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Biomedical subjects

C Milanese

Publications and source records attributed to C Milanese.

At least 55 records · Page 3Linked to original sources
PubMed1997

Development and pharmacological characterization of a modified procedure for the measurement of carbonic anhydrase activity.

Carbonic anhydrases (CAs) are a family of zinc metalloenzymes of molecular mass 30-60 kDa; seven different isoenzymes belong to this family (Okuyama et al., 1992, Proc Natl Acad Sci USA 89:1315-1319). They may be broadly recognized according to the efficiency with which they catalyze the reversible interconversion of CO2 and HCO3-, and they differ in physicochemical properties, in sensitivity to various inhibitors and in their subcellular localization; cytoplasmic (CA I, CA II, CA III, and CA VII), cell-surface membrane (CA IV), and mitochondrial (CA V) and secretory (CA VI) isoenzymes have been described. Several methods are reported in the literature for the measure of CA enzymatic activity; they may be broadly divided into two categories: those based on the measure of pH variation (pH-stat and colorimetric assays) (Wu et al., 1993, J Ocular Pharm 9:97-108; Maren, 1991, Molec Pharmacol 41:419-426) and the ones in which CO2 production is measured through pCO2 sensors (Botrè and Botrè, 1990, Anal Biochem 185:254-264).

Acetazolamide↗
PubMed1997

Reduced adhesion of PBMNCs to endothelium in methylprednisolone-treated MS patients: preliminary results.

Methylprednisolone (MP) is a synthetic steroid commonly used in the treatment of multiple sclerosis (MS) relapses. It has a wide spectrum of activities on immune cells: it might also act by preventing mononuclear cell/endothelium adhesion. We studied adhesion phenomena between cultured human umbilical vein endothelial cells (HUVECs) and PBMNCs (CD45+, CD14+) from 6 MS patients treated in vivo with MP. We also studied fluctuations in CD11a and CD18 levels on lymphocytes and monocytes, as well as changes in serum sICAM-1 and sVCAM-1 concentrations. After MP treatment, PBMNCs adhesion to endothelium decreased at 3 h, while it went back to baseline levels at 24 h. A tendency to increase in both CD11a and CD18 on the surface of lymphocytes was detected, while an increase in serum sVCAM-1 was seen at 3 h.

Adult↗
PubMed1996

Inhibition of inflammatory cytokine production and protection against endotoxin toxicity by benzydamine.

The present study was designed to assess the effect of N,N-dimethyl-3-[(1-benzyl-1H-indazol-3-yl)ossi]-1-propana mine (benzydamine) on in vivo and in vitro production of inflammatory cytokines. Benzydamine inhibited tumour necrosis factor-alpha (TNF-alpha) production in vitro by human lipopolysaccharide-stimulated monocytes with an ED50 of approximately 25 microM (12 donors). Under the same conditions, benzydamine had modest or no effect on production of interleukin (IL-1), IL-6 and IL-8. Inhibition of TNF-alpha production was not restricted to LPS in that similar results were obtained using inactivated streptococci. Inhibition of TNF production was associated with a modest (about 30% at 50 microM, 7 donors) reduction of mRNA. A similar inhibition of TNF-alpha production was also detected with mouse peritoneal macrophages. With mouse cells benzydamine also substantially inhibited IL-1 production in vitro. In vivo treatment with benzydamine (40 mg/kg s.c.) protected mice against LPS lethality. Protection against septic shock was observed when benzydamine was administered before or concomitantly with LPS. Protection against LPS toxicity was associated with a marked reduction of serum levels of TNF-alpha and IL-1 beta, whereas IL-6 was unaffected. Inhibition of inflammatory cytokine production may play a role in the anti-inflammatory activity of benzydamine and provide suggestions for novel therapeutic applications.

Animals↗
PubMed1996

Low serum interleukin-10 levels in multiple sclerosis: further evidence for decreased systemic immunosuppression?

Serum interleukin 10 (IL10) levels were assessed in patients with multiple sclerosis who were either in a stable or active clinical condition. The levels were compared with values in healthy controls. Lower IL10 levels than in controls were seen in multiple sclerosis patients, regardless of clinical disease activity. Low IL10 levels were also seen in patients with systemic lupus erythematosus. No clear-cut relationships emerged between IL10 levels and those of tumour necrosis factor alpha and transforming growth factor beta, or between IL10 and lymphocyte subsets in peripheral blood.

Adult↗
PubMed1996

Does a placebo-effect exist in clinical trials on multiple sclerosis? Review of the literature.

To verify whether the outcome in placebo-treated MS patients actually corresponds to that expected on the basis of the natural history and pretrial evolution of the disease, we here review the results of clinical trials conducted according to a placebo-controlled, randomized design, regardless of the experimental therapy used. The frequency of relapse in remitting-relapsing patients decreases during follow-up, and disability in progressive cases increases more slowly than before enrollment. These data should be borne in mind when evaluating the impact of experimental drugs on the natural course of the disease.

Clinical Trials as Topic↗
PubMed1996

A flow-cytometric method to evaluate drug antiaggregating effect on rat neutrophils.

Neutrophils are one of the first cellular populations to become involved in inflammatory processes and some features of the response to inflammatory stimuli can be partially reproduced in vitro by treatment with chemotactic peptides such as N-formyl-methionyl-leucyl-phenylalanine (FMLP). Nonsteroidal antiinflammatory drugs (NSAIDs), such as indomethacin, are known to interfere in vitro with human and rat neutrophil functions and to inhibit FMLP-induced aggregation. In this article we define the scatter parameters of rat neutrophils and demonstrate that flow-cytometric analysis of these cells can be used to analyze the inhibiting action of drugs in an in vitro model of aggregation. We show, in fact, that indomethacin at 100 microM (p < 0.05) and 200 microM (p < 0.01) is able to significantly reduce rat neutrophil aggregation. These results confirm the data obtained by light transmittance aggregometry and indicate that cytometric analysis of aggregation phenomena is a technique suitable for the screening of antiaggregating drugs.

Animals↗
PubMed1996

IFNB in MS.

Explore the source record for details and available documents.

Humans↗
PubMed1995

Analysis of HLA-class II DQA1, DQB1, DRB1 and DPB1 in Italian multiple sclerosis patients.

We studied the allelic constitution at the HLA class II DQA1, DQB1, DRB1 and DPB1 in 94 Italian multiple sclerosis (MS) patients and 98 controls. No significant increase in the frequency of DR2 alleles was detected among MS patients, as previously observed both in European and some Italian studies. A slight increase was found for the DQA1*0301 and DQB1*0602 alleles in the MS patients. No significant association was found with the glutamine residue at position 34 of the DQ alpha chain, which was noted previously in MS patients from northern Europe.

Alleles↗
PubMed1995

Evidence of linkage between susceptibility to multiple sclerosis and HLA-class II loci in Italian multiplex families.

To verify whether multiallelic polymorphisms belonging to HLA class II genes are linked to multiple sclerosis (MS) in the Italian population, we studied 28 multiplex MS families originating from different areas of Italy. Allelic characterization was carried out by analysis of RFLPs and oligonucleotide typing. Evidence supporting the existence of linkage between MS susceptibility and the HLA class II loci DRB1, DQA1 and DQB1 was provided using two non-parametric tests, affected sib-pair analysis, and affected-pedigree-member (APM) analysis. The APM analysis also suggested the existence of genetic heterogeneity for the HLA class II loci and MS susceptibility in our series. Linkage disequilibrium between MS susceptibility and the haplotype DRB1*1501,DQA1*0102,DQB1*0602 was demonstrated by applying the transmission linkage disequilibrium test to our families. Finally, lod score analysis suggests that in our Italian families, MS susceptibility is conferred by HLA class II alleles according to a low-penetrance autosomal recessive mode of inheritance.

Alleles↗
PubMed1994

A new monoclonal antibody (5D3-F7) which recognizes human monocyte-chemotactic protein-1 but not related chemokines. Development of a sandwich ELISA and in situ detection of producing cells.

Chemokines are a superfamily of structurally related cytokines involved in leukocyte recruitment in normal and neoplastic tissues. The availability of non-cross-reacting reagents specific for each member of the C-C and C-X-C family is important for careful characterization of their in vitro and in vivo production and relevance. Here we describe a novel, highly specific, mAb against monocyte chemotactic protein-1 (MCP-1). The 5D3-F7 mAb (IgG1,kappa) recognizes human recombinant and natural MCP-1 in ELISA, immunoprecipitation and immunoblot analysis. As a source of natural MCP-1 we used the 8387 human sarcoma line which produces spontaneously MCP-1 and responds to TNF with increased expression and release. The 5D3-F7 mAb inhibited the chemotactic activity of MCP-1 for monocytes. Using the 5D3-F7 mAb and a polyclonal rabbit anti-MCP-1 serum, a sandwich ELISA was developed. In both the direct and the sandwich ELISA, the 5D3-F7 mAb recognized human MCP-1, but not the closely related C-C chemokines MCP-1, MCP-2, MCP-3, MIP-1 alpha, and RANTES and the C-X-C chemokines IL-8, gro alpha and NAP-2. In culture supernatants the sensitivity of the sandwich ELISA was approximately equal to 30 pg/ml. The sandwich ELISA permitted detection of MCP-1 in resting or cytokine-stimulated endothelial, mesothelial and Kaposi's sarcoma cells. Preliminary immunohistochemical analysis revealed production of MCP-1 by macrophage-like cells at sites of inflammation. The 5D3-F7 mAb provides a novel, highly specific reagent with which to investigate the in vitro and in vivo production and role of MCP-1.

Antibodies, Monoclonal↗
PubMed1994

The myelin basic protein gene is not a major susceptibility locus for multiple sclerosis in Italian patients.

To verify whether multiallelic polymorphism adjacent to the gene encoding for myelin basic protein is associated with or linked to multiple sclerosis in Italians, we studied 54 sporadic patients, 55 control subjects and 18 families with two or more affected individuals. Allelic typing was carried out by analysis of fragment length polymorphisms after DNA amplification by the polymerase chain reaction. The presence of linkage with the disease was tested according to either autosomal dominant or autosomal recessive modes of inheritance, and with or without the introduction of liability classes accounting for the age of the individuals. Furthermore sib-pair analysis was performed in 11 siblings. No evidence for association or linkage between the myelin basic protein gene polymorphism and multiple sclerosis was found. Our data indicate that in the Italian population the myelin basic protein gene does not play a major role in conferring genetic susceptibility to multiple sclerosis, and suggest that the latter is a heterogeneous phenomenon, possibly influenced by the different ethnic origin of the populations which have been investigated.

Adult↗
PubMed1994

Serial evoked potentials in multiple sclerosis bouts. Relation to steroid treatment.

Serial recordings of multimodal sensory (visual, acoustic and somatosensory) evoked potentials were made in 19 relapsing-remitting Multiple Sclerosis patients enrolled in a clinical trial designed to evaluate the efficacy of dexamethasone versus high- and low-dose methylprednisolone in acute multiple sclerosis bouts. Electrophysiological and clinical evaluations were performed at the onset of therapy and until 6 months after the end of treatment. Using an arbitrary Evoked Potentials score that takes into account both latency and waveform alterations, we found a positive correlation between evoked potentials and clinical disability scores. Furthermore, different electrophysiological profiles were detected in the three therapeutic subgroups. Evoked potentials may be useful for monitoring acute Multiple Sclerosis bouts and evaluating the effect of therapy.

Adult↗
PubMed1994

Double-blind trial of dexamethasone versus methylprednisolone in multiple sclerosis acute relapses.

The efficacy of dexamethasone (DX) and methylprednisolone (MP) at high (HD) and low (LD) dose in acute multiple sclerosis (MS) relapses was evaluated by a double-blind trial in 31 patients followed for 1 year. DX and HDMP were similarly efficacious in promoting recovery, while LDMP was ineffective in the short-term outcome and was followed by an early clinical reactivation. The different outcomes seem to be related to different immunomodulating effects, mainly on cerebrospinal fluid (CSF) IgG synthesis and on peripheral blood and CSF CD4+ lymphocyte subsets.

Adult↗
PubMed1994

Immunological monitoring and clinical evaluation in cyclophosphamide-treated progressive multiple sclerosis patients.

15 patients with progressive Multiple Sclerosis (MS) were treated by an induction Cyclophosphamide (CFX) schedule followed by bimonthly boosters over one year. Of the 14 patients who completed the trial, only two displayed worsening of neurological status. Cerebrospinal fluid (CSF) total protein, IgG/Alb ratio and IgG index decreased at 6 months and were still low at the end of the treatment. Pretreatment percentages of PB CD8+ and CD4+CD45RA+ lymphocytes were lower in MS patients in comparison with healthy controls. However, no significant shifts occurred in percentage of PB CD4+, CD4+CD45RA+, CD4+CD45RA-, CD8+ lymphocytes of MS patients during immunosuppressive treatment. The lack of changes in the investigated immunological parameters contrasts with the good clinical response and the low rate of serious side effects during this treatment schedule.

Antigens, CD↗
PubMed1994

High-dose methylprednisolone treatment-induced changes in immunological parameters in progressive MS patients.

The effect of High-Dose Methylprednisolone (HD-MP) treatment on Peripheral Blood (PB) and Cerebrospinal Fluid (CSF) immune parameters was investigated in 9 patients with relapsing-progressive Multiple Sclerosis (MS). Short-time effects included reduction of the percentage of CD3+, CD4+ and CD4+CD45RA+ PB lymphocytes and increase in CD3-CD56+ cells. At the end of the treatment, only increase in PB CD19+ and in CSF CD8+, CD8+CD28+ and decrease of CSF CD4+CD45RA- and serum IL2R levels were observed. No changes in CD11a+CD4+, CD18+CD14+ PB cells were observed after treatment. The results further stress the complex and multifaceted action of HD-MP on immune parameters.

Adult↗
PubMed1993

A double blind study on azathioprine efficacy in multiple sclerosis: final report.

Forty patients, affected by multiple sclerosis with remitting-relapsing or progressive course, were included in a double blind study of treatment with azathioprine (2 mg/kg/day) lasting 3 years. The mean changes on the Expanded Disability Status Scale and in the survival analysis show a trend in favour of azathioprine both in slowing disease progression and reducing relapse frequency. These findings, repeatedly observed in similar trials, indicate that azathioprine should be used in the treatment of multiple sclerosis.

Actuarial Analysis↗
PubMed1993

Presence of T-cell subset abnormalities in newly diagnosed cases of multiple sclerosis and relationship with short-term clinical activity.

Abnormalities of T-cell subsets in patients with multiple sclerosis are well known; in order to assess whether immunological abnormalities are relevant in the pathogenesis of the disease after its clinical onset, peripheral blood lymphocyte subsets (CD3+, CD4+, CD4(+)-CD45RA+, CD4+CD45RA-, CD8+, CD8+CD57+, CD57+, CD25+) were analysed serially in 25 patients at the first clinical episode suggestive of inflammatory demyelinating disease and in an equal number of age- and sex-matched controls. During the follow-up period (12-18 months, mean 14) 6 of 25 patients presented new relapses: in this subgroup of patients, significant changes in CD4+ ratio (% CD4+CD45RA-/%CD4+CD45RA+) were detected in comparison both with healthy controls and with clinically stable patients. Patients clinically stable at follow-up did not display immunological abnormalities, regardless of the presence or absence of cerebrospinal fluid and/or magnetic resonance imaging alterations consistent with multiple sclerosis. These findings suggest a possible prognostic role of early T-cell subset imbalance in multiple sclerosis.

Adolescent↗