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Biomedical subjects

M Marangi

Publications and source records attributed to M Marangi.

At least 19 recordsLinked to original sources

Alpha-defensin increase in peripheral blood mononuclear cells from patients with hepatitis C virus chronic infection.

The alpha-defensin genes promoter regions contain a putative nuclear factors of activated T cells (NFAT)-binding site and it is known that hepatitis C virus (HCV) core protein activates the interleukin (IL)-2 gene transcription through the NFAT pathway. The aims of this study were to investigate if HCV affects the alpha-defensin expression in peripheral human mononuclear cells (PBMCs) and to evaluate the existence of a correlation between alpha-defensins and liver damage in patients with chronic hepatitis C. Ninety patients with chronic hepatitis C, 30 with chronic hepatitis B and 25 healthy controls were enrolled. Alpha-defensins were identified and quantified in PBMCs by mass spectrometry, enzyme-linked immunosorbent assay, antibacterial activity and mRNA levels. PBMCs from three patients and controls were stimulated with HCV core protein, hepatitis B virus core antigen and the alpha-defensin mRNAs level was quantified. We found that HCV core protein activates in vitro the alpha-defensin transcription. Alpha-defensin levels in patients with chronic hepatitis C (mean +/- SD = 1.103 +/- 0.765 ng/10(6) cells), chronic hepatitis B (0.53 +/- 0.15) and healthy controls (0.217 +/- 0.09) resulted significantly different (P < 0.001). In patients with chronic hepatitis C, the alpha-defensin levels and antibacterial activity correlate with the liver fibrosis. Our data suggest that HCV induces alpha-defensin expression. The high linear correlation of alpha-defensin levels with advancing fibrosis makes the measure of these peptides a reliable marker to evaluate fibrosis stage.

Adult↗

The impact of the Catholic Jubilee in 2000 on infectious diseases. A case-control study of giardiasis, Rome, Italy 2000-2001.

Mass gatherings are believed to increase the transmission of infectious diseases although surveillance systems have shown a low impact. The Catholic Jubilee was held in Rome, Italy in 2000. We conducted a case-control study to analyse the risk factors of giardiasis among residents. All diseases reported to the laboratory surveillance system from January 2000 to May 2001 were compared with hospital controls concurrently selected in the same season as cases and frequency-matched for age and birth country. Fifty-two cases (44.1%) and 72 controls were enrolled. In the multivariable analysis factors associated with giardiasis among adults were: travelling abroad (OR 24.2, P>0.01), exposure to surface water (OR 4.80, P=0.05), high educational level (OR 3.8, P=0.03). Having a maid from a high-prevalence country was independently associated (OR 2.3) although not statistically significant. This is the only exposure that changed during the Jubilee.

Adolescent↗

Expression modes of urinary N-acetyl-beta-D-glucosaminidase in patients with chronic renal insufficiency.

BACKGROUND: Urinary N-acetyl-beta-D-glucosaminidase (NAG) activity has emerged as potentially useful early marker of renal tubular injury. This activity is usually evaluated in random urine samples and is related to urinary creatinine concentration. Reports about the lack of correlation between NAG activity of 24-h urines and activity of random urine samples in some clinical and experimental situations led us to study the correlation existing between different procedures for expressing urinary NAG in patients with chronic renal insufficiency. METHODS: Thirty samples of 24-h urine and 30 random urine samples from chronic renal insufficiency patients were collected. The activity of urinary NAG was examined fluorimetrically. RESULTS: The following correlations were observed: (1) r = 0.431 (P = 0.017) for activity in random urine samples and total activity in 24-h urines); (2) r = 0.281 (P = 0.005) for activity in random samples and activity, expressed as U/l, in 24-h urines. CONCLUSIONS: The data show that collection of urine excreted over the whole day and evaluation of total daily excretion of NAG seems the method of choice, at least for patients with chronic renal insufficiency.

Acetylglucosaminidase↗

A case of loiasis in Rome.

BACKGROUND: Owing to the increase of an immigrant population and of Italian citizens travelling for tourism or on business, it is nowadays possible to observe clinical pictures characteristic of tropical regions, often with indistinct symptoms. One of these is Loa loa infestation, or loiasis, a form of filariasis caused by Loa loa and transmitted by the Chrysops fly. We present the case of a male immigrant from Cameroon. Characteristic symptoms were intense xerosis, mostly of the third inferior part of the legs, intensely pruritic, with numerous lesions from scratching. No benefit was obtained by emollient topics, anti-acarus and systemic antihistamines. METHOD: Serum samples and Giemsa, haematoxylin, haematoxylin + Giemsa concentration-on-membrane stains, have evidenced the presence of Loa loa microfilariae. RESULTS: A diagnosis for L. loa (loiasis) infestation was made. CONCLUSIONS: At the beginning of the migration phenomenon, particularly from Africa, Italian physicians, especially dermatologists, were eagerly looking for 'tropical' diseases; this approach can be defined as 'Salgari's syndrome' from the name of the Italian novelist who, though never travelling out of Italy, had perfectly described environments and habits typical of far away countries. Now, conversely, we have to avoid the opposite approach of considering real tropical diseases as related to social or psychological difficult conditions.

Adult↗

Are elevated levels of soluble ICAM-1 a marker of chronic graft disease in heart transplant recipients?

Positivity for circulating intercellular adhesion molecule-1 (ICAM-1) in heart transplant recipients has been claimed to predict the development of coronary artery disease and risk of graft failure. Soluble ICAM-1 were evaluated in 32 heart transplant recipients. Five of these patients, who had undergone transplantation several years before, were positive for soluble ICAM-1 but did not present any clinical sign of graft rejection. Furthermore, although heart graft coronary disease was diagnosed in 15 of the 32 patients, they did not show significantly higher titres of soluble ICAM-1 compared to the remaining patients. These findings suggest that major caution is necessary when considering ICAM-1 positivity as a marker of graft disease.

Adult↗

Conditionally adherent growth of serum-independent CHO cells for automated drug screening and biopharmaceutical production.

SSF3 is a CHO cell line adapted for growth in protein-free medium. It grows in suspension unless serum-derived attachment factors such as vitronectin are added to the medium. Serum-independent cell lines, which adhere to the substrate after induction with dexamethasone or constitutively, were created by transfection with a human vitronectin gene under control of the mouse mammary tumor-virus promoter. Substrate attachment and SSF3VN-cell spreading could be prevented with an RGD peptide (arginine-glycine-aspartic acid) confirming that attachment is mediated by an intregrin receptor. Hormone-inducible attachment could be blocked by glucocorticoid antagonist promegestone. All steps in the isolation of stable transfected SSF3VN cell lines could be done in a chemically defined medium avoiding the risk of introduction of serum-derived infectious agents. SSF3VN cells could be grown in protein-free medium in solid-phase large-scale bioreactors. Application in microplates as used in high-throughput screening was demonstrated in an assay of Ca(2+) release from internal stores induced by agonist-binding to recombinant human metabotropic glutamate receptor hmGluR1b.

Animals↗

The relationship between glycated hemoglobin and polymorphonuclear leukocyte leukotriene B4 release in people with diabetes mellitus.

In order to evaluate polymorphonuclear leukocyte (PMN) activity in diabetes mellitus, leukotriene B4 (LTB4) levels were measured in sixty patients, 31 affected with Type 1 diabetes mellitus and 29 affected with Type 2 diabetes mellitus. The LTB4 levels (12.1+/-0.2 pg/100 microl) in diabetic patients were higher compared to those of the control group (7.9+/-0.1 pg/100 microl) (P < 0.001), and remained significantly higher (P < 0.001) (12.8+/-0.2 pg/100 microl) than in the control group (11.0+/-0.2 pg/100 microl) after stimulation with calcium ionophore. A significant and positive correlation between glycated hemoglobin and LTB4 was demonstrated (P < 0.001, r = 0.80). This study demonstrates that in diabetic patients there is a PMN activation and that this activation is correlated to glycated hemoglobin level.

Adult↗

Role of group B streptococcal capsular polysaccharides in the induction of septic arthritis.

The ability of different serotypes of group B streptococci (GBS) to induce septic arthritis in mice was compared. Types II, III, IV, V, VI and VII GBS were investigated. A highly capsulate strain of type III GBS, COH1, and its mutants, COH1-11 (lacking capsular sialic acid) and COH1-13 (non-capsulate), obtained by transposon insertional mutagenesis, were used to assess the role of type-specific polysaccharide on the induction of arthritis. At an intravenous dose of 10(7) cfu/mouse, reference strains of types II, III, IV, VI and VII and type III strain COH1 induced arthritis with an incidence ranging from 70 to 90%. For type V and strain COH1-11, 10(8) cfu/mouse was required to obtain a 50% incidence of arthritis; lesions were not evident with strain COH1-13. The presence of the capsule played a major role in the induction of GBS septic arthritis. The presence and amount of sialic acid in capsular polysaccharide influenced the incidence of articular lesions. The bacterial dose affected the manifestations of arthritis; the less virulent strains of GBS also induced articular lesions when an adequate number of micro-organisms reached the joints.

Animals↗

High-level expression in insect cells and purification of secreted monomeric single-chain Fv antibodies.

We have constructed a recombinant baculovirus encoding an anti-(phenyl-oxazolone) single-chain Fv antibody (anti-phOx-scFv) fused to the baculovirus GP67 secretion signal sequence, 6 liters of Sf9 insect cells were infected with this virus at a multiplicity of infection of one and cultured in a bioreactor for 72 h. The dialyzed supernatant was subjected to cation exchange chromatography at pH 6.0 followed by size exclusion chromatography on a Sephadex G100 superfine matrix. This rapid protocol resulted in the isolation of monomeric scFv with a purity of greater than 98%. The final yield was 32 mg/l (10(9) cells/l). Partial amino-terminal sequencing revealed that the GP67 signal sequence was completely removed upon secretion. The dissociation constant of the scFv monomers is about 1 x 10(-4) M. By competitive ELISA scFv dimers yielded a half maximum inhibitory concentration of 3.4 x 10(-7 M which matches the earlier measured Kd for the anti-phOx-scFv (3.2-5.3 x 10-7 M. Marks et al. (1991) J. Mol. Biol. 222, 581-597: Marks et al. (1992) Bio/Technology 10, 779-783). This method is readily scaled up for the preparation of scFv antibodies in high yield and purity obviating any affinity chromatography and/or refolding steps by exploitation of insect cell expression as an efficient alternative to E. coli expression.

Animals↗

Evidence for the existence of ganglioside molecules in the antigen of Entamoeba histolytica.

Gangliosides were found to be present in Entamoeba histolytica. They were extracted from lyophilized trophozoites of the pathogenic strain HM-1:IMSS and purified by high performance thin-layer chromatography. Two resorcinol-positive bands, comigrating with GM2 and GD1a were demonstrated, revealing the existence of ganglioside molecules in Entamoeba histolytica. The GM2 content, determined as lipid-bound sialic acid, was 1.5 micrograms/10(8) amoebae, the content of the GD1a comigrating band was 0.32 microgram/10(8) amoebae. The identity of the GM2 comigrating band was confirmed by TLC immunostaining, using the monoclonal anti-GM2 antibody GMB28. Furthermore, six out of ten anti-amoeba positive sera selectively reacted with the GM2 comigrating band, as revealed by immunostaining on TLC plates. Absorption tests revealed that preincubation of anti-amoeba positive sera with standard GM2 was followed by a significant decrease in the reaction with amoeba trophozoites by indirect immunofluorescence. These results demonstrate that a GM2 comigrating component of Entamoeba histolytica may be one of the antigens responsible for the appearance of circulating antibodies in patients with amoebiasis.

Absorption↗

A new serological technique, time-resolved fluoroimmunoassay (TRFIA), for the immunological diagnosis of urinary schistosomiasis.

The application of a new serological method, time-resolved fluoroimmunoassay (TRFIA), is described for the diagnosis of urinary schistosomiasis. A chelate of lanthanides (europium) with a long fluorescent life-time is used as label. The intensity of fluorescence is measured after a delay selected to eliminate almost completely the background fluorescence, which decays rapidly. TRFIA was compared with an established method, enzyme linked immunosorbent assay (ELISA). Using sera from proven cases of Schistosoma haematobium infection, 98.1% of the samples were positive by TRFIA and 86.5% by ELISA. Sera from patients infected with helminths other than schistosomes produced only 1.5% of false positives with TRFIA, compared with 12.3% by ELISA. TRFIA is more sensitive and specific than ELISA.

Enzyme-Linked Immunosorbent Assay↗

Influence of sexual hormones on solid tumor growth and ATP C+ cell replication.

Solid tumors obtained by implanting ATP C+ cells subcutaneously into the back of BALB/cf/Had/Se substrain mice developed more rapidly in males than females. Experiments conducted on gonadectomized animals demonstrated that female gonads inhibit the development of this tumor. Studies on ATP C+ cells cultured in vitro with various doses of male and female hormones showed that mainly progesterone, but to a lesser extent also estradiol, significantly inhibit cell proliferation with respect to testosterone. The treatment of normal and gonadectomized animals with testosterone and progesterone confirms the results obtained in vitro. Analyses carried out on ATP C+ cells do not reveal presence of estradiol and progesterone receptors.

Animals↗