Search PubMed⌕ Search

Biomedical subjects

M Milella

Publications and source records attributed to M Milella.

At least 55 records · Page 3Linked to original sources
PubMed1995

Cross-linking of alpha 4 beta 1 and alpha 5 beta 1 fibronectin receptors enhances natural killer cell cytotoxic activity.

Cell/extracellular matrix interactions mediated by integrins regulate differentiation, migration, and effector functions of immune system components. Human NK cells express alpha 4 beta 1 and alpha 5 beta 1 integrins, which mediate their binding to fibronectin (FN). We have investigated the ability of FN and its beta 1 integrin receptors to modulate NK cytotoxicity. Our data show that the presence of immobilized FN significantly augments the spontaneous cytotoxic activity of in vitro cultured human NK cells against several NK-susceptible, but not NK-resistant, target cells; Ab-dependent cytotoxicity against Ab-coated P815 target cells and the redirected lysis of anti-CD16 hybridomas are also enhanced in the presence of FN. Solid-phase-bound anti-human beta 1, or its F(ab')2 fragment, anti-alpha 4 and anti-alpha 5 mAbs, all consistently enhance Ab-dependent cytotoxicity against Ab-coated murine target cells. The 120-kDa (alpha 5 beta 1-binding), but not the 40-kDa (alpha 4 beta 1-binding), FN fragment fully reproduced the enhancing effect observed with the entire molecule. Our data also demonstrate that alpha 4 beta 1 and alpha 5 beta 1 cross-linking on NK cells induces an increase of intracellular Ca2+ concentration that is abrogated by EGTA, thus suggesting that the capacity to mobilize Ca2+ is involved in the coactivating role of alpha 4 beta 1 and alpha 5 beta 1 FN receptors in the cytotoxic functions of NK cells.

Antibodies, Monoclonal↗
PubMed1995

Stimulation of protein tyrosine phosphorylation by interaction of NK cells with fibronectin via alpha 4 beta 1 and alpha 5 beta 1.

Recent evidence indicates that integrins do transduce signals that are involved in the regulation of a number of cellular processes. We have shown previously that human NK cells express alpha 4 beta 1 and alpha 5 beta 1 integrins, which mediate their adhesion to fibronectin (FN). Here we investigate whether cross-linking of beta 1 FN receptors on human NK cells stimulates tyrosine kinase activation. Our results indicate that cross-linking of beta 1 integrins on NK cells, either freshly isolated from PBL or generated from 10-day coculture of nonadherent PBMC with an irradiated EVB+ lymphoblastoid B cell line (long-term activated NK cells), altered the tyrosine phosphorylation pattern. In particular, we found stimulation of tyrosine phosphorylation of two proteins migrating with an apparent mass of 105 and 115 kDa, which was not observed after CD16 engagement. Phosphorylation of pp105-115 was already observed at 1 min, raised maximal values at 3 to 5 min, and persisted until 20 min after stimulation. Tyrosine phosphorylation of pp105-115 was also observed upon engagement of alpha 4 beta 1 and alpha 5 beta 1 FN receptors either with mAb directed against the alpha subunits or after NK cell adhesion to FN or its 120- and 40-kDa fragments. Pretreatment of NK cells with the tyrosine kinase inhibitor herbimycin A resulted in marked decrease of phosphorylation stimulated through alpha 4 beta 1 and alpha 5 beta 1 integrins, indicating that ligation of FN receptors on NK cells activates tyrosine kinase(s). Overall our results suggest that beta 1 integrins on NK cells play a major role as signaling molecules in the regulation of NK cell functions.

Animals↗
PubMed1995

Treatment with natural IFN of hepatitis C patients with or without antibodies to recombinant IFN.

The purpose of this study was to evaluate if a retreatment with natural human interferon alpha could lead to a recovery of the therapeutic responsiveness in non-responder patients with chronic hepatitis C who did (9 cases) or did not (14 cases) develop anti-interferon neutralizing antibodies while on treatment with recombinant interferon alpha2a. During retreatment, no patient developed detectable levels of neutralizing antibodies to natural interferon. At the end of retreatment, 6/9 positive patients showed a complete response to natural interferon therapy, while only 1/14 negative patients had a partial response. These data suggest that a second course of treatment with the natural preparation may be useful in patients who failed to respond to an earlier course of recombinant interferon, particularly the anti-interferon positive patients who showed hepatitis reactivation after an initial response concomitantly with antibody appearance. Conversely, a second course of natural interferon therapy might be useless in cases in which the lack of response is not associated to antibody development.

Antibodies↗
PubMed1994

Long term activation of natural killer cells results in modulation of beta 1-integrin expression and function.

Integrin expression and function is largely modulated by cell activation. Here we provide evidence that long term activation of human NK cells results in a marked modulation of beta 1-integrin expression and adhesive functions. By flow cytometry and immunochemical analysis we have detected induction of alpha 1 beta 1 and alpha 2 beta 1, increased expression of alpha 4 beta 1 and alpha 5 beta 1, and decline of alpha 6 beta 1 on CD3-CD56+ NK cells generated from 10-day coculture of nonadherent PBMC with irradiated RPMI 8866 EBV+ lymphoblastoid B cell line. Adhesion assays performed on extracellular matrix-coated plates showed that, unlike fresh NK cells, long term-activated NK cells bind to native collagen I via alpha 2 beta 1 and to heat-denatured collagen I in an RGD-dependent manner, although they lose the ability to bind to laminin. In regard to the adhesion to FN, no major quantitative changes are observed after long term NK cell activation. However, whereas alpha 4 beta 1 and alpha 5 beta 1 completely mediate the adhesion of fresh NK cells to fibronectin, binding of activated NK cells is only partially beta 1-dependent and seems to involve also non-beta 1-integrin(s) recognizing and RGD sequence. The modulation of beta 1-integrin expression and the acquisition of new adhesive properties on long term-activated NK cells may be relevant for their traffic and tissue localization during inflammation and immune response.

Amino Acid Sequence↗
PubMed1994

A randomized phase II study comparing sequential versus simultaneous chemo-radiotherapy in patients with unresectable locally advanced squamous cell cancer of the head and neck.

BACKGROUND: Single-modality radiotherapy is still considered standard treatment for patients with locally advanced unresectable cancer of the head and neck. As treatment outcome is poor, attempts to integrate chemotherapy into the overall management of these patients are ongoing. PATIENTS AND METHODS: A randomized study was undertaken to compare a sequential with a simultaneous chemoradiotherapy program. Between February 1986 and February 1991, 93 eligible patients with locally advanced unresectable cancer of the head and neck were stratified by WHO PS, T and N class and primary site and then randomized to receive either three courses of neoadjuvant chemotherapy with cisplatin (100 mg/m2 i.v. d 1) and 5-fluorouracil 1000 mg/m2/days 1-5 by continuous i.v. infusion every 3 weeks prior to definitive conventional radiotherapy of 65-70 Gy (sequential treatment), or cisplatin 100 mg/m2 on days 1, 22, 43 given simultaneously for the duration of the same conventional radiotherapy (simultaneous treatment). RESULTS: At the end of the entire treatment 18 complete responses (47%) in the sequential-treatment arm and 18 (41%) in the simultaneous treatment arm were obtained. No statistically significant differences in the 5-yr progression-free survival, in the median time to loco-regional and distant progression and in the 5-yr overall survival were observed. Leukopenia was more frequent in the simultaneous than in the sequential arm (p = 0.03), whereas alopecia (p = 0.008) and phlebitis (p < 0.0001) were more frequent in the sequential-treatment arm. A better compliance was associated with the concomitant treatment, with 87% of the patients completing the entire radiotherapy program versus 63% of those in the sequential arm (p = 0.01). CONCLUSIONS: In the present study, the two treatment arms showed similar activity (complete response, progression-free and overall survival rates). Compliance to treatment was better in the concomitant arm. These data suggest that concomitant chemo-radiation therapy might be considered an option in unresectable locally advanced cancer of the head and neck. Phase III studies are needed in order to establish the superiority of this combination of cisplatin and radiotherapy versus radiotherapy alone.

Adult↗
PubMed1994

Chemoembolization of hepatocellular carcinoma in patients who undergo liver transplantation.

PURPOSE: To determine the efficacy of chemoembolization (CHE) for hepatocellular carcinoma (HCC) in patients with a cirrhotic liver. MATERIALS AND METHODS: Twenty-one patients with cirrhosis and 33 HCC nodules received a mean of 2.4 cycles of CHE before orthotopic liver transplantation (OLT). Iodized oil plus doxorubicin (13 patients) or mitoxantrone (eight patients) was injected into the hepatic artery. Twelve (57%) of the patients underwent terminal embolization with gelatin sponge pledgets. After OLT, the percentage of tumor necrosis was assessed in each HCC nodule. RESULTS: Twelve (36%) of 33 HCC nodules responded completely to CHE (necrosis > 90%). Successful treatment was statistically significantly related to nodular diameter greater than 2 cm (P = .03), solitariness (P = .0009), hypervascularity (P = .001), and good retention of iodized oil (P = .05). Seven (33%) of the patients had major complications; no death occurred in association with CHE. All patients survived OLT; in three patients (14%), HCC recurred a median of 21 months after OLT. CONCLUSION: CHE seems to be an effective treatment for HCC in patients with liver cirrhosis who await OLT.

Angiography, Digital Subtraction↗
PubMed1994

The Italian experience with SH U 508 A (Levovist) in breast disease.

The authors report their experience with color Doppler US of breast disease with a new parenteral contrast agent--i.e., SH U 508 A (Levovist)--in an international multicenter experimental trial. Over a 9-month period, 17 patients with 19 solid breast lesions were examined. A baseline color Doppler study was performed on every patient, together with a second exam after injecting the contrast agent into a peripheral vein to investigate possible changes in the depiction of tumor vessels, inside and around the lesions. The 19 nodules included 16 infiltrating ductal carcinomas, 2 fibroadenomas and 1 benign phylloid tumor. Levovist improved the depiction of tumor vessels in all cases and helped detect new vascular signals which had been missed on baseline images. The increase in color Doppler signal was graded as marked and mild: in 68.75% of ductal carcinomas and in the benign phylloid tumor signal increase was marked, while in 31.25% of carcinomas and in fibroadenomas signal increase was mild. All lesions exhibited vascular signals both inside and around the lesion; many afferent poles were also depicted. Our results demonstrate that conventional color Doppler semiology changes with the use of a contrast agent and pave the way for further studies on larger series of cases to assess new diagnostic criteria to differentiate breast lesions according to color Doppler signal patterns.

Adult↗
PubMed1994

US contrast enhancement with intra-arterial CO2 injection in the staging of hepatocellular carcinomas.

Liver US contrast enhancement was obtained with the intrahepatic arterial injection of CO2 in 45 hepatocellular carcinoma patients. Before digital subtraction angiography (DSA) all the patients underwent conventional US and CT of the liver; 20 patients also underwent CT during arterial portography (CTAP). Liver US contrast enhancement with CO2 exhibited the same sensitivity as DSA (92%) in demonstrating hypervascular HCCs and proved superior to the other diagnostic procedures -- 85% for CT and 78% for conventional US. DSA combined with echocarbography, with its 2D demonstration of liver anatomy, improved the correct staging of HCCs, for better treatment planning.

Angiography↗
PubMed1993

Color-Doppler ultrasound in ovarian masses: anatomo-pathologic correlation.

BACKGROUND AND AIMS: In the radiologic assessment of ovarian masses, the major difficulty consists in the late recognition and lack of parameters for a differential diagnosis between benign and malignant lesions, especially in the post-menopause when the incidence of cancer is higher. The use of a transvaginal probe and the color-Doppler examination have recently improved the study of the female pelvis. This study is aimed to verify the possibility of the color-Doppler imaging to differentiate between malignant and benign ovarian lesions during transvaginal echographies, on the basis of the qualitative and quantitative characteristics of the vascular pattern of the ovarian lesions. RESULTS: Twenty-six expansive ovarian lesions were studied: 8/26 showed no vascular signals and were considered benign as confirmed at histology. In the remaining lesions with some vascularization, the resistance index (RI) was evaluated: those with RI > 0.40 were considered benign, those with RI < 0.40 malignant. In 8/9 benign lesions and 7/9 malignant neoplasms, the results of color-Doppler were coherent with histology. The results showed a sensibility of 87.5% and a specificity of 88.8% for the transvaginal examination. CONCLUSIONS: The main advantages of the color-Doppler transvaginal examination are: the high frequency of visualization of the ovaries, even in postmenopausal patients; the definition of small lesions; the visualization of small parenchymal vessels, both physiologic and pathologic, and their quantitative analysis. The importance of the RI cutoff was critical for the differential diagnosis between benign and malignant lesions; we think that a cutoff of 0.50, instead of 0.40 proposed by other authors, would be far more appropriate.

Adult↗
PubMed1993

Long-term response to interferon therapy in chronic hepatitis B: importance of hepatitis B virus heterogeneity.

The long-term therapeutic efficacy of alpha IFN and the influence of preC variants on the type of response were evaluated in 25 patients with chronic hepatitis B, 14 HBeAg and 11 antiHBe positive patients, treated with alpha IFN and monitored for at least four years after discontinuing therapy. In both groups of patients, serum HBV-DNA became frequently undetectable by DNA dot blot during treatment, suggesting that alpha IFN has an antiviral effect both on HBeAg and antiHBe positive chronic carriers. However, long term follow up showed that the loss of viral DNA in antiHBe carriers was only transient, because all responder patients relapsed from 1 to 48 months after IFN withdrawal. In the HBeAg positive carriers, selection for preC mutants was observed at the end of follow up in 2 patients who seroconverted to antiHBe and remained viremic. Both the frequent occurrence of reactivations in antiHBe compared to HBeAg carriers, and the association of IFN therapy with preC mutant virus selection during long term post-treatment follow up observed in this study, indicate that preC variants are more resistant to IFN therapy than preC wild type HBV. Our data suggest therefore, that IFN therapy may be less frequently able to induce a permanent remission in patients infected with preC mutants.

Alanine Transaminase↗
PubMed1993

Neutralizing antibodies to recombinant alpha-interferon and response to therapy in chronic hepatitis C virus infection.

Forty-seven patients with chronic hepatitis C were treated with recombinant interferon alpha-2a (rIFN alpha 2a) given subcutaneously in a standard dose of 3 MU thrice weekly for 12 months. Stored baseline sera and monthly samples during treatment were assayed for anti-interferon neutralizing antibodies using the antiviral neutralization bioassay against 5 IU of rIFN alpha 2a. During therapy, 15 of 47 patients (31.9%) developed detectable levels of neutralizing antibodies within 2-8 months after starting treatment. After 12 months of therapy, 26 of 32 antibody-negative patients (81.3%) showed normalization or marked reduction of ALT levels compared to 4 of 15 (26.6%) who developed anti-IFN neutralizing antibodies (p = 0.0009). Four patients demonstrated antiviral response during treatment even in the presence of low levels or late occurrence of neutralizing antibodies. Six of the seven patients who had disease reactivation after an initial response developed high titers of neutralizing antibodies. Our results suggest that reactivation of chronic hepatitis C before completion of therapy seems to be an obvious consequence of anti-IFN neutralizing antibody formation.

Alanine Transaminase↗
PubMed1992

Development of antibody to hepatitis C virus (HCV) in acute and chronic non-A, non-B post-transfusion hepatitis.

The antibody to hepatitis C virus (anti-HCV) was measured by an immunoassay in 507 serum samples from 94 patients with acute and chronic post-transfusion non-A, non-B hepatitis (NANB) and in 436 healthy blood donors. Anti-HCV was found in 70.8 of patients with acute hepatitis, in 78.2 with chronic hepatitis, and in 1.4 of healthy blood donors. In acute hepatitis, anti-HCV appeared in the serum from 4 to 34 weeks after transfusion and from 1 to 30 weeks after the onset of the overt disease. Three patients with resolving hepatitis (21%) and 2 who developed chronic hepatitis (10%) lost anti-HCV during a mean follow-up period of 28 months. Among the 36 patients with chronic hepatitis, 2 (6%) lost anti-HCV after 12 months and 8 years respectively. These data indicate that in recent years HCV has been the major etiologic agent of acute and chronic transfusion-associated hepatitis (TAH) in our geographical area. The late appearance of anti-HCV from the onset of clinical and biochemical signs of acute hepatitis in more than 70% of patients limits the diagnostic utility of this assay for an earlier serological diagnosis of acute NANB hepatitis. Additional studies are required to determine the diagnostic significance of this antibody in chronic NANB hepatitis.

Adult↗
PubMed1992

Hepatitis B virus genomes that cannot synthesize pre-S2 proteins occur frequently and as dominant virus populations in chronic carriers in Italy.

The heterogeneity of hepatitis B virus (HBV) pre-S sequences coding for envelope proteins was tested by DNA amplification and direct sequencing of viral genomes from sera of 22 unselected chronic carriers resident in Southern Italy. The sequences of the dominant viral genome populations from 15 carriers were very similar to known published "wildtype" HBV genomes and showed no deletions. In contrast, in the HBV populations of six patients, deletions in the pre-S region, mainly clustered at the amino terminal end of the pre-S2 region, were found. Four of these mutant genome populations and those from another patient cannot express pre-S2 proteins due to deletions or a mutation of the translation initiation codon. Emergence of the pre-S mutant viruses either during the natural course of infection or after interferon treatment was found in follow-up sera of one and two patients, respectively. These data indicate a high prevalence of pre-S mutant viruses which cannot express pre-S2 and normal-size pre-S1 proteins. This has important implications for the usefulness of diagnostic pre-S protein assays and possibly for interferon treatment and the efficacy of new vaccines containing pre-S proteins.

Amino Acid Sequence↗
PubMed1992

Anti-HBe-positive chronic hepatitis B with HBV-DNA in the serum response to a 6-month course of lymphoblastoid interferon.

Eighteen heterosexual HBsAg carriers with anti-HBe- and HBV-DNA-positive chronic hepatitis B (CHB) were randomly assigned to receive human lymphoblastoid interferon (ly-IFN) at a dose of 5 MU/m2 i.m. three times a week for 6 months (ten cases) or no treatment (eight cases). All patients were followed for 24 months after IFN discontinuation and received a second liver biopsy. During the 6 months of treatment all patients had a progressive reduction of serum HBV-DNA levels, and at the end of therapy nine out of ten were HBV-DNA-negative and had normal ALT values. None of the untreated patients became persistently HBV-DNA-negative or showed significant variations of ALT levels. During the post-treatment follow-up, from 1 to 17 months after ly-IFN discontinuation, eight of the nine responders (89%) had recurrent or persistent reappearance of HBV-DNA in the serum and reactivation of the liver disease activity, with an ALT peak in four of them. On the post-trial liver biopsy seven of the eight relapsed patients showed persistence of HBcAg reactivity with no significant difference in the percentage of positive cells with respect to the pre-treatment liver specimen. Histological features improved in four treated patients, worsened in one untreated case and were unchanged in the remaining patients. These results indicate that ly-IFN shows a transient antiviral effect in the therapy of anti-HBe- and HBV-DNA-positive CHB. The 6-month treatment regimen employed in this study seems insufficient for eradicating the replicating virus from the liver cells in the majority of patients and consequently does not appear to prevent HBV reactivation after IFN discontinuation.

Adult↗
PubMed1991

Lack of HBV and HDV replicative activity in HBsAg-positive intravenous drug addicts with immune deficiency due to HIV.

The effects of HIV infection on HBV and HDV replication and liver damage were evaluated by comparing the findings from 48 anti-HIV-positive HBsAg chronic carriers with those from 22 matched anti-HIV-negative subjects. The state of HBV/HDV infection was also related to the degree of immunodeficiency of the anti-HIV-positive patients. Most patients were intravenous drug addicts (IVDA) (84.2%); male homosexuals represented only a small proportion (7.1%). Serum HBV-DNA was detected more frequently in anti-HIV-negative than in anti-HIV-positive patients (50% vs. 35%) despite evidence of HDV replication in the anti-HIV-negative group (P = 0.02). Seroconversion from ongoing to inactive HBV infection occurred in 45% of anti-HIV-negative patients as well as in 23% of anti-HIV-positive patients (P = ns). The difference in severity of liver damage between the two groups was not statistically significant (P = 0.84). Furthermore, in the anti-HIV-positive subjects, HBV and/or HDV activity was detected in 63% of patients with mild immunodeficiency (CDC groups II and III with a total CD4 count greater than 400/mm3) and also in 75% of ARC-AIDS patients (CDC groups IV A-IV C) (P = ns). Severe hepatic disease occurred in subjects with CD4 counts above or below 400/mm3 (13 vs. 6, respectively). In conclusion, the data do not demonstrate that HBV or HDV infections are modified by HIV. The epidemiological background of the patients investigated and the extensive spread of hepatitis viruses in Italy before the appearance of HIV may account for the lack of relationship between HIV and HBV/HDV infections.

Adult↗