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S Riva

Publications and source records attributed to S Riva.

At least 109 records · Page 6Linked to original sources
PubMed1988

Presence of transcription signals in two putative DNA replication origins of human cells.

We describe the purification and cloning of human DNA replicated at the onset of S phase in HL60 cells synchronized with aphidicolin. A survey of the overall structural properties of these sequences did not show any distinctive features except for an enrichment in Cot0 DNA. The two longer fragments were completely sequenced and studied in more detail. Both were shown to contain transcriptional signals associated with promoters and/or enhancers, such as the binding sites of Sp1, T antigen and nuclear factor III. In one instance, a binding site for a known cellular transcription factor (USF/MLTF) was located inside the sequence by footprinting. Accordingly, by CAT assay and Northern blot, the same sequence was shown to contain an active promoter. The significance of these findings with respect to the role of transcription in initiation of DNA replication at the origin is discussed. None of the tested fragments exhibited autonomously replicating sequence (ARS) activity in transfected cells. The problems connected with the detection of ARS activity in human cells are critically examined.

Aphidicolin↗
PubMed1988

Circular dichroism and fluorescence of polyethylene glycol-subtilisin in organic solvents.

Subtilisin Carlsberg has been made soluble in organic solvents such as dioxane and acetonitrile by covalent linking to polyethylene glycol. Far-ultraviolet circular dichroism and intrinsic protein fluorescence have shown that subtilisin dissolved in dioxane, in which the enzyme is active and highly stable, maintains the native secondary structure as well as the native microenvironment for tyrosyl residues. In acetonitrile subtilisin undergoes conformational changes that cause enzyme inactivation and precipitation.

Acetonitriles↗
PubMed1988

cDNA cloning of human hnRNP protein A1 reveals the existence of multiple mRNA isoforms.

Protein A1 is one of the major component of mammalian ribonucleoprotein particles (hnRNP). Human protein A1 cDNA cloning and sequencing revealed the existence of at least two protein isoforms. Among the cDNAs examined, sequence differences were found both in the structural portion, leading to aminoacid changes (Tyr to Phe or Arg to Lys) and in the non translated 3'-region where two T-stretches of different length were observed. Interestingly one of the aminoacid substitutions falls into a consensus sequence common to many RNA binding proteins. Northern blot analysis of poly A+ RNAs from five human tissues revealed two mRNA forms of 1500 and 1900 n due to alternative polyadenylation. Analysis of genomic DNA showed at least 30 A1-specific sequences, some of which correspond to processed pseudogenes. These results suggest that protein A1 is encoded by a multigene family.

Amino Acid Sequence↗
PubMed1988

Lack of tolerance development during chronic ibopamine administration to patients with congestive heart failure.

Beta-adrenergic agonists can progressively lose their efficacy during chronic therapy in patients with heart failure. Ibopamine is a new dopamine derivative, active on dopaminergic and beta-adrenergic receptors, whose hemodynamic activity has been acutely demonstrated. To assess whether any attenuation of its efficacy occurs, the variations of the cardiac output induced during chronic therapy were monitored by impedance cardiography in 15 patients with dilated cardiomyopathy who showed a significant increase of the cardiac output (20.7 +/- 10.0%) after acute ibopamine administration. The efficacy of ibopamine was also assessed after 6 and 12 months of therapy by echocardiography, exercise testing, and 24-hour dynamic electrocardiogram (EKG) monitoring. The cardiac output response to ibopamine did not show any significant attenuation (range 15% to 19%) in the evaluations at 1, 2, 3, 4, 8, and 12 months of therapy. No significant change, was noted, after 6 and 12 months, in the exercise capacity (505 vs. 602 and 604 seconds) and the fractional shortening (16.2 vs. 18.3 and 18.5) without any change of the diastolic diameter. Ventricular arrhythmias were significantly reduced after 6, but not 12, months of therapy. No significant change in the New York Association (NYHA) functional class was noted at 6 and 12 months of therapy (2.4 +/- 5 vs. 2.3 +/- 7 and 2.4 +/-0.6, respectively). Our results show that ibopamine can maintain its hemodynamic activity even during chronic therapy.

Adult↗
PubMed1988

Combined hemodynamic overload of the left and right ventricles as a possible cause of interventricular septum preponderance in high blood pressure.

We tested whether overload of the two ventricles may be associated with a preponderance of interventricular septum in patients with high blood pressure. The rationale is that the septum is shared by the greater and lesser circulation and that in hypertension the latter shows the same qualitative hemodynamic alterations as the former. Among 65 hypertensive patients, 40 (group 1) showed (echo) posterior wall thickness within the mean +/- 1 SD of 62 normal subjects, and 25 (group 2) had a posterior wall thickness exceeding the mean + 1 SD of the normal population. Both groups were subdivided into subgroups A and B, which included patients whose ventricular septum was similar to (within the mean + 1 SD) and thicker than (exceeding the mean + 1 SD) the posterior wall thickness in the corresponding group, respectively. Resting differences in systemic and pulmonary pressure and vascular resistance among subgroups 1A, 1B, and 2A were not significant; however, in subgroup 2B these variables exceeded those in the other subgroups to a significant extent. During cold pressor testing (CPT) the levels reached and the changes in pressure and resistance from baseline values in both circuits were much greater in subgroups B than in subgroups A. The baseline plasma norepinephrine value showed a trend toward an increase from subgroup 1A to 1B and from subgroup 2A and 2B; during CPT norepinephrine invariably changed and in subgroups B it rose significantly more than in subgroups A. It was not determined whether this caused the hemodynamic overload in subgroups B.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

The reproducibility of average ambulatory, home, and clinic pressures.

The reproducibility of ambulatory, home, and clinic blood pressures was compared in 13 untreated mildly hypertensive and 14 normotensive subjects. Each subject had two sets of daily ambulatory recordings, home self-measured readings (over 6 days), and clinic measurements taken 2 weeks apart. Comparisons over the 2 weeks within and among the methods of measurements were made using a repeated-measures analysis of variance. The results showed that there was no consistent average change in the ambulatory or home pressures and no change in clinic diastolic pressures, but the clinic systolic pressure of the hypertensive subjects dropped 6 mm Hg (p less than 0.05), while that of the normotensive subjects showed no significant change. Test-retest correlations of each of the three methods were similar in magnitude, indicating a similar level of reliability. Test-retest correlations of the ambulatory standard deviations, however, were low, indicating a low reliability of this measure of daily pressure variability. These results suggest that the reproducibility of ambulatory pressures may be as good or better than that of home or clinic measurements. They also suggest that the average ambulatory pressure may be preferable as the measurement in clinical trials, since it may be less influenced by measurement anxiety, particularly in hypertensive subjects.

Activities of Daily Living↗
PubMed1988

[Coronary angiographic study of patients with non-Q myocardial infarct: correlations with the initial electrocardiographic aspect and prognosis].

The purpose of our study was to evaluate, during the early phase, the coronary anatomy of the patients who suffered from an acute non-Q myocardial infarction (non-Q MI) and to correlate it with the ECG findings and with clinical evolution. We studied 76 patients (pts.), mean age 56 +/- 9 years, who had their a first non-Q MI (reinfarctions non included). They all underwent angiographic examination within an average period of 18 days (range 5-30 days). In the whole group of patients it is worthwhile noting: A) the elevated occurrence of left main lesions (11% of pts.) and multivessel disease (60%); B) the great percentage (41%) of patients with open infarct-related vessel but with very critical residual stenosis (above 90% of total lumen); C) the presence of collateral vessels in almost all the pts. (91%) with totally occluded infarct-related artery. During the 12 month follow-up, in all groups was a high occurrence (above 70%) of coronary events (postinfarction angina, reinfarction, aortocoronary by pass or PTCA). Furthermore, it is possible to identify a subgroup of patients presenting ST-segment depression with very unfavourable coronary anatomy (80% had multivessel disease, 30% of which had a left main critical stenosis) and high prognostic risk (90% occurrence of coronary events). In the subgroup with ST-segment elevation there was an elevated percentage (65%) of open infarct-related vessel, but with an important residual stenosis. Considering the advantages of revascularization interventions in these high risk patients with extensive residual jeopardized myocardium, we conclude that it is important that all patients with non-Q MI undergo early coronary angiography.

Adult↗
PubMed1987

Characterization of human DNA sequences synthesized at the onset of S-phase.

We developed a method of enrichment for DNA replicated at the onset of S-phase in synchronized human HL60 cells. About 200 such sequences were cloned. The analysis of this selected DNA sample showed that: 1) the cloned DNA fragments derive from a limited number (750-1500) of replicons; 2) there is no extensive homology between different DNA fragments; 3) they are not significantly enriched in highly repeated sequences; 4) they are enriched in snap-back (Cot = o) DNA. The sequence of the longest fragment revealed the presence of numerous signals collected in a few hundred nucleotides: 1) homology with the origin of replication of human Papovaviruses usually associated with potential stem-loop structures; 2) binding sites for known transcription factors and for another nuclear factor; 3) potential binding sites for the chromosome "scaffold".

Base Sequence↗
PubMed1987

Improvements of Western blotting to detect monoclonal antibodies.

A comparison of the effects of different factors on the sensitivity of Western blotting technique to detect monoclonal antibodies is described. The major improvements were obtained by: A) renaturating the antigen in the gel before transferring it in carbonate buffer at pH 10 onto nitrocellulose and B) using alkaline-phosphatase-conjugated second antibody instead of peroxidase-conjugated second antibody.

Animals↗
PubMed1987

Large-scale purification of hnRNP proteins from HeLa cells by affinity chromatography on ssDNA-cellulose.

A purification procedure for proteins which bind heterogeneous nuclear RNA (hnRNP proteins) is described. The procedure, which entails standard chromatographic fractionations (single-stranded DNA cellulose, hydroxyapatite) and detection with specific antibodies, allows a large-scale preparation of these proteins and the partial separation of different polypeptides. By this method, polypeptides of higher molecular mass (53-55 kDa) can be purified, which are structurally and antigenically related to the 'canonical' hnRNP core proteins (34-43 kDa) that constitute the 40S hnRNP complexes. We also show that HeLa cells contain a protease that cleaves hnRNP core proteins to discrete smaller polypeptides of 22-28 kDa. Such protease, which has been partially purified, appears to copurify extensively with some of the hnRNP proteins.

Cellulose↗
PubMed1987

[Incidence and clinical significance of involvement of the right ventricle in acute inferior myocardial infarction].

UNLABELLED: The purpose of our study was to evaluate, with noninvasive procedures, the incidence and the clinical picture of right ventricular involvement in patients with acute transmural inferior myocardial infarction. Our study group was constituted of 107 consecutive patients admitted to our Coronary Care Unit within 10 hours from the onset of symptoms; in every patient a standard 12-leads ECG and the precordial leads V3R and V4R were obtained at admission in CCU and then every 12 hours. 80 patients underwent B-mode echocardiographic evaluation within 36 hours and in 93 patients a myocardial scintigraphy was performed, between the 48th and 72nd hour from the onset of chest pain, 1-2 hours after injection of Tc-99m-pyrophosphate. RESULTS: 45 patients (42.1%) had ECG positive for right ventricular infarction, 49 patients (51.6%) had positive Tc-99m-pyrophosphate scintigraphy and 24 patients (30%) positive echocardiography. By using the positivity of ECG and another method at least, patients were separated into 2 groups: group A (associated inferior and right infarction) was constituted of 45 patients, and group B (isolated inferior infarction) was constituted of 62 patients. In group A we noted a higher incidence of hypotension (systolic blood pressure less than 100 mmHg) and oliguria (less than 30 ml/h)- p less than 0.01-, of 2nd and 3rd A-V blocks-p less than 0.001- and primary ventricular fibrillation -p less than 0.01. The incidence of parossistic atrial fibrillation, severe bradycardia or SA blocks and of mortality was not statistically different between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1987

Autoantibodies to purified nuclear proteins related to DNA metabolism during ageing and in SLE patients.

In this study the specificity of circulating autoantibodies in ANA+ aged donors, ANA- donors and SLE patients was investigated by immunoblotting on total nuclear proteins and by ELISA on purified nuclear proteins, possibly related to DNA metabolism, such as DNA polymerase alpha, DNA-dependent ATPase, DNA Topoisomerase I, ssDBP, hnRNP, HMG and histones. Immunoblotting showed that sera from ANA+ aged donors present fewer antibodies to nuclear proteins, especially to those between 21,000 and 45,000, molecular weight (MW), than sera from SLE patients. When the specificity of antisera was further studied on purified nuclear proteins, it was found that the majority of sera from SLE patients react with most of the proteins tested, whereas sera from ANA+ aged donors mainly react with DNA polymerase alpha, DNA-dependent ATPase, DNA Topoisomerase I and histones. In addition, sera from a few ANA- donors also reacted with certain purified nuclear proteins in a statistically significant age-related manner.

Adolescent↗
PubMed1986

Mammalian single-stranded DNA binding protein UP I is derived from the hnRNP core protein A1.

Antibodies induced against mammalian single-stranded DNA binding protein (ssDBP) UP I were shown to be cross-reactive with most of the basic hnRNP core proteins, the main constituents of 40S hnRNP particles. This suggested a structural relationship between both groups of proteins. Using the anti-ssDBP antibodies, a cDNA clone (pRP10) was isolated from a human liver cDNA library in plasmid expression vector pEX1. By DNA sequencing this clone was shown to encode in its 949 bp insert the last 72 carboxy terminal amino acids of the ssDBP UP I. Thereafter, an open reading frame continued for another 124 amino acids followed by a UAA (ochre) stop codon. Direct amino acid sequencing of a V8 protease peptide from hnRNP core protein A1 showed that this peptide contained at its amino terminus the last 11 amino acids of UP I followed by 19 amino acids which are encoded by the open reading frame of cDNA clone pRP10 immediately following the UP I sequence. This proves that ssDBP UP I arises by proteolysis from hnRNP core protein A1. This finding must lead to a re-evaluation of the possible physiological role of UP I and related ssDBPs. The formerly assumed function in DNA replication, although not completely ruled out, should be reconsidered in the light of a possible alternative or complementary function in hnRNA processing where UP I could either be a simple degradation product of core protein A1 (as a consequence of controlling the levels of active A1) or may continue to function as an RNA binding protein which has lost the ability to interact with the other core proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1985

Single stranded DNA binding proteins derive from hnRNP proteins by proteolysis in mammalian cells.

As we have previously demonstrated, mammalian single stranded DNA binding proteins (ssDBP) and heterogeneous nuclear RNA binding proteins (hnRNP proteins) are antigenically and structurally related. In this paper we show that ssDBP are specific proteolytic products of hnRNP core proteins. Proteolysis can be observed in crude extract, both total and nuclear and is not inhibited by the most commonly used protease inhibitors. Such phenomenon can be observed in HeLa cells, human fibroblasts and calf thymus extracts. A trypsin-like protease that cleaves purified hnRNP proteins to give ssDBP of Mr = 24-28 Kd can be purified from HeLa cells. A precursor-product relationship can be established between hnRNP core proteins type A and an ssDBP of 24 Kd (UP1).

Animals↗