Role of Intestinal Bacteria in Gliadin-Induced Changes in Intestinal Mucosa: Study in Germ-Free Rats

10 pages, 6 figures.[Background and Aims]: Celiac disease (CD) is a chronic inflammatory disorder of the small intestine that is induced by dietary wheat gluten proteins (gliadins) in genetically predisposed individuals. The overgrowth of potentially pathogenic bacteria and infections has been suggested to contribute to CD pathogenesis. We aimed to study the effects of gliadin and various intestinal bacterial strains on mucosal barrier integrity, gliadin translocation, and cytokine production.[Methodology/Principal Findings]: Changes in gut mucosa were assessed in the intestinal loops of inbred Wistar-AVN rats that were reared under germ-free conditions in the presence of various intestinal bacteria (enterobacteria and bifidobacteria isolated from CD patients and healthy children, respectively) and CD-triggering agents (gliadin and IFN-γ) by histology, scanning electron microscopy, immunofluorescence, and a rat cytokine antibody array. Adhesion of the bacterial strains to the IEC-6 rat cell line was evaluated in vitro. Gliadin fragments alone or together with the proinflammatory cytokine interferon (IFN)-γ significantly decreased the number of goblet cells in the small intestine; this effect was more pronounced in the presence of Escherichia coli CBL2 and Shigella CBD8. Shigella CBD8 and IFN-γ induced the highest mucin secretion and greatest impairment in tight junctions and, consequently, translocation of gliadin fragments into the lamina propria. Shigella CBD8 and E. coli CBL2 strongly adhered to IEC-6 epithelial cells. The number of goblet cells in small intestine increased by the simultaneous incubation of Bifidobacterium bifidum IATA-ES2 with gliadin, IFN-γ and enterobacteria. B. bifidum IATA-ES2 also enhanced the production of chemotactic factors and inhibitors of metalloproteinases, which can contribute to gut mucosal protection.[Conclusions]: Our results suggest that the composition of the intestinal microbiota affects the permeability of the intestinal mucosa and, consequently, could be involved in the early stages of CD pathogenesis.This work was supported by grants 310/07/0414, 303/08/0367, P304/10/P406 of the Grant Agency of the Czech Republic; IAA500200801, IAA500200710, KJB50020094 of the Academy of Sciences; AV CR-C.S.I.C. 09/10, Project 2B06155 of the Ministry of Education; and Institutional Research Concept AVOZ50200510. This work was also supported by grants 2006CZ0030 and 2008CZ0023 from Consejo Superior de Investigaciones Científicas (CSIC, Spain) and AGL2008-01440/ALI and Consolider Fun-C-Food CSD2007-00063 from the Spanish Ministry of Science and Innovation. The scholarship to G. De Palma from Junta de Ampliación de Estudios - Consejo Superior de Investigaciones Científicas (JAE-CSIC; Spain) is fully acknowledged.Peer reviewe

Gut microbiota: Role in pathogen colonization, immune responses, and inflammatory disease

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/138242/1/imr12567.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/138242/2/imr12567_am.pd

Rozhovor Britských listů 315. Bezbariérový přístup nemáme. Na invalidy se kašle [Britské listy Interview 315. We do not have access for the disabled people. Everyone ignores them here.]

"In every high rise here at this housing estate, there lives at least one disabled person consigned to a wheelchar. And these are people who have been living here for years," says Marie Cinová from the North Bohemian border town of Šluknov, Czech Republic. Ms. Cinová has a disabled son. She herself is disabled, as is her husband, a cardiac waiting for a heart operation in hospital now. Ms Cinová washes her son at the sink in the kitchen because the wheelchair will not fit through the bathroom door. She needs the door to be re-built but no one will help her - nor will help come for the other disabled people. Maybe her fellow citizens should help her financially? But should they be doing what is undoubtedly the duty of the state? Her account is IBAN CZ752700 0000 0021 1375 4545. This interview was broadcast by the Czech cable TV station Regionalnitelevize.cz from Friday 4th September 2020

Postawy pacjentów chorych na cukrzycę wobec ich choroby

Background. Education of diabetic patients is integral to effective treatment. Material and methods. In the presented work, we studied Eastern Slovakian patients with diabetes type 2 with or without insulin treatment. We focused on their diabetic self-care and compared their attitudes, education on their disease, and treatment provided by professionals. There were 411 patients in the insulin-treated group. We used the standardized Diabetes Care Profile questionnaire (DCP). Results. The groups had very different attitudes toward diabetic self-care. Patients with provided professional self-care education scored higher in all areas of diabetic care. Appropriate education influenced knowledge and consequently the management and attitudes of diabetic patients toward their disease. Educated patients scored higher in the categories ‘Self-care ability’, ‘Importance of care’, ‘Self-care adherence’, ‘Diet adherence’, ‘Medical barriers’, ‘Exercise barriers’, ‘Monitoring barriers’ and ‘Understanding practice’ (p < 0.01). Patients who had not received diabetes education presented higher scores in emotional areas, i.e., negative and positive attitudes toward diabetes mellitus (p < 0.01). Conclusions. We concluded that it is beneficial for a structured e ducational process to be integrated in diabetes treatment.Wprowadzenie. Edukacja pacjentów chorych na cukrzycę stanowi integralną część ich skutecznego leczenia. Materiał i metody. W prezentowanym badaniu porównano postawy w zakresie samodzielnej profilaktyki cukrzycy w dwóch grupach pacjentów cierpiących na cukrzycę typu 2, która wymaga insulinoterapii, w odniesieniu do szkoleń edukacyjnych dotyczących ich choroby oraz leczenia prowadzonego przez specjalistów. Badaną grupę stanowiło 411 pacjentów cierpiących na cukrzycę typu 2 leczoną za pomocą insuliny, którzy pochodzą ze wschodniej części Słowacji. Zastosowano standardowy kwestionariusz DCP (ang. Diabetes Care Profile), wykorzystywany do oceny czynników społecznych i psychologicznych wśród cukrzyków. Wyniki. Postawy dwóch porównywanych grup pacjentów wobec cukrzycy różniły się istotnie w zakresie samodzielnej profilaktyki. Pacjenci z zapewnionym profesjonalnym szkoleniem dotyczącym samodzielniej profilaktyki uzyskiwali wyższe oceny we wszystkich obszarach związanych z monitorowaniem i leczeniem cukrzycy. Prawidłowy proces edukacyjny ma wpływ na poziom wiedzy, a tym samym na działania podejmowane przez pacjentów i ich postawy wobec choroby. Pacjenci, którzy otrzymali odpowiednie przeszkolenie, lepiej kontrolowali sfery takie jak „Umiejętności dot. samodzielnej profilaktyki”, „Znaczenie samodzielnej profilaktyki”, „Przestrzeganie zasad samodzielnej profilaktyki”, „Przestrzeganie diety”, „Ograniczenia natury medycznej”, „Ograniczenia w wykonywaniu ćwiczeń”, „Ograniczenia w monitorowaniu” oraz „Rozumienie procedur” (p < 0,01). Z kolei pacjenci nieposiadający odpowiedniej wiedzy prezentowali lepsze wyniki w obszarach związanych ze sferą emocjonalną – negatywnym i pozytywnym nastawieniu do cukrzycy (p < 0,01). Wnioski. W kontekście uzyskanych wyników, zintegrowanie ustrukturyzowanego procesu edukacyjnego z leczeniem osób chorujących na cukrzycę wydaje się konieczne

Cytokine array analysis of rat intestinal loops washes.

<p>Layout of the arrays (A), cytokine profiles from loops treated with PBS (control) (B), gliadin (C), gliadin+IFN-γ (D), <i>B. bifidum</i> IATA-ES2+gliadin+IFN-γ (E), <i>E. coli</i> CBL2+gliadin+IFN-γ (F), and <i>E. coli</i> CBL2+<i>B. bifidum</i> IATA-ES2+gliadin+IFN-γ (G). The data are expressed as relative levels of selected cytokines (percentage of positive controls). Cytokine-induced neutrophil chemoattractant (CINC)-2 and -3, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-3α, nerve growth factor β-(NGF), tumor necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF). The signal intensity was measured using the LAS-1000 luminescence detector (Fujifilm, Tokyo, Japan).</p

Effects of bacterial strains and gliadin on goblet cells.

<p>Histological staining of PAS-positive goblet cells in rat intestinal loops exposed to: <i>B. bifidum</i> IATA-ES2+gliadin (200 µg) (A), IFN-γ (225 U) (B), <i>Shigella</i> CBD8+gliadin (C), <i>Shigella</i> CBD8+gliadin +IFN-γ (D), <i>E. coli</i> CBL2+gliadin (E) and <i>E. coli</i> CBL2+gliadin+IFN-γ (F). Bacteria were applied at 10⁶/loop. Changes in goblet cells are expressed as medians and interquartile ranges (25% to 75%) of the number of PAS-positive goblet cells/100 epithelial cells (G). These values were for <i>B. bifidum</i> IATA-ES2 (39, 35–41), <i>E. coli</i> CBL2 (38, 35–41) and <i>Shigella</i> CBD8 (25, 20–27) when applied alone to the loops. Different letters (a–e) indicate statistically significant differences between medians as calculated by Mann-Whitney U test (P<0.05). Identical letters correspond to non-significant differences. The separate dots or asterisks indicate outliers. The pictures were obtained for the specimens viewed under an Olympus BX 40. Scale bar, 50 µm.</p

Adhesion of different bacterial strains to IEC-6 cells.

<p>The highest percentage of adhered bacteria was observed for <i>E. coli</i> CBL2 and <i>Shigella</i> CBD8. The differences between tested bacterial strains, as well as the effect of simultaneously added gliadin fragments were non-significant as established by applying the Mann-Whitney U-test. Data are expressed as medians and interquartile ranges (25% to 75%) of adhesion of four independent experiments. None of the differences was found to be statistically significant (P<0.05). The separate dot indicates an outlier.</p