The effects of interleukin-10 depletion in vivo on the immune response to Porphyromonas gingivalis in a murine model

Background: The immune response to Porphyromonas gingivalis in the mouse abscess model is known to be dependent upon CD4 T-cell activation and the regulatory role of cytokines. The role of interleukin-10 (IL-10) in this mouse model was examined in vivo. Methods: One-week-old, female BALB/c mice were divided into 4 groups. Groups 1 and 2 were given intraperitoneal (ip) injections of phosphate buffered saline (PBS) weekly for 5 weeks. Group 3 was given an ip injection of rat immunoglobulin. Group 4 was injected with rat anti-IL-10 antibodies. At week 6, group 1 was sham-immunized with PBS, and groups 2, 3, and 4 were injected with P gingivalis lipopolysaccharide (Pg-LPS) weekly for 2 weeks. One week after the final immunization, delayed-type hypersensitivity (DTH) was assessed by footpad swelling to Pg-LPS. The level of serum antibodies to Pg-LPS and IFN-gamma (IFN-gamma) was determined by enzyme-linked immunosorbent assay. Dorsal abscess formation induced by the injection of viable P gingivalis was examined daily for 30 days. Results: The footpad swelling of the anti-IL-10-treated group (group 4) was significantly higher than that of groups 1 to 3. Similarly, the serum IFN-gamma level in group 4 was much higher than that of the other experimental groups. There was no significant difference in serum IgG antibodies to Pg-LPS in any of the experimental groups. However, the level of IgM antibodies in group 4 mice was significantly lower than that in groups 2 and 3. In addition, serum IgG1 was suppressed in group 4 mice, while IgG2a antibodies were raised. However, there was no difference observed between the levels of IgG2b and IgG3 antibodies in any group of mice. The lesions in sham-immunized mice (group 1) persisted for 30 days, and those in group 2 and 3 were undetected by day 18 and 20, respectively. In sharp contrast, lesions in group 4 had healed completely by day 13. Conclusions: This study has shown that IL-10 depletion in vivo in P gingivalis LPS-induced immune response in mice led to an elevated DTH response, an increase in serum IFN-gamma levels, and raised levels of IgG and IgG2a antibodies. Treatment with anti-IL-10 antibodies resulted in suppressed IgG I and IgM responses and a more rapid healing of abscesses than in non-IL-10-depleted mice. These results suggest that IL-10 depletion in Pg-LPS-induced immune response in mice may lead to a Th1-like immune response and provide strong protection against a subsequent challenge with live P gingivalis in an abscess model.

Finding gold in our shadow: psychotherapy and the shadow syndrome

Drawing on the work of Carl Jung and Robert Bly, Yaro Starak explores how 'shadow work' can be used within a Gestalt group therapy environment to uncover the 'gold in our shadow bags' - hidden inner strengths and resources that we were previously unaware of that may lead to healing and transformation. (editor abstract)

Sox18 is transiently expressed during angiogenesis in granulation tissue of skin wounds with an identical expression pattern to Flk-1 mRNA

Sox18 encodes a member of the Sry-related high mobility group box (SOX) family of developmental transcription factors. Examination of Sox18 expression during embryogenesis has shown that Sox18 is expressed transiently in endothelial cells of developing blood vessels, and mutations in Sox18 have been found to underlie the mouse vascular and hair follicle mutant ragged. In this study we have examined the expression of Sox18 in angiogenesis during wound healing. Full-thickness skin wounds were created in mice, and subsequent expression of vascular endothelial growth factor (VEGF), the VEGF receptor Flk-1, alpha1 (iv) collagen (Col4a1), and Sox18 were studied using in situ hybridization. As has been previously reported, VEGF was expressed predominantly in the keratinocytes at the wound margins. Sox18 expression was found Rye days after wounding during capillary sprouting in granulation tissue and persisted through the proliferative phase of healing, but was not detected in fully epithelialized wounds 21 days after wounding. Sox18 mRNA expression was detected in capillaries within the granulation tissue and showed an identical pattern of distribution to Flk-1 and Col4a1 mRNA expression in endothelial cells. Immunostaining with a polyclonal anti-Sox18 antibody showed SOX18 protein localized in capillary endothelial cells within the granulation tissue. capillaries in the subcutaneous tissue of unwounded skin showed no Sox18 expression. Sox18 may therefore represent a transcription factor involved in the induction of angiogenesis during wound healing and tissue repair, but not in the maintenance of endothelial cells in undamaged tissue.

Laminin 10/11: an alternative adhesive ligand for epidermal keratinocytes with a functional role in promoting proliferation and migration

We have investigated the expression and function of the isoforms of laminin bearing the alpha(5) chain, i.e. laminin-10/11 in neonatal and adult human skin. By immunostaining human skin derived from a variety of anatomic sites, we found that the laminin-alpha(5) chain is expressed abundantly in the basement membrane underlying the interfollicular epidermis and the blood vessels in the dermis. Interestingly, while the expression level of the well-studied laminin-5 isoform did not change significantly with age, laminin-10/11 (a5 chain) appeared to decrease in the basement membrane underlying the epidermis, in adult skin. In contrast, the levels of laminin-10/11 in the basement membrane underlying blood vessels remained unchanged in neonatal vs. adult skin. Importantly, in vitro cell adhesion assays demonstrated that laminin-10/11 is a potent adhesive substrate for both neonatal and adult keratinocytes and that this adhesion is mediated by the alpha(3)beta(1), and alpha(6)beta(4) integrins. Adhesion assays performed with fractionated basal keratinocytes showed that stem cells, transit amplifying cells and early differentiating cells all adhere to purified laminin-10/11 via these receptors. Further, laminin-10/11 provided a proliferative signal for neonatal foreskin keratinocytes, adult breast skin keratinocytes, and even a human papillomavirus type-18 transformed tumorigenic keratinocyte cell line in vitro. Finally, laminin-10/11 was shown to stimulate keratinocyte migration in an in vitro wound healing assay. These results provide strong evidence for a functional role for laminin-10/11 in epidermal proliferation during homeostasis, wound healing and neoplasia.

The role of CD4(+) cells in vivo on the induction of the immune response to Porphyromonas gingivalis in mice

Background: It has previously been suggested that CD4(+) T cells play a pivotal role in regulating the immune response to periodontal pathogens. The aim of the present study therefore was to determine delayed type hypersensitivity (DTH), spleen cell proliferation, serum and splenic anti-Porphyromonas gingivalis antibody levels, and lesion sizes following challenge with viable P. gingiualis in CD4-depleted BALB/c mice immunized with P. gingiualis outer membrane proteins (OMP). Methods: Four groups of BALB/c mice were used. Groups 1 and 2 were injected intraperitoneally (ip) with saline for 3 consecutive days and then weekly throughout the experiment. Groups 3 and 4 were injected ip with rat immunoglobulin and a monoclonal rat anti-mouse CD4 antibody, respectively. Two days later, group 1 mice were injected ip with saline only, while all the other groups were immunized ip with P. gingiualis OMP weekly for 3 weeks. One week later following the last immunization of OMP, 3 separate experiments were conducted to determine: 1) the DTH response to P. gingiualis OMP by measuring footpad swelling; 2) the levels of antibodies to P. gingiualis in serum samples and spleen cell cultures using an enzyme-linked immunosorbent assay, as well as spleen cell proliferation after stimulation with OMP; and 3) the lesion sizes after a subcutaneous challenge with viable P. gingiualis cells. Results: In CD4(+) T-cell-depleted mice (group 4), the DTH response and antigen-stimulated cell proliferation were significantly suppressed when compared to groups 2 and 3. Similarly, the levels of serum and splenic IgM, IgG, and all IgG subclass antibodies to P. gingiualis OMP were depressed. Delayed healing of P. gingivalis-induced lesions was also observed in the CD4(+) T-cell-depleted group. Conclusions: This study has shown that depletion of CD4(+) T cells prior to immunization with P. gingiualis OMP led to the suppression of both the humoral and cell-mediated immune response to this microorganism and that this was associated with delayed healing. These results suggest that the induction of the immune response to P. gingiualis is a CD4(+) T-cell-dependent mechanism and that CD4(+) T cells are important in the healing process.

Cooperative fishing interactions between Aboriginal Australians and dolphins in eastern Australia

Published eyewitness accounts and stories form Aboriginal Australians are used to provide an overview of the geographical extent and characteristics of cooperative fishing between Aboriginal Australians and dolphins in eastern Australia. These sources indicate that cooperative fishing was geographically widespread in eastern Australia, involved both bottlenose dolphins and orcas, and had a significance (emotional and Spiritual) to Aboriginal people beyond the acquisition of food. These fishing interactions represent both context and precedent for the economic and emotional-objectives of contemporary human-dolphin interactions such as dolphin provisioning. (C) 2002 International Society for Anthrozoology.

The morals of metaphysics: Kant's 'Groundwork' as intellectual paideia

To approach philosophy as a way of working on the self means to begin not with the experience it clarifies and the subject it discovers, but with the acts of self‐transformation it requires and the subjectivity it seeks to fashion. Commenting on the variety of spiritual exercises to be found in the ancient schools, Pierre Hadot remarks that: Some, like Plutarch’s ethismoi, designed to curb curiosity, anger or gossip, were only practices intended to ensure good moral habits. Others, particularly the meditations of the Platonic tradition, demanded a high degree of mental concentration. Some, like the contemplation of nature as practiced in all philosophical schools, turned the soul toward the cosmos, while still others—rare and exceptional—led to a transfiguration of the personality, as in the experiences of Plotinus. We also saw that the emotional tone and notional content of these exercises varied widely from one philosophical school to another: from the mobilization of energy and consent to destiny of the Stoics, to the relaxation and detachment of the Epicureans, to the mental concentration and renunciation of the sensible world among the Platonists.1 While successfully applied to ancient philosophy,2 this approach has not been widely exploited in the history of philosophy more broadly. There is, however, at least one study of medieval metaphysics in these terms,3 and there are some important discussions of early modern Stoicism and Epicureanism.4 And a recent study of Hume shows the fruitfulness of the approach for Enlightenment philosophy.5 It is all the more surprising then that there seems to have been no serious attempt to approach Kant’s moral philosophy in this way.

Erotic Literature in Nineteenth-Century Bali

Two nineteenth-century Balinese genres in which the erotic predominates are epic kakawin poetry and tutur (religious manuals) on sexual yoga. The article points to the strong intertextual links between these diverse genres. Through their focus on practical sexual matters and on the pursuit of sexual pleasure as integral to spiritual growth, tutur and kakawin also offer insight into notions of gender and sexuality in nineteenth-century Bali.

Cadherin-directed actin assembly: E-cadherin physically associates with the Arp2/3 complex to direct actin assembly in nascent adhesive contacts

Cadherin cell adhesion molecules are major determinants of tissue patterning which function in cooperation with the actin cytoskeleton [1-4]. In the context of stable adhesion [1], cadherin/catenin complexes are often envisaged to passively scaffold onto cortical actin filaments. However, cadherins also form dynamic adhesive contacts during wound healing and morphogenesis [2]. Here actin polymerization has been proposed to drive cell surfaces together [5], although F-actin reorganization also occurs as cell contacts mature [6]. The interaction between cadherins and actin is therefore likely to depend on the functional state of adhesion. We sought to analyze the relationship between cadherin homophilic binding and cytoskeletal activity during early cadherin adhesive contacts. Dissecting the specific effect of cadherin ligation alone on actin regulation is difficult in native cell-cell contacts, due to the range of juxtacrine signals that can arise when two cell surfaces adhere [7]. We therefore activated homophilic ligation using a specific functional recombinant protein. We report the first evidence that E-cadherin associates with the Arp2/3 complex actin nucleator and demonstrate that cadherin binding can exert an active, instructive influence on cells to mark sites for actin assembly at the cell surface.

Dental and skeletal pathology in lungfish jaws and tooth plates

Many lungfish of the tooth plated lineage, both fossil and living, are affected by dental and skeletal pathologies including dental caries, abscesses and cysts within the bone or tooth plate, osteopenia, bone hypertrophy, and malocclusion. These conditions, while influenced in part by structural relationships of soft and hard tissues in the tooth plates, jaw bones and surrounding oral tissues, can also be used as indicators of the kind of environment inhabited by the fish. The disease processes have specific structural consequences, related either to the pathology or to attempts to heal the damage, and usually alter the form and function of the tooth plate or bone. Consequently they can be distinguished from postmortem diagenetic or taphonomic effects, which alter the structure in less specific ways and show no sign of healing. Dental caries, the most common pathological condition in dipnoan dentitions, is recognisable in lungfish from the Devonian of Western Australia, the Tertiary of South Australia and the Northern Territory and from living lungfish in south east Queensland. Other pathologies have a more sporadic occurrence.

Tissue engineering for bone regeneration using differentiated alveolar bone cells in collagen scaffolds

Regeneration of osseous defects by a tissue-engineering approach provides a novel means of treatment utilizing cell biology, materials science, and molecular biology. In this study the concept of tissue engineering was tested with collagen type I matrices seeded with cells with osteogenic potential and implanted into sites where osseous damage had occurred. Explant cultures of cells from human alveolar bone and gingiva were established. When seeded into a three-dimensional type I collagen-based scaffold, the bone-derived cells maintained their osteoblastic phenotype as monitored by mRNA and protein levels of the bone-related proteins including bone sialoprotein, osteocalcin, osteopontin, bone morphogenetic proteins 2 and 4, and alkaline phosphatase. These in vitro-developed matrices were implanted into critical-size bone defects in skulls of immunodeficient (SCID) mice. Wound healing was monitored for up to 4 weeks. When measured by microdensitometry the bone density within defects filled with osteoblast-derived matrix was significantly higher compared with defects filled with either collagen scaffold alone or collagen scaffold impregnated with gingival fibroblasts. New bone formation was found at all the sites treated with the osteoblast-derived matrix at 28 days, whereas no obvious new bone formation was identified at the same time point in the control groups. In situ hybridization for the human-specific Alu gene sequence indicated that the newly formed bone tissue resulted from both transplanted human osteoblasts and endogenous mesenchymal stem cells. The results indicate that cells derived from human alveolar bone can be incorporated into bioengineered scaffolds and synthesize a matrix, which on implantation can induce new bone formation.

Benzedeiras de Maruípe: uma prática de cuidado humano em extinção

Estudo realizado com benzedeiras de uma área de saúde do município de Vitória - ES, objetivando identificá-las, conhecer suas histórias de vida e o interesse das mesmas em articularem-se com os profissionais das unidades básicas de saúde locais. Por se tratar de uma região marcada pela violência advinda do tráfico de drogas, tornou-se impossível identificar o universo dessas mulheres, face à impossibilidade de acesso a alguns desses bairros; assim posto, nossa amostra ficou limitada a cinco benzedeiras. A coleta de material do estudo se deu através de entrevistas e observações registradas em um diário de campo. O material transcrito e os apontamentos do diário de campo possibilitaram a narrativa de inspiração cartográfica deste estudo. Essas benzedeiras são mulheres entre 64 a 88 anos de idade, residem em locais inóspitos e em moradias humildes. Algumas benzem apenas crianças, outras todos aqueles que as procuram, inclusive para benzimento de seus animais. Nenhuma delas cobra e tão pouco aceita agradecimento pela atenção prestada, pois segundo elas, o agradecimento deve ser dirigido a Deus. São mulheres humildes, todas moradoras antigas da área, ora reconhecidas como importantes pelo dom que têm, ora rechaçadas como demoníacas por grupos religiosos. No tocante a uma aproximação com as equipes locais de saúde, todas as benzedeiras se mostraram avessas à ideia, no entendimento de que tal aproximação significaria uma demanda de benzimentos aumentada e obrigatória, o que contraria a lógica da atenção prestada pelas mesmas, que só benzem de acordo com a conveniência: sentindo-se bem, praticam o benzimento; estando desvitalizadas, evitam benzer. Por se tratar de mulheres idosas, as benzedeiras encontram-se ameaçadas de extinção, visto que aprender o oficio não tem sido objeto de interesse das novas gerações.

Angola, jeje e ketu : memórias e identidades em casas e nações de candomblé na Região Metropolitana da Grande Vitória (ES)

Angola, jeje e ketu: Memórias e identidades em casas e nações de candomblé na Região Metropolitana da Grande Vitória (ES), que é o tema desta dissertação, requer adentrar em análises de categorias nativas do povo de santo, e, em seguida, passar a questões teóricas sobre esses temas. Na cidade de Serra encontram-se as quatro casas de santo onde a pesquisa de campo foi realizada com três iyalorixás e um babalorixá, que dividem suas memórias e experiências religiosas compondo um exercício teórico sobre a história e a formação do candomblé no Espírito Santo. Nesse estado, que não é referência dessa religião, a mesma encontra-se em ascensão. A preocupação dos integrantes das comunidades de terreiros é transformar parte das tradições orais em produção escrita. Tendo em vista os processos políticos de reconhecimento legal da diversidade cultural, o debate se deu em torno de hibridização e múltiplas formas de identidade. O universo encantador e mágico do candomblé é composto pelos toques dos atabaques, danças rituais e fé em forças da natureza. Os “filhos da diáspora africana” trazidos para o Brasil eram de várias regiões da África, o que nos permite entender a diversidade cultural que marca esses grupos. Em função do sincretismo entre as próprias religiões de matriz africana e delas com o catolicismo e as doutrinas espiritualistas, essas religiões encontram-se de norte a sul do país. Este encontro de crenças e rituais é tão evidente que já não dizemos religiões “africanas” e sim religiões “afro-brasileiras”. O candomblé, desde o seu surgimento, vem sendo criado e recriado pela transmissão de suas tradições e ritos. A tradição oral nas comunidades de terreiro é um dos elementos demarcadores da construção da sua identidade, a partir de uma organização interna e do aprendizado hierarquicamente transmitido pelos depositários do saber, seguindo uma ordem de senioridade de iniciação, os antigos são detentores dos saberes e segredos. Por ser uma religião iniciática, o aprendizado ocorre permanentemente, em especial o da língua ritual, onde o exercício e o contato levam a transmissão cultural. O povo de santo reconstrói uma ligação com uma comunidade imaginada que remonta a África e desenvolve relações de parentesco ficcional entre os membros das comunidades de terreiro e forma uma família de santo e de axé.

Para uma sócio-história da língua Guarani no Espírito Santo : uma análise sob a perspectiva sociolinguística

Estima-se que restam hoje cerca de 50 mil índios Guarani no Brasil, que se situam, principalmente, na faixa litorânea que vai desde os estados do sul até o território capixaba, o Espírito Santo. Considerando que essa comunidade se mantém bilíngue, o presente trabalho objetiva discutir se, na situação de contato entre o Guarani e o Português, a primeira língua está ou não cedendo lugar à segunda. Para alcançar esse objetivo, foi formado um banco de dados de fala por meio de entrevistas realizadas nas aldeias, que versaram sobre as tradições históricas, a família, a religião, a economia e o meio ambiente – aspectos considerados por eles como as principais armas de resistência desse povo. A análise tomou por base os pressupostos da Sociolinguística/Contato Linguístico, com teóricos como Weinreich (1953), Fishman (1968; 1972), Appel e Muysken (1996), Coulmas (2005) e outros, que discutem temas pertinentes à pesquisa em questão: o contato linguístico e a manutenção/substituição de línguas minoritárias. Acredita-se que, apesar do contato com o português pela venda de artesanatos, pela mídia e pela atuação da escola e sua ação integralizadora, prevista pelo Estatuto do Índio, o Guarani mantém a sua língua materna - ainda que estigmatizada - devido à forte religiosidade que norteia todo o seu modo de vida. Ele entende a palavra como um dom e confere a ela um poder mítico de conexão com o mundo espiritual, o que, ao mesmo tempo, confere extrema importância à língua minoritária e favorece a sua preservação, enquanto marca importante da cultura e identidade desse povo.

Particulate bioglass in the regeneration of alveolar bone in dogs: clinical, surgical and radiographic evaluations

Bone loss, either by trauma or other diseases, generates an increasing need for substitutes of this tissue. This study evaluated Bioglass as a bone substitute in the regeneration of the alveolar bone in mandibles of dogs by clinical, surgical and radiological analysis. Twenty-eight adult dogs were randomly separated into two equal groups. In each animal, a bone defect was created on the vestibular surface of the alveolar bone between the roots of the fourth right premolar tooth. In the treated group, the defect was immediately filled with bioglass, while in the control, it remained unfilled. Clinical evaluations were performed daily for a week, as well as x-rays immediately after surgery and at 8, 14, 21, 42, 60, 90 and 120 days post-operative. Most animals in both groups showed no signs of inflammation and wound healing was similar. Radiographic examination revealed a gradual increase of radiopacity in the region of the defect in the control group. In the treated group, initial radiopacity was higher than that of adjacent bone, decreasing until 21 days after surgery. Then it gradually increased until 120 days after surgery, when the defect became undetectable. The results showed that Bioglass integrates into bone tissue, is biocompatible and reduced the period for complete bone regeneration.