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.

Inferring animal husbandry strategies in coastal zones through stable isotope analysis: new evidence from the Flemish coastal plain (Belgium, 1st-15th century AD)

In a proof-of-concept study, Britton et al. (2008) demonstrated that the isotopic composition of halophytic plants can be traced in the skeletal tissues of their animal consumers. Here we apply the method to domestic herbivore remains (n = 303) from nine archaeological sites in or near the Flemish coastal plain (Belgium), where, prior to embankments, salt-marshes offered extensive pasture grounds for domestic herbivores. The sites span a period of ∼1500 years (Roman to late medieval period), during which the coastal landscape was progressively transformed from little managed wetlands to a fully embanked polder area. The bulk collagen data show variations between sites and over time, which are consistent with this historical framework and are interpreted as reflecting environmental change and differences in animal management in the coastal plain throughout the late Holocene. The study demonstrates the immense value of faunal stable isotope analysis for characterising coastal husbandry strategies beyond the means of traditional zooarchaeological techniques.

Efficient animal-serum free 3D cultivation method for adult human neural crest-derived stem cell therapeutics

Due to their broad differentiation potential and their persistence into adulthood, human neural crest-derived stem cells (NCSCs) harbour great potential for autologous cellular therapies, which include the treatment of neurodegenerative diseases and replacement of complex tissues containing various cell types, as in the case of musculoskeletal injuries. The use of serum-free approaches often results in insufficient proliferation of stem cells and foetal calf serum implicates the use of xenogenic medium components. Thus, there is much need for alternative cultivation strategies. In this study we describe for the first time a novel, human blood plasma based semi-solid medium for cultivation of human NCSCs. We cultivated human neural crest-derived inferior turbinate stem cells (ITSCs) within a blood plasma matrix, where they revealed higher proliferation rates compared to a standard serum-free approach. Three-dimensionality of the matrix was investigated using helium ion microscopy. ITSCs grew within the matrix as revealed by laser scanning microscopy. Genetic stability and maintenance of stemness characteristics were assured in 3D cultivated ITSCs, as demonstrated by unchanged expression profile and the capability for self-renewal. ITSCs pre-cultivated in the 3D matrix differentiated efficiently into ectodermal and mesodermal cell types, particularly including osteogenic cell types. Furthermore, ITSCs cultivated as described here could be easily infected with lentiviruses directly in substrate for potential tracing or gene therapeutic approaches. Taken together, the use of human blood plasma as an additive for a completely defined medium points towards a personalisable and autologous cultivation of human neural crest-derived stem cells under clinical grade conditions.

Novel glycopolymer hydrogels as mucosa-mimetic materials to reduce animal testing

Glycopolymer hydrogels capable of mimicking mucosal tissue in mucoadhesion testing have been designed. Liquid formulations containing mucoadhesive polymers were found to be retained on these tissues to the same extent as ex vivo gastric mucosa, when using a dynamic method of assessing mucoadhesion.

Human versus animal: contrasting decomposition dynamics of mammalian analogues in experimental taphonomy

Taphonomic studies regularly employ animal analogues for human decomposition due to ethical restrictions relating to the use of human tissue. However, the validity of using animal analogues in soil decomposition studies is still questioned. This study compared the decomposition of skeletal muscle tissues (SMTs) from human (Homo sapiens), pork (Sus scrofa), beef (Bos taurus), and lamb (Ovis aries) interred in soil microcosms. Fixed interval samples were collected from the SMT for microbial activity and mass tissue loss determination; samples were also taken from the underlying soil for pH, electrical conductivity, and nutrient (potassium, phosphate, ammonium, and nitrate) analysis. The overall patterns of nutrient fluxes and chemical changes in nonhuman SMT and the underlying soil followed that of human SMT. Ovine tissue was the most similar to human tissue in many of the measured parameters. Although no single analogue was a precise predictor of human decomposition in soil, all models offered close approximations in decomposition dynamics.

Samba, a Xenopus hnRNP Expressed in Neural and Neural Crest Tissues

RNA binding proteins regulate gene expression at the posttranscriptional level and play important roles in embryonic development. Here, we report the cloning and expression of Samba, a Xenopus hnRNP that is maternally expressed and persists at least until tail bud stages. During gastrula stages, Samba is enriched in the dorsal regions. Subsequently, its expression is elevated only in neural and neural crest tissues. In the latter, Samba expression overlaps with that of Slug in migratory neural crest cells. Thereafter, Samba is maintained in the neural crest derivatives, as well as other neural tissues, including the anterior and posterior neural tube and the eyes. Overexpression of Samba in the animal pole leads to defects in neural crest migration and cranial cartilage development. Thus, Samba encodes a Xenopus hnRNP that is expressed early in neural and neural crest derivatives and may regulate crest cells migratory behavior. Developmental Dynamics 238:204-209, 2009. (C) 2008 Wiley-Liss, Inc.

Implante endoscópico no esôfago distal de polimetilmetacrilato para controle do refluxo gastrofágico : estudo experimental em modelo animal

XIXA doença do refluxo gastroesofágico (DRGE) comumente afeta o esôfago e provavelmente é a condição mais prevalente no segmento alto do trato gastrointestinal, acometendo entre 5 e 45% da população ocidental. A terapêutica atual para essa doença além de medidas gerais e dietéticas, inclui tratamento farmacológico e/ou cirurgia. Ambos podem ser eficazes, mas apresentam elevado custo financeiro. O tratamento com fármacos pode apresentar baixa aderência medicamentosa e a cirurgia tem baixa, mas não desprezível, morbidade e mortalidade. Idealmente o tratamento da DRGE deveria ser eficaz, com baixo risco e com baixo custo. Objetivos: 1. Desenvolver um modelo experimental em suínos para o estudo do Refluxo Gastroesofágico através da Pressão e do Volume de Vazão Gástricos; 2. Avaliar a eficácia do implante endoscópico de PMMA ao nível do Esfíncter Esofágico Inferior (EEI) para aumentar a Pressão de Vazão Gástrica, o Volume de Vazão Gástrico e a Pressão Basal do EEI; 3. Descrever as reações histológicas associadas ao implante de PMMA. Material e Métodos: Suínos da raça Large White, do sexo feminino com 8 semanas de vida foram estudados no experimento. Foi realizada manometria do esfíncter esofágico inferior com registro da pressão basal com cateter de perfusão com água e técnica de retirada lenta. Calculou-se, também, a extensão do EEI. Após, gastrostomia era realizada com colocação intragástrica da extremidade distal de uma sonda de Foley com três vias e um cateter de pHmetria esofágica introduzido via oral com o sensor distal posicionado 5 cm acima do bordo superior de esfíncter esofágico inferior. Iniciava-se, XXentão, a infusão contínua no estômago de uma solução de HCl a 0,02N com medida e registro simultâneos da Pressão e do Volume de Vazão Gástricos e do pH esofágico. Definiu-se a Pressão de Vazão Gástrica (PVG) e o Volume de Vazão Gástrico quando houve brusca e sustentada acidificação do esôfago distal (pH<3). Após, introduzia-se um tubo metálico (Tubo Introdutor ou TI) via oral e, em seguida, o endoscópio, seguindo ambos até o esôfago distal. Cateter de nylon com agulha calibre 16 era introduzido pelo tubo introdutor e o PMMA implantado na área correspondente ao EEI com uma pistola dosadora volumétrica que permitia a injeção de volumes prédeterminados em 3 ou 4 pontos da submucosa (total por ponto = 0,73 ml de PMMA). As medidas de PVG, VVG e pressão basal do EEI foram repetidas após 28 dias, sacrificando-se os animais e removendo-se esôfago médio e distal, junção esofagogástrica, fundo e corpo gástricos para estudo histológico. Previamente ao experimento, três projetos pilotos foram desenvolvidos. O primeiro designado como “Projeto Piloto: Refluxo Gastroesofágico por pHmetria de 24H” (RGE 24H) buscou a via de acesso transnasal para registro de pHmetria por 24 horas. No segundo, designado “Projeto Piloto: Esofagostomia”, para confirmação de refluxo gastroesofágico espontâneo em suínos da raça Large White, utilizou-se anestesia com associação de tiletamina 125 mg e zolazepam 125 mg em combinação com um sedativo, a xilazina a 2% . A pHmetria foi mantida por 24 horas. Os dados descritos por Kadirkamanathan et al. não foram reproduzidos no nosso laboratório. Optou-se então pela realização de um terceiro projeto, “Projeto Piloto: Gastrostomia” para induzir refluxo gastroesofágico através de um modelo experimental e testar a reprodutibilidade da Pressão de Vazão Gástrica. Obteve-se sucesso. Resultados: Trinta e sete animais foram estudados em 60 intervenções no Laboratório de motilidade experimental. O projeto piloto “RGE:24h” foi abandonado após perda de 5 animais pela anestesia com halotano e, solucionada esta questão, a sistemática infecção do tecido celular subcutâneo nasal, dificultando a manutenção do cateter de phmetria em outros 5 animais. No “Projeto Piloto: Esofagostomia”, em cinco animais estudados (cada animal em duas ocasiões diferentes) não se reproduziram os achados de refluxo espontâneo. No terceiro projeto “Projeto Piloto: Gastrostomia” quatro animais foram estudados em dois momentos diferentes obtendo-se sucesso e reprodutibilidade XXIdos dados. Criado o modelo experimental, quatorze suínos foram submetidos ao experimento com implante endoscópico de PMMA com os seguintes resultados: A média dos pesos com 8 semanas de idade foi 14,98 ± 2,43 e 28 dias após o implante, 20,26 ± 3,68. O ganho ponderal foi considerado normal para a espécie. A Pressão de Vazão Gástrica média no dia 1 foi 8,08 mmHg e no dia 28, 10,69mmHg (Teste t de Student: t = 2,72 gl = 13 p = 0,017). Os Volumes de Vazão Gástricos médios foram: 392,86 ml para o dia 1 e 996,71 ml no dia 28 (teste t de Student: t = 11,66 gl = 13 p< 0,001). A Pressão Basal do EEI e o comprimento do esfíncter, não apresentaram diferenças estatisticamente significativas. PMMA foi identificado como depósitos de grandes vacúolos no tecido intersticial ao exame histológico da junção esofagogástrica associado a histiócitos, plasmócitos e presença de células gigantes tipo Langhans indicando reação tecidual de corpo estranho em todos os animais. Fibrose e macrófagos com vacúolos intracelulares estiveram presentes em menor freqüência. Conclusões: 1. O modelo experimental, em suínos, desenvolvido viabilizou o estudo do Refluxo Gastroesofágico através da Pressão de Vazão Gástrica e Volume de Vazão Gástrico; 2. O implante de PMMA, no presente estudo, aumentou a Pressão de Vazão e o Volume de Vazão Gástricos, mas não a Pressão Basal do EEI, nem tampouco aumentou o seu comprimento; 3. Depósito de PMMA implantado e evidência de processo inflamatório crônico e reação tecidual de corpo estranho foi encontrada no local do implante de PMMA. Macrófagos com vacúolos intracelulares e fibrose foram encontrados com menos freqüência.

Fluorescence spectroscopy for the detection of tongue carcinoma - validation in an animal model

The efficacy of fluorescence spectroscopy to detect squamous cell carcinoma is evaluated in an animal model following laser excitation at 442 and 532 nm. Lesions are chemically induced with a topical DMBA application at the left lateral tongue of Golden Syrian hamsters. The animals are investigated every 2 weeks after the 4th week of induction until a total of 26 weeks. The right lateral tongue of each animal is considered as a control site (normal contralateral tissue) and the induced lesions are analyzed as a set of points covering the entire clinically detectable area. Based on fluorescence spectral differences, four indices are determined to discriminate normal and carcinoma tissues, based on intraspectral analysis. The spectral data are also analyzed using a multivariate data analysis and the results are compared with histology as the diagnostic gold standard. The best result achieved is for blue excitation using the KNN (K-nearest neighbor, a interspectral analysis) algorithm with a sensitivity of 95.7% and a specificity of 91.6%. These high indices indicate that fluorescence spectroscopy may constitute a fast noninvasive auxiliary tool for diagnostic of cancer within the oral cavity. (C) 2008 Society of Photo-Optical Instrumentation Engineers.

Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues

Dezoito bovinos foram inoculados com Toxoplasma gondii e distribuídos aleatoriamente em três grupos de seis bovinos cada: GI (2,5x10(5) oocistos da cepa P), GII (5,0x10(6) taquizoítos da cepa RH) e GIII (controle). Exames clínicos, sorológicos e parasitêmicos foram realizados. Pesquisas do parasito, por meio da bioprova e pela técnica de Reação em Cadeia pela Polimerase (PCR), foram realizadas no sêmen e em fragmentos de musculatura esquelética, linfonodos, cérebro, retina, baço, fígado, pulmão, testículo, epidídimo e vesícula seminal. Amostras de sangue e sêmen foram colhidas nos dias -2, -1, 1, 3, 5, 7, 14 e, semanalmente, até o 84º dia pós-infecção (DPI). Os bovinos inoculados (GI e GII) apresentaram hipertermia do 3º ao 16º DPI. Anticorpos contra T. gondii foram detectados (IFI) no 5º DPI (1:16), em ambos grupos inoculados (oocistos e taquizoítos), atingindo picos de 1:4096 no 7º DPI. Surtos parasitêmicos ocorreram em todos os bovinos infectados, principalmente do 7º ao 28º DPI, independente da cepa e inóculo utilizados. O bioensaio revelou a presença do parasito em amostras seminais dos bovinos infectados com oocistos (GI) e taquizoítos (GII), em diversas datas experimentais, entre o 7º e 84º DPI. Parasitismo tissular por T. gondii foi diagnosticado por meio da bioprova e pela técnica da PCR, em vários fragmentos de tecidos e/ou órgãos. Os achados sugerem a possibilidade da ocorrência da transmissão sexual do T. gondii na espécie bovina.

Influence of implants with different sizes and configurations installed immediately into extraction sockets on peri-implant hard and soft tissues: an experimental study in dogs

Aim: To study the influence on the healing of soft and hard peri-implant tissues when implants of different sizes and configurations were installed into sockets immediately after tooth extraction.Material and methods: Transmucosal cylindrical implants, 3.3 mm in diameter in the control sites, and conical 5 mm in diameter in the test sites, were installed into the distal socket of the fourth mandibular premolars in dogs immediately after tooth extraction. After 4 months, the hard and soft tissue healing was evaluated histologically. Results: All implants were integrated in mineralized mature bone. Both at the test and control sites, the alveolar crest underwent resorption. The buccal bony surface at the implant test sites (conical; 3.8 mm) was more resorbed compared with the control sites (cylindrical; 1.6 mm). The soft tissue dimensions were similar in both groups. However, in relation to the implant shoulder, the peri-implant mucosa was located more apically at the test compared with the control sites.Conclusion: The present study confirmed that the distance between the implant surface and the outer contour of the buccal alveolar bony crest influenced the degree of resorption of the buccal bone plate. Consequently, in relation to the implant shoulder, the peri-implant mucosa will be established at a more apical level, if the distance between the implant surface and the outer contour of the alveolar crest is small.

Transcriptome architecture across tissues in the pig

Background: Artificial selection has resulted in animal breeds with extreme phenotypes. As an organism is made up of many different tissues and organs, each with its own genetic programme, it is pertinent to ask: How relevant is tissue in terms of total transcriptome variability? Which are the genes most distinctly expressed between tissues? Does breed or sex equally affect the transcriptome across tissues?Results: In order to gain insight on these issues, we conducted microarray expression profiling of 16 different tissues from four animals of two extreme pig breeds, Large White and Iberian, two males and two females. Mixed model analysis and neighbor - joining trees showed that tissues with similar developmental origin clustered closer than those with different embryonic origins. Often a sound biological interpretation was possible for overrepresented gene ontology categories within differentially expressed genes between groups of tissues. For instance, an excess of nervous system or muscle development genes were found among tissues of ectoderm or mesoderm origins, respectively. Tissue accounted for similar to 11 times more variability than sex or breed. Nevertheless, we were able to confidently identify genes with differential expression across tissues between breeds (33 genes) and between sexes (19 genes). The genes primarily affected by sex were overall different than those affected by breed or tissue. Interaction with tissue can be important for differentially expressed genes between breeds but not so much for genes whose expression differ between sexes.Conclusion: Embryonic development leaves an enduring footprint on the transcriptome. The interaction in gene x tissue for differentially expressed genes between breeds suggests that animal breeding has targeted differentially each tissue's transcriptome.

Free radical production by azomethine H: Effects on pancreatic and hepatic tissues

The antimalarial properties of azomethine H represent the basis for its use as a chemotherapeutic agent. This work was carried out in order to verify the biological side effects of azomethine H and to clarify the contribution of reactive oxygen species (ROS) in this process. It was shown that azomethine H increased serum activities of amylase, alanine transaminase (ALT) and the TEARS concentrations, in rats. No changes were observed in glutathione peroxidase and catalase activities. The drug-induced tissue damage might be due to superoxide radicals (O-2(.-)), since Cu-Zn superoxide dismutase activities were increased by azomethine I-I treatment. This study allows tentative conclusions to be drawn regarding which reactive oxygen metabolites play a role in azomethine H activity. We concluded that (O-2(.-)) maybe produced as a mediator of azomethine H action.

Acellular dermal tissue study: an ultrastructural evaluation of human and porcine derived tissues in a rat model

The purpose of this study was to evaluate the host response of a human and a porcine derived acellular dermal tissue (ADT) implanted in the subcutaneous tissue of a rat model. Two subcutaneous pockets were surgically created along the dorsal midline of 25 rats (5 rats/group). The human ADT was placed superiorly and the porcine ADT, inferiorly. The animals were sacrificed at 07, 15, 30, 60 and 180 postoperative days (PO) and the ADTs and surrounding soft tissues were assessed for ultrastructural evaluation by transmission electron microscopy. The ultrastructural findings were similar in both materials. Normal collagen and elastic fibers bundles were observed during all experimental moments, as well as macrophages presenting cytoplasmic enlargements digesting cellular portions after 15 PO. From 30 until 180 PO, vacuolar structures filled with an amorphous, electron-transparent substance, were present inside and outside the fibroblasts. Both human and porcine ADT showed similar pattern of ultrastructural response when implanted in the subcutaneous tissue of rats. The porcine ADT appears as a good alternative to be used as a biomaterial.