NemaFootPrinter: a web based software for the identification of conserved non-coding genome sequence regions between C. elegans and C. briggsae

BACKGROUND: NemaFootPrinter (Nematode Transcription Factor Scan Through Philogenetic Footprinting) is a web-based software for interactive identification of conserved, non-exonic DNA segments in the genomes of C. elegans and C. briggsae. It has been implemented according to the following project specifications: a) Automated identification of orthologous gene pairs. b) Interactive selection of the boundaries of the genes to be compared. c) Pairwise sequence comparison with a range of different methods. d) Identification of putative transcription factor binding sites on conserved, non-exonic DNA segments. RESULTS: Starting from a C. elegans or C. briggsae gene name or identifier, the software identifies the putative ortholog (if any), based on information derived from public nematode genome annotation databases. The investigator can then retrieve the genome DNA sequences of the two orthologous genes; visualize graphically the genes' intron/exon structure and the surrounding DNA regions; select, through an interactive graphical user interface, subsequences of the two gene regions. Using a bioinformatics toolbox (Blast2seq, Dotmatcher, Ssearch and connection to the rVista database) the investigator is able at the end of the procedure to identify and analyze significant sequences similarities, detecting the presence of transcription factor binding sites corresponding to the conserved segments. The software automatically masks exons. DISCUSSION: This software is intended as a practical and intuitive tool for the researchers interested in the identification of non-exonic conserved sequence segments between C. elegans and C. briggsae. These sequences may contain regulatory transcriptional elements since they are conserved between two related, but rapidly evolving genomes. This software also highlights the power of genome annotation databases when they are conceived as an open resource and the possibilities offered by seamless integration of different web services via the http protocol. Availability: the program is freely available a

Prosthetic strap system for simplified ventral hernia repair. Its results in a porcine experimental model.

Abstract: Introduction Aiming to achieve a simplified ventral hernia repair, a proprietary shaped mesh was experimentally tested in the porcine model. The mesh is structured with a large central body and radiating straps. The friction of the straps passing through the tissues are hypothesized to be adequate to maintain the mesh in place during tissue ingrowth, avoiding classic point fixation while ensuring a wide coverage of the abdomen. Methods The mesh, having 6 radial straps, was placed using a sublay preperitoneal technique in 4 pigs. All straps were passed laterally through the abdominal wall and exteriorized from the skin. The straps were trimmed at the level of the skin, allowing the stumps to recoil into the subcutaneous space. The animal were euthanized at 1 and 4 months to determine the integration of the straps. Results Macroscopically all 24 straps were firmly incorporated within the abdominal wall. The tension free placement of the mesh by using the straps was effective. The friction of the straps passing through the tissues was adequate to keep the mesh well orientated. No dislocation of the implants was observed. The strap system also allowed a broader coverage of the abdominal wall, far beyond the wound opening. Conclusions The described arm system of the described implant seems to be effective in eliminating point fixation of the mesh. The fixation arms seemed to have ensured the mesh stayed orientated in all animals. A very wide lateral mesh placement was accomplished, assuring sufficient defect overlap when shrinkage occurs

Polybutylene succinate artificial scaffold for peripheral nerve regeneration

Regeneration and recovery of nerve tissues are a great challenge for medicine, and positively affect the quality of life of patients. The development of tissue engineering offers a new approach to the problem with the creation of multifunctional artificial scaffolds that act on various levels in the damaged tissue, providing physical and biochemical support for the growth of nerve cells. In this study, the effects of the use of a tubular scaffold made of polybutylene succinate (PBS), surgically positioned at the level of a sciatic nerve injured in rat, between the proximal stump and the distal one, was investigated. Scaffolds characterization was carried out by scanning electron microscopy and X-ray microcomputed tomography and magnetic resonance imaging, in vivo. The demonstration of the nerve regeneration was based on the evaluation of electroneurography, measuring the weight of gastrocnemius and tibialis anterior muscles, histological examination of regenerated nerves and observing the recovery of the locomotor activity of animals. The PBS tubular scaffold minimized iatrogenic trauma on the nerve, acting as a directional guide for the regenerating fibers by conveying them toward the distal stump. In this context, neurotrophic and neurotropic factors may accumulate and perform their functions, while invasion by macrophages and scar tissue is hampered

OBTAINING MESENCHYMAL STEM CELLS FROM ADIPOSE TISSUE OR MURIN ORIGIN: EXPERIMENTAL STUDY.

The aim of this study was to isolate and characterize rat adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order to evaluate their proliferative potential and their ability to different cell types. AD-MSCs and Derived Mesenchymal Stem Cells (BM-MSCs) have the same characteristic in terms of plasticity. The advantage of adipose tissue is that it is an easier accessible source and it offers a large amount of MSCs by less invasive surgical tecniques. MSCs were obtained from subcutaneous adipose tissue of Wistar rats. first of all microbiological controls were made to exclude the presence of bacteria of fungi in then tissue. Adipose tissue was mechanically and enzimatically fragmented and stromal cell fraction was seeded in adherent culture flasks in DMEM 20% FBS. After 48 h the medium was replaced. Cells were characterized by evaluating:1)their ability tho adhere to the plastic; 2) the clonogenic potential by Colony Forming Unit (CFU) assay, 3) their ability to differentiate in 3 mesodermal lineages (adipocytes, osteocytes and chondrocytes). AD-MSCs are able to differentiate in adipocytes, osteocytes and chondrocytes as confirmed by oil red'O staining, von Kossa staining and histological analuysis respectively. This first characterization is essential for the second part of our study in which we are planning to use AD-MSCs in vivo to restore renal function after an induced ischemic damage in experimental animals

The effects of physical training without equipment on pain perception and balance in the elderly: a randomized controlled trials

Background: Research supports a link between exercise and falls prevention in the older population. Objectives: Our aims were to evaluate pain perception and balance skills in a group of elderly subjects and to examine the consequences of a standardized equipment-free exercise program intervention on these variables. The study utilized a randomized controlled trial method. Methods: 92 subjects were recruited from a rural Sicilian village (Resuttano, Sicily, Italy). Subjects were randomly split into two groups, an experimental group (EG; n= 49) and a control group (CG; n =43). Qualified fitness instructors delivered the standardized physical exercise program for the EG whilst the CG did not receive this exercise intervention. The Berg Balance Scale and the Oswestry Disability Index were administered in both groups before (T0) and after the intervention (T1). Results: At T1, the EG group significantly improvement in balance (p<0.0001) and pain perception (p<0.0001). No significant differences were found within the CG both in BBS and ODI, respectively. Conclusions: Our findings suggest that a 13-weeks standardized exercise equipment-free program is effective in improving balance and perception of pain in the elderly. This type of intervention can consequently provide a low cost strategy to counteract the rate of disability in elderly

The Caenorhabditis elegans Elongator Complex Regulates Neuronal α-tubulin Acetylation

Although acetylated α-tubulin is known to be a marker of stable microtubules in neurons, precise factors that regulate α-tubulin acetylation are, to date, largely unknown. Therefore, a genetic screen was employed in the nematode Caenorhabditis elegans that identified the Elongator complex as a possible regulator of α-tubulin acetylation. Detailed characterization of mutant animals revealed that the acetyltransferase activity of the Elongator is indeed required for correct acetylation of microtubules and for neuronal development. Moreover, the velocity of vesicles on microtubules was affected by mutations in Elongator. Elongator mutants also displayed defects in neurotransmitter levels. Furthermore, acetylation of α-tubulin was shown to act as a novel signal for the fine-tuning of microtubules dynamics by modulating α-tubulin turnover, which in turn affected neuronal shape. Given that mutations in the acetyltransferase subunit of the Elongator (Elp3) and in a scaffold subunit (Elp1) have previously been linked to human neurodegenerative diseases, namely Amyotrophic Lateral Sclerosis and Familial Dysautonomia respectively highlights the importance of this work and offers new insights to understand their etiology

Requirements for F-BAR Proteins TOCA-1 and TOCA-2 in Actin Dynamics and Membrane Trafficking during Caenorhabditis elegans Oocyte Growth and Embryonic Epidermal Morphogenesis

The TOCA family of F-BAR–containing proteins bind to and remodel lipid bilayers via their conserved F-BAR domains, and regulate actin dynamics via their N-Wasp binding SH3 domains. Thus, these proteins are predicted to play a pivotal role in coordinating membrane traffic with actin dynamics during cell migration and tissue morphogenesis. By combining genetic analysis in Caenorhabditis elegans with cellular biochemical experiments in mammalian cells, we showed that: i) loss of CeTOCA proteins reduced the efficiency of Clathrin-mediated endocytosis (CME) in oocytes. Genetic interference with CeTOCAs interacting proteins WSP-1 and WVE-1, and other components of the WVE-1 complex, produced a similar effect. Oocyte endocytosis defects correlated well with reduced egg production in these mutants. ii) CeTOCA proteins localize to cell–cell junctions and are required for proper embryonic morphogenesis, to position hypodermal cells and to organize junctional actin and the junction-associated protein AJM-1. iii) Double mutant analysis indicated that the toca genes act in the same pathway as the nematode homologue of N-WASP/WASP, wsp-1. Furthermore, mammalian TOCA-1 and C. elegans CeTOCAs physically associated with N-WASP and WSP-1 directly, or WAVE2 indirectly via ABI-1. Thus, we propose that TOCA proteins control tissues morphogenesis by coordinating Clathrin-dependent membrane trafficking with WAVE and N-WASP–dependent actin-dynamics

Amyotrophic lateral sclerosis in Palermo, Italy: an epidemiological study

The incidence, prevalence and natural course of ALS were determined in the population of the province of Palermo, Italy. The average annual incidence calculated for the years 1973 through 1984, was 044./100.000 inhabitants. The prevalence rate on prevalence day December 31, 1984, was 1.67/100.000 population. The male/female ratio was 1.38. The mean age at onset was 54.3±11.02. The most common clinical form was the conventional one (61.4%); the bulbar form was more frequent among females than males. The mean duration of the disease was 33.7±35.8 months. The longest duration belongs to the pseudopolyneuritic form. The median survival was 36 months: 16 months for the bulbar, 36 months for the conventional and 51 months for the pseudopolyneuritic form. © 1989 Masson Italia Periodici

A case-control study of amyotrophic lateral sclerosis

A retrospective case-control study was conducted using 46 patients affected by amyotrophic lateral sclerosis and 92 closely matched healthy controls. Cases were ascertained through typical clinical and instrumental findings. Putative risk factors (bone fractures or major trauma, exposure to domestic animals, surgical operations, disease among first degree relatives and others) were investigated anamnestically using a standard questionnaire. Using Mantel-Haenzsel estimates of the odds ratio, no association was found between amyotrophic lateral sclerosis and the investigated variables