Prosthesis grasp reflex via peripheral nerve control: an in vitro study

Here we present an economical and versatile platform for developing motor control and sensory feedback of a prosthetic hand via in vitro mammalian peripheral nerve activity. In this study, closed-loop control of the grasp function of the prosthetic hand was achieved by stimulation of a peripheral nerve preparation in response to slip sensor data from a robotic hand, forming a rudimentary reflex action. The single degree of freedom grasp was triggered by single unit activity from motor and sensory fibers as a result of stimulation. The work presented here provides a novel, reproducible, economic, and robust platform for experimenting with neural control of prosthetic devices before attempting in vivo implementation.

Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity

Foot-and-mouth disease virus (FMDV) is a significant economically and distributed globally pathogen of Artiodactyla. Current vaccines are chemically inactivated whole virus particles that require large-scale virus growth in strict bio-containment with the associated risks of accidental release or incomplete inactivation. Non-infectious empty capsids are structural mimics of authentic particles with no associated risk and constitute an alternate vaccine candidate. Capsids self-assemble from the processed virus structural proteins, VP0, VP3 and VP1, which are released from the structural protein precursor P1-2A by the action of the virus-encoded 3C protease. To date recombinant empty capsid assembly has been limited by poor expression levels, restricting the development of empty capsids as a viable vaccine. Here expression of the FMDV structural protein precursor P1-2A in insect cells is shown to be efficient but linkage of the cognate 3C protease to the C-terminus reduces expression significantly. Inactivation of the 3C enzyme in a P1-2A-3C cassette allows expression and intermediate levels of 3C activity resulted in efficient processing of the P1-2A precursor into the structural proteins which assembled into empty capsids. Expression was independent of the insect host cell background and leads to capsids that are recognised as authentic by a range of anti-FMDV bovine sera suggesting their feasibility as an alternate vaccine.

A spirochete isolated from a case of severe virulent ovine foot disease is closely related to a Treponeme isolated from human periodontitis and bovine digital dermatitis

The isolation of spirochetes from severe ovine foot disease has been reported recently by our research group. In this study we describe the preliminary classification of this spirochete based on nucleotide sequence analysis of the PCR-amplified 16S rRNA gene. Phylogenetic analysis of this sequence in comparison with other previously reported 16S rRNA gene sequences showed that the spirochete belonged to the treponemal phylotype Treponema vincentii which has been associated with bovine digital dermatitis and human periodontal disease. Further work is required to define the common virulence determinants of these closely related treponemes in the aetiology of these tissue destructive diseases. (C) 2000 Elsevier Science B.V. All rights reserved.

Genetic relatedness and phenotypic characteristics of Treponema associated with human periodontal tissues and ruminant foot disease

Treponema have been implicated recently in the pathogenesis of digital dermatitis (DID) and contagious ovine digital dermatitis (CODD) that are infectious diseases of bovine and ovine foot tissues, respectively. Previous analyses of treponemal 16S rDNA sequences, PCR-amplified directly from DID or CODD lesions, have suggested relatedness of animal Treponema to some human oral Treponema species isolated from periodontal tissues. In this study a range of adhesion and virulence-related properties of three animal Treponema isolates have been compared with representative human oral strains of Treponema denticola and Treponema vincentii. In adhesion assays using biotinylated treponemal cells, T denticola cells bound in consistently higher numbers to fibronectin, laminin, collagen type 1, gelatin, keratin and lactoferrin than did T. vincentii or animal Treponema isolates. However, animal DID strains adhered to fibrinogen at equivalent or greater levels than T denticola. All Treponema strains bound to the amino-terminal heparin l/fibrin I domain of fibronectin. 16S rDNA sequence analyses placed ovine strain UB1090 and bovine strain UB1467 within a cluster that was phylogenetically related to T vincentii, while ovine strain UB1466 appeared more closely related to T denticola. These observations correlated with phenotypic properties. Thus, T denticola ATCC 35405, GM-1, and Treponema UB1466 had similar outer-membrane protein profiles, produced chymotrypsin-like protease (CTLP), trypsin-like protease and high levels of proline iminopeptidase, and co-aggregated with human oral bacteria Porphyromonas gingivalis and Streptococcus crista. Conversely, T vincentii ATCC 35580, D2A-2, and animal strains UB1090 and UB1467 did not express CTLP or trypsin-like protease and did not co-aggregate with P. gingivalis or S. crista. Taken collectively, these results suggest that human oral-related Treponema have broad host specificity and that similar control or preventive strategies might be developed for human and animal Treponema-associated infections.

Dairy farmers’ attitudes and intentions towards improving dairy cow foot health

Dairy cow foot health is a subject of concern because it is considered to be the most important welfare problem in dairy farming and causes economic losses for the farmer. In order to improve dairy cow foot health it is important to take into account the attitude and intention of dairy farmers. In our study the objective was to gain insight into the attitude and intention of dairy farmers to take action to improve dairy cow foot health and determine drivers and barriers to take action, using the Theory of Planned Behavior. Five hundred Dutch dairy farmers were selected randomly and were invited by email to fill in an online questionnaire. The questionnaire included questions about respondents’ intentions, attitudes, subjective norms and perceived behavioral control and was extended with questions about personal normative beliefs. With information from such a framework, solution strategies for the improvement of dairy cow foot health can be proposed. The results showed that almost 70% of the dairy farmers had an intention to take action to improve dairy cow foot health. Most important drivers seem to be the achievement of better foot health with cost-effective measures. Possible barriers to taking action were labor efficiency and a long interval between taking action and seeing an improvement in dairy cow foot health. The feed advisor and foot trimmer seemed to have most influence on intentions to take action to improve dairy cow foot health. Most farmers seemed to be satisfied with the foot health status at their farm, which probably weakens the intention for foot health improvement, especially compared to other issues which farmers experience as more urgent. Subclinical foot disorders (where cows are not visibly lame) were not valued as important with respect to animal welfare. Furthermore, 25% of the respondents did not believe cows could suffer pain. Animal welfare, especially the provision of good care for the cows, was valued as important but was not related to intention to improve dairy cow foot health. The cost-effectiveness of measures seemed to be more important. Providing more information on the effects of taking intervention measures might stimulate farmers to take action to achieve improvement in dairy cow foot health.

Structure-based energetics of protein interfaces guides foot-and-mouth disease virus vaccine design

Virus capsids are primed for disassembly, yet capsid integrity is key to generating a protective immune response. Foot-and-mouth disease virus (FMDV) capsids comprise identical pentameric protein subunits held together by tenuous noncovalent interactions and are often unstable. Chemically inactivated or recombinant empty capsids, which could form the basis of future vaccines, are even less stable than live virus. Here we devised a computational method to assess the relative stability of protein-protein interfaces and used it to design improved candidate vaccines for two poorly stable, but globally important, serotypes of FMDV: O and SAT2. We used a restrained molecular dynamics strategy to rank mutations predicted to strengthen the pentamer interfaces and applied the results to produce stabilized capsids. Structural analyses and stability assays confirmed the predictions, and vaccinated animals generated improved neutralizing-antibody responses to stabilized particles compared to parental viruses and wild-type capsids.

An Alternative Section Method for Casting and Posterior Laser Welding of Metallic Frameworks for an Implant-Supported Prosthesis

Purpose: The aim of this study was to compare the accuracy of fit of three types of implant-supported frameworks cast in Ni-Cr alloy: specifically, a framework cast as one piece compared to frameworks cast separately in sections to the transverse or the diagonal axis, and later laser welded. Materials and Methods: Three sets of similar implant-supported frameworks were constructed. The first group of six 3-unit implant-supported frameworks were cast as one piece, the second group of six were sectioned in the transverse axis of the pontic region prior to casting, and the last group of six were sectioned in the diagonal axis of the pontic region prior to casting. The sectioned frameworks were positioned in the matrix (10 N(.)cm torque) and laser welded. To evaluate passive fit, readings were made with an optical microscope with both screws tightened and with only one-screw tightened. Data were submitted to ANOVA and Tukey-Kramer`s test (p < 0.05). Results: When both screws were tightened, no differences were found between the three groups (p > 0.05). In the single-screw-tightened test, with readings made opposite to the tightened side, the group cast as one piece (57.02 +/- 33.48 mu m) was significantly different (p < 0.05) from the group sectioned diagonally (18.92 +/- 4.75 mu m) but no different (p > 0.05) from the group transversally sectioned (31.42 +/- 20.68 mu m). On the tightened side, no significant differences were found between the groups (p > 0.05). Conclusions: Results of this study showed that casting diagonally sectioned frameworks lowers misfit levels of prosthetic implant-supported frameworks and also improves the levels of passivity to the same frameworks when compared to structures cast as one piece.

A novel locus for split-hand/foot malformation associated with tibial hemimelia (SHFLD syndrome) maps to chromosome region 17p13.1-17p13.3

Split-hand/foot malformation (SHFM) associated with aplasia of long bones, SHFLD syndrome or Tibial hemimelia-ectrodactyly syndrome is a rare condition with autosomal dominant inheritance, reduced penetrance and an incidence estimated to be about 1 in 1,000,000 liveborns. To date, three chromosomal regions have been reported as strong candidates for harboring SHFLD syndrome genes: 1q42.2-q43, 6q14.1 and 2q14.2. We characterized the phenotype of nine affected individuals from a large family with the aim of mapping the causative gene. Among the nine affected patients, four had only SHFM of the hands and no tibial defects, three had both defects and two had only unilateral tibial hemimelia. In keeping with previous publications of this and other families, there was clear evidence of both variable expression and incomplete penetrance, the latter bearing hallmarks of anticipation. Segregation analysis and multipoint Lod scores calculations (maximum Lod score of 5.03 using the LINKMAP software) using all potentially informative family members, both affected and unaffected, identified the chromosomal region 17p13.1-17p13.3 as the best and only candidate for harboring a novel mutated gene responsible for the syndrome in this family. The candidate gene CRK located within this region was sequenced but no pathogenic mutation was detected.

mRNA expression of corticotropin-releasing factor and urocortin 1 after restraint and foot shock together with alprazolam administration

Corticotropin-releasing factor (CRF) is expressed in the paraventricular nucleus of the hypothalamus (PVN), and act centrally to provoke stress-like autonomic and behavioral responses. Urocortins 1-3 are additional ligands to the CRF receptors 1 and 2. Ucn 1 neurons are primarily concentrated in the Edinger-Westphal (EW) nucleus and also have been associated with stress responses. It is also known that UCN 1 respond in different ways depending on the stressor presented. Benzodiazepines can act via the CRF peptidergic system and chronic administration of alprazolam does not interfere with CRF mRNA expression in the PVN, but significantly increase Ucn 1 mRNA expression in the EW. The aim of our study was to investigate the relationship between different stressor stimuli, foot shock (FS) and restraint (R), and the mRNA expression of CRF and Ucn 1 in the PVN and EW using alprazolam (A). We employed fos activation and in situ hybridization. Restraint group presented increased fos-ir and CRF mRNA expression in the PVN compared to FS group. The stress responses of R group were prevented by A. In the EW,fos-ir was higher in the FS group than in the R group, whereas Ucn 1 mRNA expression was higher in the R group than in the FS group. Alprazolam significantly increased fos-ir and Ucn 1 mRNA expression in both groups. Our results show that PVN and EW respond in different ways to the same stressors. Furthermore, EW of stressed animals replies in a complementary way comparing to PVN with the use of Alprazolam. (C) 2010 Elsevier Inc. All rights reserved.

Clinical and radiographic evaluation of a polyester prosthesis in dogs with cranial cruciate ligament rupture

Os achados clínicos e radiográficos após reparo intra-articular do ligamento cruzado cranial com prótese de poliéster, utilizando a técnica over-the-top modificada, foram avaliada em seis cães. Sete cirurgias foram realizadas devido ao acometimento bilateral em um dos animais. A avaliação clínica foi realizada ao 3º, 10º e 40º dias de pós-operatório, e a avaliação radiográfica realizada ao 5º e 24º meses após a cirurgia em cinco cães, por meio da qual se confirmou a progressão da doença articular degenerativa. A resolução dos sinais clínicos foi observada entre o 25º e 68º dias após a cirurgia, segundo avaliação realizada pelos proprietários. A função do membro operado foi considerada boa. Dois cães apresentaram desgaste e ruptura da prótese após a cirurgia. Concluiu-se que a prótese de poliéster, da forma como foi implantada neste estudo, não pode ser considerada como substituto satisfatório, uma vez que resultados superiores podem ser obtidos com ténicas menos invasivas e mais simples.

Foot-and-mouth disease in tropical wildlife

This review of foot-and-mouth disease in cloven-hoofed, free-living animals, describes the disease, the wide range of the hosts, the carrier state, and the interrelationship between disease in domestic livestock and wildlife. This information becomes even more crucial to the development of control strategies when linked to the process of pathogenesis and the epidemiology of the disease.

Genetic relatedness of commensal strains of Candida albicans carried in the oral cavity of patients' dental prosthesis users in Brazil

The aim of this study is to describe the degree of yeast-colonization in diabetic and hemodialysed-users of dental prostheses. Individuals (306) were examined using an oral rinse technique in order to evaluate the incidence of yeast-carriage, and genotype of C. albicans. Yeasts were isolated from 68.4% (91/133) individual's dental prostheses users. Dental prostheses were found to be a significant factor for the yeast colonization (P < 0.05). Overall, the intensity of carriage was higher in diabetic patients as compared with health and hemodialysed individuals (P < 0.05). The isolation rates were: C. albicans (51.7%), C. parapsilosis (20.9%), C. tropicalis (14.3%), C. glabrata (6.6%), C. krusei (3.3%), C. rugosa (1.1%), and Pichia (Pichia ohmeri, 2.2%). Ready-To-Go RAPD Analysis Beads were used and primer OPJ 6 distinguished the C. albicans isolates found in prostheses users. All the isolates were grouped into 11 RAPD profiles in four main clusters and, the average S (AB) for the entire collection of 47 C. albicans isolates were 0.779 +/- 0.178. Over 85% of isolates had a similarity level higher than or equal to 0.8 reinforcing the idea that the use of dental prostheses, independently of the host's clinical condition, probably provides the necessary conditions for these strains to gain a growth-specific advantage over others.

Prevalência de doença arterial obstrutiva periférica (DAOP) e alterações sensoriais em pacientes diabéticos tipo 2: impacto da DAOP sobre a qualidade de vida, nível de atividade física e composição corporal

Diabetes Mellitus (DM) affected approximately 171 million people in the world in the year 2000 as described by the World Health Organization (WHO). Because DM is a multisystem disease it can cause several complications especially those related to the cardiovascular system. The Peripheral Arterial Disease (PAD) of the lower limbs and the Diabetic Distal Symmetric Polyneuropathy (DDSP) can affect the DM patient causing consequences as the diabetic foot and eventually amputations. The main objective of this study was to determine the prevalence of PAD and sensorial impairment in 73 type 2 DM (DM2) patients and also assess the impact of PAD on quality of life, level of physical activity and body composition. For clinical assessment it was used: the ankle-brachial index (ABI); quantitative sensorial test for tactile sensibility (ST), pain (SD), vibration (SV); Achilles tendon reflex (RA); quality of life questionnaire (SF-36); modified Baecke physical activity questionnaire and bioelectric impedance. Prevalence of PAD in the studied population was 13.7%. ABI was inversely correlated to age (p=0,03; rhô= -0,26), diabetes duration (p=0,02; rhô= -0,28) and blood pressure (p= 0,0007; rhô= -0,33). There were lower scores for physical health summary on the SF-36 in DM2 patients; however, the presence of PAD predominantly mild did not significantly impact quality of life, body composition or physical activity level assessed by questionnaire. Fourteen patients (19.2%) present bilateral and symmetrical alterations in two or more sensorial tests compatible to DPN diagnosis. Abnormalities in ST, SD and SV were present in 27.3%, 24.6% and 8.2%; respectively. There was association of results from ST abnormalities with RA and mainly with SD, suggesting the importance of 10g monofilament use in DM2 routine assessment. In conclusion, the prevalence of PAD in subclinical DM2 was slightly higher compared to the general population and in agreement to previously published data in DM patients. The PAD severity was predominantly mild and still without repercussion on quality of life and body composition. Our study demonstrated a significant prevalence of both PAD and DPN in DM2 without previous diagnosis of these complications and indicates the necessity of early preventive and therapeutic interventions for this population