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1.
Benef Microbes ; 9(3): 389-399, 2018 Apr 25.
Article in English | MEDLINE | ID: mdl-29633636

ABSTRACT

Current research into original therapies to treat intestinal inflammation is focusing on no-drug therapies. KLD is a mixture of krill oil (KO), probiotic Lactobacillus reuteri (LR), and vitamin D (VitD3). The aim of this study was to assess in vitro and in vivo the potential cooperative effects of KLD in reducing gut inflammation. Colorectal adenocarcinoma cell lines, CACO2 and HT29, and C57BL/6 mice were used for in vitro and in vivo analyses, respectively. Cells were exposed to cytomix (interferon gamma + tumour necrosis factor alpha (TNF-α)) to induce inflammation or co-exposed to cytomix and KO, LR and VitD3 alone or to cytomix and KLD. Animals were treated for 7 days with dextran sodium sulphate (DSS) to induce colitis or with DSS and KLD. In vitro assays: F-actin expression was analysed by immunofluorescence; scratch test and trans-epithelial electric resistance test were performed to measure wound healing; adhesion/invasion assays of adhesive and invasive Escherichia coli (AIEC) bacteria were made; mRNA expression of TNF-α, interleukin (IL)-8 and vitamin D receptor (VDR) was detected by quantitative PCR. In vivo assays: body weight, clinical score, histological score and large intestine weight and length were estimated; mRNA expression of TNF-α, IL-1ß, IL-6, IL-10 by quantitative PCR; VDR expression was detected by quantitative PCR and immunohistochemistry. In vitro: KLD restores epithelial cell-cell adhesion and mucosal healing during inflammation, while decreases the adhesiveness and invasiveness of AIEC bacteria and TNF-α and IL-8 mRNA expression and increases VDR expression. In vivo: KLD significantly improves body weight, clinical score, histological score and large intestine length of mice with DSS-induced colitis and reduces TNF-α, IL-1ß and IL-6 mRNA levels, while increases IL-10 mRNA and VDR levels. KLD has significant effects on the intestinal mucosa, strongly decreasing inflammation, increasing epithelial restitution and reducing pathogenicity of harmful commensal bacteria.


Subject(s)
Anti-Inflammatory Agents/administration & dosage , Colitis/therapy , Drug Synergism , Limosilactobacillus reuteri/growth & development , Oils/administration & dosage , Probiotics/administration & dosage , Vitamin D/administration & dosage , Animals , Anti-Inflammatory Agents/pharmacology , Bacterial Adhesion , Body Weight , Cell Line , Colitis/chemically induced , Colitis/pathology , Cytokines/analysis , Dextran Sulfate/administration & dosage , Dextran Sulfate/toxicity , Disease Models, Animal , Euphausiacea , Histocytochemistry , Humans , Mice, Inbred C57BL , Models, Biological , Oils/pharmacology , Probiotics/pharmacology , Treatment Outcome , Vitamin D/pharmacology
2.
Clin Endocrinol (Oxf) ; 83(5): 656-62, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26201937

ABSTRACT

OBJECTIVE: The aim of this study was to evaluate the metabolic effects of fatty pancreas (nonalcoholic fatty pancreas disease - NAFPD) in a group of obese paediatric patients with nonalcoholic fatty liver disease (NAFLD). METHODS: We included 121 consecutive children with echographic evidence of hepatic steatosis. All patients underwent to abdominal ultrasound to evaluate pancreatic echogenic pattern. We divided the patients into two groups on the basis of the presence of fatty pancreas. In all patients liver function tests, lipid and gluco-insulinemic profile were evaluated. A selected subset of patients (67) underwent to liver biopsy. RESULTS: Of these 121 patients, 58 showed NAFPD and 63 patients exhibited a normal pancreatic echogenic pattern. No differences were found in age, transaminases serum levels, lipid profile and pancreatic enzymes between the two groups. The patients with NAFPD had a significantly higher z-BMI, fasting insulin, insulin resistance (HOMA-IR) and lower ISI respect to the group without fatty pancreas. The patients with fatty pancreas showed a more advanced form of liver disease, with higher values of fibrosis, ballooning and NAS score with respect to the group without NAFPD. CONCLUSIONS: Our study demonstrated that NAFPD is a frequent condition in obese paediatric patients affected by NAFLD. Our data suggest that pancreatic fat should not be considered an inert accumulation of fat, but as an additional factor able to affect glucose metabolism and severity of liver disease, increasing the risk of develop metabolic syndrome.


Subject(s)
Non-alcoholic Fatty Liver Disease/epidemiology , Pancreatic Diseases/epidemiology , Adiposity , Adolescent , Anthropometry , Biopsy , Cytokines/blood , Female , Humans , Italy/epidemiology , Liver/pathology , Male , Metabolic Syndrome/complications , Non-alcoholic Fatty Liver Disease/blood , Non-alcoholic Fatty Liver Disease/complications , Non-alcoholic Fatty Liver Disease/diagnosis , Pancreatic Diseases/blood , Pancreatic Diseases/diagnosis , Pancreatic Diseases/etiology
3.
Neurogastroenterol Motil ; 26(2): 196-204, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24304324

ABSTRACT

BACKGROUND: Neuroimmune interactions and inflammation have been proposed as factors involved in sensory-motor dysfunction and symptom generation in adult irritable bowel syndrome (IBS) patients. In children with IBS and healthy controls, we measured ileocolonic mast cell infiltration and fecal calprotectin and evaluated the relationships between these parameters and abdominal pain symptoms and stooling pattern. METHODS: Irritable bowel syndrome patients diagnosed according to Pediatric Rome III criteria and healthy controls kept a 2-week pain/stooling diary. Ileocolonic mucosal mast cells (MC) and MC in close proximity to nerve fibers (MC-NF) were identified immunohistochemically and quantified. Fecal calprotectin concentration was measured. KEY RESULTS: 21 IBS patients and 10 controls were enrolled. The MC-NF count was significantly higher in the ileum (p = 0.01), right colon (p = 0.04), and left colon (p < 0.001) of IBS patients compared with controls. No differences in fecal calprotectin concentration were noted. Abdominal pain intensity score correlated with ileal MC count (r(s) = 0.47, p = 0.030) and right colon MC-NF count (r(s) = 0.52, p = 0.015). In addition, children with IBS with >3 abdominal pain episodes/week had greater ileal (p = 0.002) and right colonic (p = 0.01) MC counts and greater ileal (p = 0.05) and right colonic (p = 0.016) MC-NF counts than children with less frequent pain. No relationship was found between MC and MC-NF and fecal calprotectin or stooling pattern. CONCLUSIONS & INFERENCES: Mast cells-nerve fibers counts are increased in the ileocolonic mucosa of children with IBS. Mast cells and MC-NF counts are related to the intensity and frequency of abdominal pain.


Subject(s)
Abdominal Pain/etiology , Irritable Bowel Syndrome/etiology , Neuroimmunomodulation , Abdominal Pain/immunology , Abdominal Pain/pathology , Adolescent , Child , Child, Preschool , Female , Humans , Intestines/pathology , Irritable Bowel Syndrome/diagnosis , Irritable Bowel Syndrome/immunology , Irritable Bowel Syndrome/pathology , Leukocyte L1 Antigen Complex/analysis , Male , Mast Cells/pathology , Nerve Fibers/pathology
4.
Aliment Pharmacol Ther ; 35(3): 327-34, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22150569

ABSTRACT

BACKGROUND: Intestinal microbiota manipulation, one of the pathogenetic components of inflammatory bowel disease (IBD), has become an attractive therapy for ulcerative colitis (UC). AIM: To assess in children with active distal UC the effectiveness of Lactobacillus (L) reuteri ATCC 55730 enema on inflammation and cytokine expression of rectal mucosa. METHODS: A total of 40 patients (median age: 7.2 years range 6-18) with mild to moderate UC were enrolled in a prospective, randomised, placebo-controlled study. They received an enema solution containing 10(10) CFU of L. reuteri ATCC 55730 or placebo for 8 weeks, in addition to oral mesalazine. Clinical endoscopic and histological scores as well as rectal mucosal expression levels of IL-10, IL-1ß, TNFα and IL-8 were evaluated at the beginning and at the end of the trial. RESULTS: Thirty-one patients accomplished the trial (17 males, median age 13 year, range 7-18). Mayo score (including clinical and endoscopic features) decreased significantly in the L. reuteri group (3.2 ± 1.3 vs. 8.6 ± 0.8, P < 0.01) compared with placebo (7.1 ± 1.1 vs. 8.7 ± 0.7, NS); furthermore, histological score significantly decrease only in the L. reuteri group (0.6 ± 0.5 vs. 4.5 ± 0.6, P < 0.01) (placebo: 2.9 ± 0.8 vs. 4.6 ± 0.6, NS). At the post-trial evaluation of cytokine mucosal expression levels, IL-10 significantly increased (P < 0.01) whereas IL-1ß, TNFα and IL-8 significantly decreased (P < 0.01) only in the L. reuteri group. CONCLUSIONS: In children with active distal ulcerative colitis, rectal infusion of L. reuteri is effective in improving mucosal inflammation and changing mucosal expression levels of some cytokines involved in the mechanisms of inflammatory bowel disease.


Subject(s)
Colitis, Ulcerative/therapy , Enema/methods , Limosilactobacillus reuteri , Probiotics/administration & dosage , Administration, Rectal , Adolescent , Child , Cytokines/metabolism , Double-Blind Method , Female , Humans , Intestinal Mucosa/metabolism , Male , Statistics as Topic , Treatment Outcome
5.
Aliment Pharmacol Ther ; 32(8): 1017-22, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20937047

ABSTRACT

BACKGROUND: Patients with ulcerative colitis often receive thiopurines as immunomodulators (IMs) to maintain remission and avoid corticosteroids. If unresponsive or intolerant to these agents, patients are treated with methotrexate, an antimetabolite never assessed in paediatric ulcerative colitis. AIM: To describe the experience with methotrexate in children with ulcerative colitis. METHODS: Thirty-two patients (median age 13.9 years) received methotrexate. Pediatric Ulcerative Colitis Activity Index (PUCAI) and use of corticosteroids were the main outcomes evaluated at baseline and at 3, 6 and 12 months. RESULTS: Indications to methotrexate were azathioprine unresponsiveness in 18 patients, azathioprine intolerance/toxicity in 10 and spondyloarthropathy in four. Response or remission was achieved in 72%, 63% and 50% of patients at 3, 6 and 12 months respectively. Mean PUCAI were 49.5 ± 23.3 at baseline and 32.9 ± 21.9, 29.5 ± 21.8 and 29.4 ± 19.9 at 3, 6 and 12 months respectively (P: 0.03). At the beginning of methotrexate, 16 patients (50%) received corticosteroids that were discontinued in 13 of them (81%) by 6 months. At the end of the study, 11 patients (33%) needed short courses of corticosteroids for disease relapse. CONCLUSIONS: Methotrexate may be useful in treating children with ulcerative colitis, although large, controlled trials are warranted to define better its effectiveness.


Subject(s)
Anti-Inflammatory Agents/therapeutic use , Antimetabolites, Antineoplastic/therapeutic use , Colitis, Ulcerative/drug therapy , Methotrexate/therapeutic use , Adolescent , Adrenal Cortex Hormones/therapeutic use , Child , Female , Humans , Longitudinal Studies , Male , Remission Induction , Retrospective Studies
6.
Int J Radiat Biol ; 82(5): 339-46, 2006 May.
Article in English | MEDLINE | ID: mdl-16782651

ABSTRACT

PURPOSE: The possibility of genotoxicity of radiofrequency radiation (RFR) applied alone or in combination with x-rays was investigated in vitro using several assays on human lymphocytes. The chosen specific absorption rate (SAR) values are near the upper limit of actual energy absorption in localized tissue when persons use some cellular telephones. The purpose of the combined exposures was to examine whether RFR might act epigenetically by reducing the fidelity of repair of DNA damage caused by a well-characterized and established mutagen. METHODS: Blood specimens from 14 donors were exposed continuously for 24 h to a Global System for Mobile Communications (GSM) basic 935 MHz signal. The signal was applied at two SAR; 1 and 2 W/Kg, alone or combined with a 1-min exposure to 1.0 Gy of 250 kVp x-rays given immediately before or after the RFR. The assays employed were the alkaline comet technique to detect DNA strand breakage, metaphase analyses to detect unstable chromosomal aberrations and sister chromatid exchanges, micronuclei in cytokinesis-blocked binucleate lymphocytes and the nuclear division index to detect alterations in the speed of in vitro cell cycling. RESULTS: By comparison with appropriate sham-exposed and control samples, no effect of RFR alone could be found for any of the assay endpoints. In addition RFR did not modify any measured effects of the x-radiation. CONCLUSIONS: This study has used several standard in vitro tests for chromosomal and DNA damage in Go human lymphocytes exposed in vitro to a combination of x-rays and RFR. It has comprehensively examined whether a 24-h continuous exposure to a 935 MHz GSM basic signal delivering SAR of 1 or 2 W/Kg is genotoxic per se or whether, it can influence the genotoxicity of the well-established clastogenic agent; x-radiation. Within the experimental parameters of the study in all instances no effect from the RFR signal was observed.


Subject(s)
Cell Phone , Chromosome Aberrations/radiation effects , Chromosomes, Human/radiation effects , Lymphocytes/pathology , Lymphocytes/radiation effects , Microwaves/adverse effects , Adult , Cells, Cultured , Chromosomes, Human/genetics , Dose-Response Relationship, Radiation , Female , Humans , Male , Middle Aged , Mutagenicity Tests , Radiation Dosage
7.
Bioelectromagnetics ; 25(8): 613-9, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15515032

ABSTRACT

The question whether extremely low frequency magnetic fields (ELFMFs) may contribute to mutagenesis or carcinogenesis is of current interest. In order to evaluate the possible genotoxic effects of ELFMFs, human blood cells from four donors were exposed in vitro for 48 h to 50 Hz, 1 mT uniform magnetic field generated by a Helmholtz coil system. Comet assay (SCGE), sister chromatid exchanges (SCE), chromosome aberrations (CAs), and micronucleus (MN) test were used to assess the DNA damage. ELF pretreated cells were also irradiated with 1 Gy of X-ray to investigate the possible combined effect of ELFMFs and ionizing radiation. Furthermore, nuclear division index (NDI) and proliferation index (PRI) were evaluated. Results do not evidence any DNA damage induced by ELFMF exposure or any effect on cell proliferation. Data obtained from the combined exposure to ELFMFs and ionizing radiation do not suggest any synergistic or antagonistic effect.


Subject(s)
Chromosomes, Human/radiation effects , Cytogenetic Analysis/methods , Electromagnetic Fields/adverse effects , Lymphocytes/radiation effects , Mutagenicity Tests/methods , Radiation, Nonionizing , Adult , Cell Proliferation/radiation effects , Cells, Cultured , Chromosome Aberrations/radiation effects , Humans , Lymphocytes/pathology , Male , Radiation Dosage , Reproducibility of Results , Sensitivity and Specificity
8.
Bioelectromagnetics ; 25(1): 41-8, 2004 Jan.
Article in English | MEDLINE | ID: mdl-14696052

ABSTRACT

In the past, epidemiological studies indicated a possible correlation between the exposure to ELF fields and cancer. Public concern over possible hazards associated with exposure to extremely low frequency magnetic fields (ELFMFs) stimulated an increased scientific research effort. More recent research and laboratory studies, however, have not been able to definitively confirm the correlation suggested by epidemiological studies. The aim of this study was to evaluate the effects of 50 Hz magnetic fields in human blood cells exposed in vitro, using several methodological approaches for the detection of genotoxicity. Whole blood samples obtained from five donors were exposed for 2 h to 50 Hz, 1 mT uniform magnetic field generated by a Helmholtz coil system. Comet assay, sister chromatid exchanges (SCE), chromosome aberrations (CA), and micronucleus (MN) tests were used to assess DNA damage, one hallmark of malignant cell transformation. The effects of a combined exposure with X-rays were also evaluated. Results obtained do not show any significant difference between ELFMFs exposed and unexposed samples. Moreover, no synergistic effect with ionizing radiation has been observed. A slight but significant decrease of cell proliferation was evident in ELFMFs treated samples and samples subjected to the combined exposure.


Subject(s)
Chromosome Aberrations/radiation effects , Chromosomes, Human/radiation effects , DNA/radiation effects , Electromagnetic Fields , Leukocytes/radiation effects , Micronuclei, Chromosome-Defective/radiation effects , Mutagenicity Tests/methods , Sister Chromatid Exchange/radiation effects , Adult , Cells, Cultured , Comet Assay , Dose-Response Relationship, Radiation , Humans , Male , Micronucleus Tests , Radiation Dosage , Single-Blind Method
9.
Cancer ; 92(9): 2484-92, 2001 Nov 01.
Article in English | MEDLINE | ID: mdl-11745306

ABSTRACT

BACKGROUND: The Ku protein is a tightly associated heterodimer, comprised of 70-kilodalton (kD) and 86-kD subunits, that forms the DNA-dependent protein kinase (DNA-PK) complex together with the 470-kD DNA-PKcs catalytic subunit, and is involved mainly in DNA double-strand breaks (DSBs) repair. The objective of the current study was to investigate the expression and DNA-binding activity of the Ku protein in fresh tissues from patients with bladder carcinoma and to compare it with that in nontumor tissues obtained from the same organ. Moreover, the DNA-binding activity of Ku was assessed after exposure of the tumor cells to 1 or 2 grays (Gy) of X-rays. Furthermore, the level of phosphorylated Ku was analyzed in both the nuclear and cytoplasmic compartment of normal tissue after exposure to 2 Gy of X-rays. METHODS: The expression and DNA-binding activity of Ku protein were assessed in tumor samples from patients who all were diagnosed with transitional cell carcinoma (TCC) of the bladder using Western blot analysis and the electrophoretic mobility shift assay, respectively. RESULTS: Enhanced Ku activity and expression were found in tumor tissue compared with normal tissue for each patient. Moreover, variations in Ku activity were found in a dose-dependent manner after the tumor cells were exposed to 1 or 2 Gy of X-rays. A decrease in phosphorylated Ku in the cytoplasm and a parallel increase in the nucleus of normal tissue cells were observed after exposure to X-rays. CONCLUSIONS: The results of the current study suggest a possible role of Ku in regulating the DNA-PK activity of DSBs repair in bladder tumors.


Subject(s)
Antigens, Nuclear , Carcinoma, Transitional Cell/genetics , DNA Helicases , DNA-Binding Proteins/biosynthesis , Gene Expression Regulation, Neoplastic , Nuclear Proteins/biosynthesis , Protein Serine-Threonine Kinases/metabolism , Urinary Bladder Neoplasms/genetics , Aged , Aged, 80 and over , Blotting, Western , Carcinoma, Transitional Cell/pathology , DNA Adducts , DNA Probes , DNA Repair , DNA-Activated Protein Kinase , Electrophoresis, Polyacrylamide Gel , Female , Gene Expression Regulation, Neoplastic/radiation effects , Humans , Ku Autoantigen , Male , Middle Aged , Protein Serine-Threonine Kinases/biosynthesis , Radiation Injuries , Tumor Cells, Cultured , Urinary Bladder Neoplasms/pathology
10.
Radiat Environ Biophys ; 40(2): 125-9, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11484783

ABSTRACT

The Semipalatinsk region (Kazakhstan Republic) has been affected by extensive radioactive contamination due to more than 450 nuclear tests of which almost 100 were exploded in the atmosphere. The present results refer to cytogenetic assessments in a study cohort of the population of Dolon, a settlement located on the NE boundary of the nuclear weapon test site, which was exposed to elevated doses of ionising radiation primarily due to the first Soviet nuclear test in 1949. Conventional cytogenetic analyses were carried out on 21 blood samples from individuals (more than 50 years old) living in Dolon since the very beginning of nuclear testing. A matched control group included 20 individuals living in non-contaminated areas. Higher frequencies of chromosome aberrations were found in the Dolon cohort compared to the control group, even though they remain within the range of the background levels reported for large normal human population studies on elderly individuals.


Subject(s)
Chromosome Aberrations , Environmental Exposure/adverse effects , Nuclear Warfare , Adult , Aged , Case-Control Studies , Female , Humans , Kazakhstan , Male , Middle Aged
11.
Radiat Prot Dosimetry ; 94(4): 335-45, 2001.
Article in English | MEDLINE | ID: mdl-11499437

ABSTRACT

Dose-response curves were measured for the induction of chromosomal aberrations in peripheral blood lymphocytes after acute exposure in vitro to 60Co gamma rays. Blood was obtained from four different healthy donors, and chromosomes were either observed at metaphase, following colcemid accumulation, or prematurely condensed by calyculin A. Cells were analysed in three different Italian laboratories. Chromosomes 1, 2, and 4 were painted, and simple-type interchanges between painted and non-painted chromosomes were scored in cells exposed in the dose range 0.1-3.0 Gy. The chemical-induced premature chromosome condensation method was also used combined with chromosome painting (chromosome 4 only) to determine calibration curves for high dose exposures (up to 20 Gy X rays). Calibration curves described in this paper will be used in our laboratories for biological dosimetry by fluorescence in situ hybridisation.


Subject(s)
Chromosome Aberrations , Chromosome Disorders , In Situ Hybridization, Fluorescence , Lymphocytes/radiation effects , Adult , Calibration , Chi-Square Distribution , Cobalt Radioisotopes , Dose-Response Relationship, Radiation , Gamma Rays , Humans , Male , Radiation Dosage
12.
Mutat Res ; 401(1-2): 193-7, 1998 Jun 05.
Article in English | MEDLINE | ID: mdl-9639704

ABSTRACT

As a result of the activities of the first Soviet plutonium production reactor, large territories of the Southern Urals were exposed to radioactive contamination. Three different incidents occurring between 1948 and 1967 lead to major exposure. A total of 280,000 people residing on the contaminated territories were exposed both to external and internal contamination particularly due to the long-lived radionuclides 137Cs and 90Sr. The highest doses were received by 28,000 people living on the Techa riverside villages. In the present paper 15 presumably exposed children coming from the Muslyumovo village on the Techa river have been analyzed using conventional cytogenetic procedure in order to assess a radiation-induced damage. The data obtained have been compared to a group of matched unexposed controls. The results show a statistical difference between the two cohorts which suggests a possible residual contamination representing a continuous hazard for the new generations.


Subject(s)
Blood Cells/radiation effects , Chromosome Aberrations , Nuclear Reactors , Radioactive Hazard Release , Cesium Radioisotopes , Child , Cohort Studies , Environmental Pollution , Female , Humans , Male , Metaphase , Russia , Strontium , USSR
13.
Mutat Res ; 395(2-3): 249-54, 1997 Dec 12.
Article in English | MEDLINE | ID: mdl-9465937

ABSTRACT

In a previous paper we reported that a group of children exposed to ionizing radiation following the Chernobyl accident exhibited an appreciable number of chromosome breaks and rearrangements reflecting the persistence of a radiation-induced damage. The results suggested that the children were still exposed to radioactive contamination through consumer foodstuff and life styles. In the present paper, 31 exposed children have been considered together with a control group of 11 children with the aim to confirm previous results. All children underwent whole-body counter (WBC) measures and conventional cytogenetic analysis. The frequency of chromosome aberrations detected by conventional cytogenetics in the group of children chronically exposed to low doses of ionizing radiation resulted in significant differences with respect to the control group. The present work suggests that, for these groups of children, even if the frequency of aberrations is very low and the observation of statistically significant differences is consequently a problem, a persistently abnormal cytogenetic picture is still present several years after the accident.


Subject(s)
Chromosome Aberrations , Lymphocytes/radiation effects , Radioactive Hazard Release , Child , Female , Humans , Male , Nuclear Reactors , Power Plants , Radiation Dosage , Republic of Belarus , Ukraine
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