The obesity gene and colorectal cancer risk: a population study in Northern Italy.

BACKGROUND: Representing the second cause of cancer-related death after lung cancer in men and breast cancer in women, colorectal cancer (CRC) is a major health problem in Italy. Obesity is reckoned to favor CRC; however, the underlying mechanisms are unclear. Recently, a single nucleotide polymorphism (SNP) in the fat mass and obesity associated (FTO) gene was found to be significantly associated with obesity. AIMS: To establish whether the FTO SNP rs9939609 may represent a risk factor for CRC and adenoma in the Italian population. PATIENTS AND METHODS: 1,037 subjects were enrolled in the study and divided in 3 groups: CRC (341 pts., M/F=197/144, mean age=65.17±11.16 years), colorectal adenoma (385 pts., M/F=247/138, mean age=62.49±13.01 years), healthy controls (311 pts., M/F=150/161, mean age=57.31±13.84 years). DNA was extracted from whole blood, and stored frozen for rs9939609 genotyping by real-time PCR. RESULTS: The frequency of the obesity-associated mutated A allele (AA+AT) on the FTO gene was 69.77% among controls, and 71.85% and 65.71% respectively among CRC and polyp patients. Compared to control subjects the AA+AT genotype had no significant effect on the risk for either CRC (OR=1.106; CI 95%=0.788-1.550; p=0.561) or colorectal adenomas (OR=0.830; CI 95%=0.602-1.144; p=0.255). We did not observe any association between the AA genotype and CRC/polyp localization and age at diagnosis. As measured in a patient subset, carriership of the risk alleles did not reflect in a significantly altered BMI. CONCLUSION: The obesity-linked FTO variants do not play a significant role in modulating the colorectal cancer risk in the Italian population.

Transglutaminase 2 expression is significantly increased in cyclosporine-induced gingival overgrowth.

Cyclosporine A is a potent immunosuppressant used to prevent organ transplant rejection and treat various autoimmune diseases. However, cyclosporine A can also induce gingival overgrowth, which is characterized by increased extracellular matrix due to an altered balance between collagen synthesis and degradation. This study proposed to verify whether trans-glutaminase 2, an enzyme thought to be responsible for the assembly and remodelling of extracellular matrix, plays any role in the pathogenesis of cyclosporine A-induced gingival overgrowth. Cyclosporine A-induced gingival overgrowths were collected from 21 liver transplant patients and case-controlled with 20 non-hyperplastic gingival biopsies from healthy patients who had previous periodontal treatment. In both groups, the presence and tissue distribution of transglutaminase 2 were determined by immunohistochemistry and analyzed in comparison with the tissue morphology and expression of lymphocyte-related antigens (CD3 and CD20) and a vessel-related marker (CD34). Transglutaminase 2 expression showed a significant increase (2.6-fold) in the stromal component of cyclosporine A-treated patients compared with controls (p<0.001), which suggested that transglutaminase 2 had a role in the pathogenesis of the disease. Further studies should investigate the therapeutic effect of anti-transglutaminase 2 drugs (putrescine or 1,4-diamino-butane) in these patients.

Spelt and emmer flours: characterization of the lactic acid bacteria microbiota and selection of mixed starters for bread making.

AIMS: This study aimed at characterizing the lactic acid bacteria microbiota and selecting mixed endogenous starters to be used for sourdough fermentation of spelt or emmer flours. METHODS AND RESULTS: Identification of lactic acid bacteria was carried out by partial sequencing of the 16S rRNA, recA, 16S/23S rRNA spacer region and pheS genes. Spelt flour showed the largest biodiversity, while Lactobacillus plantarum dominated in emmer flour. Isolates were subjected to RAPD-PCR analysis and screened based on the kinetics of growth and acidification, quotient of fermentation and liberation of free amino acids (FAA) during sourdough fermentation. After selection, mixed starters were used according to a two-step fermentation process. Wheat flour was fermented by the same starters. Spelt and emmer sourdoughs had slightly higher pH than wheat sourdoughs but titratable acidity, concentration of FAA and phytase activity were higher. Specific volume and crumb grain of emmer and, especially, spelt breads approached those of wheat breads. Sensory analysis confirmed the suitability of spelt and emmer for bread making. CONCLUSIONS: The sourdough biotechnology was indispensable to completely exploit the potential of spelt and emmer flours. SIGNIFICANCE AND IMPACT OF THE STUDY: Results filled up the lack of knowledge on the lactic acid bacteria microbiota and technological performances of spelt and emmer flours.

Synthesis of gamma-aminobutyric acid by lactic acid bacteria isolated from a variety of Italian cheeses.

The concentrations of gamma-aminobutyric acid (GABA) in 22 Italian cheese varieties that differ in several technological traits markedly varied from 0.26 to 391 mg kg(-1). Presumptive lactic acid bacteria were isolated from each cheese variety (total of 440 isolates) and screened for the capacity to synthesize GABA. Only 61 isolates showed this activity and were identified by partial sequencing of the 16S rRNA gene. Twelve species were found. Lactobacillus paracasei PF6, Lactobacillus delbrueckii subsp. bulgaricus PR1, Lactococcus lactis PU1, Lactobacillus plantarum C48, and Lactobacillus brevis PM17 were the best GABA-producing strains during fermentation of reconstituted skimmed milk. Except for L. plantarum C48, all these strains were isolated from cheeses with the highest concentrations of GABA. A core fragment of glutamate decarboxylase (GAD) DNA was isolated from L. paracasei PF6, L. delbrueckii subsp. bulgaricus PR1, L. lactis PU1, and L. plantarum C48 by using primers based on two highly conserved regions of GAD. A PCR product of ca. 540 bp was found for all the strains. The amino acid sequences deduced from nucleotide sequence analysis showed 98, 99, 90, and 85% identity to GadB of L. plantarum WCFS1 for L. paracasei PF6, L. delbrueckii subsp. bulgaricus PR1, L. lactis PU1, and L. plantarum C48, respectively. Except for L. lactis PU1, the three lactobacillus strains survived and synthesized GABA under simulated gastrointestinal conditions. The findings of this study provide a potential basis for exploiting selected cheese-related lactobacilli to develop health-promoting dairy products enriched in GABA.

Comparison of the compositional, microbiological, biochemical, and volatile profile characteristics of nine Italian ewes' milk cheeses.

Nine Italian ewes' milk cheeses were compared for compositional, microbiological, biochemical, and volatile profile characteristics. Mean values for the gross composition were rather similar among cheeses. The lowest pH values were found for cheeses that used primary starters. At the end of ripening, cheeses made from raw milk contained >6.0 log10 cfu/g of nonstarter lactic acid bacteria. Several species of lactobacilli were identified, but Lactobacillus plantarum and Lactobacillus paracasei were dominant. Random amplified polymorphic DNA-PCR analysis showed the biodiversity among the strains, and in several cases a relationship with the cheese of provenance. Cheeses differed mainly for secondary proteolysis, as shown by the principal component analysis applied to reversed-phase fast protein liquid chromatography data of the pH 4.6-soluble fractions and by determination of the free AA. A total of 113 volatile components were identified in the Italian Pecorino cheeses by solid-phase microextraction coupled with gas chromatography-mass spectrometry analysis. The volatile profiles of the 9 cheeses differed significantly. Quantitatively, alcohols were the most abundant chemical class for some cheeses, whereas ketones were the most abundant for other cheeses. Esters and carboxylic acids were largely found. Specific volatile components seemed to distinguish specific cheeses.

[The role of the pathological anatomy museum: past glory, present crisis, and future prospects]. / Ruolo del Museo di Anatomia Patologica: glorie passate, crisi attuale e prospettive future.

Scientific Museum and in particular pathology Museum were in the past an important landmark for Pathology and in general for Medicine. At present they are in a critical stage due to the technological progress, because they are considered by now obsolete compared to other didactic supports, so as for example CD-ROM or virtual presentations. In opposite these old collections are still very precious for Medicine, because they show old diseases, now disappeared or very rare, thanks to the therapeutic and diagnostic progresses. Furthermore they are very useful for teaching of Pathology to medical students and also to laboratory's technicians. The specimens may be used also for organisation of exhibitions, in graduate courses or medical updating courses, for a better popularisation of Anatomy or of some diseases of large environmental or social impact. For all these reasons we propose the creation of renewed museum structures and a protocol for the restoration of the specimens, which are often damaged owing to time and negligence.

Ileal perforation due to ileocecal endometriosis: a case with an unusual clinical and pathological presentation.

A 34-year-old woman, was admitted to the emergency room of our hospital with a history of symptoms of abdominal pain dating back several years and transitorily related to the menstrual cycle. These had worsened in the days prior to admission. Radiological and clinical examinations detected no signs of peritonitis. During exploratory operative laparoscopy, opted for because of persistence and intensification of the abdominal pain, diffuse peritonitis was found due to an ileal perforation immediately upstream of an ileocaecal mass causing a precaecal stenosis. Laparoscopy enabled us to diagnose the preoperatively undetected complication, to perform a through peritoneal lavage and, following minimal conversion by laparotomy, to perform ileocaecal resection, thereby limiting the severity of the surgical trauma. The definitive pathological diagnosis was ileocaecal endometriosis with signs of transmural fistulisation and the presence of endometrial glandular structures in one of the lymph nodes around the lesion. Perforation is a rare complication on those segments of the intestine most often affected by endometriosis, such as the colon and appendix. However, it is even more unusual when it affects the ileum and no other cases have been reported in the literature. Its genesis is attributed to late diagnosis. The pathological findings highlight the particular characteristics of this case. We would stress the pre-, intra- and postoperative diagnostic difficulties encountered and the importance of a thorough anamnestic assessment when making differential diagnoses in women of child-bearing age with abdominal or pelvic pain and perimenstrual symptoms.

White blood cell DNA adducts, smoking, and NAT2 and GSTM1 genotypes in bladder cancer: a case-control study.

We conducted a case-control study on 114 bladder cancer patients and 46 hospital controls. DNA adducts were measured in WBCs by 32P postlabeling and showed no association with smoking habits and the glutathione-S-transferase M1 genotype. A strong association between adduct levels and the N-acetyltransferase (NAT2) genotype was found (P = 0.0002). The NAT2 genotype was associated in a nonstatistically significant way to the case-control status (odds ratio, 1.6; 95% confidence interval, 0.8-3.2). In a logistic regression model, the log of DNA adduct levels was associated in a highly significant way to the risk of bladder cancer (regression coefficient, 0.75; P = 0.0006), independently of smoking habits. Using the median of DNA adducts (RAL, 0.3) as a cutoff point, the odds ratio for the risk of bladder cancer was 4.1 (age-adjusted; 95% confidence interval, 1.9-9.0). Our study suggests that sources other than tobacco smoke contribute to the formation of aromatic DNA adducts in WBCs. The role of WBC-DNA adducts in predicting bladder cancer is still to be clarified.

4-Aminobiphenyl-DNA adducts and p53 mutations in bladder cancer.

Epidemiologic studies have suggested that smokers of air-cured tobacco (rich in arylamines) are at higher risk of bladder cancer than smokers of flue-cured tobacco. The risk has been shown to be modulated by the N-acetyltransferase genotype. We analyzed the biopsies of 45 patients with bladder cancer. p53 mutations were sought by direct sequencing, and 4-aminobiphenyl-DNA adducts were measured by negative ion gas chromatography-mass spectrometry. 4-Aminobiphenyl-DNA adducts were higher in smokers of air-cured tobacco and in current smokers, but no relationship with the number of cigarettes smoked was found. Adducts were higher in more advanced histologic grades of tumors. No pattern was evident for p53 mutations. Seven of 9 mutations occurred in grade 3 tumors. No association was found between 4-ABP adducts and GSTM1 or NAT2 genetic polymorphisms.

[Squamous carcinoma of the gallbladder. Anatomo-clinical study of a case with reference to oncogene expression]. / Carcinoma squamoso della colecisti. Studio anatomo-clinico di un caso con riferimento all'espressione di oncogeni.

Squamous cell carcinoma of the gallbladder is a rare tumor with an ill-defined clinical course. We report a case of pure squamous carcinoma of the gallbladder with a rapidly fatal outcome. The tumor cells were positive for oncogenes P53, c-Erb B2 and Ki 67. This feature has no comparison with other reported cases of squamous carcinoma of the gallbladder but it is in accordance with oncogene expression in adenocarcinomas.

Exposure to aromatic amines and ras and c-erbB-2 overexpression in bladder cancer.

We have analyzed the bladder biopsies of six bladder cancer patients exposed to high levels of 2-naphthylamine and benzidine, 11 unexposed bladder cancer patients, six subjects with benign conditions of the bladder, and 16 healthy subjects. Immunohistochemical analysis of the p21 and p185 protein products, for overexpression of ras and c-erbB-2 oncogenes, was performed. Overexpression of ras was found in four of six exposed cancer patients, 3 of 11 unexposed cancer patients, zero of six benign disease patients, and zero of 16 healthy subjects. The odds ratio for ras overexpression, comparing exposed with unexposed cases, was 5.3 (90% confidence interval 0.6 to 64). Overexpression of c-erB-2 was apparently not associated with occupational exposure.

Effects of some psychosocial variables on different disease manifestations in 112 cadets: a longitudinal study.

One-hundred and twelve cadets attending the 141st training course for reserve officers of the 'Scuola Transmission' of the Italian Army were administered, 10 days after incorporation, a battery of personality tests and measures of stressful events in the preceding year. Test scores were considered both individually and grouped into factors. During the 6 months of the course all disease episodes for each cadet were recorded. Total episodes infections and traumas were considered. A significantly higher number of total episodes and, specifically, of infections was present in subjects reporting a higher number of stressful events, in interaction with attitudes towards parent figures, hardiness, loneliness, and an alienation factor. A younger age of subjects also appeared predictive of a higher number of total episodes and infections, as a main effect and in interaction with attitudes to mother, hardiness, and alienation. Very few effects were obtained for traumas, suggesting that the effects of variables are mediated through a biological route rather than through behaviour, as would be the case if traumas were involved. No effect was shown by measures of stressful events alone. Results are discussed in the light of a reconsideration of the notion of stressful events.

Psychosocial factors and clinical evolution in HIV-1 infection: a longitudinal study.

The aim of the study was to assess the relationship between the initial psychosocial situation and the probability of later symptom development in HIV-1 infection. One hundred HIV-1 seropositive subjects, 79 in Stage III (LAS) and 21 in Stage II (asymptomatic), were examined both immunologically (CD4+, Skin Test) and psychologically (test battery). Follow-up at 6 and 12 months involved clinical and immunological reassessment of subjects, who were then classified as fully symptomatic (S, Stage IV) or unchanged (U). The two groups were compared through ANOVA on initial psychosocial measures, while stepwise logistic multiple regression was employed to assess the predictive value of psychosocial measures on clinical and immunological evolution. Psychosocial measures most clearly showing an association with clinical evolution were Denial/Repression attitudes (negatively) and Fighting Spirit (positively), whereas aspects of Hardiness and Social Support showed an effect in interaction with initial CD4+ levels. No stable results were obtained on immunologic evolution. The two groups (U and S) did not show significant differences on other independent variables, with the exception of age.

Dual rearrangement of immunoglobulin and T-cell receptor gene in a case of T-cell hairy-cell leukemia.

We report a case of T-cell hairy-cell leukemia with a dual rearrangement of Ig- and T-cell receptor genes. The cytochemical, transmission electron microscopy, and surface antigens data (CD3+, CD8+, CD11+, HLA-DR+, CD19-, CD20-) were consistent with a T-cell hairy-cell leukemia. Molecular analysis according to Southern revealed a dual rearrangement of immunoglobulin heavy-chain (JH) and T-cell receptor beta (TcR beta) chain genes. Our findings suggest that the coexistence of JH and TcR gene rearrangements, frequently detected in acute leukemia, may also be observed in hematologic malignancies derived from more differentiated cells.

The human myeloma cell line LP-1: a versatile model in which to study early plasma-cell differentiation and c-myc activation.

The characteristics of a human cell line (LP-1) derived from the peripheral blood of a patient with IgG-lambda myeloma in leukemic transformation are described. The cells resemble immature plasma cells in that they exhibit a membrane phenotype that is intermediate between late B lymphocytes and plasma cells, even though they secrete IgG-lambda chains. Treatment of LP-1 cells with 12-0 tetradecanoylphorbol-13-acetate (TPA) or pokeweek mitogen (PWM) induces the appearance of surface markers and ultrastructural features typical of mature plasma cells but does not affect their proliferative activity. Molecular analysis of the cell line showed an increased expression of the c-myc protooncogene and the presence of abnormally sized transcripts. Conventional cytogenetics and pulsed-field gel electrophoresis showed no structural rearrangements of the c-myc gene, suggesting that the abnormal c-myc expression may be due to point mutations or small deletions within the gene. The LP-1 cell line is a useful model in which to study the process of B-cell maturation; such study may lead to the uncovering of unusual mechanisms of c-myc activation. Furthermore, the LP-1 cell is a potential partner in the generation of human hybridomas.

[Percutaneous needle biopsy of the lung. Critical review of 496 cases]. / Agobiopsia percutanea polmonare. Revisione critica di 496 casi.

Between February 1979 and December 1986, 618 lung biopsies were performed on 496 patients (122 with double puncture, using Tru-cut and 19-20 G needles). Overall diagnostic accuracy was 84.5%; the 77 patients examined in the past 18 months only underwent thin needle (19-20 G) biopsies--which were carried out by an experienced radiologist--and more sophisticated cyto-histological techniques were employed. Such a procedure gave better results (sensitivity 94.1%, accuracy 94.8%) than did the use of larger caliber needles in the past years; moreover, no complications occurred in these patients. In conclusion, 19-20 G needles are to be preferred to bigger ones, provided that the operator be experienced and cyto-histology be correctly performed.

A combined glucagonoma and VIPoma syndrome. First pathologic and clinical report.

We report a case of pancreatic tumour metastatic to the liver in a patient with insulin-treated diabetes, anaemia, cheilitis, necrolytic migratory erythema, hypokalemia and chronic watery diarrhea, a picture suggesting combined glucagonoma and VIPoma syndromes. Immunocytochemistry of a biopsied hepatic metastatic nodule revealed both glucagon and vasoactive intestinal peptide (VIP) positive cells. Increased plasma glucagon and VIP levels were detected (values of 900 pmol/l and 277 pmol/l respectively). This is the first reported case showing not only immunocytochemical, but also clinical evidence of the combined secretion of these hormones.

Selective growth response to IL-3 of a human leukaemic cell line with megakaryoblastic features.

A new human leukaemic cell line (M-O7) with the phenotypic characteristics of CFU-mega is described. Its cells are positive for T200 leucocyte common antigen (LCA) and negative with MAbs recognizing T and B cells and mature myelomonocytic antigens. In contrast, they react with MAbs recognizing antigenic determinants common to multi-lineage (CD13, CD33, CD34) and to bipotent erythromegakaryoblastic (CD36, H25) haemopoietic precursors, and with MAbs specific for platelet glycoproteins (CD41w, CD42w). A small proportion (10%) of the cells were large and multinucleated, and on electron-microscopy examination showed peripheral splitting of platelet-like cytoplasm particles. When transferred to a serum-free Iscove modified Dulbecco's medium supplemented with human insulin and transferrin, M-O7 cells stop proliferating. Of the haemopoietic growth factors tested for their ability to restore the proliferative activity of this quiescent population, only rH IL-3 proved effective. Moreover, it also increased the cloning efficiency in methylcellulose more than any other CSFs. The M-O7 cell line may provide a valuable tool for the biological assay of IL-3, and a model for biochemical studies of the megakaryocytic lineage.

Dual secretory and myoepithelial differentiation in the transplantable R3230AC rat mammary carcinoma.

The hormone-responsive R3230AC mammary carcinoma, serially transplantable in Fisher rats, shows striking functional and morphological similarities to the normal mammary gland. We have studied its cellular composition by both light and electron microscopy, employing markers of myoepithelial and epithelial cells. We identified two cell types: the major cellular component corresponded to epithelial milk-protein secreting cells, while a second component showed immunocytochemical and ultrastructural characteristics of the myoepithelial cells. These cells were positive with a monoclonal antibody detecting alpha smooth muscle actin. The dual differentiation which normally occurs in breast ducts is therefore reproduced in a malignant experimental tumor. The coexistence of neoplastic cell populations, divergent in morphology and function, that persist in a tumor despite many transplant generations, leads to reconsideration of the relationship between cellular differentiation and malignant transformation.