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.

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.

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.

G1 heavy chain disease: clinicopathological, ultrastructural and immunochemical study of a new case.

Clinicopathological ultrastructural, and immunohistochemical findings of a new case of IgG1 heavy chain disease are reported in detail. The abnormal protein lacks the VH and CH1 region with sequence starting at 225 residue. The main pathologic feature was plasmacytic tumor of the lymph nodes with B-cell immunoblastic sarcoma patterns. Neoplastic diffusion to other organs was also present. Plasmacytic neoplastic cells have also been found in the bone marrow. The ultrastructure of the noeplastic cells was characterized by more or less abundant plasmacytic-like endoplasmic reticulum with very frequent peculiar whorled configurations. Immunohistochemical methods revealed the abnormal protein production by neoplastic cells in different stages of differentiation. From this case and from the data of the literature it is concluded that the gamma HCD is due to a neoplastic proliferation of lymphoplasmacytic cells whereas the reticulum cells are never involved.

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.

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.

Osteomalacic myopathy in a case of diffuse nodular lipomatosis of the small bowel.

The case of a 57-year-old woman is described with a two months history of proximal muscle weakness and pain, marked hypotrophy and brisk reflexes. Clinical investigation demonstrated normal serum CK, myopathic EMG and osteomalacia. Muscle biopsy showed type II fibre atrophy and mitochondrial alterations without inclusions. Further examinations including a jejunal biopsy revealed malabsorption accounting for osteomalacia. At autopsy diffuse nodular lipomatosis of the small bowel was detected (Acta neurol. belg., 1982, 82, 65-71).

[Cellular immunity in two familial cases of thyroid carcinoma (author's transl)]. / Aspetti dell'immunità cellulare in due casi di carcinoma tiroideo nella stessa famiglia

Cellular immunity in two brothers with thyroid carcinoma in a family presenting pathological thyroid changes is investigated. In these brothers a low PHA lymphocyte blast-transformation and an elevated number of surface membrane Ig bearing lymphocytes were observed. These data are discussed in relation to the question of immunological defects which frequently are present in carcinoma patients.

[Glomerulonephritis and sarcoidosis: study of a case]. / Glomerulonefrite e sarcoidosi: studio di un caso

A case of rapidly progressing glomerulonephritis accompanied by sarcoidosis is presented. The immunological pathogenesis of the underlying disease was shown by the positivity of the reactions for anti-IgA and anti-betal C/beta 1 A globulins and the presence of thick deposits in the mesexential matrix and some parts of the thickened basal membrane. Since generalised sarcoidosis is regarded as the expression of an immunological reaction to an unknown, diffuse antigen, this type of glomerulonephritis may be associated with the particular immunopathological condition observed in sarcoidosis.

[Adenopathies in chronic myeloid leukemia (CML). Study of 161 cases]. / Adenopatie nella leucemia mieloide cronica (LMC). Studio su 161 casi.

161 cases of CML have been studied. Clinically significant adenopathies were present in 3,2% of the patients at the moment of diagnosis, and in the subsequent course they appeared in 7% of them. The behaviour of adenopathies showed to be unrelated to: --splenomegaly; --blastic metamorphosis in the peripheral blood or in the marrow (which they often preceded from 3 to 26 months); --hematological sensibility to cytostatic therapy; and furthermore they often acted as the most important clinical and therapeutic problem. From the cyto-histological point of view three features have been observed: 1) blastic metamorphosis in a lymphnode showing features of myeloid metaplasia; 2) blastic invasion in a lymphnode without any sign of myeloid metaplasia; 3) malignant lymphoma. Cytological examination of imprints and ultrastructural studies, besides the usual histological investigations, proved to be useful for the definition of the above mentioned features. In lymphomatous forms, together with the study of the cariotype and the research of the Ph' chromosome, the performance of immunocytological investigations is also necessary.

The role of platelet-activating factor in experimental immune complex pathology.

Tissue localization of immune complexes (IC) after infusion of synthetic platelet-activating factor (PAF) in rabbits was studied. Bovine serum albumin (BSA) was injected intravenously either before or after the infusion of synthetic PAF, later followed by the administration of anti-BSA antibodies over a 30-min period. Control rabbits received both BSA and anti-BSA antibodies, followed by lyso-PAF or saline-BSA instead of PAF. The infusion of PAF induced structural alterations in the heart, lung and kidney which were consistent with an increased vascular permeability. Deposits of BSA, IgG, and C3 were found in the heart, lung, and kidney of rabbits infused with PAF but not in control rabbits. In the liver of PAF-infused rabbits, immune deposits were only infrequently observed, whereas they were constantly present in the cardiac valves, thoracic aorta and at the bifurcation of the renal artery as they were in the control rabbits. These results suggest that PAF favors the localization of IC in the heart, lung and kidney vessels but does not influence the formation of immune deposits at the sites of turbulence of the blood flow.

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.

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.

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.

Distribution of two tumor-associated antigens in gastric lesions as detected by two monoclonal antibodies on tissue sections.

Two monoclonal antibodies antitumor-associated antigens, B1.1 and B72.3, have been used with the immunoperoxidase technique on tissue sections to study gastric carcinomas, dysplasia, intestinal metaplasia, and adenomas. B1.1 reacts with carcinoembryonic antigen (CEA); B72.3 reacts with a 220-400 kd glycoprotein present in colon, breast, and other carcinomas. CEA was found in 89% (34 of 38) and B72.3 antigen in 92% (35 of 38) of carcinomas. In half of these more than 50% of tumor cells were positive. The normal epithelium was usually negative or sporadically positive in a few cells. In dysplastic areas and adenomas the number of cells that were positive for the two antigens was greater than in normal epithelium and smaller than in carcinomas. In intestinal metaplasia B72.3 antigen was almost always present, whereas CEA was sometimes undetectable. Both the antigens proved to be good markers of neoplastic versus normal cells. The presence of B72.3 antigen in addition to CEA in dysplasia, adenomas, and intestinal metaplasia adds further evidence of their close relationship with gastric cancer.

Human platelet cationic proteins bind to rat glomeruli, induce loss of anionic charges and increase glomerular permeability.

The binding of human platelet cationic proteins (HuPlt CP) to rat renal cortex in vitro and in vivo, the loss of glomerular polyanions (GPA) and the increase in glomerular permeability were studied. HuPlt CP were purified by sequential cation-exchange chromatography and chromatofocusing, by which these proteins were shown to be highly cationic in nature (pI 10.5) and mainly composed of three molecular species of 55.60 kD, 40.45 kD, and 10 kD as studied by gel permeation in high pressure liquid chromatography and SDS-polyacrylamide gel electrophoresis. Binding of HuPlt CP to glomerular capillary walls (GCW), mesangium and to peritubular capillaries of the rat renal cortex was demonstrated by immunofluorescence, using a specific goat anti-HuPlt CP antiserum, after incubation of the sections with HuPlt CP in vitro and after injection of HuPlt CP in vivo. This interaction was ionic in nature, since treatment of sections with heparin abrogated the binding of HuPlt CP to glomerular structures. The glomerular deposits of HuPlt CP were associated with the loss of GPA as revealed by colloidal iron staining (light microscopy) in both in vitro and in vivo experiments and by ruthenium red staining (electron microscopy) in in vivo studies. After the injection of native ferritin, the increase in glomerular permeability produced by an infusion of HuPlt CP was observed by the increased ratio of counted particles within the glomerular basement membrane with respect to controls. The binding of HuPlt CP to GCW and the loss of GPA was consistent with the interpretation that HuPlt CP may increase glomerular permeability due to the neutralization of GPA.