Pesquisas sobre a imunidade da Framboesia tropica no homem: observações feitas em 33 superinoculações e 7 reinoculações

Como resultado das 40 experiências relatadas neste trabalho, as seguintes conclusões podem ser tiradas: 1) Dos 2 até aos 8 meses de molestia há na bouba uma grande resistência á superinoculação (13 experiências negativas em 15). Nas 2 experiências positivas, foram obtidas lesões atípicas (pianides): a) Tal resistência parece independer da presença de lesões boubaticas cutaneas e se fez sentir mesmo em casos com a lesão inicial exclusiva. b) Dentro desse período de molestia, tal resistência pode desaparecer com o tratamento: 2 doentes tratados aos 6 meses e reinoculados adquiriram bouba em tempo normal. Porém, um caso tratado aos 7 meses e reinoculado, desenvolveu uma pianide. Êste fato parece mostrar que essa resistência depende principalmente da presença da infecção ativa ou latente, raramente traduzindo uma imunidade no verdadeiro sentido do têrmo. c) No caso de emprego de homo-virus, essa resistência prolongou-se até um ano d molestia, havendo apenas um caso duvidoso em 10 inoculações. 2) Do 10º mês ao 4º ano de molestia, em 8 superinoculações observou-se uma resistência parcial que se traduziu de 2 modos, quanto à natureza da lesão atípica que se obteve no ponto inoculado; "lesão frustra papulo-eritematosa" (7 meses); e lesão semelhante ás "pianides" da infecção natural (1 vez), a qual permaneceu localizada sem manifestações metastaticas, até 4 meses de observação. a) Também este estado de resistência parcial, parece independer da presença de lesões boubaticas aparentes. b) Com o tratamento esse estado parece não se modificar: um doente tratado nesse período da molestia, reagiu à inoculação de modo semelhante, embora sem nenhuma manifestação clínica e com a R. Wa. negativa. Inegavelmente, essas "lesões frustras" e também as "pianides" obtidas, representam respostas de organismos que obtiveram vantagens na luta com a doença, possuindo um certo grau de imunidade, que é muitíssimo maior no caso das primeiras. 3) Depois do 5º ano de molestia, a resposta á superinoculação traduziu um estado de maior sensibilidade do organismo infectado. Observou-se no ponto inoculado uma reação precoce papulo-eritemato-ulcerosa, francamente necrotica e destrutiva, ao mesmo tempo que se verificou exacerbação das lesões dos pacientes, com grande infartamento gangliomar satélite. Até 18 meses depois, em um caso, a lesão obtida permaneceu localizada sem manifestações generalizadas. a) O tratamento não modificou tal estado. Doentes tratados ( e parcial ou totalmente curados), reagiram da mesma maneira à reinoculação dentro desse período da molestia. Apenas em um caso de mais de 5 anos (nº 40) não se obteve a lesão ulcero-necrotica. Era o único dos 13 experimentados que não tinha nem tivera lesões ulcero-gomoides destrutivas. por outro lado, 2 pacientes apresentando lesões gomo-ulcerativas, mas tendo menos de 3 anos de molestia, não deram a lesão ulcerativo-necrotica em resposta à superinoculação. (Experiências ns. 38 e 39). Interessante é que esta lesão ulcerativo-necrotica contém treponemas embora raros, e em evolução pode tomar o carater das lesões destrutivas gomo-ulcerativas peculiares ao chamado "período terciario" da doença, com as quais também se assemelha histopatológicamente. Sob o ponto de vista imunológico, esta lesão representa um estado de maior sensibilidade do organismo para o agente infeccioso. 4) Como o tratamento precoce perturba o desenvolvimento da imunidade, sob o ponto de vista epidemiológico, seria aconselhável aguardar o período terminal do chamado secundarismo, isto é da fase de generalização boubatica, para tratar os pacientes em Postos, hospitais ou ambulatórios pois, tais doentes poderiam se reinfestar uma vez retornados ao fóco. Claro que em campanhas terapeuticas profilaticas, as lesões "abertas" primo-secundarias, devem ser rapidamente eliminadas uma vez que são as mais contagiantes, por mais ricas em germes. 5) Existe na framboesia trópica uma verdadeira imunidade além de uma simples resistência á superinoculação devido a presença da infecção ativa ou latente. Com efeito, pacientes tratados em determinado período da molestia e curados clinica e sorologicamente, mostraram resistência parcial á reinoculação, reagindo de modo semelhante a outros do mesmo período de molestia e não tratados. 6) A imunidade na framboesia tropica se manifesta seja como uma resistência á superinoculação ou reinoculação seja como uma modificação da lesão boubatica inicial, seja, finalmente, como uma resistência á generalização da doença. 7) Os resultados das esperiências sugerem que as diferentes manifestações cutaneas da molestia são condicionadas até certo ponto pelo estado imunitario do organismo infectado. 8) Os diferentes gráus de imunidade, encontrados na framboesia trópica, estão até certo ponto relacionados com o tempo de doença. Porém, são atingidos mais ou menos ràpidamente, segundo o organismo infectado e, talvez segundo a virulência do treponema, do mesmo modo como os chamados "secundarismo" e 'terciarismo" da doença.

Prevalence of Clostridium spp. and Clostridium difficile in children with acute diarrhea in São Paulo city, Brazil

Species of Clostridium are widely distributed in the environment, inhabiting both human and animal gastrointestinal tracts. Clostridium difficile is an important pathogen associated with outbreaks of pseudomembranous colitis and other intestinal disorders, such as diarrhea. In this study, the prevalence of Clostridium spp. and C. difficile, from hospitalized children with acute diarrhea, was examined. These children were admitted to 3 different hospitals for over 12 months. Eighteen (20%) and 19 (21%) stool specimens from children with (90) and without (91) diarrhea respectively, were positive to clostridia. Only 10 C. difficile strains were detected in 5.5% of the stool samples of children with diarrhea. None healthy children (without diarrhea) harbored C. difficile. From these 10 C. difficile, 9 were considered as toxigenic and genotyped as tcdA+/tcdB+ or tcdA-/tcdB+, and 1 strain as nontoxigenic (tcdA-/tdcB-). They were detected by the citotoxicity on VERO cells and by the multiplex-polymerase chain reaction. Thirty clinical fecal extracts produced minor alterations on VERO cells. The presence of C. difficile as a probable agent of acute diarrhea is suggested in several countries, but in this study, the presence of these organisms was not significant. More studies will be necessary to evaluate the role of clostridia or C. difficile in diarrhoeal processes in children.

Neutrophil transepithelial migration: role of toll-like receptors in mucosal inflammation

The symptomatic phases of many inflammatory diseases are characterized by migration of large numbers of neutrophils (PMN) across a polarized epithelium and accumulation within a lumen. For example, acute PMN influx is common in diseases of the gastrointestinal system (ulcerative colitis, Crohn's disease, bacterial enterocolitis, gastritis), hepatobiliary system (cholangitis, acute cholecystitis), respiratory tract (bronchial pneumonia, bronchitis, cystic fibrosis, bronchiectasis), and urinary tract (pyelonephritis, cystitis). Despite these observations, the molecular basis of leukocyte interactions with epithelial cells is incompletely understood. In vitro models of PMN transepithelial migration typically use N-formylated bacterial peptides such as fMLP in isolation to drive human PMNs across epithelial monolayers. However, other microbial products such as lipopolysaccharide (LPS) are major constituents of the intestinal lumen and have potent effects on the immune system. In the absence of LPS, we have shown that transepithelial migration requires sequential adhesive interactions between the PMN beta2 integrin CD11b/CD18 and JAM protein family members. Other epithelial ligands appear to be abundantly represented as fucosylated proteoglycans. Further studies indicate that the rate of PMN migration across mucosal surfaces can be regulated by the ubiquitously expressed transmembrane protein CD47 and microbial-derived factors, although many of the details remain unclear. Current data suggests that Toll-like receptors (TLR), which recognize specific pathogen-associated molecular patterns (PAMPs), are differentially expressed on both leukocytes and mucosal epithelial cells while serving to modulate leukocyte-epithelial interactions. Exposure of epithelial TLRs to microbial ligands has been shown to result in transcriptional upregulation of inflammatory mediators whereas ligation of leukocyte TLRs modulate specific antimicrobial responses. A better understanding of these events will hopefully provide new insights into the mechanisms of epithelial responses to microorganisms and ideas for therapies aimed at inhibiting the deleterious consequences of mucosal inflammation.

Epithelial cell signaling responses to enterohemorrhagic Escherichia coli infection

Enterohemorrhagic Escherichia coli, including the serotype O157:H7 that is most commonly identified with human disease, cause both sporadic cases and outbreaks of non-bloody diarrhea and hemorrhagic colitis. In about 10% of infected subjects, the hemolytic uremic syndrome (hemolytic anemic, thrombocytopenia, and acute renal failure) develops, likely as a consequence of systemic spread of bacterial-derived toxins variously referred to as Shiga-like toxin, Shiga toxin, and Verotoxin. Increasing evidence points to a complex interplay between bacterial products - for example, adhesins and toxins - and host signal transduction pathways in mediating responses to infection. Identification of critical signaling pathways could result in the development of novel strategies for intervention to both prevent and treat this microbial infection in humans.

Epithelial effects of proteinase-activated receptors in the gastrointestinal tract

The intestinal epithelium plays a crucial role in providing a barrier between the external environment and the internal milieu of the body. A compromised mucosal barrier is characteristic of mucosal inflammation and is a key determinant of the development of intestinal diseases such as Crohn's disease and ulcerative colitis. The intestinal epithelium is regularly exposed to serine proteinases and this exposure is enhanced in numerous disease states. Thus, it is important to understand how proteinase-activated receptors (PARs), which are activated by serine proteinases, can affect intestinal epithelial function. This review surveys the data which demonstrate the wide distribution of PARs, particularly PAR-1 and PAR-2, in the gastrointestinal tract and accessory organs, focusing on the epithelium and those cells which communicate with the epithelium to affect its function. PARs have a role in regulating secretion by epithelia of the salivary glands, stomach, pancreas and intestine. In addition, PARs located on subepithelial nerves, fibroblasts and mast cells have important implications for epithelial function. Recent data outline the importance of the cellular site of PAR expression, as PARs expressed on epithelia may have effects that are countered by PARs expressed on other cell types. Finally, PARs and their ability to promote epithelial cell proliferation are discussed in terms of colon cancer.

Characterization of Shigella spp. by antimicrobial resistance and PCR detection of ipa genes in an infantile population from Porto Velho (Western Amazon region), Brazil

The incidence of Shigella spp. was assessed in 877 infants from the public hospital in Rondônia (Western Amazon region, Brazil) where Shigella represents the fourth cause of diarrhea. Twenty-five isolates were identified: 18 were Shigella flexneri, three Shigella sonnei, three Shigella boydii and one Shigella dysenteriae. With the exception of S. dysenteriae, all Shigella spp. isolated from children with diarrhea acquired multiple antibiotic resistances. PCR detection of ipa virulence genes and invasion assays of bloody diarrhea and fever (colitis) were compared among 25 patients testing positive for Shigella. The ipaH and ipaBCD genes were detected in almost all isolates and, unsurprisingly, all Shigella isolates associated with colitis were able to invade HeLa cells. This work alerts for multiple antibiotic resistant Shigella in the region and characterizes presence of ipa virulence genes and invasion phenotypesin dysenteric shigellosis.

Trichuris trichiura in a post-Colonial Brazilian mummy

Trichuris trichiura is a soil-transmitted helminth which is prevalent in warm, moist, tropical and subtropical regions of the world with poor sanitation. Heavy whipworm can result either in Trichuris dysenteric syndrome - especially in children - or in a chronic colitis. In heavy infections, worms can spread proximally and may cause ileitis. Here we provide first microscopic evidence for a T. trichiura adult worm embedded in the rectum of a post-Colonial Brazilian adult mummy. During Colonial and post-Colonial times, many European chroniclers described a parasitic disease named Maculo whose symptomatology coincides with heavy helminthiasis. Based on our findings and on comparison of ancient textual evidence with modern description of heavy whipworm, we feel confident in considering that the two syndromes are expressions of the same pathological condition.

Clostridium difficile ribotypes in humans and animals in Brazil

Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies aboutC. difficile polymerase chain reaction (PCR) ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficileribotypes isolated from humans and animals in Brazil. Seventy-six C. difficile strains isolated from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf were distributed into 24 different PCR ribotypes. Among toxigenic strains, PCR ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4%) and eight (10.5%) samples, respectively. Fourteen different PCR ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.

Tratamento cirúrgico do linfoma gástrico primário

OBJETIVO: o objetivo deste estudo retrospectivo foi analisar os resultados de 25 doentes com linfoma gástrico primário operados com intenção curativa. MÉTODO: os dados foram obtidos pela revisão dos prontuários e contato com os doentes ou familiares. A doença foi estadiada pelo sistema Ann Arbor modificado por Musshoff e Schmidt-Vollmer e a classificação histológica utilizada foi o sistema de Kiel. O esquema de radioterapia utilizado foi o CHOP e a radioterapia aplicada foi de 2000 a 4000 cGy. RESULTADOS: os sintomas e sinais clínicos assemelhavam-se aos da doença péptica ulcerosa ou do carcinoma gástrico Obteve-se o diagnóstico pré-operatório pela biópsia endoscópica em três casos e a exploração cirúrgica foi necessária para o diagnóstico nos restantes. No pré-operatório, sete doentes (30,4%) foram submetidos ao mielograma, que foi normal. Todos os pacientes foram submetidos à ressecção (12 gastrectomias subtotais e 13 gastrectomias totais) com retirada dos linfonodos regionais. Dez doentes (40%) receberam tratamento complementar (quimioterapia e/ou radioterapia). O estadiamento foi significativamente mais avançado nas lesões fundocárdicas e nos mais idosos e a sobrevida média foi de 31,5 meses. CONCLUSÕES: nesta série, as variáveis que influenciaram significativamente os índices de sobrevida foram a idade e o estádio avançados, o tamanho da lesão maior que 6,0cm e a realização do tratamento adjuvante pós-operatório (p< 0,05). Estes resultados sugerem que a ressecção completa da lesão com linfonodos adjacentes, acompanhada de tratamento adjuvante, constitui a melhor abordagem do linfoma gástrico primário ressecável.

Enterohemorrhagic Escherichia coli O157: H7 from healthy dairy cattle in Mid-West Brazil: occurrence and molecular characterization

Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 represents the major Shiga toxin-producing E. coli (STEC) strain related to large outbreaks and severe diseases such as hemorrhagic colitis (HC) and the potentially lethal hemolytic uremic syndrome (HUS). The aim of this study was to report the occurrence and molecular characterization of O157:H7 isolates obtained by rectal swab from 52 healthy dairy cattle belonging to 21 farms in Mid-West of Brazil. Detection of 16SrRNA, stx1, stx2, rfbO157, fliCh7, eae, ehxA, saa, cnf1, chuA, yjaA and TSPE4.C2 genes was performed by PCR. The isolates were further characterized by serotyping. Two hundred and sixty E. coli isolates were obtained, of which 126 were characterized as STEC. Two isolates from the same cow were identified as serotype O157:H7. Both isolates presented the stx2, eae, ehxA, saa and cnf1 virulence factor genes and the chuA gene in the phylogenetic classification (virulent group D), suggesting that they were clones. The prevalence of O157:H7 was found to be 1.92% (1/52 animals), demonstrating that healthy dairy cattle from farms in the Mid-West of Brazil are an important reservoir for highly pathogenic E. coli O157:H7.

The Shiga toxin 2 B subunit inhibits net fluid absorption in human colon and elicits fluid accumulation in rat colon loops

Shiga toxin (Stx)-producing Escherichia coli (STEC) colonizes the large intestine causing a spectrum of disorders, including watery diarrhea, bloody diarrhea (hemorrhagic colitis), and hemolytic-uremic syndrome. It is estimated that hemolytic-uremic syndrome is the most common cause of acute renal failure in infants in Argentina. Stx is a multimeric toxin composed of one A subunit and five B subunits. In this study we demonstrate that the Stx2 B subunit inhibits the water absorption (Jw) across the human and rat colonic mucosa without altering the electrical parameters measured as transepithelial potential difference and short circuit current. The time-course Jw inhibition by 400 ng/ml purified Stx2 B subunit was similar to that obtained using 12 ng/ml Stx2 holotoxin suggesting that both, A and B subunits of Stx2 contributed to inhibit the Jw. Moreover, non-hemorrhagic fluid accumulation was observed in rat colon loops after 16 h of treatment with 3 and 30 ng/ml Stx2 B subunit. These changes indicate that Stx2 B subunit induces fluid accumulation independently of A subunit activity by altering the usual balance of intestinal absorption and secretion toward net secretion. In conclusion, our results suggest that the Stx2 B subunit, which is non-toxic for Vero cells, may contribute to the watery diarrhea observed in STEC infection. Further studies will be necessary to determine whether the toxicity of Stx2 B subunit may have pathogenic consequences when it is used as a component in an acellular STEC vaccine or as a vector in cancer vaccines.

Association among genetic predisposition, gut microbiota, and host immune response in the etiopathogenesis of inflammatory bowel disease

Inflammatory bowel disease (IBD), which includes Crohn's disease (CD) and ulcerative colitis (UC), is a chronic disorder that affects thousands of people around the world. These diseases are characterized by exacerbated uncontrolled intestinal inflammation that leads to poor quality of life in affected patients. Although the exact cause of IBD still remains unknown, compelling evidence suggests that the interplay among immune deregulation, environmental factors, and genetic polymorphisms contributes to the multifactorial nature of the disease. Therefore, in this review we present classical and novel findings regarding IBD etiopathogenesis. Considering the genetic causes of the diseases, alterations in about 100 genes or allelic variants, most of them in components of the immune system, have been related to IBD susceptibility. Dysbiosis of the intestinal microbiota also plays a role in the initiation or perpetuation of gut inflammation, which develops under altered or impaired immune responses. In this context, unbalanced innate and especially adaptive immunity has been considered one of the major contributing factors to IBD development, with the involvement of the Th1, Th2, and Th17 effector population in addition to impaired regulatory responses in CD or UC. Finally, an understanding of the interplay among pathogenic triggers of IBD will improve knowledge about the immunological mechanisms of gut inflammation, thus providing novel tools for IBD control.

Chronic granulomatous disease: why an inflammatory disease?

Chronic granulomatous disease is a primary immunodeficiency caused by mutations in the genes encoding subunits of the phagocytic NADPH oxidase system. Patients can present with severe, recurrent infections and noninfectious conditions. Among the latter, inflammatory manifestations are predominant, especially granulomas and colitis. In this article, we systematically review the possible mechanisms of hyperinflammation in this rare primary immunodeficiency condition and their correlations with clinical aspects.

Classical and recent advances in the treatment of inflammatory bowel diseases

Crohn's disease (CD) and ulcerative colitis (UC) are intestinal disorders that comprise the inflammatory bowel diseases (IBD). These disorders have a significant effect on the quality of life of affected patients and the increasing number of IBD cases worldwide is a growing concern. Because of the overall burden of IBD and its multifactorial etiology, efforts have been made to improve the medical management of these inflammatory conditions. The classical therapeutic strategies aim to control the exacerbated host immune response with aminosalicylates, antibiotics, corticosteroids, thiopurines, methotrexate and anti-tumor necrosis factor (TNF) biological agents. Although successful in the treatment of several CD or UC conditions, these drugs have limited effectiveness, and variable responses may culminate in unpredictable outcomes. The ideal therapy should reduce inflammation without inducing immunosuppression, and remains a challenge to health care personnel. Recently, a number of additional approaches to IBD therapy, such as new target molecules for biological agents and cellular therapy, have shown promising results. A deeper understanding of IBD pathogenesis and the availability of novel therapies are needed to improve therapeutic success. This review describes the overall key features of therapies currently employed in clinical practice as well as novel and future alternative IBD treatment methods.