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Biomedical subjects

F Cipriani

Publications and source records attributed to F Cipriani.

At least 19 recordsLinked to original sources

Automation of sample mounting for macromolecular crystallography.

A standard sample holder and vial for cryocooled macromolecular crystals has been defined for use with robotic sample changers. This SPINE standard sample holder is a modified version, with added features and specifications, of sample holders in common use. In particular, the SPINE standard meets the precision required for automatic sample exchange and includes a cap that is identified by a two-dimensional datamatrix code as well as an optional vial. At the ESRF, the sample holder standard is in use with the EMBL/ESRF/BM14 robotic sample changer (SC3) which is installed on eight beamlines. The SC3 can hold up to 50 crystals stored in five baskets. A datamatrix reader in the SC3 ensures safe management of the sample flow and facilitates fully automatic screening and characterization of samples. Tools for handling and transporting 50 samples in a dry shipping dewar have been developed. In addition to the SC3, the SPINE sample holder is currently compatible with a number of other robotic sample changers.

Automation↗

High-throughput sample handling and data collection at synchrotrons: embedding the ESRF into the high-throughput gene-to-structure pipeline.

An automatic data-collection system has been implemented and installed on seven insertion-device beamlines and a bending-magnet beamline at the ESRF (European Synchrotron Radiation Facility) as part of the SPINE (Structural Proteomics In Europe) development of an automated structure-determination pipeline. The system allows remote interaction with beamline-control systems and automatic sample mounting, alignment, characterization, data collection and processing. Reports of all actions taken are available for inspection via database modules and web services.

Data Collection↗

Dehydroepiandrosterone increases synaptosomal glutamate release and improves the performance in inhibitory avoidance task.

Dehydroepiandrosterone (DHEA) exerts multiple effects in the rodent central nervous system (CNS), mediated through its nongenomic actions on several neurotransmitter systems, increasing neuronal excitability, modulating neuronal plasticity and presenting neuroprotective properties. It has been demonstrated that DHEA is a potent modulator of GABA(A), NMDA and Sigma receptors. In the present study, we investigated the effect of DHEA on (i) basal- and K(+)-stimulated l-[(3)H]glutamate release from synaptosomes (both in vitro and ex vivo), (ii) synaptosomal l-[(3)H]glutamate uptake (in vitro), and (iii) an inhibitory avoidance task (in vivo). The results indicated that DHEA in vitro increased glutamate release by 57%, and its intracerebroventricular infusion increased the basal-[(3)H]glutamate release by 15%. After 30 min of intraperitoneal administration, DHEA levels in the serum or CSF increased 33 and 21 times, respectively. Additionally, DHEA, intraperitoneally administrated 30 min before training, improved memory for inhibitory avoidance task. Concluding, DHEA could improve memory on an inhibitory avoidance task, perhaps due to its ability to physiologically strength the glutamatergic tonus by increasing glutamate release.

Animals↗

Oligothiophene isothiocyanates as a new class of fluorescent markers for biopolymers.

The regioselective synthesis of fluorescent oligothiophene isothiocyanates is described. The isothiocyanates were reacted with bovine serum albumin (BSA) following standard procedures and the optical properties of the oligothiophene-BSA conjugates were analyzed as a function of oligomer concentration, time, and irradiation power. The oligothiophene-BSA conjugates were chemically very stable and their photoluminescence characteristics persisted unaltered for several months. Photoluminescence data relative to the conjugate of an oligothiophene-S,S-dioxide isothiocyanate with monoclonal anti-CD8 antibody are reported. No fluorescence quenching was observed following the binding of the isothiocyanate to the antibody and the conjugate displayed high chemical stability and photostability.

Antibodies, Monoclonal↗

Plasma levels of antioxidant vitamins and cholesterol in a large population sample in central-northern Italy.

BACKGROUND: In a population-based multicenter case-control study of diet, life-style, and gastric cancer a large series of adults, aged 30-75 years (mean 58.9 years), were randomly sampled from the general population in 3 areas of Central-Northern Italy. AIM OF THE STUDY: To evaluate the relationship between plasma levels of antioxidant vitamins and cholesterol, and sociodemographic characteristics, life-style factors, and dietary intake of selected nutrients in a sample of the Italian population. METHODS: A fasting blood sample was available for 945 subjects (553 men, 392 women). The plasma concentrations of ascorbic acid, carotene, retinol, alpha-tocopherol, and cholesterol were determined by a centralized laboratory. All participants answered to a detailed questionnaire collecting information on sociodemographic and anthropometric characteristics, smoking, alcohol drinking, and dietary habits. Covariance analysis models, with post hoc Dunnett tests, including terms for age, sex, study center, and period of blood drawing, were used for selected multiple-way comparisons of mean values of plasma nutrients. RESULTS: Mean plasma values of retinol were higher among men while women had higher levels of plasma carotene, ascorbic acid, alpha-tocopherol, and cholesterol. Plasma carotene levels showed an inverse association with body mass index, alcohol consumption, and smoking and a positive association with social class. Carotene concentrations were higher in plasma samples obtained in spring/summer, while ascorbic acid levels were higher in autumn/winter. Partial correlation co-efficients between plasma vitamin levels showed a strong correlation between carotene and ascorbic acid (0.69 in men; 0.74 in women), between carotene and alpha-tocopherol (0.44; 0.37), and between alpha-tocopherol and ascorbic acid (0.45; 0.41). Plasma alpha-tocopherol and retinol correlated with plasma cholesterol. On the other hand, plasma carotene and ascorbic acid were correlated with their estimated dietary intakes, while the intakes of other nutrients, as expected, correlated rather poorly with the respective plasma concentrations. CONCLUSIONS: Socio-economic factors, life-style, and specific nutrient intake, in addition to gender, are related to nutrient plasma levels in Italian adults and may provide specific suggestions for the prevention of chronic diseases.

Adult↗

Protein microcrystals and the design of a microdiffractometer: current experience and plans at EMBL and ESRF/ID13.

There is a growing demand for the examination of protein microcrystals at third-generation synchrotron sources. After successful pilot experiments at EMBL/ESRF, which proved that protein microcrystals are often suitable for data collection, operation of the microfocus beamline ID13 was made more user-friendly and suitable for macromolecular crystallography experiments. Given the excellent quality of the beamline microfocusing optics, the key element for successful experiments becomes the handling and visualization of microcrystals. To address this, a microdiffractometer has been designed to allow maximum precision combined with ease of usage and is currently under construction.

Cryoprotective Agents↗

Alcohol-related mortality in Italy.

This study evaluated the impact of alcohol drinking habits on mortality in Italy during the 1980-1990 period. Alcohol Attributable Fractions for a list of Alcohol-Related Diseases were assessed from national and international medical literature and then applied to national mortality data according to 5 y age groups, sex, and place of residence for each year of the 1980-1990 period. Mortality rates and 95% Confidence Intervals were standardized with the direct method. Years of potential life lost (YPLL) were calculated for 1990. 18,033 Italian residents died during 1990 from causes related to alcohol drinking, representing 3.3% of general mortality (males 4.6%; females 2.0%) which corresponded to over 200,000 YPLL up to age 70 y. Chronic diseases account for 65% of alcohol-related mortality. Age-adjusted death rates declined in both genders during 1980-1990. Geographic differences in death rates are well evident. Alcohol drinking is a strong determinant of mortality in Italy. The variability of alcohol-related death rates across the country and the comparison with the results from other populations suggest that health consequences, mainly liver damage, of alcohol intake in Italy may be affected by other factors than alcohol itself. The need for further methodological efforts to improve alcohol attributable fraction estimates is recommended.

Adolescent↗

Neutron Laue diffractometry with an imaging plate provides an effective data collection regime for neutron protein crystallography.

Neutron quasi-Laue diffraction data (2 A resolution) from tetragonal hen egg-white lysozyme were collected in ten days with neutron imaging plates. The data processing Laue software, LAUEGEN, developed for X-ray Laue diffractometry, was adapted for neutron diffractometry with a cylindrical detector. The data analysis software, X-PLOR, was modified and used for the refinement of hydrogen atoms, and the positions of 960 hydrogen atoms in the protein and 157 bound water molecules, were determined. Several examples are given of the methods used to identify hydrogen atoms and water molecules.

Animals↗

A neutron image plate quasi-Laue diffractometer for protein crystallography.

An instrument which is based on image plate technology has been constructed to perform cold neutron Laue crystallography on protein structures. The crystal is mounted at the center of a cylindrical detector which is 400mm long and has a circumference of 1000mm, with gadolinium oxide-containing image plates mounted on its exterior surface. Laue images registered on the plate are read out by rotating the drum and translating a laser read head parallel to the cylinder axis, giving a pixel size of 200 microm x 200 microm and a total read time of 5 minutes. Preliminary results indicate that it should be possible to obtain a complete data set from a protein crystal to atomic resolution in about two weeks.

Animals↗

[Etiological epidemiology of gastric tumors].

Italy still ranks high among European countries for gastric cancer (GC) incidence and mortality. The main determinants of GC are nutritional: a dietary pattern "at high risk" is typically characterized by a large component of salted, preserved, monotonous and traditional foods, opposed to a "low risk" pattern, with a high-consumption of fresh fruit and vegetables and a varied diet. "Low risk" diet effect is supposed to be linked to its component in antioxidant micronutrients, which could positively interfere in the natural history of the disease. Other relevant identified factors are represented by cigarette smoking, some occupational exposures to dust and the infection with Helicobacter pylori. This last factor has been identified in several international studies, but its role in Italy needs further research.

Alcohol Drinking↗

[Etiological epidemiology of colorectal tumors].

We reviewed the main results of colon cancer (CC) epidemiologic studies, according to data published in the 1973-1994 period, with a particular mention to dietary factors and to differences with Italian findings. Meat (mostly, red meat), animal fats and high energy intake not counterbalanced by sufficient physical activity seem to be the most consistent risk factors for CC. On the contrary, the vegetarian based diet seems to reduce the risk of CC. Although interestingly, the relevance to CC of other life-style and diet-related factors (alcohol intake, smoking habits, processing and cooking methods, occupation, drugs, personal medical and reproductive history) must be better defined and requests further investigations. More recently, genetic studies are clarifying the hereditary risk of CC. Several colon carcinogenesis hypotheses have been proposed, but general agreement on the most reliable is still lacking. Authors argue that in the next future, new acquirements could emerge from metabolic polymorphism studies, possibly reconciling the biological significance of individual susceptibility and environmental factors to CC incidence.

Alcohol Drinking↗

Reproductive history and gastric cancer among post-menopausal women.

A total of 339 post-menopausal women with gastric cancer (GC) and 515 population controls were interviewed in a multicenter study in Italy for information on reproductive history, dietary habits and other socio-demographic variables. After adjusting for these factors, GC risk was significantly higher among women reporting menopause below age 45, while no effect was shown for age at menarche, parity, age at first livebirth, and number of abortions. When age at menopause was considered together with age at menarche for an index of fertility years, GC risk showed a significant inverse relation to duration of fertility. These results suggest a protective role for endocrine or other factors related to late age at menopause or to length of fertility. These findings may be relevant to explain the fact that rates of GC reported among females are lower than among males around the world, and suggest the need for further research into hormonal and other related risk factors.

Age Distribution↗

Occupational exposures as risk factors for gastric cancer in Italy.

Occupational associations with gastric cancer were investigated in a multicenter case-control study in Italy involving interviews with 640 histologically confirmed male cases and 959 controls, randomly selected from the resident populations of the study areas. From information on the three jobs each person held the longest, risks were evaluated according to employment in 35 occupations (ever or 21+ years) and to estimated exposure (ever or 21+ years) to six chemicals using a job-exposure matrix. All risk estimates were adjusted by personal, demographic, and dietary variables identified as gastric-cancer risk factors in previous analyses. The only significantly increased risk was observed for sailors, seamen, and allied groups (ever employed: odds ratio [OR] = 2.9; 95 percent confidence interval [CI] = 1.1-8.0; 21+ years: OR = 3.1, CI = 0.8-13). Nonsignificant increases after 21+ years of employment were observed for forestry workers, miners, and janitors and cleaners. Crude ORs were elevated significantly among farmers, but adjusting for demographic and lifestyle factors largely eliminated the association: a nonsignificant 30 percent excess risk remained for farm laborers, but there was no rise in risk among long-term farm laborers and no excess among farm owners. Application of the job-exposure matrix revealed excess risks of borderline significance associated with potential exposure to mineral dusts and nitrogen oxides. For subjects with 21+ years of potential exposure, nonsignificantly increased risks were related to mineral dusts, asbestos, fertilizers, and nitrosamines.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Image plate detectors for macromolecular neutron diffractometry.

Neutron diffraction studies of macromolecules require large position sensitive detectors. It is proposed that such a device can be based on image plate technology, which relies on re-usable photostimuable phosphors, combined with a neutron to gamma-ray converter. Design parameters such as the best wavelength for the neutron radiation and the optimum sample to detector distance are discussed, and a design for a cylindrical detector is outlined. Presently a prototype of such a detector is being built, and the very first test-recording of an X-ray diffraction pattern from a protein crystal is presented.

Journal Article↗

Mitotic activity in colorectal mucosa of healthy subjects in two Italian areas with different dietary habits.

The proliferative activity was evaluated in colorectal biopsies of 39 healthy subjects living in two distinct geographical areas, Trieste in northern and Florence in central Italy. Subjects living in Trieste had a significantly higher mitotic activity compared with subjects living in Florence (mitoses/cells counted x 100 were 0.17 +/- 0.04 in Trieste and 0.089 +/- 0.02 in Florence). The results of a dietary questionnaire also showed that subjects in Trieste consumed significantly fewer starches, fibers, nitrites, and proteins. However, no correlation was evident between the consumption of these nutrients and intestinal proliferation.

Adult↗

A case-control study of cancers of the gastric cardia in Italy.

In a case-control study of gastric cancer (GC) in high-risk and low-risk areas in Italy, 923 GCs were reviewed by one pathologist and classified according to anatomic site. There were 68 (7.4%) cancers occurring in the gastric cardia. Compared to other GCs, cardia cancer tended to occur more often in males (sex ratio 2.8 vs 1.7) and as intestinal or unclassified histologic types. Nutritional factors for cardia tumours resembled those of other GCs, showing inverse associations with the consumption of raw vegetables, citrus and other fresh fruit, and ascorbic acid, and positive associations with the intake of traditional soups and meat, protein and cholesterol, and preference for salty foods. Cigarette smoking and wine consumption were unrelated to cardia cancer risk, and there was only a weak association with total alcohol intake. Cardia tumours showed a greater familial occurrence of GC than did other sites, with a 7-fold increase in risk for those reporting two first-degree relatives with GC. The authors discuss these findings in view of the rising incidence of adenocarcinomas of the cardia and lower oesophagus that has been reported in some western countries.

Adult↗

A case-control study of gastric cancer and diet in Italy. III. Risk patterns by histologic type.

In a case-control study of gastric cancer (GC) in high-risk and low-risk areas of Italy, 923 GCs were systematically categorized by one pathologist according to the Lauren classification distinguishing 2 main histologic types, intestinal (55%) and diffuse (23%). Intestinal types outnumbered diffuse types by a 3 to 1 margin in high-risk regions in the north-central part of the country, while both types occurred at nearly equal rates in low-risk areas. Intestinal types also occurred relatively more frequently at older ages and among males. Relative risks of each type of GC were evaluated in relation to dietary and other data obtained from interviews with the cancer patients and controls. The risk patterns for intestinal and diffuse types were remarkably similar. Increased risks of both types were associated with high intake of meat, salted/dried fish, seasoned cheeses and traditional soups, while decreased risks of both types were found among heavy consumers of fresh vegetables and fruits. Correspondingly similar patterns were seen with indices of nutrients, with risks of both intestinal and diffuse GC rising with animal protein intake and declining with consumption of vitamins C and E. Both types were inversely related to socio-economic status, and neither was associated with cigarette smoking. A familial history of GC was reported more frequently by patients with each type than by controls, although the highest risk was for unclassified GC, a group of poorly differentiated and medullary carcinomas accounting for 15% of all GCs in this study. The findings suggest that, despite differences in geographic and demographic patterns, the intestinal and diffuse types of GC have etiologic factors in common.

Adult↗

Reproducibility of histologic classification of gastric cancer.

A panel review of histologic specimens was carried out as part of a multi-centre case-control study of gastric cancer (GC) and diet. Comparisons of diagnoses of 100 GCs by six pathologists revealed agreement in histologic classification for about 70-80% of the cancers. Concordance was somewhat higher when using the Lauren rather than the Ming or World Health Organization classification systems. Histologic types from reading biopsy tissue agreed with those derived from surgical specimens for 65-75% of the 100 tumours. Intra-observer agreement in histologic classification, assessed by repeat readings up to 3 years apart by one pathologist, was 95%. The findings indicate that, although overall concordance was good, it is important to standardise diagnoses in multi-centre epidemiologic studies of GC by histologic type.

Case-Control Studies↗