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Biomedical subjects

R Chiesa

Publications and source records attributed to R Chiesa.

At least 109 records · Page 6Linked to original sources
PubMed1993

Neurotoxicity of a prion protein fragment.

The cellular prion protein (PrPC) is a sialoglycoprotein of M(r) 33-35K that is expressed predominantly in neurons. In transmissible and genetic neurodegenerative disorders such as scrapie of sheep, spongiform encephalopathy of cattle and Creutzfeldt-Jakob or Gerstmann-Sträussler-Scheinker diseases of humans, PrPC is converted into an altered form (termed PrPSc) which is distinguishable from its normal homologue by its relative resistance to protease digestion. PrPSc accumulates in the central nervous system of affected individuals, and its protease-resistant core aggregates extracellularly into amyloid fibrils. The process is accompanied by nerve cell loss, whose pathogenesis and molecular basis are not understood. We report here that neuronal death results from chronic exposure of primary rat hippocampal cultures to micromolar concentrations of a peptide corresponding to residues 106-126 of the amino-acid sequence deduced from human PrP complementary DNA. DNA fragmentation of degenerating neurons indicates that cell death occurred by apoptosis. The PrP peptide 106-126 has a high intrinsic ability to polymerize into amyloid-like fibrils in vitro. These findings indicate that cerebral accumulation of PrPSc and its degradation products may play a role in the nerve cell degeneration that occurs in prion-related encephalopathies.

Amino Acid Sequence↗
PubMed1993

Alcohol drinking among adolescents from town and mountainous areas in north Italy.

Alcohol consumption was investigated among 9th and 13th grade high school students in the town of Brescia (an urban area) and in the Local Health Unit of Breno, a mountainous area in the province of Brescia. A total of 1364 students from Brescia and 1373 from Breno completed an anonymous questionnaire during school time. In both groups alcohol consumption was greater among 13th graders (modal age: 18 years) and males than 9th graders (modal age: 14 years) and females. Beer was the favourite alcoholic beverage, but wine was the most widely consumed daily beverage: among males, about 8% in grade 9 and 15% in grade 13 drank at least one glass of wine every day. The majority of both 9th and 13th graders had more than one type of alcoholic beverage a week, although many females drank beer only. Total alcohol consumption was higher among the mountainous area students than urban students, especially 9th graders, and greater differences were observed in the consumption of beer with respect to wine. The proportion of male students from Brescia and Breno who consumed more than 6 alcohol units a week were, respectively, 17% and 25% in grade 9 and 36% and 42% in grade 13. A higher prevalence of drunkenness was found among the students from the mountainous area with respect to the urban students, especially males. Among 13th grade males, 42% of students from Brescia and 60% from Breno claimed one or more episodes of drunkenness during the year prior to the interview.

Adolescent↗
PubMed1993

Three dimensional time-of-flight magnetic resonance angiography in carotid artery surgery: a comparison with digital subtraction angiography.

Three dimensional time-of-flight magnetic resonance angiography (MRA) and intra-arterial digital subtraction angiography (DSA), considered the gold-standard technique, were compared for pre- and postoperative evaluation of the carotid bifurcation. Images of suitable quality of 194 bifurcations were available with both techniques. Percentage stenosis was graded into 5 groups: A = 0-29%, B = 30-59%, C = 60-89%, D = 90-99%, E = 100% (occlusion). Preoperative MRA (126 bifurcations) overgraded the stenosis in 14 cases and undergraded it in 11. The two techniques agreed in 101 cases and the extent of misgrading was never more than one category. Regression analysis showed a good correlation between the two methods. Severe ulceration was better identified by DSA. As far as the surgical indication was concerned, MRA had a sensitivity of 92.6% and a specificity of 98.3%. In 68 operated cases, postoperative MRA and intraoperative completion angiography showed a satisfactory endarterectomy with no residual stenosis in any of the cases. In conclusion, MRA seems an accurate modality for imaging of carotid bifurcations. Significant limitations still exist for an adequate demonstration of intracranial circulation.

Adult↗
PubMed1993

Apoptosis mediated neurotoxicity induced by chronic application of beta amyloid fragment 25-35.

To investigate whether and how amyloid-beta protein (A beta) is involved in the neurodegenerative changes characteristic of Alzheimer's disease (AD), primary hippocampal neurones from foetal rat brain were exposed acutely and chronically to micromolar concentrations of a synthetic peptide homologous to residues 25-35 of A beta (beta 25-35). A single application of this peptide (25-100 microM) was ineffective but when the neuronal cultures were exposed to beta 25-35 (25-100 microM) repeatedly every two days for ten days, cell survival was dramatically reduced. The structural changes and the DNA fragmentation of cells chronically exposed to the peptide suggested that neuronal death occurred by apoptosis. Furthermore, beta 25-35 showed the intrinsic ability to polymerize into amyloid-like fibrils in vitro. These results confirm the potential pathogenic role of A beta in AD, and indicate that amyloid fibrils may induce neuronal death through a specific programmed process.

Amino Acid Sequence↗
PubMed1993

Molecular changes during the photooxidation of alpha-crystallin in the presence of uroporphyrin.

Singlet oxygen reacts preferentially with three amino acids in proteins, His, Trp and Met. In order to study the specific molecular events that result from such oxidations, calf alpha-crystallin was photooxidized in the presence of uroporphyrin and the reactions were investigated by high performance liquid chromatography peptide mapping using a photodiode array detector followed by fast atom bombardment mass spectrometry (FAB-MS). From these studies, the following conclusions can be inferred: (1) Upon photooxidation residue Met-68 of the B chain is oxidized to Met sulfoxide, whereas residue Trp-60 remains intact. (2) Two of the 16 His residues in alpha-crystallin are photooxidized with an apparent pKa of ca 7.0 (3) FAB-MS analysis suggests that residue Lys-166 close to the C-terminal end of the A chain forms a cross-link with the His-7 residue close to the N-terminal end of the A chain. This may be either an inter- or intramolecular cross-link.

Amino Acid Sequence↗
PubMed1993

Adhesion properties of plasma sprayed hydroxylapatite coatings for orthopaedic prostheses.

Using Air Plasma Spraying (APS) and Vacuum Plasma Spraying (VPS) techniques, hydroxylapatite (HA) and mixtures of HA and titanium (Ti) were deposited on a Ti6A14V alloy (and on an AISI 316L steel) subjected to different surface treatments. The deposits were investigated for their crystallinity, thickness, and adhesion properties. Higher adhesion values were obtained with VPS rather than with APS. By utilising VPS, the deposition conditions were selected in order to achieve crystallinity values between 70 and 90%. The adhesion results depend on the crystallinity (increasing with its decrease), on the thickness (decreasing slightly with its increase) and especially on the surface finish of the metallic substrate. A porous Ti precoat was more effective than either chemical etching in HCl or sandblasting; sandblasting being the least effective. In particular, the double deposits consisting of a porous Ti precoat and a successive layer of HA proved to be most interesting for their higher adhesion properties and for their capability of providing primary stability due to the presence of the HA and secondary stability, in the case of its reabsorption, due to the porous metal.

Alloys↗
PubMed1993

[Mammographic screening of the health district 38, hospital of Gardone Val Trompia (Brescia - Lombardy). Results of the first 2 rounds].

The authors report the results of the first two rounds of a mammographic screening program by the Health District of Gardone Val Trompia (Brescia) including women 50 to 69 years old. 9,650 women were invited for round one and 6,415 (66.5%) attended; 9,656 were invited for round two and 6,552 (67.8%) attended. Fifty-four cancers were found in round one (detection rate: 8.4/1000): 11.1% of them were in situ carcinomas, 63% were T1 and 24.1% were T2 lesions, or higher. In round two, 32 lesions were detected (detection rate: 4.9/1000): 15.6% of them were in situ carcinomas, 78.1% were T1 and 3.2% were T2 lesions. In the positive cases axillary lymph nodes involvement was observed in 24.1% of cases in round one and was unknown in 27.8%, but the figures decreased to 6.2% and 9.4%, respectively, in round two. Effectiveness indicators were always observed.

Aged↗
PubMed1992

Monoclonal antibodies to human DNA topoisomerase I and the two isoforms of DNA topoisomerase II: 170- and 180-kDa isozymes.

Several monoclonal antibodies of different isotypes specific to human DNA topoisomerase I, to 170- and 180-kDa DNA topoisomerase II isozymes, were produced and characterized. The specificity of monoclonal antibodies was confirmed by comparison with polyclonal antibodies by Western blot, by immunoprecipitation of enzyme activity, and by immunoprecipitation of DNA topoisomerases with characterized polyclonal antisera. Morphological studies performed by immunofluorescence indicate that the three groups of monoclonal antibodies (MoAbs) stain the nucleus with characteristic patterns, which can be compared with those obtained with polyclonal antibodies. In particular the MoAbs to the 100-kDa DNA topoisomerase I stain the nucleolus and the nucleoplasm; the MoAbs to 170- and 180-kDa DNA topoisomerase II give completely distinct intranuclear patterns: those to the 170-kDa protein stain mainly the nucleoplasm, whereas those to the 180-kDa protein stain only the nucleolus. The two DNA topoisomerase II isozymes clearly exhibit fluctuations in their expression during cell growth: the 170-kDa isozyme is more abundant during the logarithmic phase of growth, while the 180-kDa isozyme is mainly present during the plateau phase of growth.

Antibodies, Monoclonal↗
PubMed1992

The role of transcranial Doppler in carotid artery surgery.

In order to evaluate the usefulness of transcranial Doppler (TCD) in carotid artery surgery, 90 patients who underwent carotid endarterectomy were studied. Transcranial Doppler was performed preoperatively with digital compression of the carotid artery and intraoperatively at the time of cross-clamping, together with routine EEG monitoring. The percentage residual middle cerebral artery velocity (%MCAV) during carotid artery compression or cross-clamping was considered. The intraoperative %MCAV at cross-clamping was compared with the values obtained in the preoperative examination during the compression test and with the EEG data; the EEG records were blindly reviewed by a neurologist and divided into three groups: group A, normal; group B, moderate anomalies and group C, major anomalies. There were no significant differences between preoperative %MCAV (compression test) and intraoperative %MCAV (cross-clamping) by separate analysis of the data related to systolic, diastolic and mean velocity. The systolic, diastolic, mean %MCAV and the Gosling index after cross-clamping were then compared with the EEG data: no significant differences were seen between groups A and B; on the other hand, in all patients with major EEG anomalies (group C) the %MCAV at TCD was reduced to 0. In conclusion, preoperative TCD associated with carotid artery compression appears a safe and useful way of identifying patients at risk of cerebral ischaemia during carotid artery cross-clamping. Patients with no residual flow at intraoperative TCD need to be considered as presenting a very high risk of ischaemia and in this group the use of a shunt is mandatory.

Aged↗
PubMed1992

[Factors associated with alcohol consumption in the youth of the province of Brescia].

We investigated factors related to adolescent alcohol consumption by administering an anonymous questionnaire to 2737 9th and 13th grade high-school students in the province of Brescia. A total of 1364 students coming from an urban area (the town of Brescia with about 200,000 inhabitants) and 1373 students from a mountainous area (Breno, Local Health Unit N. 37 in the Lombardy Region with 83,000 inhabitants) were interviewed. The associations between alcohol drinking and some demographic, socio-economic, behavioural and environmental variables were assessed by computing the correspective odds ratios (OR) by fitting logistic regression models for ordinal response variables ("cumulative odds" models). The results showed that the following variables were positively associated with alcohol consumption: 1) residence in a mountainous area, especially as regards spirits consumption; 2) experimental and regular smoking; 3) peer alcohol drinking (best friend and partner). Parents' daily alcohol consumption was associated with wine drinking, but not with either spirits consumption or frequency of drunkenness among students. A slight inverse relationship was also found between students' alcohol intake and their knowledge of the health hazards of alcohol, but it was not significant for all groups of students.

Adolescent↗
PubMed1991

Phosphorylation of alpha-crystallin B in Alexander's disease brain.

The phosphorylation of alpha-crystallin B was studied in homogenates of autopsy samples of brain tissue from patients with Alexander's disease, a condition characterized by over-expression of this protein. After incubation in the presence of [gamma-32P]ATP and cAMP the homogenates were analyzed by two-dimensional electrophoresis, (isoelectric focusing followed by SDS-PAGE). Three major polypeptides having the same molecular weight as bovine lens alpha-crystallin B and pIs 7.1, 6.9 and 6.7 were detected in the Coomassie blue stained gels. These three polypeptides were recognized by an alpha-crystallin B-specific antiserum in Western blots. The polypeptides with pIs 7.1 and 6.7 co-migrated in isoelectric focusing gels with bovine lens alpha B and its phosphorylated form alpha Bp, respectively. Radioautography of the two-dimensional gels demonstrated the presence of 32P in the most acidic polypeptide. The results demonstrate the occurrence of alpha B phosphorylation in Alexander's disease brain tissue.

Blotting, Western↗
PubMed1991

Identification of calmodulin-sensitive Ca(2+)-transporting ATPase in the plasma membrane of bovine corneal epithelial cell.

ATP-dependent Ca2+ uptake was characterized in a plasma membrane enriched fraction obtained from the bovine corneal epithelium. This uptake essentially represented intravesicular accumulation because 72% of the Ca2+ content was releasable following exposure to 10(-6) M A23187. The substrate and Ca2+ requirements for maximal transport activity were similar to those described in the red blood cell because: (1) exogenous calmodulin (3 microM) significantly decreased the apparent Km for Ca2+ to 0.31 microM and increased the rate of Ca2+ uptake; (2) a hydroxylamine labile Ca(2+)-dependent phosphoenzyme intermediate was identified with an apparent molecular size of 140 kDa; (3) Ca(2+)-dependent binding of 125I-labelled calmodulin to this protein was demonstrated which could be antagonized with a calmodulin antagonist, trifluoperazine. These results show that the plasma membrane contains an ATP-dependent Ca2+ transporter. However, its relationship to a previously described high affinity form of Ca(2+)-stimulated Mg(2+)-dependent ATPase is not apparent because their [Mg2+] requirements to elicit maximal activity differed by two orders of magnitude.

Adenosine Triphosphate↗
PubMed1991

Photooxidation of specific residues in alpha-crystallin polypeptides.

Singlet oxygen is a biologically important, photochemically generated species that preferentially oxidizes His, Trp, and Met residues of protein molecules. Calf alpha-crystallin was photooxidized with use of meso-tetra(p-sulfonatophenyl)porphyrin (TPPS) and uroporphyrin (UP) as singlet oxygen generators. The effects of photooxidation were monitored by analyzing the changes in alpha-crystallin peptide maps obtained by reversed-phase HPLC using a photodiode array absorbance detector. The reaction led to the loss of six specific peptides, five of which contained photooxidizable residues. Peptides containing His-97 and His-154 from the A chain and Met-68 from the B chain are preferentially photooxidized, suggesting that those residues have access to singlet oxygen. Trp residues in the N-terminal region are converted to NFK, whereas Trp-60 in the B chain is not photooxidized strongly suggesting that the former are close to the surface of alpha-crystallin while the latter Trp residue is buried. Only one peptide that is lost from the peptide maps does not contain a photooxidizable group; however, this peptide does contain an apparently undigested Lys residue. It is suggested that it forms a cross-link with a photooxidized His residue.

Amino Acid Sequence↗
PubMed1991

[Vascular access for chronic hemodialysis in diabetic patients].

The aim of this study carried out on 33 uremic diabetic patients submitted to chronic hemodialytic treatment was to assess the kind of complications related to the vascular approach used as well as their short- and long-term incidences. Out of the 46 anastomoses prepared, 39 were arteriovenous fistulae according to Brescia-Cimino and 7 were PTFE grafts. The actuarial survival rate was 88%, 79%, and 63% after one, two and four years, respectively. The most frequent compliance was thrombosis. Our experience demonstrates that the distal arteriovenous fistula may be considered a valid vascular access for hemodialysis also in diabetic patients.

Adult↗
PubMed1991

A six-amino acid deletion in basic fibroblast growth factor dissociates its mitogenic activity from its plasminogen activator-inducing capacity.

A recombinant deletion mutant of the 155-amino acid form of human basic fibroblast growth factor (bFGF), lacking amino acid residues 27-32 (Lys-Asp-Pro-Lys-Arg-Leu), was expressed in Escherichia coli and purified to homogeneity by heparin-Sepharose affinity chromatography. When maintained in the presence of an equimolar concentration of soluble heparin, the bFGF mutant (M1-bFGF) is as potent as bFGF in stimulating cell proliferation in normal and transformed fetal bovine aortic endothelial cells, in adult bovine aortic endothelial cells, and in NIH 3T3 fibroblasts. However, under the same experimental conditions, M1-bFGF is at least 100 times less efficient than bFGF in stimulating plasminogen activator (PA) production in endothelial cells, as assayed by chromogenic PA assay, SDS/PAGE zymography, and Northern blot analysis of urokinase-type PA mRNA. In the presence of heparin, M1-bFGF binds to bFGF plasma membrane receptors present on endothelial cells in a manner undistinguishable from bFGF. It also induces the same tyrosine phosphorylation pattern when added to NIH 3T3 cells. The data suggest that the PA-inducing activity of bFGF may depend upon a functional domain that differs from those involved in the mitogenic activity of the growth factor and that the binding of bFGF to its plasma membrane receptor may not be sufficient to induce urokinase-type PA production in endothelial cells.

Amino Acid Sequence↗
PubMed1991

Factor(s) required by EBV transformed lymphocytes to grow under limiting dilution conditions.

IL-6 was demonstrated to promote growth of EBV transformed lymphocytes. However IL-6 was ineffective at promoting growth of EBV transformed lymphocytes cultured at the single cell level under limiting dilution conditions. On the contrary, HECS, which is known to contain IL-6, supported very efficiently the growth of 1-2 EBV transformed cells. When IL-6 was removed from HECS, by using specific antibodies, no reduction in HECS activity was observed, indicating that probably more than one growth factor is required to support the growth of EBV transformed cells cultured at very low cell numbers in the absence of feeder cells.

Cell Division↗