[A particularly active chemotherapeutic agent in infections of the urinary tracts].
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Cystitis↗
Biomedical subjects
R De Santis
Publications and source records attributed to R De Santis.
[The "Müller test" in endocrine diseases].
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Endocrine System Diseases↗
Isolation of influenza A(H3N2) virus with "O"-->"D" phase variation.
We report the isolation of two influenza A(H3N2) virus strains which were unable, in the first passages in MDCK cell culture, to agglutinate chicken erythrocytes, though reacting with guinea pig and turkey red blood cells. This observation demonstrates that the occurrence of this phenomenon is not exclusive to influenza A(H1N1) viruses, as previously reported. In order to investigate the molecular basis of this phenomenon, we analysed the nucleotide sequence of the HA-1 region, presumed to be involved in the switch of haemagglutination properties, from the virus present in the original samples and in the corresponding strains isolated and cultivated in MDCK cells and in embryonated eggs. The substitution of amino acid 138 (Ala-->Ser) in MDCK cells could be related to the change in haemagglutination characteristics.
Amino Acid Sequence↗
Role of cross-linking agents in determining the biochemical and pharmacokinetic properties of Mgr6-clavin immunotoxins.
Several immunotoxins (ITs) were synthesized by the attachment of clavin, a recombinant toxic protein derived from Aspergillus clavatus, to the monoclonal antibody Mgr6 that recognizes an epitope of the gp185(HER-2) extracellular domain expressed on breast and ovarian carcinoma cells. Conjugation and purification parameters were analyzed in an effort to optimize the antitumor activity and stability of the ITs in vivo. To modulate the in vitro and in vivo properties of the immunotoxins, different coupling procedures were used and both disulfide and thioether linkages were obtained. Unhindered and hindered disulfide with a methyl group linkage ethyl S-acetyl 3-mercaptopropionthioimidate ester hydrochloride (AMPT) or ethyl S-acetyl 3-mercaptobutyrothioimidate ester hydrochloride (M-AMPT) were obtained by reaction with recombinant clavin, while the monoclonal antibody Mgr6 was derivatized with ethyl 3-[(4-carboxamidophenyl)dithio]propionthioimidate ester hydrochloride (CDPT). To achieve higher hindrance (a disulfide bond with a geminal dimethyl group), Mgr6 was derivatized with the N-hydroxysuccinimidyl 3-methyl-3-(acetylthio)butanoate (SAMBA) and clavin with CDPT. To evaluate the relevance of the disulfide bond in the potency and pharmacokinetic behavior of the ITs, a conjugate consisting of a stable thioether bond was also prepared by derivatizing Mgr6 with the N-hydroxysuccinimidyl ester of iodoacetic acid (SIA) and clavin with AMPT. The immunotoxins were purified and characterized using a single-step chromatographic procedure. Specificity and cytotoxicity were assayed on target and unrelated cell lines. The data indicate that the introduction of a hindered disulfide linkage into ITs has little or no effect on antitumor activity and suggest that disulfide cleavage is essential for activity; indeed, the intracellularly unbreakable thioether linkage produced an inactive IT. Analysis of IT stability in vitro showed that the release of mAb by incubation with glutathione is proportional to the presence of methyl groups and increases exponentially with the increase in steric hindrance. Analysis of the pharmacokinetic behavior of ITs in Balb/c mice given intravenous bolus injections indicated that ITs with higher in vitro stability were eliminated more slowly; i.e., the disulfide bearing a methyl group doubled the beta-phase half-life (from 3.5 to 7.1 h) compared with that of the unhindered, while a geminal dimethyl protection increased the elimination phase to 24 h. The thioether linkage showed its intrinsic stability with a beta-phase half-life of 46 h. The thioether linkage also increased the distribution phase from 17 to 32 min. The in vitro characteristics and in vivo stability of Mgr6-clavin conjugates composed of a methyl and dimethyl steric hindered disulfide suggest clinical usefulness.
Animals↗
Monoclonal suppressor T-cell factor displaying V H restriction and fine antigenic specificity.
The production of stable T-cell clones is essential for the study of T-cell-derived, specific immunoregulatory products and of specific T-cell receptors. T-cell clones have been established by radiation leukaemia virus (RadLV)-induced transformation of suppressor T lymphocytes specific for hen egg white lysozyme (HEL). We report here that culture supernatant obtained from these T-cell clones can, when injected into mice, specifically suppress the anti-HEL antibody response. This monoclonal T-cell product suppresses the antibody response induced by HEL and human lysozyme, but not that induced by ring-necked pheasant egg white lysozyme (REL), thus displaying fine antigenic specificity probably restricted to an epitope involving phenylalanine at amino acid residue 3, present in the N-terminal region of HEL and shared by human lysozyme but absent in REL. The suppression induced by this monoclonal T-cell product is restricted by both H-2 and Igh-1 genes whereas anti-HEL antibodies bearing a predominant idiotype are induced in all mice strains tested, irrespective of their H-2 haplotype or Igh-1 allotype.
Animals↗
Serous cutaneous glands of the western spade-foot toad Pelobates cultripes (Amphibia, Anura): an ultrastructural study on adults and juveniles.
The venom glands of the western spade-foot toad Pelobates cultripes were studied under light and electron microscopes. The glands exhibit the structural patterns usual in anurans, including the typical secretory syncytium. The peripheral cytoplasm contains a single row of nuclei and secretory organelles related to proteosynthesis. The inner cytoplasm is filled with large vesicles holding a thin product which originates from the merging of smaller ones containing a thicker material derived from the Golgi apparatus. The appearance and maturation of P. cultripes venom have been compared with patterns of biosynthesis and secretory evolution described in serous cutaneous glands of several anuran species. Following these criteria, the traditional trends in the terminology and classification of serous glands in anuran skin are discussed and reviewed.
Amphibian Venoms↗
A non-hormonal therapeutic alternative in cervico-vaginal dystrophies.
The pharmaco therapeutic effect of polydeoxyribonucleotide vaginal suppositories in two dosages (PDRN 1,65 mg and PDRN 5 mg)* on vaginal dystrophy was studied. 40 women, aged more than forty, in surgical or physiological climacteric with cervico-vaginal dystrophy were examined. The study was carried out according to the double-blind method, randomized, between groups. Both treatments produced important ameliorations of subjective symptomatology and objective signs. No significant difference emerged between the two preparations. PDRN was proved to be a valid alternative to local hormonal therapies.
Administration, Topical↗
[Case of an odontoma caused by an impacted tooth. Report of a clinical case treated with human fibrin glue (Tissucol) and lyophilized bone (Kieler)].
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Adolescent↗
[Melituric picture in a group of myasthenics].
On the ground of preceding researches carried out about myodystrophias, the authors studied the melituric symptomatology in both a group of myasthenic patients and a group of control. The chromatographic analysis and the dosage of some urinary carbohydrates, showed that there are not statistically significant differences between the group of myasthenic patients and that of controls.
Adult↗
Prostaglandin E2 bladder instillation for the treatment of hemorrhagic cystitis after allogeneic bone marrow transplantation.
BACKGROUND: Hemorrhagic cystitis (HC) is a major complication of high-dose cyclophosphamide therapy used in the preparative regimen for allogeneic or autologous bone marrow transplantation. Several viruses (adenovirus, cytomegalovirus and polyomavirus BK) have also been implicated in the etiology of HC. No one established method of treatment is as yet available. MATERIALS AND METHODS: HC developed in 10 patients after allogeneic bone marrow transplantation and was BK viruria-associated in all cases. All patients were treated with instillations of prostaglandin E2 (PGE2) directly into the bladder. RESULTS: A complete resolution of hematuria within a short time (5 +/- 1 days) was observed in all cases; in 4/10 patients urine cleared within 24 hours of the initial treatment. Intravesical PGE2 therapy caused no systemic circulatory or respiratory problems, although bladder spasms occurred in all patients. CONCLUSIONS: Intravesical prostaglandin E2 instillation appears to be an effective treatment for hemorrhagic cystitis in bone marrow transplant patients; further studies are required to assess the actual role of BK virus in the pathogenesis of HC in bone marrow transplant patients.
Administration, Intravesical↗
Production and characterisation of a recombinant single-chain anti ErbB2-clavin immunotoxin.
We generated a recombinant immunotoxin, named scFv(MGR6)-Cla, composed of the Fv region of an anti ErbB2 monoclonal antibody (MGR6) fused to clavin, a type 1 ribosome-inactivating protein (RIP) from Aspergillus clavatus. ErbB2 is a tyrosine kinase receptor which is overexpressed in most adenocarcinomas; clavin is a 17 kDa ribonuclease which inhibits protein synthesis by inactivating ribosomes. A recombinant DNA construct containing the cDNA of the single chain Fv fragment (scFv) of the MGR6 antibody fused to the clavin cDNA, was expressed at high levels in Escherichia coli as an insoluble fusion protein containing an N-terminal affinity tag of six consecutive histidine residues. Inclusion bodies were denatured and the recombinant fusion protein was purified under denaturing conditions by single-step purification using immobilised metal ion affinity chromatography (IMAC). The purified immunotoxin was renatured at high yield and histidine tag removed by digestion with enterokinase. The purity of the immunotoxin obtained after refolding was confirmed by SDS-PAGE, RP-HPLC, GPC-HPLC and N-terminal sequence analysis. Cell-free protein synthesis inhibition and binding assays showed that both clavin and scFv(MGR6) maintained their properties after refolding.
Antibodies, Monoclonal↗