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Biomedical subjects

E Ferrari

Publications and source records attributed to E Ferrari.

At least 307 records · Page 17Linked to original sources
PubMed1987

Effects of long-term treatment (4 years) with pentoxifylline on haemorheological changes and vascular complications in diabetic patients.

Twenty-one insulin-dependent and 30 non-insulin-dependent diabetic patients were treated over 48 months with pentoxifylline ('Trental' 400) 1200 mg/day orally. All patients had haemorheological alterations and vascular complications. A marked improvement in erythrocyte deformability and a reduction in plasma fibrinogen levels was already evident after 6 months of therapy; these improvements were maintained throughout the 48 months of the study and were independent of short-term and long-term glycometabolic changes. The normalization of blood rheology pattern was associated with a significant decrease in total urinary protein excretion rate and in urinary albumin excretion rate. An improvement in both microvascular, i.e., retinopathy and nephropathy, and macrovascular, i.e. ischaemic heart disease and peripheral occlusive arterial disease, complications was demonstrated after the long-term trial with pentoxifylline. No side-effects occurred during the observation period. These data suggest that pentoxifylline may have an important role in both the treatment of diabetic haemorheological changes and renal disorders and in the prevention of accompanying degenerative vascular complications.

Adult↗
PubMed1986

Cloning and expression of a cDNA encoding a catalytically active fragment of calf thymus DNA polymerase alpha.

A calf thymus cDNA expression library was constructed in the EcoRI site of lambda gt11 and probed with an antibody raised against calf thymus DNA polymerase alpha. Three classes of antibody-reactive clones were isolated. The largest class carried a 1.9 kilobase calf cDNA insert and expressed a 165-175 kilodalton beta-galactosidase:calf fusion protein which displayed DNA polymerase activity. The characteristic responses of the polymerase activity to alpha-specific inhibitors and antibodies identified the 1.9 kilobase cDNA as a sequence specifically derived from the structural gene encoding the pol alpha catalytic core.

Base Sequence↗
PubMed1986

Cytochemistry of circulating lymphocytes in diabetes mellitus with and without retinopathy and in newly diagnosed type I (insulin dependent) diabetes.

Cytochemical studies have been performed on peripheral blood lymphocytes of 68 diabetic subjects, with various conditions of metabolic control, and 15 newly diagnosed insulin-dependent diabetic patients. 20 patients of the 1 group had diabetic retinopathy. In diabetic patients periodic acid Schiff positivity, acid phosphatase, and N-acetyl-beta-glucosaminidase activities of lymphocytes are fairly impaired, particularly in insulin-dependent diabetes. Concerning the alpha-naphthyl-acetate-esterase activity, the percentage of positive cells with coarse granules is significantly reduced (p less than 0.001) in diabetic patients as compared to controls, without difference related to age and sex. These abnormalities are more evident in patients with poor glyco-metabolic control. In patients with newly diagnosed insulin-dependent diabetes we have found a further decrease in alpha-naphthyl-acetate-esterase activity, and an increase in acid phosphatase and N-acetyl-beta-glucosaminidase activities. Cyto-enzymatic activities are not significantly different in subjects with diabetic retinopathy. The results of peripheral lymphocyte enzymatic activities in diabetics could be related to a depression of the cell-mediated immunity and could enhance the infections risk of these patients. Furthermore our data show an altered immunological balance in subjects with newly diagnosed type I diabetes.

Acid Phosphatase↗
PubMed1986

Identification of the pleiotropic sacQ gene of Bacillus subtilis.

The sacQ gene of Bacillus subtilis, a pleiotropic gene affecting the expression of a number of secreted gene products, has been identified as a small 46-amino-acid polypeptide. The increased expression of this polypeptide in strains carrying the sacQ36 allele, or in strains carrying the sacQ gene on a high copy plasmid, appears to be responsible for the phenotype of higher levels of proteases seen in these strains. A deletion of the sacQ gene had no apparent phenotype, indicating that it is not an essential gene.

Alleles↗
PubMed1986

Effect of stage 0 sporulation mutations on subtilisin expression.

Subtilisin expression as a function of growth and sporulation was determined using a presubtilisin-beta-galactosidase gene fusion. An approximately 500-base-pair region upstream of the subtilisin gene and including the first eight codons of the presubtilisin protein was fused at the eighth codon of beta-galactosidase in the integrative vector pJF751. This gene fusion does not carry a signal sequence, and therefore its synthesis is uncoupled from maturation of presubtilisin. The fusion protein gene was integrated into a variety of recipient strains to test for the effect of various mutations on the initial rate of presubtilisin-beta-galactosidase synthesis. Among the spo0 mutations tested, the spo0A mutations showed a strong, 10-fold decrease in the rate of beta-galactosidase synthesis. This effect of the spo0A mutations was not evident when the presubtilisin-beta-galactosidase fusion was present on a multicopy plasmid. The sacU mutation, which was known to increase the extracellular level of levansucrase and proteases, was found to increase the synthesis of the presubtilisin-beta-galactosidase gene fusions 7-fold, and the hpr mutations were shown to increase the rate of presubtilisin-beta-galactosidase gene fusions 17-fold, indicating that these mutations influence either transcription or translation of the presubtilisin gene. However, the effect of these mutations was only observed in the stationary phase of growth, indicating they did not render synthesis constitutive. By using multicopy plasmids and an integrated gene fusion, it was shown that there is likely to be a titratable repressor controlling subtilisin synthesis.

Bacillus subtilis↗
PubMed1985

Characterization of the spo0A locus and its deduced product.

The highly pleiotropic stage 0 sporulation locus of Bacillus subtilis, spo0A, has been cloned in bacteriophage lambda, subcloned in plasmids, and sequenced. The locus was found to code for a protein of 29,691 Da. Analysis of the in vivo transcripts from this region by nuclease S1 protection experiments located the start and stop of transcription of the locus. The transcription start site was preceded by a promoter resembling sigma 37-dependent promoters. Two mutations originally assigned to a second locus, spo0C, in this region because of their weakly pleiotropic phenotypes were cloned and sequenced. The mutations were found to be different missense alterations in the same base of the 10th codon preceding the carboxyl end of the Spo0A protein. These results, along with the finding that mutations in the spo0A gene product [Hoch, J. A., Trach, K., Kawamura, F. & Saito, H. (1985) J. Bacteriol. 161, 552-555] suppress the requirement for spo0B, spo0E, and spo0F gene products in transcription from sigma 28-dependent promoters, suggest that the Spo0A protein interacts directly with the transcription machinery to effect the initiation of sporulation. The deduced amino acid sequence of the Spo0A protein was highly related to that of the OmpR regulatory protein of Escherichia coli.

Amino Acid Sequence↗
PubMed1985

Molecular cloning of a Bacillus subtilis gene involved in spore outgrowth.

A lambda Charon 4A derivative carrying the outB gene of Bacillus subtilis has been identified by transformation of a B. subtilis mutant temperature-sensitive in spore outgrowth. The cloned region is a single EcoRI fragment 14 kb in length. In addition to outB, the cloned DNA includes at least part of the amyE and aroI loci.

Bacillus subtilis↗
PubMed1985

Demonstration of rabies viral antigen in paraffin tissue sections: comparison of the immunofluorescence technique with the unlabeled antibody enzyme method.

The immunofluorescence technique and the peroxidase-antiperoxidase method were used to demonstrate rabies antigen in a retrospective study on formalin-fixed, paraffin-embedded brain tissues from 34 naturally infected wild and domestic animals. Rabies was confirmed with immunofluorescent staining on fresh brain tissue at the time of necropsy of the animals. There was a perfect correlation (serial sections from a given brain area were always positive by both methods), but the peroxidase-antiperoxidase technique was preferred, since no trypsin digestion was required. Twenty six of the 34 animals were immunohistochemically positive and had encephalitis, and in 21 of these 26, the hematoxylin and eosin-stained sections contained detectable intracytoplasmic inclusion bodies in at least 1 brain area. Of the remaining 8 animals (with no inflammatory lesions), 7 were positive for rabies antigen and 2 had no inclusion bodies. Rabies antigen was apparent in 62% of the brain areas in which inclusion bodies were not found in the corresponding hematoxylin and eosin stained sections. Thus, together with the inclusion body positive areas, which were all immunohistochemically positive, it was possible to diagnose rabies in a total 84% of the areas examined. Both techniques greatly facilitate the diagnosis of rabies and may be a reliable help to the diagnostic pathologist when only formalin-fixed tissues are available. However, the methods should not be considered substitutes for the immunofluorescence technique and the mouse inoculation test with fresh brain tissue.

Animals↗
PubMed1985

[Pentoxifylline and arterial hypertension in diabetes mellitus: long-term results in randomized groups].

This follow-up study concerns the use of pentoxifylline (PTX) as antihypertensive drug in randomized diabetic groups. A significant reduction of systolic and diastolic arterial blood pressure has been obtained in diabetic patients treated with PTX alone or with PTX associated to conventional antihypertensive therapy. The normalization of arterial blood pressure was associated to an improvement of erythrocyte filterability and to a reduction of proteinuria. Therefore, pentoxifylline can be considered a good pharmacological approach in the treatment of diabetic hypertension and in the prevention of diabetic renal disease.

Diabetes Complications↗
PubMed1985

Diabetic retinal vascular complications and erythrocyte filtrability; results of a 2-year follow-up study with pentoxifylline.

A long-term trial with oral pentoxifylline ("Trental" 400) in a dosage of 1200 mg per day was carried out in 70 diabetic patients with and without vascular afflictions. A statistically significant improvement in erythrocyte filtrability and a statistically significant reduction in fibrinogen levels were observed after 2 months of therapy; the significant improvement in both parameters was maintained throughout the 24 months of treatment. Blood rheology normalization was followed by a marked reduction in proteinuria and albumin excretion rate. After pentoxifylline administration, an improvement in both microvascular and macrovascular afflictions was demonstrated. The results of this study suggest that pentoxifylline is an effective drug for the long-term treatment of diabetic vascular complications and that it may also possibly depress or prevent the progress of the disease.

Adult↗
PubMed1984

Replacement of the Bacillus subtilis subtilisin structural gene with an In vitro-derived deletion mutation.

The entire subtilisin structural gene from Bacillus subtilis I168 has been cloned, and its nucleotide sequence has been determined. When expressed on a high-copy-number shuttle vector, a fivefold increase in serine protease activity was observed. The DNA sequence of the gene is 80% homologous to the Bacillus amyloliquefaciens subtilisin structural gene, and the translated mature coding sequence is 85% homologous to the published protein sequence of subtilisin BPN'. The chloramphenicol resistance determinant of a plasmid integrated at the subtilisin locus was mapped by PBS1 transduction and was found to be linked to glyB (83%) and argC (60%), but not with metC or purB . The chromosomal locus containing the wild-type subtilisin allele was replaced with an in vitro-derived allele of the gene (delta apr-684) that contained a 684-base-pair deletion. The technique used for introducing the deletion is a variation of the gene replacement methods used in Saccharomyces cerevisiae and Escherichia coli. When used in B. subtilis, deletion mutants could be directly screened among the transformants. Physiological characterization of the delta apr-684 mutation revealed no discernable effect on the formation of heat-resistant endospores, but strains carrying the mutation produced only 10% of wild-type serine protease activity. A model is presented that outlines the pathway for plasmid integration and deletion formation in B. subtilis.

Bacillus subtilis↗
PubMed1984

Cloning of the neutral protease gene of Bacillus subtilis and the use of the cloned gene to create an in vitro-derived deletion mutation.

The neutral protease gene of Bacillus subtilis has been cloned, and its nucleotide sequence has been determined. The cloned gene was used to create an in vitro-derived deletion mutation, which was used to replace the wild-type copy of the gene. This deletion, in combination with a deletion of the alkaline protease gene, completely abolished protease production. The loss of the proteases had no detectable effect on growth, morphology, or sporulation.

Amino Acid Sequence↗