact assessments that go beyond simple carcass identification at wind farms.

A clinical survey on the prevalence and types of cheek teeth disorders present in 400 Zamorano-Leonés and 400 Mirandês donkeys (Equus asinus)

Dental disease is now recognised as a major but often unrecognised disorder of equids, including horses and donkeys. However, very few large clinical studies have documented the prevalence and type of dental disease present in different equid populations and no dental studies have been reported in Zamorano-Leonés or Mirandês donkeys, two endangered donkey breeds. Clinical and detailed oral examinations were performed in 400 Mirandês and 400 Zamorano-Leonés donkeys in Portugal and Spain. It was found that just 4.5 per cent had ever received any previous dental care. Cheek teeth (CT) disorders were present in 82.8 per cent of these donkeys, ranging from a prevalence of 29.6 per cent in the <2.5-year-old group to 100 per cent in the >25-year-old group. These CT disorders included enamel overgrowths (73.1 per cent prevalence but with just 6.3 per cent having associated soft tissue injuries), focal overgrowths (37.3 per cent), periodontal disease (23.5 per cent) and diastemata (19.9 per cent). Peripheral caries was present in 5.9 per cent of cases, but inexplicably, infundibular caries was very rare (1.3 per cent prevalence); this may have been due to their almost fully foraged diet. The high prevalence of enamel overgrowths in these donkeys, most which never received concentrates, also raises questions about the aetiology of this disorder. This very high prevalence of CT disorders, especially in older donkeys, was of great welfare concern in some cases and emphasises the need for routine dental care in these cases on welfare grounds and in order to help preserve these unique breeds.

ERBB2 in cat mammary neoplasias disclosed a positive correlation between RNA and protein low expression levels: a model for erbB-2 negative human breast cancer.

Human ERBB2 is a proto-oncogene that codes for the erbB-2 epithelial growth factor receptor. In human breast cancer (HBC), erbB-2 protein overexpression has been repeatedly correlated with poor prognosis. In more recent works, underexpression of this gene has been described in HBC. Moreover, it is also recognised that oncogenes that are commonly amplified or deleted encompass point mutations, and some of these are associated with HBC. In cat mammary lesions (CMLs), the overexpression of ERBB2 (27%–59.6%) has also been described, mostly at the protein level and although cat mammary neoplasias are considered to be a natural model of HBC, molecular information is still scarce. In the present work, a cat ERBB2 fragment, comprising exons 10 to 15 (ERBB2_10–15) was achieved for the first time. Allelic variants and genomic haplotype analyses were also performed, and differences between normal and CML populations were observed. Three amino acid changes, corresponding to 3 non-synonymous genomic sequence variants that were only detected in CMLs, were proposed to damage the 3D structure of the protein. We analysed the cat ERBB2 gene at the DNA (copy number determination), mRNA (expression levels assessment) and protein levels (in extra- and intra protein domains) in CML samples and correlated the last two evaluations with clinicopathological features. We found a positive correlation between the expression levels of the ERBB2 RNA and erbB-2 protein, corresponding to the intracellular region. Additionally, we detected a positive correlation between higher mRNA expression and better clinical outcome. Our results suggest that the ERBB2 gene is post-transcriptionally regulated and that proteins with truncations and single point mutations are present in cat mammary neoplastic lesions. We would like to emphasise that the recurrent occurrence of low erbB-2 expression levels in cat mammary tumours, suggests the cat mammary neoplasias as a valuable model for erbB-2 negative HBC.