Tea catechins induce crosstalk between signaling pathways and stabilize mast cells in ulcerative colitis.

It is well documented that nutraceuticals, in general, and Green tea catechins, in particular, possess a potential therapeutic value in inflammatory bowel diseases (IBD) due to their anti-oxidative and anti-inflammatory effects. This study aimed to investigate the possible mechanism of action of catechins in a rat model of colitis induced by 2.4.6 trinitrobenzene sulfonic acid (TNBS). Thirty-five young adult Sprague-Dawley rats were divided into four groups: normal control (n=5), catechins (n=9), TNBS (n=9) and TNBS plus catechins (n=12) treated. Catechin in the form of Epigallocatechin-3-gallate (EGCG) was administered daily by intraperitoneal injection, 1 week before the induction date of UC. Biopsies of the descending colon were collected on days 3, 10 and 17, and partly frozen for molecular studies or fixed for light microscopy. The status of intestinal tissue alterations and mast cells number were also assessed, as well as the mRNA expressions of IL-6, TNF-a and NF-kB, and determination of ROS expression. Histological data depicted a significant amelioration in the TNBS- and EGCG-treated rats compared to the non-treated animals. Catechin expressed strong anti-inflammatory and anti-oxidant effects, ameliorated ulcerative colitis and stabilized mast cells. The mechanism of action occurred basically through the NF-kB pathway and possibly through a crosstalk with other pathways.

The immunohistochemical peptidergic expression of leptin is associated with recurrence of malignancy in laryngeal squamous cell carcinoma.

Leptin is a peptide that plays a key role in the control of satiety, energy expenditure, food intake and various reproductive processes. In the last years, the expression of leptin had been found in malignant cells of various origins. The aim of this study is to evaluate leptin expression in human laryngeal squamous cell carcinoma (SCC) and to investigate its possible role in predicting prognosis. Leptin expression was determined by immunohistochemistry in pathological and healthy tissue specimens from 24 patients with laryngeal SCC. Specimens were stained with an anti-leptin antibody. All measurements were performed using a computer-based image analysis system and scale of staining intensity was determined. All tumoural specimens showed significant immunoreactivity for leptin compared to healthy tissues (p ≤ 0.05), but showed different immunoreactivity that was related to clinicopathological features. High leptin expression was not significantly related with TNM, histological grading (HG) or advanced (III and IV) clinical stage (p > 0.05). Recurrence of malignancy was found to be significantly related with high expression of leptin by Spearman's rank correlation test (r = 0.59; p = 0.002), Fisher's test (p = 0.017) and Kaplan- Meier product-limit estimate (Log-rank test, p ≤ 0.05). In particular, multivariate logistic regression analysis showed that recurrences were significantly related with nodal involvement, HG and leptin expression (p ≤ 0.05). These preliminary results suggest that leptin may be a valuable parameter for predicting prognosis in laryngeal SCC.

Corynebacterium glucuronolyticum causing genitourinary tract infection: Case report and review of the literature.

Corynebacterium species are increasingly recognized as opportunistic pathogens. A growing number of taxonomic studies has yielded a description of numerous new Corynebacterium species, such as those related to the urogenital tract, with Corynebacterium glucuronolyticum found to be rarely involved in genitourinary tract infections, particularly in male individuals. In this report, we describe a urethritis case caused by C. glucuronolyticum in a 37-year-old, apparently healthy male, who complained mild pain in the lower abdomen, with several urinary symptoms. While urethral and semen specimens did not yield positive results for microbiological evaluation, cultures of urine samples revealed the monomicrobial growth on blood-containing media of tiny colonies after 24 h of incubation, clearly evident only after 48 h of incubation under CO2-enriched atmosphere. Colonies were identified as C. glucuronolyticum both by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and 16S rRNA gene sequencing. Oral ciprofloxacin gradually led to clinical improvement and, finally, to a complete recovery, in accordance with microbiological findings. In spite of its infrequent detection, C. glucuronolyticum might be a potential urogenital pathogen in males more commonly that what believed, perhaps due to slow growth leading to underrecognition; we suggest therefore to consider the organism in the differential diagnostics of bacterial diseases of the urinary tract.

Lactobacillus casei and bifidobacterium lactis supplementation reduces tissue damage of intestinal mucosa and liver after 2,4,6-trinitrobenzenesulfonic acid treatment in mice.

Probiotics (PB) are living microorganisms that act as a commensal population in normal intestines and confer numerous beneficial effects on the host. The introduction of probiotics in the treatment of inflammatory bowel disease (IBD) prolongs remission. The aim of this study was to investigate the intestinal and hepatic effects of PB supplementation in an experimental IBD model in mice induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). In the first step of the experimental procedure, CD-1 male mice, 5 to 6 weeks old, were randomly divided into 3 groups and inoculated intrarectally with, respectively, saline, alcohol, or TNBS to assess the experimental IBD model. In the second step, mice treated, or not, with TNBS inoculation, were treated with PB (Lactobacillus Casei, Bifidobacterum Lactis) for 1, 2 or 3 weeks, on a daily basis. Large bowel (colon and rectum) and liver were processed for histological alterations, according to a scoring system. Large bowel was also assessed for apoptosis by TUNEL assay. TNBS induced, as expected, severe damage and inflammation in the large bowel, including nuclear alterations and apoptosis, and, to a lesser extent, to the liver. Administration of PB determined significant reduction of both histological alterations and apoptosis. PB administration in advance protects from inflammation. In conclusion, supplementation with Lactobacillus casei, Bifidobacterum lactis PB is able to ameliorate the colitis by reversing the histological changes caused by TNBS in mice. Experimentation in human subjects in needed to prove their efficacy in reducing histological alterations that may be present in subjects with IBD.

A novel biotechnology product for the degradation of biofilm-associated polysaccharides produced by Streptococcus mutans.

In this study we evaluated the activity of ABR preparation, a first-in-class agent obtained through fermentation process by genetically unmodified Bacillus spp., in breaking down polysaccharide produced by Streptococcus mutans, primary coloniser of tooth surface and abundant in dental biofilms. Our results showed that ABR preparation is able in degrading sugars formed by S. mutans, both in broth culture and onto teeth surface. Its activity is not influenced by the presence of saliva, commercial mouthwashes or oral disinfectants. ABR preparation has the potential to remove preformed plaque and counteract its development, thus offering conservative control of gingival and periodontal disease.

First report of an acute purulent maxillary sinusitis caused by Pseudomonas aeruginosa secondary to dental implant placement in an immunocompetent patient.

STUDY DESIGN: In this case report, we present maxillary Pseudomonas aeruginosa sinusitis in an immunocompetent patient who underwent an autologous bone transplant for the insertion of dental implants. RESULTS: The infection was eradicated after removal of the dental implants and long-term antibiotic therapy. CONCLUSION: Despite the infection resolution, severe complications were observed with important legal consequences.

Antibacterial and anti-biofilm effects of cathelicidin peptides against pathogens isolated from cystic fibrosis patients.

Six different cathelicidin-derived peptides were compared to tobramycin for antibacterial and anti-biofilm effects against S. aureus, P. aeruginosa, and S. maltophilia strains isolated from cystic fibrosis patients. Overall, SMAP-29, BMAP-28, and BMAP-27 showed relevant antibacterial activity (MIC(50) 4-8µg/ml), and in some cases higher than tobramycin. In contrast, indolicidin, LL-37, and Bac7(1-35) showed no significant antimicrobial activity (MIC(50)>32µg/ml). Killing kinetics experiments showed that in contrast to tobramycin the active cathelicidin peptides exert a rapid bactericidal activity regardless of the species tested. All three peptides significantly reduced biofilm formation by S. maltophilia and P. aeruginosa strains at 1/2× MIC, although at a lower extent than tobramycin. In addition, BMAP-28, as well as tobramycin, was also active against S. aureus biofilm formation. Preformed biofilms were significantly affected by bactericidal SMAP-29, BMAP-27 and BMAP-28 concentrations, although at a lesser extent than tobramycin. Overall, our results indicate the potential of some cathelicidin-derived peptides for the development of novel therapeutic agents for cystic fibrosis lung disease.

Consequences of metaphase II oocyte cryopreservation on mRNA content.

INTRODUCTION: We studied the consequences of freezing/thawing processes on mRNA contents in MII oocytes after slow-freezing/rapid thawing (SF/RT) and vitrification/warming (V/W) protocols, and compared the results to fresh MII oocytes. We quantified the nuclear transcript mRNA responsible for the translation of proteins belonging either to trans-regulatory protein family or to functional structural proteins such as proteins involved in DNA structural organization (NAP1L1, TOP1, H1F0H1), chromosomal structure maintenance (SMC, SCC3, RAD21, SMC1A, SMC1B, STAG3, REC8), mitochondrial energetic pathways (ATP5GJ, SDHC), cell cycle regulation and processes (CLTA, MAPK6, CKS2) and staminal cell potency-development competence stage (DPPA3, OCT4, FOXJ2). MATERIAL AND METHODS: Surplus MII oocytes were donated from patients in IVF cycles and divided in three groups of 15 oocytes. Group 1 was comprised of non-cryopreserved oocytes and Groups 2 and 3 underwent SF/RT and V/W procedures, respectively. RESULTS: There was an overall decrease of mRNA extracted from cryopreserved oocytes compared to control group. Only 39.4% of mRNA content were preserved after SF/RT while 63.3% of mRNA content were maintained after V/W. CONCLUSIONS: Oocyte cryopreservation is associated with molecular injury associated with the decrease of stored mRNA. However the V/W protocol is more conservative than SF/RT resulting in a level of mRNA sufficient to maintain biologic functions in the subsequent fertilized oocyte.

Hsp10, Hsp70, and Hsp90 immunohistochemical levels change in ulcerative colitis after therapy.

Ulcerative colitis (UC) is a form of inflammatory bowel disease (IBD) characterized by damage of large bowel mucosa and frequent extra-intestinal autoimmune comorbidities. The role played in IBD pathogenesis by molecular chaperones known to interact with components of the immune system involved in inflammation is unclear. We previously demonstrated that mucosal Hsp60 decreases in UC patients treated with conventional therapies (mesalazine, probiotics), suggesting that this chaperonin could be a reliable biomarker useful for monitoring response to treatment, and that it might play a role in pathogenesis. In the present work we investigated three other heat shock protein/molecular chaperones: Hsp10, Hsp70, and Hsp90. We found that the levels of these proteins are increased in UC patients at the time of diagnosis and decrease after therapy, supporting the notion that these proteins deserve attention in the study of the mechanisms that promote the development and maintenance of IBD, and as biomarkers of this disease (e.g., to monitor response to treatment at the histological level).

Exposure to flaxseed during lactation does not alter prostate area or epithelium height but changes lipid profile in rats.

UNLABELLED: Flaxseed intake has increased owing to beneficial effects to health and prevention of diseases. Provided that it's an important source of lignan, a phytoestrogen, the present study aimed at evaluating the possible effect of the intake of this seed during lactation upon prostate, sexual hormones and lipidic profile of the offspring in adult life. MATERIAL AND METHODS: 16 female Wistar rats were used. After delivery, they were divided into two different groups to receive one of the following diets during lactation: Control group (CG), with a casein based diet and Flaxseed group (FG), with a flaxseed based diet containing 25% flaxseed. At weaning, male pups received commercial chow until adult life (170 days old), when they were sacrificed. RESULTS: No differences were perceived concerning offspring food intake and body weight at 170 days. There was a reduction in total cholesterol levels (FG = 45.71 +/- 8.96 mg/dL; CG = 63.43 +/- 15.69 mg/dL, p = 0.02) and triglycerides (FG = 54.29 +/- 11.10 mg/dL; CG = 79.86 +/- 25.68 mg/dL, p = 0.03). Also, no alterations were observed in prostatic morphology, testosterone or estradiol levels in the two groups analyzed. CONCLUSION: Flaxseed intake during lactation did not produce histological alterations in prostatic alveolus or in sexual hormones, but programmed to a reduction in lipid profile in adult life with decreased cardiovascular risk.

Exposure to flaxseed during lactation does not alter prostate area or epithelium height but changes lipid profile in rats / Exposición a la linaza durante la lactación no cambia el área o altura epitelial de la próstata pero cambia el perfil lipídico en ratones

Flaxseed intake has increased owing to beneficial effects to health and prevention of diseases. Provided that it's an important source of lignan, a phytoestrogen, the present study aimed at evaluating the possible effect of the intake of this seed during lactation upon prostate, sexual hormones and lipidic profile of the offspring in adult life. Material and methods: 16 female Wistar rats were used. After delivery, they were divided into two different groups to receive one of the following diets during lactation: Control group (CG), with a casein based diet and Flaxseed group (FG), with a flaxseed based diet containing 25% flaxseed. At weaning, male pups received commercial chow until adult life (170 days old), when they were sacrificed. Results: No differences were perceived concerning offspring food intake and body weight at 170 days. There was a reduction in total cholesterol levels (FG = 45.71 ± 8.96 mg/dL; CG = 63.43 ± 15.69 mg/dL, p = 0.02) and triglycerides (FG = 54.29 ± 11.10 mg/dL; CG = 79.86 ± 25.68 mg/dL, p = 0.03). Also, no alterations were observed in prostatic morphology, testosterone or estradiol levelsin the two groups analyzed. Conclusion: Flaxseed intake during lactation did not produce histological alterations in prostatic alveolus or in sexual hormones, but programmed to a reduction in lipid profile in adult life with decreased cardiovascular risk (AU)

El consumo de la linaza ha aumentado debido a los efectos beneficiosos para la salud y la prevención de enfermedades. Siendo una importante fuente de lignanos, un fitoestrógeno, el presente estudio evaluó los efectos de la administración de esta semilla durante la lactación en la próstata, las hormonas sexuales y perfil lipídico de loshijos en la vida adulta. Material y métodos: Fueron utilizados 16 ratones Wistar hembras. Después del parto fueron divididas en dos grupos recibiendo durante la lactación las siguientes dietas: Grupo Control (GC), con ración a base de caseína y Grupo Linaza (GL), con ración a base de caseína conteniendo 25% de semilla de linaza. En el destete, las crías machos pasaron a recibir ración comercial hasta la edad adulta, cuando fueron muertos a los 170 días de vida. Resultados: No fueron verificadas diferencias sobre el consumo alimentario y peso corporal de los animales a los 170 días. Hubo una reducción en los niveles de colesterol total (GL = 45,71 ± 8,96 mg/dL; GC = 63,43 ± 15,69 mg/dL, p = 0,02) y triglicéridos (GL = 54,29 ± 11,10 mg/dL; GC = 79,86 ± 25,68 mg/dL, p = 0,03). Además, no se observaron alteraciones en la morfología de la próstata, la testosterona o los niveles de estradiol en los dos grupos analizados. Conclusión: La administración de la semilla durante la lactación no promueve alteraciones histológicas en los alvéolos de próstata o en las hormonas sexuales, pero programado para una reducción en el perfil lipídico en la vida adulta con una disminución del riesgo cardiovascular (AU)

Subinhibitory concentrations of moxifloxacin decrease adhesion and biofilm formation of Stenotrophomonas maltophilia from cystic fibrosis.

Stenotrophomonas maltophilia is an emerging nosocomial bacterial pathogen that is currently isolated with increasing frequency from the airways of cystic fibrosis (CF) patients. In this study the effect of subinhibitory concentrations (subMICs) of moxifloxacin on adhesion, biofilm formation and cell-surface hydrophobicity of two strains of S. maltophilia isolated from CF patients were evaluated. Adhesion and biofilm formation assays were carried out on polystyrene and quantified by colony counts. Cell-surface hydrophobicity was determined by a test for adhesion to n-hexadecane. Moxifloxacin at 0.03x and 0.06x MIC caused a significant decrease in adhesion and biofilm formation by both strains tested. A significant reduction in cell-surface hydrophobicity following exposure to subMICs of moxifloxacin was observed for one strain only. The results of the present study provide an additional rationale for the use of moxifloxacin in CF patients and more generally in biofilm-related infections involving S. maltophilia.

Quick and reliable galactomannan detection in crude minced lung specimens from haematological patients with suspected invasive fungal infection: results from a case series.

Invasive aspergillosis (IA) is the leading direct or contributory cause of death in patients with haematological malignancies. Early diagnosis remains difficult and often elusive due the heterogeneity of clinical presentations and the low sensitivity of both histological examination and cultures of specimens obtained from patients at risk. We report two cases of IA, both of which lacked both histological and cultural evidence of IA from pulmonary specimens. In both patients, detection of galactomannan (GM) by enzyme immunoassay (EIA) on pulmonary tissue homogenates led to the diagnosis of IA, which was confirmed by Aspergillus DNA (real time PCR). In conclusion, we provide preliminary evidence that lung homogenates may be prepared for GM EIA assays, which may contribute to quick diagnosis of IA on otherwise negative samples. We feel that our results open up the opportunity of a prospective and comparative evaluation of this diagnostic technique.

Influence of temperature on biofilm formation by Listeria monocytogenes on various food-contact surfaces: relationship with motility and cell surface hydrophobicity.

AIMS: To assess the ability of Listeria monocytogenes to form biofilm on different food-contact surfaces with regard to different temperatures, cellular hydrophobicity and motility. METHODS AND RESULTS: Forty-four L. monocytogenes strains from food and food environment were tested for biofilm formation by crystal violet staining. Biofilm levels were significantly higher on glass at 4, 12 and 22 degrees C, as compared with polystyrene and stainless steel. At 37 degrees C, L. monocytogenes produced biofilm at significantly higher levels on glass and stainless steel, as compared with polystyrene. Hydrophobicity was significantly (P < 0.05) higher at 37 degrees C than at 4, 12 and 22 degrees C. Thirty (68.2%) of 44 strains tested showed swimming at 22 degrees C and 4 (9.1%) of those were also motile at 12 degrees C. No correlation was observed between swimming and biofilm production. CONCLUSIONS: L. monocytogenes can adhere to and form biofilms on food-processing surfaces. Biofilm formation is significantly influenced by temperature, probably modifying cell surface hydrophobicity. SIGNIFICANCE AND IMPACTS OF THE STUDY: Biofilm formation creates major problems in the food industry because it may represent an important source of food contamination. Our results are therefore important in finding ways to prevent contamination because they contribute to a better understanding on how L. monocytogenes can establish biofilms in food industry and therefore survive in the processing environment.

Molecular characterization of virulence determinants of Stenotrophomonas maltophilia strains isolated from patients affected by cystic fibrosis.

Stenotrophomonas maltophilia is an emerging nosocomial bacterial pathogen which is currently isolated with increasing frequency from the airways of cystic fibrosis (CF) patients. In this study 13 S. maltophilia strains (11 isolated from the airways of independent CF patients, and two non-CF respiratory reference strains) have been characterized for the expression of several virulence-associated factors. In particular, the ability to form biofilm on abiotic surfaces has been determined and correlated with different features, such as motility, adherence and the ability to invade A549 respiratory epithelial cells. Moreover, the presence of a flagellum-associated gene as well as that of the StmPr1 gene, which encodes an extracellular protease, have been determined by Southern blot hybridization. Our data indicate that the different degree of biofilm formation exhibited by the 11 CF isolates does not correlate with motility, ability to adhere to and invade A549 cells, or with the presence of flagella. On the other hand, among the CF isolates the StmPr1 gene was found only in two strains, both able to establish chronic lung infections in CF patients. Moreover, only four of the strains analyzed show a temperature-independent antibiotic-resistance profile, suggesting either a different origin of these strains or an intervening adaptation to host tissues.

Management and triage of women with human papillomavirus infection in follow-up for low-grade cervical disease: association of HPV-DNA and RNA-based methods.

Type-specific persistent infection with Human Papillomavirus (HPV) is a significant risk factor for the development of cervical diseases. Persistent infection could be further refined by a sequencing approach to detect early cervical lesions that are at high risk of developing an invasive squamous cervical cancer. The aim of the present study is to investigate the clinical utility of detecting mRNA transcripts of HPV oncogenes E6/E7 by using a Real-time NASBA technology (mRNA test) and to identify women with low-grade cytological disease but with an increased risk of developing high-grade cervical abnormalities or invasive squamous cervical cancer. Our preliminary results show that E6/E7 is detected in only a subset of HR-HPV-positive cases. Since viral persistence is considered to be the true precursor of neoplastic progression, only the detection of E6/E7 mRNA can identify the infection which is more likely to persist and induce neoplasia in future. For these reasons we believe that this test would be useful for the characterization of women with HR-HPV DNA positivity who should be effectively treated because at high-risk of developing a high grade cervical lesion or an invasive squamous cervical cancer.

Effect of environmental factors on biofilm formation by clinical Stenotrophomonas maltophilia isolates.

The influence of environmental factors (temperature, aerobiosis-anaerobiosis, static-dynamic conditions, pH) was determined on biofilm formation by 51 S. maltophilia clinical isolates. The strains produced more biofilm at 32 degrees C than at 37 or 18 degrees C. Aerobic and 6% CO2 atmosphere yielded comparable biofilm amounts, higher than under anaerobic conditions. Biofilm production was not affected by static vs. agitated culture conditions. Biofilm production at pH 7.5 and 8.5 was comparable but significantly higher than at pH 5.5. The capacity of individual strains to form biofilm and thus contribute to the severity of some diseases is influenced by host traits and environmental conditions at the site of infection, and play an important role in the pathogenesis of biomaterial-related disease caused by S. maltophilia.

Serum and mucosal cytokine profiles in patients with active Helicobacter pylori and ischemic heart disease: is there a relationship?

This study is designed to investigate, for the first time, circulating and gastric mucosal levels of IL1-alpha, IL-6, IL-8 and TNF-alpha in patients with ischemic heart disease (IHD) and matched controls, according to the presence or absence of active Helicobacter pylori infection. Furthermore, in order to evaluate whether modified lipid profile was associated to an increased cardiovascular risk, this was determined in the same groups. Cytokine levels were measured using ELISA in 58 patients with IHD and 52 controls. Active H. pylori infection was assessed if either culture of H. pylori or rapid urease test gave a positive result. Our findings indicate increasing cytokine mucosal levels in H. pylori-positive patients compared to H. pylori-negative subjects. However, the increase was statistically significant only for IL-6 and TNF-alpha in the gastric mucosa of IHD patients. In H. pylori-positive controls, IL-8 mucosal levels positively correlated with both IL1-alpha (r = 0.98; P = 0.0003) and IL-6 (r = 0.83; P = 0.03) levels. Circulating cytokine levels were comparable in IHD and healthy subjects, regardless of H. pylori status. There were no correlations between mucosal and circulating cytokine levels. Active H. pylori infection was not associated with a modified lipid profile in either controls or IHD patients, although ApoAI levels were significantly higher in H. pylori-positive controls compared to those H. pylori-negative. Taken together, the results of the present study provide evidence that active H. pylori infection may play a role as a trigger factor in the pathophysiology of IHD by inducing an inflammatory cascade concentrated on gastric mucosa.