Genomic organization and single-nucleotide polymorphism map of desmuslin, a novel intermediate filament protein on chromosome 15q26.3

Background

Congenital fibrosis of the extraocular muscles type 1 (CFEOM1) is an autosomal dominant eye movement disorder linked to the pericentromere of chromosome 12 (12p11.2 - q12). Sarcospan is a member of the dystrophin associated protein complex in skeletal and extraocular muscle and maps to human chromosome 12p11.2. Mutations in the genes encoding each of the other components of the skeletal muscle sarcospan-sarcoglycan complex (α - δ sarcoglycan) have been shown to cause limb girdle muscular dystrophy (LGMD2C-F). To determine whether mutations in the sarcospan gene are responsible for CFEOM1 we: (1) attempted to map sarcospan to the CFEOM1 critical region; (2) developed a genomic primer set to directly sequence the sarcospan gene in CFEOM1 patients; and (3) generated an anti-sarcospan antibody to examine extraocular muscle biopsies from CFEOM1 patients.

Results

When tested by polymerase chain reaction, sarcospan sequence was not detected on yeast or bacterial artificial chromosomes from the CFEOM1 critical region. Sequencing of the sarcospan gene in CFEOM1 patients from 6 families revealed no mutations. Immunohistochemical studies of CFEOM1 extraocular muscles showed normal levels of sarcospan at the membrane. Finally, sarcospan was electronically mapped to bacterial artificial chromosomes that are considered to be outside of the CFEOM1 critical region.

Conclusions

In this report we evaluate sarcospan as a candidate gene for CFEOM1. We have found that it is highly unlikely that sarcospan is involved in the pathogenesis of this disease. As of yet no sarcospan gene mutations have been found to cause muscular abnormalities.     

The case for studying tadpole autecology,with comments on strategies to study other small,fast-moving animals in nature

Two of the most fundamental questions in tadpole biology, also applicable to most small, under-studied organisms are: (1) ‘Why are they built the way they are?’ and (2) ‘Why do they live where they do?’ Regrettably, despite significant progress in most aspects of tadpole biology, the answers to these questions are not much better now than they were in the last century. We propose that an autecological approach, that is the careful observation of individuals and how they interact with the environment, is a potential path towards a fuller understanding of tadpole ecomorphology and evolution. We also discuss why more attention should be given to studying atypical tadpoles from atypical environments, such as torrential streams, water-filled cavities of terrestrial plants and wet rock surfaces neighbouring streams. Granted, tadpoles are rare in these settings, but in those unusual habitats the physical environments can be well described and characterized. In contrast, the more common ponds where tadpoles are found are typically too structurally complex to be easily delineated. This makes it difficult to know exactly what individual tadpoles are doing and what environmental parameters they are responding to. Our overall thesis is that to understand tadpoles we must see exactly what they are doing, where they are doing it, and how they are doing it. This takes work, but we suggest it is feasible and could greatly advance our understanding of how anuran larvae have evolved. The same strategies for studying tadpoles that we encourage here can be applied to the study of many other small and fast-moving animals.     

Endothelial nitric oxide synthase,vascular integrity and human exceptional longevity

Aging is the sum of the deleterious changes that occur as time goes by. It is the main risk factor for the development of cardiovascular disease, and aging of the vasculature is the event that most often impacts on the health of elderly people. The “free-radical theory of aging” was proposed to explain aging as a consequence of the accumulation of reactive oxygen species (ROS). However, recent findings contradict this theory, and it now seems that mechanisms mediating longevity act through induction of oxidative stress. In fact, calorie restriction ? a powerful way of delaying aging ? increases ROS accumulation due to stimulation of the basal metabolic rate; moreover, reports show that antioxidant therapy is detrimental to healthy aging. We also now know that genetic manipulation of the insulin-like-growth-factor-1/insulin signal (IIS) has a profound impact on the rate of aging and that the IIS is modulated by calorie restriction and physical exercise. The IIS regulates activation of nitric oxide synthase (eNOS), the activity of which is essential to improving lifespan through calorie restriction, as demonstrated by experiments on eNOS knockout mice. Indeed, eNOS has a key role in maintaining vascular integrity during aging by activating vasorelaxation and allowing migration and angiogenesis. In this review, we will overview current literature on these topics and we will try to convince the reader of the importance of vascular integrity and nitric oxide production in determining healthy aging.     

Production,purification and partial characterisation of a novel laccase from the white-rot fungus <Emphasis Type="Italic">Panus tigrinus</Emphasis> CBS 577.79

Extracellular laccase from Panus tigrinus CBS 577.79 was produced in a bubble-column reactor using glucose-containing medium supplemented with 2,5-xylidine under conditions of nitrogen sufficiency. The main laccase isoenzyme was purified to apparent homogeneity by ultra-filtration, anion-exchange chromatography and gel filtration that led to a purified enzyme with a specific activity of 317 IU (mg protein)⁻¹ and a final yield of 66%. Laccase was found to be a monomeric protein with a molecular mass of 69.1 kDa, pI of 3.15 and 6.9% N-glycosylation of the high mannose type. Temperature and pH optima were 55°C and 3.75 (2,6-dimethoxyphenol as substrate). At 50 and 60°C, the enzyme half-lives were 281 and 25 min, respectively. The P. tigrinus laccase oxidized a wide range of both naturally occurring and synthetic aromatic compounds: the highest catalytic efficiencies were for 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonic) acid and 2,6-dimethoxyphenol (5.99 × 10⁶ and 3.07 × 10⁶ M⁻¹ s⁻¹, respectively). Catalytic rate constants for typical N–OH redox mediators, such as 1-hydroxybenzotriazole (2.6 s⁻¹), violuric acid (8.4 s⁻¹) and 2,2,6,6-tetramethylpiperidin-N-oxide radical (7.8 s⁻¹), were found to be higher than those reported for other high redox potential fungal laccases.     

Stoned olive pomace fermentation with Pleurotus species and its evaluation as a possible animal feed

The use of stoned olive pomace (SOP) as an unconventional feedstuff for livestock is limited by its inherently low crude protein (CP) content and by the presence of anti-nutritional compounds such as phenols. Aim of this study was to assess whether solid-state fermentation of SOP with selective lignin-degrading fungi might ameliorate nutritional properties of the waste. Incubation of SOP, mixed (25%, w/w) with various conventional feedstuffs (i.e., wheat bran, wheat middlings, barley grains, crimson clover, wheat flour shorts and field beans), with Pleurotus ostreatus and Pleurotus pulmonarius led to significant CP increases, ranging from 7 to 29%, and marked removal (from ca. 50–90%) of phenols after 6 weeks. Both species, however, led to moderate delignification associated with significant consumption of hemicelluloses. Consequently, no improvements of both organic matter digestibility (OMD) and net energy of SOP–feedstuff mixtures occurred after the fungal colonization.     

"Positive biology": the centenarian lesson

ABSTRACT: The extraordinary increase of the elderly in developed countries underscore the importance of studies on ageing and longevity and the need for the prompt spread of knowledge about ageing in order to satisfactorily decrease the medical, economic and social problems associated to advancing years, because of the increased number of individuals not autonomous and affected by invalidating pathologies.Centenarians are equipped to reach the extreme limits of human life span and, most importantly, to show relatively good health, being able to perform their routine daily life and to escape fatal age-related diseases. Thus, they are the best example of extreme longevity, representing selected people in which the appearance of major age-related diseases, such as cancer, and cardiovascular diseases among others, has been consistently delayed or escaped. To discuss the relevance of genetics and life style in the attainment of longevity, five papers mostly focused on Italian centenarians have been assembled in this series. The aim is to realize, through a" positive biology" approach (rather than making diseases the central focus of research, "positive biology" seeks to understand the causes of positive phenotypes, trying to explain the biological mechanisms of health and well-being) how to prevent and/or reduce elderly frailty and disability.