Are the dietary habits of treated individuals with celiac disease adherent to a Mediterranean diet?

BACKGROUND AND AIMS: The only treatment for celiac disease (CD) is strict, lifelong adherence to a gluten-free (GF) diet. To date, there are contrasting data concerning the nutritional adequacy of GF products and diet. There have been no studies that have assessed the adherence of individuals with CD to a Mediterranean diet (MD), a protective dietary regimen against major non-communicable diseases (NCDs). Therefore, we examined the adherence to an MD of a group of Italian individuals with CD and compared it with that of a healthy control group. METHODS AND RESULTS: In a cross-sectional study, a sample of individuals with CD and a group of healthy subjects were included. The dietary habits of all participants were recorded using a validated food frequency questionnaire, and the adherence to an MD was determined using the Italian Mediterranean Index. Typical Mediterranean food consumption was not significantly different between individuals with CD and the healthy participants, except for fruits (P = 0.017). However, individuals with CD consumed significantly higher amounts of potatoes (P = 0.003) and red and processed meat (P = 0.005) than healthy participants. The resulting mean Italian Mediterranean Index was significantly higher in healthy participants than in individuals with CD (P < 0.001). CONCLUSION: The results raise questions concerning the food choices of individuals with CD, suggesting the need of encouraging them to make better food choices more in line with an MD, which would improve their nutritional status and better protect them from NCDs at long term. PROTOCOL REGISTRATION: ClinicalTrials.gov (ID NCT01975155) on November 4 2013.

Use of a proline-specific endopeptidase to reintroduce gluten in patients with non-coeliac gluten sensitivity: A randomized trial.

BACKGROUND: A gluten-free diet (GFD) is the main therapy for non-coeliac gluten sensitivity (NCGS). However, the availability of novel enzymes with the ability to digest gluten could represent a therapeutic opportunity for NCGS patients to avoid a GFD. AIMS: To evaluate the controlled reintroduction of gluten with or without the endopeptidase P1016 in NCGS patients. METHODS: This is a randomized, double-blind, placebo-controlled monocentric study, Registered under ClinicalTrials.gov Identifier no. NCT01864993. Gluten was reintroduced incrementally over a 3-week period under nutritional control. NCGS patients were randomized into two groups and administered P1016 or placebo during gluten reintroduction. We evaluated symptoms (visual analogue scale, VAS), quality of life (SF-36) and mental health symptoms (SCL-90) on a weekly basis. RESULTS: We enrolled a total 23 patients who were allocated to a placebo group (n = 11, age 38.4 ± 2.9) or an intervention group (n = 12, age 39.5 ± 3.1). No effect of P1016 on symptoms was found. During gluten reintroduction, patients reported a significant increase in abdominal pain and a worsening of stool consistency. Furthermore, no differences were found between the groups regarding SCL-90 and SF-36 scores. CONCLUSIONS: Our results demonstrate a lack of effect of P1016 in the management of NCGS patients and the possible reintroduction of gluten.

In situ hybridization analysis of interstitial C-heterochromatin in marker chromosomes of two human melanomas.

Two distinct marker chromosomes, presenting with intercalated C- and distamycin A-Dapi-positive regions, were observed in a metastatic and a primary melanoma. To establish the origin of these heterochromatic sequences, we performed in situ hybridization analysis using specific probes for human repetitive DNA. The marker of the primary melanoma, m2, a der 16 chromosome resulting from the translocation of the 1q12-qter segment to band q23 of chromosome 16, showed specific hybridization with Sau3A but not with EcoRI sequences at the interstitial C-band. Thus the origin of this region from the normal chromosome 1 pericentromeric heterochromatin, containing both EcoRI and Sau3A sequences, could be established. On the other hand, the marker of the metastatic melanoma, m1, a der 1 chromosome showing an abnormally banded region inserted between 1q11 and 1q21-qter, failed to give any hybridization signals at the C- and distamycin A-Dapi-positive band when the same EcoRI and Sau3A probes were used. Furthermore, no hybridization was observed using either a probe for SatIII-specific sequences (QP23), mapping to chromosome 9 heterochromatic block, or LS6BB, a ribosomal DNA probe. From these data we speculate that more complex molecular rearrangements may have occurred during the transposition of heterochromatin from its original site to m1. The heterochromatin change found in m1 may be related to advanced stages of malignancy.

GKLF in thymus epithelium as a developmentally regulated element of thymocyte-stroma cross-talk.

Gut-enriched Krüppel-like factor (GKLF) is a transcriptional regulator expressed in differentiated epithelia. We identified GKLF transcript as a regulated element in thymic epithelium of recombinase-deficient mice during thymus development induced by anti-CD3 antibody injection. This treatment recapitulates the organogenetic process depending on productive rearrangement of T cell receptor (TCR) beta gene with thymocytes expansion and acquisition of the CD4+8+ double positive phenotype. In wildtype mice, GKLF is expressed very early in embryogenesis and becomes intensely up-regulated in thymus epithelium at day 18 of gestation when TCR beta expressing cells have selectively expanded and express both CD4 and CD8. The results presented here suggest that thymocytes may regulate GKLF transcriptionally in the cortical epithelium at the developmental check-point controlled by TCR beta gene rearrangement. Furthermore, GKLF expression in hematopoietic stroma might suggest the thus far uncharacterised participation of this factor in hematopoiesis.

Early and delayed reproductive death in human cells exposed to high energy iron ion beams.

The aim of this research was to determine the biological effectiveness for early and delayed effects of high energy, high linear energy transfer (LET) charged particles. Survival and delayed reproductive death were measured in AG1522 human fibroblast cells exposed to Fe-ion beams of energies between 0.2 and 1 GeV/n, 0.97 GeV/n Ti-ion and 0.49 GeV/n Si-ion beams. The cells were irradiated at the HIMAC accelerator in Chiba, Japan (0.2 and 0.5 GeV/n Fe and 0.49 GeV/n Si) and at the NASA Space Radiation Laboratory in Brookhaven, USA (1 GeV/n Fe and 0.97 GeV/n Ti ions). The dose-effect curves were measured in the dose range between 0.25 and 2 Gy. For comparison cells were exposed to 60Co gamma rays. Analysis of the dose-effect curves show that all the heavy ion beams induce inactivation and delayed reproductive death more effectively than 60Co gamma rays. The only exception is the 0.2 GeV/n Fe-ion beam at low doses. The progeny of the irradiated cells show delayed damage in the form of reproductive death with all the heavy ion beams with the 1 GeV/n Fe-ion beam being the most effective. The relative biological effectiveness at low doses of the iron beams is highest for LET values between 140 and 200 keV/micrometers with values of 1.6 and 3 for early and delayed reproductive death, respectively. Analysis of the fluence-effect curves shows that the cross-sections for early and delayed inactivation increase with increasing LET up to 442 keV/micrometers.

High incidence of chromosomal lesions involving C-heterochromatin in four human melanoma lines.

Cytogenetic analysis of early in vitro cultures derived from human melanomas, two primary tumors (Me 10538, Me 1402) and two metastatic lesions in the same patient (Me 665/1, Me 665/2) showed non-random involvement of C-heterochromatin in clonal chromosome rearrangements. Marker chromosomes with C- and DA-Dapi-positive bands were identified in one of the metastases, Me 665/1 (m1) and in the two primary tumors, Me 10538 (m2) and Me 1402 (m3). C-positive fragments predominated in the other metastasis, Me 665/2, which lacked C-regions intercalated in rearranged chromosomes, and were also detected with appreciable frequency in the Me 665/1 and Me 1402 cells. The frequencies of marker chromosomes and their mean number per cell allowed m2 and m1 to be considered as early markers of tumor formation and m3 as a marker of tumor progression. Dissection of chromosome structure, including the origin of the intercalated C-band, has so far been achieved only with the m2 chromosome of the primary tumor Me10538. This was the only cell line which displayed few C-fragments and a narrow chromosomal distribution with a well defined mode. A gradient of malignancy could be detected in the four cell lines, by local and disseminated tumor growth in xenotransplanted mice, with the two primary melanomas 10538 and the 1402 cells at the lowest and upper extremes. This gradient closely parallels the increase in cytogenetic heterogeneity and C-heterochromatin lesions from the 10538 to the 1402 cells.

Role of heterochromatin variation in the instability of a marker chromosome during tumor progression.

Karyotypic evolution of the poorly metastasizing tumorigenic RSV-transformed B77-3T3 fibroblast line was investigated both in highly metastasizing clones (selected by growth in hard agar) and in spontaneous metastases. Analysis of structural chromosome aberrations associated with the transition from the nonmetastatic to the metastatic phenotype was focused on a readily identifiable marker chromosome (A), displaying an extracentromeric heterochromatic region as a main feature promoting genetic instability. Well-defined changes in the structure of this marker were observed, both in vitro and in vivo, and invariably involved C-heterochromatic variation. In the metastatic clones, a specific rearrangement of the A chromosome was selected. This structural variant (B) showed two extracentromeric C-positive regions and probably originated from duplication of the segment of A included between the centromere and the internal C-band. On the other hand, selection of a modified form of chromosome A, not displaying the interpolated C-heterochromatin, had occurred in the extremely rare B77-3T3 spontaneous metastases. The connection among heterochromatin variants, genetic instability, and chromosome aberrations is discussed.

Cytogenetic instability in a family with gastric cancer recurrence.

An index case with a congenital malformation syndrome enabled detection of a family that had a previous history of spontaneous abortuses and recurrence of neoplasia through three generations. Cytogenetic analysis performed on lymphocytes from 11 subjects in the second and third generation showed karyotypic alterations in both tumor bearers and apparently normal subjects. Chromosome variations consisted of: spontaneous chromosome fragility; chromosome translocations; polymorphisms in the heterochromatic regions in chromosomes Y, #1, #16, #22. The inheritance pattern of all chromosome rearrangements and heteromorphisms observed was established starting with the second generation, and the contribution of specific individuals was identified. Although the relationship between chromosomal instability and predisposition to gastric cancer does not appear to be coincidental, no specific chromosome alteration in normal somatic cells was shared by all members of the family who developed or are at risk of developing tumors.

Mosaicism in the C-banded region of chromosome 1 in cancer families.

Chromosome 1 C-band length mosaicism was detected in lymphocytes from six tumor patients and one healthy subject belonging to three families with a high incidence of cancer. In all cases the variant cell population showed a decreased amount of C-heterochromatin in one chromosome, whereas the C-banded pattern of the homolog was identical to that of the nonvariant cell population. A family tendency to unequal mitotic crossing over, possibly leading to C-band heteromorphism, may explain the high frequency of detection of C-heterochromatin mosaicism in cancer family members (seven of 13 cases studied). The possible role of heterochromatin in inducing cancer has been widely discussed. The special feature of the acquired C-band variants vis-à-vis the inherited ones is that they mark intrinsic genetic instability that may result, through multiple mechanisms, in increased susceptibility to malignancy.

Malformation syndrome with t(2;22) in a cancer family with chromosome instability.

A de novo unbalanced t(2;22)(q37;q11.2) [corrected], resulting in the deletion of the 22pter-q11 and 2q37-qter regions, was observed in a 12-year-old girl born with a congenital malformation syndrome and later displaying signs of neurologic impairment. Some of the clinical signs observed appear to overlap those found in subjects monosomic in the 22q11 region affected by the DiGeorge syndrome. The chromosomal rearrangement observed may be related to a familial cytogenetic instability that also gives rise to sustained cancer predisposition.

Multicolor FISH in chronic lymphocytic leukemia. An interphase study of patients with early-onset disease.

Trisomy 12 and deletions of 13q14.2 and 14q32 are the most common chromosome abnormalities in patients with B-chronic lymphocytic leukemia (B-CLL), but whether specific chromosomal defects influence the course of B-CLL is still a matter of discussion. The aim of our study was to assess the possible correlation between cytogenetic findings and clinical characteristics. Thirty patients with previously untreated early-onset B-CLL were recruited. The incidence of trisomy 12, and observations of 13q14.2 and 14q32 was analyzed in unstimulated bone marrow cells by means of multicolor interphase FISH. No correlation was found between trisomy 12 and the patients' clinical characteristics. The analysis of the patients with trisomy 12 and observations of 13q14.2 and 14q32 revealed heterogeneity of the leukemic cell population, thus indicating that these chromosomal abnormalities are probably a secondary event in CLL leukemogenesis. The finding of RB1 gene nullisomy and 14q32 deletions in patients at an advanced clinical stage suggests a possible correlation between these rearrangements and disease progression. Multicolor FISH analysis in B-CLL provides important diagnostic, clinical, and prognostic information that may help in assessing prognosis and making treatment decisions.

Trisomy 4 leading to duplication of a mutated KIT allele in acute myeloid leukemia with mast cell involvement.

A G-->T transversion at nucleotide 2467 of the c-KIT gene leading to Asp816-->Tyr (D816Y) substitution in the phosphotransferase domain has been previously identified in a patient with rapidly progressing AML-M2 and mast cell involvement; the patient's blasts had a 47,XY, +4,t(8;21)(q22;q22) karyotype. Herein we confirm the simultaneous presence of both major chromosomal changes by multicolor fluorescence in situ hybridization (FISH) on interphase CD34+ mononuclear cells. By setting up culture leukemic blasts, spontaneous differentiation of adherent cells with mast-cell like features was proved by histochemical and immunoenzymatic analyses. Fluorescence in situ hybridization evidence of trisomy 4 confirmed the origin of differentiated cells from the leukemic blasts. Semiquantitative polymerase chain reaction (PCR) and phosphoimage densitometry of wild-type and mutated KIT alleles on bone marrow blasts made it possible to demonstrate that chromosome 4 trisomy led to a double dosage of the mutated KIT allele. This finding, and that of trisomy 7 and MET mutation in hereditary renal carcinoma represent the only cases of human tumors in which an increased number of chromosomes carrying an oncogene activated by point mutation have been detected.

Three-way and two-way rearrangements involving chromosomes 10, 2, 5 and 5, 2 in two marker chromosomes of a human melanoma cell line.

Two marker chromosomes (mar1 and mar2), provided with two closely spaced heterochromatic bands, were observed in the 14932 cell line established from a human metastatic melanoma. Fluorescence in situ hybridization (FISH) with the alphoid sequence p82H common to all human centromeres showed strong signals over the double C-bands of mar1 and mar2. These were recognized by a chromosome 2-specific alphoid probe, although chromosome in situ suppression (CISS) hybridization with a chromosome 2 library failed to reveal any painting along mar1 and mar2. The centromere of mar1 was identified by a chromosome 10-specific alphoid sequence and the marker chromosome was decorated from pter to a region proximal to the interpolated C-band by a chromosome 10 library. The centromere of mar2 could not be recognized by any chromosome-specific alphoid probe, but the whole mar2 was decorated by a chromosome 5 library. This library also painted the distal q arm of mar1, which was not painted by the chromosome 10 library, as well as a small band proximal to the double C-band. Identification of the two marker chromosomes reveals their common origin and indicates a role for chromosomes 2, 5 and 10 in the genesis and/or progression of the 14932 melanoma. Alteration to the chromosome-specific alphoid sequence in the centromere of mar2 provides evidence for rearrangement of constitutive heterochromatin alphoid sequences in human tumours.

Expression of the c-ski proto-oncogene in human melanoma cell lines.

The aim of this study was to establish whether the expression of proto-oncogene c-ski in melanoma might be related to alterations of chromosome 1q involving the native location of the gene. Six melanoma cell lines, including two carrying marker chromosomes derived from breakage at 1q12-q21, were studied. Expression of c-ski was observed in all cell lines, with very high levels in five of them. However no alteration in c-ski structure or dosage was found in any of the melanoma cell lines, including those with non-random breakpoints near the gene. c-ski Transcripts were detected in cell cultures from normal melanocytes, but at a much lower level than that observed in melanoma cell lines. Transcripts of c-myb and the beta-NGF gene were not detectable in any of the melanoma cell lines, whereas sis- and epidermal growth factor (EGF) receptor gene-specific transcripts were present in two and four melanoma cell lines, respectively. The constant expression of c-ski in the melanoma-derived cell lines at a level of expression much higher than that of normal melanocytes suggests that this proto-oncogene may play a role in melanocyte transformation.

Liability to chromosome damage in lymphocytes of "cancer family" subjects: a study of spontaneous and induced chromosomal fragility.

Spontaneous chromosomal fragility was detected in seven tumor patients and one healthy member from two families with a high recurrence of cancer. Major chromosome lesions, such as terminal deletions and rearranged chromosomes, were found at levels significantly higher than those reported for control individuals. The prevalence of these aberrations in comparison to minor ones (chromosome gaps and chromatid breaks) in this group of patients seems to indicate that the fragility observed is the end-point of a process of chromosomal instability, which may have already been brought to expression. Study of other parameters of genetic instability in the most unstable karyotypes showed that the chromosome damage observed was neither paralleled by abnormal SCE frequency nor sustained by defective DNA repair mechanisms or expression of inherited or constitutional fragile sites. As all the subjects investigated here had previously been shown to display intraindividual variations in the C-banded region of chromosome 1, it is possible that spontaneous fragility and acquired C-heterochromatin polymorphism may be markers that, combined with chromosomal instability, create genetic predisposition to cancer.

Differential effectiveness of solar UVB subcomponents in causing cell death, oncogenic transformation and micronucleus induction in human hybrid cells.

PURPOSE: (1). To determine the biological effectiveness of two solar ultraviolet (UVB) spectra with different lower wavelength thresholds for oncogenic transformation and micronucleus induction in CGL1 cells; (2). to investigate whether the action spectra for short- and long-term effects are similar; and (3). to investigate possible links between transformation and other delayed effects. MATERIALS AND METHODS: Two spectra were derived from a solar UV simulator by using two filters: the first transmitted radiation with lambda > 284 nm, the second with lambda > 293 nm. The resulting spectra have the same UVA, but different UVB components (lambda between 284 and 320 nm, 19 W m(-2), and lambda between 293 and 320 nm, 13 W m(-2)). CGL1 cells were irradiated with 466 J m(-2) with lambda > 284 nm and 1582 J m(-2) with lambda > 293 nm. These doses were approximately equilethal. The endpoints examined were oncogenic transformation, and centromere-positive and -negative micronucleus frequencies in the directly irradiated cells and in transtheir progeny. RESULTS: At equilethal doses, the oncogenic transformation frequency in the directly irradiated cells was greater by a factor of at least 7 for lambda > 284 nm irradiation compared with lambda > 293 nm. The micronucleus induction frequency was also significantly higher with the lambda > 284 spectrum. Consistent with our previous findings, no delayed micronucleus formation was found in the progeny of cells exposed to lambda > 293 nm, while a threefold elevation above controls was seen in the progeny of cells exposed to lambda > 284 nm irradiation. This was also the case for formation of micronuclei with a centromere. CONCLUSIONS: It was found that: (1). for equilethal doses the lambda > 284 nm spectrum was more biologically effective than the lambda > 293 nm spectrum for induction of oncogenic transformation and micronucleus formation; and (2). the higher effectiveness of the lambda > 284 nm spectrum found at equilethal doses for delayed effects in the progeny of irradiated cells resembles that found for transformation. The results suggest that the UVB action spectrum for cell killing is different from that of some delayed effects, and from that of transformation.

High spontaneous chromosomal damage in lymphocytes from patients with hereditary megaduodenum.

Chromosomal aberration and micronucleus assays were used to investigate the extent of cytogenetic damage in peripheral blood lymphocytes from four patients in two unrelated families with hereditary megaduodenum. The frequencies of total chromosomal aberrations, which significantly correlated with those of micronuclei, were higher in the patients than in sex- and age-matched controls, with no overlapping between the two groups. The considerable chromosomal fragility in patients with hereditary megaduodenum may be a genotypic marker for preclinical diagnosis predictive of increased cancer risk.

Fibronectin, laminin in hybrids of Rous sarcoma virus transformed and normal mouse fibroblasts.

Hybrid clones derived from the fusion of normal and Rous sarcoma virus-transformed 3T3 fibroblasts were analyzed for fibronectin and laminin pattern of expression in order to find a possible correlation with tumorigenicity. Both organization in the pericellular matrix and secretion into the culture media were investigated by immunofluorescence and ELISA techniques. No significant difference in fibronectin or laminin release was found among hybrid clones exhibiting different levels of tumorigenicity. In contrast, distinctive immunofluorescence patterns of concomitant presence of low levels of fibronectin and high levels of laminin were constantly observed in all the tumorigenic clones.

Plasma oxidative stress biomarkers, nitric oxide and heat shock protein 70 in trained elite soccer players.

The physiological response to the physical exercise involves a number of changes in the oxidative balance and in the metabolism of some important biological molecules, including nitric oxide (NO) and heat shock proteins (Hsp 70). With the aim to optimise previous laboratory diagnostic panels, we measured the plasma concentration of reactive oxygen metabolites (ROMs), total antioxidant status (TAS), glutathione reductase (GR) activity, and NO and Hsp 70 levels in 44 elite, antioxidant-supplemented and trained soccer players and in 15 sedentary controls. Although no statistically significant difference between athletes and controls was detected in the plasma level of ROMs and TAS, soccer players showed a significantly higher plasma GR activity, NO and Hst 70 levels than those of sedentary controls. These findings suggest that the measuring of relatively novel biomarkers in sport medicine, like GR, NO and Hsp 70, in addition to the well-known and reliable assays (d-ROMs test and TAS) may be useful to a clinician to better assess and evaluate the benefits of training and/or supplementation programs.