Plant lectins ConBr and CFL modulate expression toll-like receptors, pro-inflammatory cytokines and reduce the bacterial burden in macrophages infected with Salmonella enterica serovar Typhimurium.

BACKGROUND: Plant lectins have long been used in biomedical research as immunomodulators against tumor cells and microbial infections. PURPOSE: To test the ability of plant lectins ConBr (Canavalia brasiliensis) and CFL (Cratylia argentea) to activate antimicrobial and immunomodulatory activities of murine peritoneal macrophages (pMØ) infected with a virulent strain of Salmonella enterica serovar Typhimurium (STm). METHODS: We incubated pMØ with non-toxic amounts of ConBr and CFL either before (preventive schedule) or after (curative schedule) exposure to STm. RESULTS: In uninfected pMØ, ConBr and CFL greatly increased levels of mRNA transcripts for IL-1ß, TNF-α and IL-6 and the inducible nitric oxide synthase (iNOs), but not IL-10 and IL-12. Exposure to naïve splenocytes of culture supernatants of pMØ previously stimulated with CFL resulted in expression of IL-12 and IFN-γ. Both preventive and curative treatment schedules significantly reduced the intracellular load of Salmonella. Experiments in infected macrophages exposed to lectins in the preventive schedule showed that mRNA transcripts for IL-6 and TNF-α were increased by CFL, whereas ConBr enhanced IL-12 (subunit p40). In the curative schedule, CFL induced significant expression of IL-12 (p40) whereas ConBr enhanced expression IL-1ß and TNF-α genes. The lectin treatments did not influence on iNOs expression in pMØ infected with STm C5 regardless of the treatment schedule. Curative treatments with CFL increased approximately 130-fold expression of TLR-4 whist expression of TLR-9 was increased by treatments with ConBr. CONCLUSION: We conclude that lectins ConBr and CFL have immunomodulatory properties that are beneficial on control of cells infected by Salmonella.

First Report of Powdery Mildew Caused by Erysiphe platani on Platanus × acerifolia in Rio Grande do Sul, Brazil.

Platanus × acerifolia (Aiton) Willd. (London planetree) is a tree commonly used as an ornamental and in the furniture industry. In the summer of 2013, powdery mildew was observed on shoots of P. × acerifolia plants in the cities of Pelotas and Canela (State of Rio Grande do Sul, Brazil). Voucher specimens (n = 2) were deposited in the Phytopathological Museum Manoel Alves Oliveira at Federal University of Pelotas. Dense white powdery masses of conidia and mycelium were observed on leaves (abaxial and adaxial surfaces), petioles, and young stems. Leaves with high disease severities (≥70%) were deformed with curved edges to the adaxial side, and they often died. Mycelia were superficial with lobed appressoria. Conidiophores were straight, sometimes curved at the base, unbranched, cylindrical, 98 to 236 µm long (137.3 ± 41.2 µm) and composed of a cylindrical foot cell 49 to 102 µm long (66.9 ± 19.5 µm) and 4.4 to 6.4 µm wide (5.3 ± 0.8 µm) followed by two to four cells. Conidia were produced singly or in short chains (two to three), without distinct fibrosin bodies, ellipsoid to ovoid and measuring 24 to 37 µm long (29.5 ± 3.2 µm) and 12 to 19 µm wide (15.2 ± 1.4 µm), often with a wrinkled appearance. Primary conidia had truncate bases and rounded apex while both base and apex were truncated in secondary conidia. Germ tubes were produced apically (pseudoidium type). Chasmothecia were not observed. Genomic DNA was used to amplify the internal transcribed spacer (ITS) region using the ITS1 and ITS4 primers. The resulting sequence (602 bp) was deposited (Accession No. KF499270) in GenBank. BLASTn searches revealed similarity of 100 and 99% with Erysiphe platani from P. orientalis L. (Accession No. JQ365943.1) and P. occidentalis L. (Accession No. JX997805.1), respectively. Phylogenetic analysis placed our sequence in a clade (99% bootstrap support) which included only other E. plantani sequences. In short, morphological and molecular approaches allowed us to identify the infecting fungus as E. platani. For Koch's postulates, 10 detached leaves were inoculated (10 to 15 conidia cm2) on their adaxial surface using an eyelash brush. Non-inoculated leaves served as control. All leaves were kept inside trays with petiole immersed in humidified cotton and maintained at 25 ± 1°C. Symptoms identical to those of the original leaves were observed 6 to 8 days after inoculation, whereas the control leaves remained symptomless. Although E. platani has been previously reported on P. × acerifolia in the city of Poços de Calda, state of Minas Gerais, Brazil (1) and on P. occidentalis in Korea (2), to our knowledge, this is the first record of E. platani on P. × acerifolia in Rio Grande do Sul, Brazil. References: (1) E. M. Inokuti et al. New Dis. Rep. 15:38, 2007. (2) Y. J. La and H. D. Shin. Plant Dis. 97:843, 2013.

Modeling pulmonary fibrosis by abnormal expression of telomerase/apoptosis/collagen V in experimental usual interstitial pneumonia

Limitations on tissue proliferation capacity determined by telomerase/apoptosis balance have been implicated in pathogenesis of idiopathic pulmonary fibrosis. In addition, collagen V shows promise as an inductor of apoptosis. We evaluated the quantitative relationship between the telomerase/apoptosis index, collagen V synthesis, and epithelial/fibroblast replication in mice exposed to butylated hydroxytoluene (BHT) at high oxygen concentration. Two groups of mice were analyzed: 20 mice received BHT, and 10 control mice received corn oil. Telomerase expression, apoptosis, collagen I, III, and V fibers, and hydroxyproline were evaluated by immunohistochemistry, in situ detection of apoptosis, electron microscopy, immunofluorescence, and histomorphometry. Electron microscopy confirmed the presence of increased alveolar epithelial cells type 1 (AEC1) in apoptosis. Immunostaining showed increased nuclear expression of telomerase in AEC type 2 (AEC2) between normal and chronic scarring areas of usual interstitial pneumonia (UIP). Control lungs and normal areas from UIP lungs showed weak green birefringence of type I and III collagens in the alveolar wall and type V collagen in the basement membrane of alveolar capillaries. The increase in collagen V was greater than collagens I and III in scarring areas of UIP. A significant direct association was found between collagen V and AEC2 apoptosis. We concluded that telomerase, collagen V fiber density, and apoptosis evaluation in experimental UIP offers the potential to control reepithelization of alveolar septa and fibroblast proliferation. Strategies aimed at preventing high rates of collagen V synthesis, or local responses to high rates of cell apoptosis, may have a significant impact in pulmonary fibrosis.

Modeling pulmonary fibrosis by abnormal expression of telomerase/apoptosis/collagen V in experimental usual interstitial pneumonia.

Limitations on tissue proliferation capacity determined by telomerase/apoptosis balance have been implicated in pathogenesis of idiopathic pulmonary fibrosis. In addition, collagen V shows promise as an inductor of apoptosis. We evaluated the quantitative relationship between the telomerase/apoptosis index, collagen V synthesis, and epithelial/fibroblast replication in mice exposed to butylated hydroxytoluene (BHT) at high oxygen concentration. Two groups of mice were analyzed: 20 mice received BHT, and 10 control mice received corn oil. Telomerase expression, apoptosis, collagen I, III, and V fibers, and hydroxyproline were evaluated by immunohistochemistry, in situ detection of apoptosis, electron microscopy, immunofluorescence, and histomorphometry. Electron microscopy confirmed the presence of increased alveolar epithelial cells type 1 (AEC1) in apoptosis. Immunostaining showed increased nuclear expression of telomerase in AEC type 2 (AEC2) between normal and chronic scarring areas of usual interstitial pneumonia (UIP). Control lungs and normal areas from UIP lungs showed weak green birefringence of type I and III collagens in the alveolar wall and type V collagen in the basement membrane of alveolar capillaries. The increase in collagen V was greater than collagens I and III in scarring areas of UIP. A significant direct association was found between collagen V and AEC2 apoptosis. We concluded that telomerase, collagen V fiber density, and apoptosis evaluation in experimental UIP offers the potential to control reepithelization of alveolar septa and fibroblast proliferation. Strategies aimed at preventing high rates of collagen V synthesis, or local responses to high rates of cell apoptosis, may have a significant impact in pulmonary fibrosis.

Efficacy of free and nanoencapsulated Eucalyptus citriodora essential oils on sheep gastrointestinal nematodes and toxicity for mice.

Herbal medicines with anthelmintic effects are alternatives for the sustainable control and prevention of disease caused by gastrointestinal parasites. The nanoencapsulation of essential oils has been proposed to enhance the absorption of their constituents and improve their efficacy. The present study aimed to evaluate the efficacy of free and nanoencapsulated Eucalyptus citriodora essential oil (EcEO) on the control of gastrointestinal nematodes of small ruminants in vitro and in vivo. Chitosan was used as a matrix for the formulation of a nanoemulsion. Chromatographic and physico-chemical analyses of EcEO were performed. Egg hatch (EHT) and larval development (LDT) tests were conducted to evaluate the effectiveness of nanoencapsulated and free EcEO on the eggs and larvae of Haemonchus contortus. Acute toxicity of free and nanoencapsulated EcEO was evaluated using mice. Finally, nanoencapsulated EcEO efficacy on the control of gastrointestinal nematodes was calculated by fecal egg count reduction test (FECRT) treating 30 sheep naturally infected with 250 mg/kg of free and nanoencapsulated EcEO. In vitro tests were analyzed by an analysis of variance (ANOVA) followed by comparison with the Tukey test. The efficacy of FECRT was calculated by the BootStreet program through arithmetic average, using the formula 100 (1-XT/XC). To compare the differences between epg, the data were transformed to log(x+1) and subjected to an ANOVA to compare the significant differences between groups by Tukey's. The level of significance was P<0.05. The free (4 mg/ml concentration) and nanoencapsulated (2mg/ml concentration) EcEO inhibited larvae hatching by 97.2% and 92.8%, respectively. Free and nanoencapsulated EcEO at 8 mg/ml inhibited larval development by 99.8% and 98.1%, respectively. In the acute toxicity test, the LD10 and LD50 of free EcEO was 1999 and 2653 mg/kg, respectively, while the LD10 and LD50 of nanoencapsulated EcEO was 1121 and 1681 mg/kg, respectively. Nanoencapsulated and free EcEO reduced FEC similarly by 40.5% and 55.9%, respectively at 10 days post-treatment. Nanoencapsulated EcEO did not obtain the expected efficacy in vivo.

Phytochemistry and Preliminary Assessment of the Antibacterial Activity of Chloroform Extract of Amburana cearensis (Allemão) A.C. Sm. against Klebsiella pneumoniae Carbapenemase-Producing Strains.

The chloroform extract of the stem bark of Amburana cearensis was chemically characterized and tested for antibacterial activity.The extract was analyzed by gas chromatography and mass spectrometry. The main compounds identified were 4-methoxy-3-methylphenol (76.7%), triciclene (3.9%), α -pinene (1.0%), ß -pinene (2.2%), and 4-hydroxybenzoic acid (3.1%). Preliminary antibacterial tests were carried out against species of distinct morphophysiological characteristics: Escherichia coli, Salmonella enterica Serotype Typhimurium, Pseudomonas aeruginosa, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus. The minimum inhibitory concentration (MIC) was determinate in 96-well microplates for the chloroform extract and an analogue of themain compound identified, which was purchased commercially.We have shown that plant's extract was only inhibitory (but not bactericidal) at the maximum concentration of 6900 µ g/mL against Pseudomonas aeruginosa and Bacillus cereus. Conversely, the analogue 2-methoxy-4-methylphenol produced MICs ranging from215 to 431 µ g/mL against all bacterial species.New antibacterial assays conducted with such chemical compound against Klebsiella pneumoniae carbapenemase-producing strains have shown similarMICresults and minimumbactericidal concentration (MBC) of 431 µ g/mL.We conclude that A. cearensis is a good source of methoxy-methylphenol compounds,which could be screened for antibacterial activity againstmultiresistant bacteria fromdifferent species.

Occurrence of Salmonella sp. and coagulase-positive staphylococci in raw eggs and Coalho cheese: comparative study between two cities of Brazil's northeast

Microbiological analyses of chicken eggs in Recife and Salvador have shown a high occurrence of Salmonella in the egg shells and yolks. Likewise, the occurrence of Salmonella plus coagulase-positive staphylococci in Coalho cheese reached alarming levels. The data revealed a significant risk of infections and intoxications from consuming these foods in the cities.

Root-Knot Disease Caused by Meloidogyne arenaria and M. javanica in Teak in São Paulo State, Brazil.

Teak (Tectona grandis Linn. F.) is one of the most important forest crops in Brazil, occupying areas in different regions, such as Goiás, Mato Grosso, Paraná, and São Paulo states. Teak wood is used for many purposes such as shipbuilding, rolling and plywood, firewood, and charcoal. In May 2011, teak symptomatic feeder root samples, exhibiting inconspicuous, small galls, were collected in the municipality of Piracicaba, São Paulo State, Brazil (22°41'46.90″S, 47°38'36.84″W). Specimens were identified through perineal patterns and esterase phenotypes of 20 adult females (1,2). Perineal patterns and esterase phenotypes were consistent with those described for Meloidogyne arenaria (Neal, 1889) Chitwood, 1949 and M. javanica (Treub, 1885) Chitwood, 1949. Perineal patterns of M. arenaria showed a low dorsal arch, compressed dorsolaterally, with lateral field marked by some forked and broken striae; no punctate markings between anus and tail terminus were observed. Perineal patterns of M. javanica were rounded, with low dorsal arch, striae smooth, lateral field distinct, clearly demarcated from striae by parallel lines. From the esterase electrophoresis we obtained A2 (Rm:1.2;1.3) and J3 (Rm:1.0;1.25;1.4) phenotypes, typical from M. arenaria and M. javanica, respectively. To our knowledge, this is the first report of M. arenaria parasitizing teak roots in Brazil and elsewhere (new host) and the first report of M. javanica infecting teak in the State of São Paulo. Previously, M. javanica was reported to be infecting teak-growing areas in the State of Mato Grosso (3). This finding has a great importance, not only by the inclusion of these parasites in teak pathological scenario, but also for predicting possible damage in plant species used in teak-based intercropping systems. References: (1) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 22:10, 1990. (2) K. M. Hartman and J. N. Sasser. 1985. Page 115 in: An Advanced Treatise on Meloidogyne. Volume II, Methodology. K. R. Barker et al., eds. North Carolina State University Graphics, Raleigh,1985. (3) R. A. Silva et al. Nematol. Bras. 27:261, 2003.

Occurrence of Salmonella sp. and coagulase-positive staphylococci in raw eggs and Coalho cheese: comparative study between two cities of Brazil's northeast.

Microbiological analyses of chicken eggs in Recife and Salvador have shown a high occurrence of Salmonella in the egg shells and yolks. Likewise, the occurrence of Salmonella plus coagulase-positive staphylococci in Coalho cheese reached alarming levels. The data revealed a significant risk of infections and intoxications from consuming these foods in the cities.

Recovery and screening for antibiotic susceptibility of potential bacterial pathogens from the oral cavity of shark species involved in attacks on humans in Recife, Brazil.

The number of incidents involving sharks and humans at beaches in Recife, on the north-eastern Brazilian coast, is among the highest worldwide. In addition, wound infections in survivors are common; but the nature and risk of the aetiological agents is unknown. In the present study, 81 potential bacterial pathogens were identified in the oral cavity of sharks involved in attacks in Recife, and were subjected to antibiotic susceptibility tests using the standardized disc-diffusion method. The majority were enterobacteria such as Enterobacter spp., Citrobacter spp., Proteus spp., Providencia alcalifaciens, Escherichia coli, Moellerella wisconcensis and Leclercia adecarboxylata. Other Gram-negative bacteria included Vibrio spp., Burkholderia cepacia, Acinetobacter spp. and Pseudomonas spp. In addition, coagulase-positive and coagulase-negative Staphylococcus spp., Enterococcus spp. and Micrococcus spp. were identified, besides Streptococcus spp. from the viridans group. Resistance was especially found in the Proteus mirabilis and Citrobacter freundii, and ranged from 4 to 6 antibiotics out of the 13 tested. Gentamicin and vancomycin were the most effective against Gram-positive cocci strains, whereas levofloxacin was fully inhibitory against Gram-positive and Gram-negative bacteria. These data are discussed in light of a retrospective evaluation of the medical records of three shark victims treated at Restauração Hospital in Recife.

Antifungal activity of the naphthoquinone beta-lapachone against disseminated infection with Cryptococcus neoformans var. neoformans in dexamethasone-immunosuppressed Swiss mice

The in vivo antifungal activity of the naphthoquinone beta-lapachone against disseminated infection by Cryptococcus neoformans was investigated. Swiss mice were immunosuppressed daily with dexamethasone (0.5 mg per mouse) intraperitoneally for 3 days, the procedure was repeated 4 days later, and the animals were then challenged intravenously with C. neoformans (10(6) CFU/mL) 1 week later. Seven days after infection, the mice were divided into groups and treated daily with beta-lapachone (10 mg/kg, iv) for 7 (N = 6) and 14 days (N = 10). Amphotericin B (0.5 mg/kg) was used as comparator drug and an additional group received PBS. Treatment with beta-lapachone cleared the yeast from the spleen and liver, and the fungal burden decreased approximately 10(4) times in the lungs and brain 14 days after infection when compared to the PBS group (P < 0.05). This result was similar to that of the amphotericin B-treated group. Protection was suggestively due to in vivo antifungal activity of this drug and apparently not influenced by activation of the immune response, due to similar leukocyte cell counts among all groups. This study highlights the prospective use of beta-lapachone for treatment of disseminated cryptococcosis.

Antifungal activity of the naphthoquinone beta-lapachone against disseminated infection with Cryptococcus neoformans var. neoformans in dexamethasone-immunosuppressed Swiss mice.

The in vivo antifungal activity of the naphthoquinone beta-lapachone against disseminated infection by Cryptococcus neoformans was investigated. Swiss mice were immunosuppressed daily with dexamethasone (0.5 mg per mouse) intraperitoneally for 3 days, the procedure was repeated 4 days later, and the animals were then challenged intravenously with C. neoformans (10(6) CFU/mL) 1 week later. Seven days after infection, the mice were divided into groups and treated daily with beta-lapachone (10 mg/kg, iv) for 7 (N = 6) and 14 days (N = 10). Amphotericin B (0.5 mg/kg) was used as comparator drug and an additional group received PBS. Treatment with beta-lapachone cleared the yeast from the spleen and liver, and the fungal burden decreased approximately 10(4) times in the lungs and brain 14 days after infection when compared to the PBS group (P < 0.05). This result was similar to that of the amphotericin B-treated group. Protection was suggestively due to in vivo antifungal activity of this drug and apparently not influenced by activation of the immune response, due to similar leukocyte cell counts among all groups. This study highlights the prospective use of beta-lapachone for treatment of disseminated cryptococcosis.

Antibacterial activity, chemical composition, and cytotoxicity of leaf's essential oil from Brazilian pepper tree (Schinus terebinthifolius, Raddi)

The antibacterial potential of leaf's essential oil (EO) from Brazilian pepper tree (Schinus terebinthifolius Raddi) against staphylococcal isolates from dogs with otitis externa was evaluated. The minimum inhibitory concentration of EO ranged from 78.1 to 1,250 fg/mL. The oil was analyzed by GC and GC/MS and cytotoxicity tests were carried out with laboratory animals.

Antibacterial activity, chemical composition, and cytotoxicity of leaf's essential oil from brazilian pepper tree (schinus terebinthifolius, raddi).

The antibacterial potential of leaf's essential oil (EO) from Brazilian pepper tree (Schinus terebinthifolius Raddi) against staphylococcal isolates from dogs with otitis externa was evaluated. The minimum inhibitory concentration of EO ranged from 78.1 to 1,250 µg/mL. The oil was analyzed by GC and GC/MS and cytotoxicity tests were carried out with laboratory animals.

Pequi is a Host for Pratylenchus zeae in the Brazilian Cerrado Region.

The Brazilian Cerrado Region has many natural resources that have high social economic interest. Pequi (Caryocar brasiliense Camb.), a native species from that area, has an edible fruit, which is highly appreciated by the local population, and also a high-quality wood. In January 2010, pequi root samples were collected near the municipality of Rio Verde, Goiás State, Brazil (17°49'25.76″S, 51°02'10.06″W). Roots were washed with tapwater, dried on absorbent paper, cut in 1-cm2 pieces, and processed for nematode extraction by the blender centrifugal flotation method (2). The specimens were identified by morphological and morphometrical characteristics of six adult females mounted in formaldehyde temporary slides (1). Morphological characters used for identification included female body, stylet, pharyngeal overlapping, pharynges, postvulval uterine sac, tail lengths, stylet knobs, number of labial rings, vulva position in relation to body length, body diameters (high body, vulval, and anus region), and the de Man's ratios (a, b, b', c, and c'). Characters measured were consistent with those described for Pratylenchus zeae Graham, 1951 (1); the labial region showed three annuli, stylet was 14.83 (±0.93) µm long, with broad, anteriorly flattened basal knobs. Vulva position was 71.66% (±0.98) of body length and spermatheca was round, small, and without sperm (males were not found). Postvulval uterine sac was short (31.3 ± 4.03 µm) and tail (26.6 ± 3.61 µm) was conoid, pointed, and unstriated. Pharyngeal overlapping length was 30.5 (±6.5) µm; pharynges were 150.83 (± 28.16) µm long. The de Man's ratios obtained were: a = 24.26 ± 2.31; b = 3.89 ± 0.69; b' = 3.08 ± 0.48; c = 17.17 ± 1.47; and c' = 2.25 ± 0.19. To our knowledge, this is the first report of P. zeae infecting pequi. It is difficult to determine the economic importance of this nematode parasite to pequi production since pequi is not yet a commercial crop in Brazil. This finding, however, has long term importance because researchers have been developing improved cultivars by combining favorable agronomic characteristics with high oilseed content for biofuel production. If these are commercialized, P. zeae could become an important pathogen in pequi plantings. References: (1) P. Castillo and N. Vovlas. Pratylenchus (Nematoda: Pratylenchidae): Diagnosis, Biology, Pathogenicity and Management. Koninklijke Brill NV, Leiden, the Netherlands, 2007. (2) W. A. Coolen and C. J. D'Herde. A Method for the Quantiative Extraction of Nematodes from Plant Tissue. State Agric. Entomol. Res. Stn. Ghent, Belgium, 1972.

Unicystic ameloblastoma in a child: a differential diagnosis from the dentigerous cyst and the inflammatory follicular cyst.

Unicystic ameloblastoma (UA) is a benign epithelial odontogenic tumor of the jaws with an aggressive potential that commonly occurs in children. This cystic odontogenic neoplasm is generally asymptomatic and found during routine radiographs. The purposes of this report were to describe a case of UA involving the crown of the unerupted right mandibular second premolar in an 11-year-old girl under orthodontic treatment, and discuss its diagnosis and radiographic and microscopic findings, emphasizing its distinction from the dentigerous cyst and the inflammatory follicular cyst.

Effect of the Escherichia coli EMO strain on experimental infection by Salmonella enterica serovar Typhimurium in gnotobiotic mice.

An experimental infection with Salmonella enterica subsp. enterica serovar Typhimurium was evaluated in gnotobiotic mice previously exposed to a plasmid-free non-pathogenic Escherichia coli (EMO strain). Mice were exposed to EMO (experimental) or not (control) 10 days before challenge with Salmonella Typhimurium (10(2) colony forming units (CFU)/mouse). Survival after challenge was higher (P < 0.05) in the experimental group (16%) than in the control animals (0%). Histopathological examination of the colon and ileum mucosa of the experimental group showed less extensive lesions such as edema, cell inflammatory infiltration and hyperemia. The epithelial cells of the mucosal surface and the production of the mucous layer were also better preserved in the experimental group. The population levels of Salmonella Typhimurium in the feces were initially 10-fold lower (P < 0.05) in the experimental groups. However, 3 days after challenge both experimental and control groups showed similar population levels ranging from 10(8) to 10(9) CFU/g of feces. The intestinal contents of total and anti-Salmonella Typhimurium sIgA were higher in the experimental groups 10 days after inoculation of E. coli EMO strain. Translocation of Salmonella Typhimurium to the spleen was 10-fold lower (P < 0.05) in the experimental group only on day 3 after infection. This was not related to an increase in the bacterial blood clearance of the animals, as shown by experimental venous challenge with E. coli B41. In conclusion, treatment of mice with E. coli EMO strain promoted a relative protection against experimental infection with Salmonella Typhimurium. This protection was not due to the reduction of the population of pathogens in the intestine but was probably related to stimulation of the immune response.

Effect of the Escherichia coli EMO strain on experimental infection by Salmonella enterica serovar Typhimurium in gnotobiotic mice

An experimental infection with Salmonella enterica subsp. enterica serovar Typhimurium was evaluated in gnotobiotic mice previously exposed to a plasmid-free non-pathogenic Escherichia coli (EMO strain). Mice were exposed to EMO (experimental) or not (control) 10 days before challenge with Salmonella Typhimurium (10² colony forming units (CFU)/mouse). Survival after challenge was higher (P < 0.05) in the experimental group (16 percent) than in the control animals (0 percent). Histopathological examination of the colon and ileum mucosa of the experimental group showed less extensive lesions such as edema, cell inflammatory infiltration and hyperemia. The epithelial cells of the mucosal surface and the production of the mucous layer were also better preserved in the experimental group. The population levels of Salmonella Typhimurium in the feces were initially 10-fold lower (P < 0.05) in the experimental groups. However, 3 days after challenge both experimental and control groups showed similar population levels ranging from 10(8) to()10(9) CFU/g of feces. The intestinal contents of total and anti-Salmonella Typhimurium sIgA were higher in the experimental groups 10 days after inoculation of E. coli EMO strain. Translocation of Salmonella Typhimurium to the spleen was 10-fold lower (P < 0.05) in the experimental group only on day 3 after infection. This was not related to an increase in the bacterial blood clearance of the animals, as shown by experimental venous challenge with E. coli B41. In conclusion, treatment of mice with E. coli EMO strain promoted a relative protection against experimental infection with Salmonella Typhimurium. This protection was not due to the reduction of the population of pathogens in the intestine but was probably related to stimulation of the immune response.

Evidence of type II pneumocyte apoptosis in the pathogenesis of idiopathic pulmonary fibrosis (IFP)/usual interstitial pneumonia (UIP).

BACKGROUND/AIMS: The pathogenesis of idiopathic pulmonary fibrosis (IPF)/usual interstitial pneumonia (UIP), a chronic and incurable human respiratory disease, is not well established. This study was designed to investigate whether the apoptosis of type II pneumocytes could be the precipitating factor in the pathogenesis of IPF. METHODS: Nineteen specimens obtained by retrospective review of the medical and pathological records of 55 patients with IPF, four normal subjects, and 10 disease control lungs were analysed. The selected specimens had normal alveoli with intervening patchy scarring of the lung parenchyma, fulfilling the pathological criteria for UIP. To identify individual cells undergoing apoptosis in the normal alveoli, electron microscopy and in situ end labelling of fragmented DNA were performed on paraffin was embedded sections using digoxigenin-11-dUTP and the enzyme terminal deoxynucleotidyl transferase. RESULTS: Apoptosis was detected in the normal alveoli of 17 of the 19 patients with IPF/UIP and was absent in the controls. Electron microscopy demonstrated apoptotic changes in type II pneumocytes. These results indicate that apoptotic type II pneumocyte death occurs in normal alveoli of IPF/UIP and could be the principal cause of several events that account for the histological, clinical, and functional alterations seen in IPF/UIP. CONCLUSIONS: In conclusion, numerous type II pneumocytes from the normal alveoli of most patients with IPF/UIP actively undergo programmed cell death. This finding may shed new light on the pathogenesis of this disease, with implications mainly for the treatment of affected patients.