Proteomic comparisons of opaque and transparent variants of <i>Streptococcus pneumoniae</i> by two dimensional-differential gel electrophoresis

Streptococcus pneumoniae (the pneumococcus) is a human pathogen, accounting for massive global morbidity and mortality. Although asymptomatic colonization of the nasopharynx almost invariably precedes disease, the critical determinants enabling pneumococcal progression from this niche to cause invasive disease are poorly understood. One mechanism proposed to be central to this transition involves opacity phase variation, whereby pneumococci harvested from the nasopharynx are typically transparent, while those simultaneously harvested from the blood are opaque. Here, we used two dimensional-differential gel electrophoresis (2D-DIGE) to compare protein expression profiles of transparent and opaque variants of 3 pneumococcal strains, D39 (serotype 2), WCH43 (serotype 4) and WCH16 (serotype 6A) in vitro. One spot comprising a mixture of capsular polysaccharide biosynthesis protein and other proteins was significantly up-regulated in the opaque phenotype in all 3 strains; other proteins were differentially regulated in a strain-specific manner. We conclude that pneumococcal phase variation is a complex and multifactorial process leading to strain-specific pathogenicity.Melissa H. Chai, Florian Weiland, Richard M. Harvey, Peter Hoffmann, Abiodun D. Ogunniyi, James C. Pato

Nasopharyngeal Colonization and Invasive Disease Are Enhanced by the Cell Wall Hydrolases LytB and LytC of Streptococcus pneumoniae

Background: Streptococcus pneumoniae is a common colonizer of the human nasopharynx and one of the major pathogens causing invasive disease worldwide. Dissection of the molecular pathways responsible for colonization, invasion, and evasion of the immune system will provide new targets for antimicrobial or vaccine therapies for this common pathogen. Methodology/Principal Findings: We have constructed mutants lacking the pneumococcal cell wall hydrolases (CWHs) LytB and LytC to investigate the role of these proteins in different phases of the pneumococcal pathogenesis. Our results show that LytB and LytC are involved in the attachment of S. pneumoniae to human nasopharyngeal cells both in vitro and in vivo. The interaction of both proteins with phagocytic cells demonstrated that LytB and LytC act in concert avoiding pneumococcal phagocytosis mediated by neutrophils and alveolar macrophages. Furthermore, C3b deposition was increased on the lytC mutant confirming that LytC is involved in complement evasion. As a result, the lytC mutant showed a reduced ability to successfully cause pneumococcal pneumonia and sepsis. Bacterial mutants lacking both LytB and LytC showed a dramatically impaired attachment to nasopharyngeal cells as well as a marked degree of attenuation in a mouse model of colonization. In addition, C3b deposition and phagocytosis was more efficient for the double lytB lytC mutant and its virulence was greatly impaired in both systemic and pulmonary models of infection. Conclusions/Significance: This study confirms that the CWHs LytB and LytC of S. pneumoniae are essential virulence factor

Bowel management for the treatment of pediatric fecal incontinence

Fecal incontinence is a devastating underestimated problem, affecting a large number of individuals all over the world. Most of the available literature relates to the management of adults. The treatments proposed are not uniformly successful and have little application in the pediatric population. This paper presents the experience of 30 years, implementing a bowel management program, for the treatment of fecal incontinence in over 700 pediatric patients, with a success rate of 95%. The main characteristics of the program include the identification of the characteristics of the colon of each patient; finding the specific type of enema that will clean that colon and the radiological monitoring of the process

Epithelial-immune cell interplay in primary Sjogren syndrome salivary gland pathogenesis

In primary Sjogren syndrome (pSS), the function of the salivary glands is often considerably reduced. Multiple innate immune pathways are likely dysregulated in the salivary gland epithelium in pSS, including the nuclear factor-kappa B pathway, the inflammasome and interferon signalling. The ductal cells of the salivary gland in pSS are characteristically surrounded by a CD4(+) T cell-rich and B cell-rich infiltrate, implying a degree of communication between epithelial cells and immune cells. B cell infiltrates within the ducts can initiate the development of lymphoepithelial lesions, including basal ductal cell hyperplasia. Vice versa, the epithelium provides chronic activation signals to the glandular B cell fraction. This continuous stimulation might ultimately drive the development of mucosa-associated lymphoid tissue lymphoma. This Review discusses changes in the cells of the salivary gland epithelium in pSS (including acinar, ductal and progenitor cells), and the proposed interplay of these cells with environmental stimuli and the immune system. Current therapeutic options are insufficient to address both lymphocytic infiltration and salivary gland dysfunction. Successful rescue of salivary gland function in pSS will probably demand a multimodal therapeutic approach and an appreciation of the complicity of the salivary gland epithelium in the development of pSS. Salivary gland dysfunction is an important characteristic of primary Sjogren syndrome (pSS). In this Review, the authors discuss various epithelial abnormalities in pSS and the mechanisms by which epithelial cell-immune cell interactions contribute to disease development and progression

Detecting Acute Otitis Media Symptom Episodes Using a Mobile App : Cohort Study

BACKGROUND: Population cohort studies are useful to study infectious diseases episodes not attended by health care services, but conventional paper diaries and questionnaires to capture cases are prone to noncompliance and recall bias. Use of smart technology in this setting may improve case finding. OBJECTIVE: The objective of our study was to validate an interactive mobile app for monitoring occurrence of acute infectious diseases episodes in individuals, independent of health care seeking, using acute otitis media (AOM) symptom episodes in infants as a case study. We were interested in determining participant compliance and app performance in detecting and ascertaining (parent-reported) AOM symptom episodes with this novel tool compared with traditional methods used for monitoring study participants. METHODS: We tested the InfectieApp research app to detect AOM symptom episodes. In 2013, we followed 155 children aged 0 to 3 years for 4 months. Parents recorded the presence of AOM symptoms in a paper diary for 4 consecutive months and completed additional disease questionnaires when AOM symptoms were present. In 2015 in a similar cohort of 69 children, parents used an AOM diary and questionnaire app instead. RESULTS: During conventional and app-based recording, 93.13% (17,244/18,516) and 94.56% (7438/7866) of symptom diaries were returned, respectively, and at least one symptom was recorded for 32.50% (n=5606) and 43.99% (n=3272) of diary days (P<.01). The incidence of AOM symptom episodes was 605 and 835 per 1000 child-years, respectively. Disease questionnaires were completed for 59% (17/29) of episodes when participants were using conventional recording, compared with 100% (18/18) for app-based recording. CONCLUSIONS: The use of the study's smart diary app improved AOM case finding and disease questionnaire completeness. For common infectious diseases that often remain undetected by health care services, use of this technology can substantially improve the accurateness of disease burden estimates

Detecting Acute Otitis Media Symptom Episodes Using a Mobile App : Cohort Study

BACKGROUND: Population cohort studies are useful to study infectious diseases episodes not attended by health care services, but conventional paper diaries and questionnaires to capture cases are prone to noncompliance and recall bias. Use of smart technology in this setting may improve case finding. OBJECTIVE: The objective of our study was to validate an interactive mobile app for monitoring occurrence of acute infectious diseases episodes in individuals, independent of health care seeking, using acute otitis media (AOM) symptom episodes in infants as a case study. We were interested in determining participant compliance and app performance in detecting and ascertaining (parent-reported) AOM symptom episodes with this novel tool compared with traditional methods used for monitoring study participants. METHODS: We tested the InfectieApp research app to detect AOM symptom episodes. In 2013, we followed 155 children aged 0 to 3 years for 4 months. Parents recorded the presence of AOM symptoms in a paper diary for 4 consecutive months and completed additional disease questionnaires when AOM symptoms were present. In 2015 in a similar cohort of 69 children, parents used an AOM diary and questionnaire app instead. RESULTS: During conventional and app-based recording, 93.13% (17,244/18,516) and 94.56% (7438/7866) of symptom diaries were returned, respectively, and at least one symptom was recorded for 32.50% (n=5606) and 43.99% (n=3272) of diary days (P<.01). The incidence of AOM symptom episodes was 605 and 835 per 1000 child-years, respectively. Disease questionnaires were completed for 59% (17/29) of episodes when participants were using conventional recording, compared with 100% (18/18) for app-based recording. CONCLUSIONS: The use of the study's smart diary app improved AOM case finding and disease questionnaire completeness. For common infectious diseases that often remain undetected by health care services, use of this technology can substantially improve the accurateness of disease burden estimates