Novel Murine Infection Models Provide Deep Insights into the “Ménage à Trois” of Campylobacter jejuni, Microbiota and Host Innate Immunity

BACKGROUND: Although Campylobacter jejuni-infections have a high prevalence worldwide and represent a significant socioeconomic burden, it is still not well understood how C. jejuni causes intestinal inflammation. Detailed investigation of C. jejuni-mediated intestinal immunopathology is hampered by the lack of appropriate vertebrate models. In particular, mice display colonization resistance against this pathogen. METHODOLOGY/PRINCIPAL FINDINGS: To overcome these limitations we developed a novel C. jejuni-infection model using gnotobiotic mice in which the intestinal flora was eradicated by antibiotic treatment. These animals could then be permanently associated with a complete human (hfa) or murine (mfa) microbiota. After peroral infection C. jejuni colonized the gastrointestinal tract of gnotobiotic and hfa mice for six weeks, whereas mfa mice cleared the pathogen within two days. Strikingly, stable C. jejuni colonization was accompanied by a pro-inflammatory immune response indicated by increased numbers of T- and B-lymphocytes, regulatory T-cells, neutrophils and apoptotic cells, as well as increased concentrations of TNF-α, IL-6, and MCP-1 in the colon mucosa of hfa mice. Analysis of MyD88(-/-), TRIF(-/-), TLR4(-/-), and TLR9(-/-) mice revealed that TLR4- and TLR9-signaling was essential for immunopathology following C. jejuni-infection. Interestingly, C. jejuni-mutant strains deficient in formic acid metabolism and perception induced less intestinal immunopathology compared to the parental strain infection. In summary, the murine gut flora is essential for colonization resistance against C. jejuni and can be overcome by reconstitution of gnotobiotic mice with human flora. Detection of C. jejuni-LPS and -CpG-DNA by host TLR4 and TLR9, respectively, plays a key role in immunopathology. Finally, the host immune response is tightly coupled to bacterial formic acid metabolism and invasion fitness. CONCLUSION/SIGNIFICANCE: We conclude that gnotobiotic and "humanized" mice represent excellent novel C. jejuni-infection and -inflammation models and provide deep insights into the immunological and molecular interplays between C. jejuni, microbiota and innate immunity in human campylobacteriosis

Ação do cepa e do ácido giberélico na frutificação da videira 'niagara rosada'

Studies were carried out to establish the effects of exogenous growth regulators on Vitis (labrusca x vinifera) 'Niagara Rosada' fruiting. The investigations were done in the Jundiaí Research Station, Agronomic Institute State of São Paulo, always using disease-free vineyards of good productivity. The morphological transformations of clusters were carried out under the following aspects: weight, length and width of cluster; number of berries; weight, length average and width average of berries; length average/width average ratio of berries; number of seeds; length and diameter of secondary rachis. That characteristics were determined at the time of maturity plus the total sugars, total acid, Maturity Index and reducing sugars in samples of all treatments. The experiment were conduced in order to determine the doses that resulted in the most beneficial effects, always using applications by immersion of the inflorescence. The experiment consisted of appplications of (2-chloroethyl) phosphonic acid (CEPA) at concentrations of 50, 100, 250, 500, 1,000 and 2,000 ppm, 14 days before flowering; treatments with gibberellic acid at concentrations of 100 and 200 ppm before full bloom, 10 days after full bloom, and both before plus after full bloom. Treatment with CEPA 100 ppm plus gibberellic acid 100 ppm before full bloom and check treatment were also used. The use of CEPA before flowering at the concentrations used, did not result in good results in 'Niagara Rosada' clusters; applications of gibberellic acid did not differ significantly from the nontreated vines under the conditions studied.Estudou-se o efeito da aplicação, por imersão, do CEPA (ácido 2-cloroetil fosfônico) e do ácido giberélico, 14 dias antes do florescimento, nas características morfológicas da panícula da videira Vitis (labrus-ca x vinifera) "Niagara Rosada". Alguns tratamentos com ácido giberélico foram concluídos com nova aplicação 10 dias após o florescimento. Neste experimento verificou-se que, aplicação do CEPA na concentração de 250 ppm resultou na formação de panículas com a maioria de características indesejáveis. o tratamento misto CEPA 100 ppm + ácido giberélico 100 ppm também promoveu o aparecimento de panículas subdesenvolvidas. Aplicação de ácido giberélico na concentração de 100 ppm em pré e pós-ílorescimento, resultou médias mais elevadas, com relação ao peso da panícula, comprimento da panícula, peso das bagas e comprimento da ráquis. Ácido giberélico na concentração de 100 ppm aplicado em pós-ílorescimento, promoveu uma tendência de aumento nas médias do tratamento quanto ao comprimento médio das bagas, largura média das bagas, largura do engaço e comprimento da ráquila. Devemos considerar porém, que os resultados obtidos não apresentaram diferenças significativas com relação ao controle, quanto às características das frutificações, nas condições de estudo