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Biomedical subjects

G Anastasi

Publications and source records attributed to G Anastasi.

At least 19 recordsLinked to original sources

Actin-associated proteins in ameloblast differentiation.

Focal contacts are systems of adherens junctions of the cell-extracellular matrix type, which allow the transfer of fundamental signals from the extracellular matrix to nuclear compartments, capable of regulating adhesion, proliferation, migration and differentiation of cells. Recently, many authors have concentrated their attention on epitheliomesenchymal interactions which guide organogenesis of dental germ, identifying numerous growth and differentiation factors and having the inner enamel epithelial cells of the enamel organ as a target. Given that the two cellular compartments in their tooth germ are separated by a basal membrane and by an extracellular matrix, which touches it, we wanted to evaluate the presence of focal contacts through the identification of talin and vinculin, proteins of the actin-associated protein complex. In this study we utilized the hemimandibles of young Wistar rats and we extracted the related odontogenic tooth organs present at their apical end. Specimens are processed with antibody against vinculin and talin. Results show that these junctional system proteins are present at the apical poles of both cellular compartments suggesting that putative epithelial-mesenchymal interactions, other than marker molecules, may use focal contacts as a system for transmission of signals.

Actins↗

AQP1 in red blood cells of uremic patients during hemodialytic treatment.

Hemodialysis influences the transport of water through the erythrocytic membrane, and induces morphologic and functional modifications. Recently water channels, called aquaporins (AQP), have been identified on the membrane of red blood cells. The aim of the present study was, therefore, to evaluate any relationships between volumetric changes in erythrocytes (MCV), plasma osmolarity and membrane expression of AQP1 in 22 uremic patients during a hemodialysis session, and compare value with those in a control group of 22 healthy volunteers. Membranal AQP1 expression was evaluated using three methods: indirect immunofluorescence under confocal microscopy, immunoenzymatic method after membrane extraction, and immunoblotting. In uremic subjects, at baseline membrane AQP1 expression was significantly lower, whereas plasma osmolality was higher than in controls. At 1 and 2 h of replacement therapy, a progressive increase was observed in erythrocytic AQP1, values similar to those in controls being attained after 3.5 h. During the session osmolality values reduced progressively, becoming significantly lower than basal values. The mean erythrocytic corpuscular volume in patients with ESRD was significantly lower than in cntrols at baseline. This value increased during hemodialysis, attaining statistical significance with respect to the basal value at 3.5 h of dialysis. Close correlations were found between plasma osmolality and AQP1 values (r = -0.930; p < 0.05), and also between MCV and plasma osmolality trend (r = -0.909; p < 0.05). There was a linear correlation (r = 0.63, p < 0.05) between plasma AVP concentrations and plasma osmolality. The variations found in plasma osmolarity during hemodialysis, may induce AQP1 expression on the membrane of intact red blood cells.

Adult↗

The CD85/LIR-1/ILT2 inhibitory receptor is expressed by all human T lymphocytes and down-regulates their functions.

The inhibitory molecule CD85/LIR-1/ILT2 has been detected previously on the surface of a small proportion of T lymphocytes. In this study, evidence is provided that, although only a fraction of CD3+ cells are stained by mAb specific for CD85/LIR-1/ILT2 on their surface, this inhibitory receptor is present in the cytoplasm of all T lymphocytes, and that it is detectable on the surface of all T cell clones by the M402 mAb. Biochemical analyses further demonstrate that CD85/LIR-1/ILT2 is present in all T clones analyzed, and that the protein is tyrosine-phosphorylated. Expression of mRNA coding for CD85/LIR-1/ILT2 has been assessed by RT-PCR. Notably, in the NKL cell line and in one T cell clone, amplification of the messenger required 30 cycles only, whereas, in other T cell clones, an amplification product was detected by increasing the number of cycles. CD85/LIR-1/ILT2 inhibits CD3/TCR-mediated activation in both CD4+ and CD8+ clones, and it down-regulates Ag recognition by CD8+ cells in a clonally distributed fashion. Addition of anti-ILT2 HP-F1 mAb in the cytolytic assay enhances target cell lysis mediated by Ag-specific CTL. This could be due to interference of the mAb with receptor/ligand interactions. In contrast, HP-F1 mAb cross-linking triggers inhibitory signals that reduce cytotoxicity. CD85/LIR-1/ILT2 also controls responses to recall Ags and, in low responders, its engagement sharply increases T cell proliferation. The inhibitory function of the molecule is also confirmed by its ability to reduce CD3/TCR-induced intracellular Ca2+ mobilization.

Antibodies, Monoclonal↗

Ultrastructural and immunohistochemical study of basal lamina of the testis in adolescent varicocele.

OBJECTIVE: To evaluate a possible involvement of the extracellular matrix (ECM) in the testes of adolescents with varicocele. DESIGN: Cross-sectional study. SETTING: University-based medical center. PATIENT(S): Twenty-four adolescents aged between 13 and 18 years underwent surgical treatment for repair of left idiopathic varicocele. INTERVENTION: A testis biopsy was performed at time of surgery. MAIN OUTCOME MEASURE(S): Transmission electron microscopy study of basal lamina and immunofluorescence studies of collagen type IV and laminin, two major components of basal lamina. RESULT(S): Transmission electron microscopy observations showed an uneven profile of the basal lamina with a variable thickness. Immunofluorescence studies showed an irregular immunofluorescent line that appeared interrupted in some observations. Collagen type IV showed some areas of strong immunostaining with other areas with reduced immunoreactivity. CONCLUSION(S): Our ultrastructural and immunohistochemical observations highlight focal damage at the level of peritubular basal lamina, but this damage is not as severe as that described in adult varicocele. Initial involvement of basal lamina could represent one of the mechanisms responsible for varicocele-induced histologic alterations of the testes.

Adolescent↗

Molecules that inhibit T-cell functions: cytochemical localization and shuttling.

Adaptive immune responses to antigens are mediated by specific receptors expressed on B cells (BCR's) and T cells (TCR's). Effector cells and memory cells are produced following a proliferative wave that accounts for clonal expansion. If not down-regulated, clonal expansion might lead to uncontrolled lymphoproliferation that would be harmful for the organism. Several mechanisms that account for the down-sizing of activated lymphocyte clones are briefly reviewed here. We next consider in detail one such mechanism that deals with the functional characterization and the immunocytochemical localization of two T-cell inhibitory molecules, namely the Cytotoxic T Lymphocyte Antigen-4 (CTLA-4) and the HP-F1 antigen, both present in all T lymphocytes. CTLA-4 and HP-F1 inhibit CD4+ T-helper cell proliferation and the lytic ability of CD8+ T-cytotoxic cells in non-specific and in antigen-specific cytolytic assays. Interestingly, a clonal distribution exists as for the ability of CTLA-4 and HP-F1 to inhibit T-cell functions. In resting and activated T cells, both molecules are largely confined in the endosomal compartment, as shown by immunofluorescence analyses. However, upon interaction of T cells with Antigen-Presenting Cells (APC's) or with target cells that must be killed, CTLA-4 molecules are transported to the plasma membrane, at the site of cell-to-cell contact where, following interaction with ligands, they trigger inhibitory signals.

Abatacept↗

[Unusual case of calcified renal fibroma].

The renal fibroma is an extremely rare event that takes its origin from parenchima, from the peri-renal tissues or from the renal capsule. A case of renal fibroma of a rarely met medullary origin is described.

Adult↗

[Validity of PSA density of the transition zone in the diagnosis of prostate cancer].

One hundred four patients (mean age 70.6 years) with prostatic specific antigen (PSA) values between 4 and 10 ng/ml (average 7.9 ng/ml), and with no suspects for neoplasia by digital rectal examination (DRE) and transrectal ultrasound (TRUS) were studied. In all patients PSA density for the entire prostate (PSAD) and PSA density for the transition zone (PSAT) were calculated. TRUS was performed using a 5 MHz probe. Prostate and transition zone volumes were obtained by ellipsoid formula. Aim of the study was to evaluate the PSAT predictivity for prostate cancer compared to the PSAD. Sixteen out of 104 patients (15.4%) had histologically confirmed prostate cancer, and 88 (84.6%) had benign prostatic hyperplasia. When cut-off for PSAD was 0.15 ng/ml/cc, specificity and sensitivity were respectively 75% and 68% with positive and negative predictive values of 54% and 17%; when cut-off for PSAT was 0.34% ng/ml/cc, sensitivity and specificity were respectively 100% and 68% with positive and negative predictive values of 60% and 18%. Our results, according to the literature data, suggest that PSAT seems to have a higher predictivity for prostate cancer than PSAD, providing an optimization for the employ of prostatic biopsy, especially for those patients with PSA values between 4 and 10 ng/ml.

Aged↗

[Usefulness of ultrasonography in the diagnosis of acute scrotal edema].

Ultrasonography has gained an important role for the diagnosis of acute and chronic scrotal lesions. We report our experience on 6 cases of acute scrotal edema. The etiology of edema was congestive cardiopathy (two patients), glomerulonephrosis (three patients) and lumboaortic lymphnode metastases from bladder transitional cell carcinoma (one patient). Ultrasound scan scrotal appearances were similar in all examined patients, and the only pathologic findings observed were "onion" like appearance and a thickened scrotal wall. Therefore, to define acute scrotal edema etiology, it isn't enough an ultrasound examination, but it's necessary to extend the ultrasound scan assessment to other organs.

Acute Disease↗

Comparative responses to three different types of interferon-alpha in patients with chronic hepatitis C.

We investigated the efficacy and tolerability of three different types of interferon-alpha, administered with the same schedule to naive patients with chronic hepatitis C. One hundred and seven patients with histologically proven chronic hepatitis C were enrolled during a period of three years and randomly divided into three groups, to receive (a) leukocyte-interferon-alpha, 6 MU three times a week for 4 months, followed by 3 MU three times a week for 8 months (Group I); (b) recombinant-IFN-alpha-2a, with the same schedule (Group II); and (c) lymphoblastoid-IFN-alpha-N1, with the same schedule (Group III). All patients were followed-up for 6 months to evaluate the long-term response. The 'Complete Response' rates at the end of treatment were: 50%, 46.1% and 41.6%, in Groups I, II and III, respectively; most patients relapsed after the end of therapy, so that the 'sustained responders' were, after 6 months of follow-up, 18.7%, 23.1% and 19.4%, respectively. Analysis of pre-treatment variables showed that age, ALT and gamma GT serum levels, as well as the prevalence of liver cirrhosis, were lower in the 'sustained responder' group. Four patients were eliminated from the study because of severe adverse events: 1, 2 and 1, in Groups I, II and III, respectively. Our results indicate a similar response rate with the three different types of interferon-alpha, although at baseline, age, serum levels of gamma globulins and the number of patients with cirrhosis-possible negative-risk factors, were higher in Group I.

Aged↗

[Procollagen III and laminin in chronic viral hepatopathies].

OBJECTIVES: We measured serum concentrations of the N-terminal peptide of type III procollagen (PIIIP) and laminin (Lam-P1) in patients with chronic viral liver disease in the various stages of the clinical course, to judge their value in assessing liver fibrogenesis, and also compared them with a number of liver function tests and histological scores of inflammation and fibrosis. METHODS: Twenty-nine patients with chronic persistent hepatitis, 39 with chronic active hepatitis and 42 with liver cirrhosis were studied. The control group was composed of 45 healthy blood donors. Serum PIIIP and Lamp-P1 were determined by radioimmunoassay; hepatic function was measured by routine assay; liver fibrosis and inflammation were graded on a 0-3 score scale. RESULTS: A significantly higher increase in serum levels of PIIIP and Lam-P1 was found in the liver cirrhosis group compared with the other groups (p < 0.05). Serum PIIIP correlated with transaminase levels in the chronic active hepatitis group and with gammaglobulin and total bilirubin in the liver cirrhosis group. A positive correlation was found only with gammaglobulin and total bilirubin in the cirrhosis group. A positive correlation was found between serum PIIIP levels and the rating scale of liver necrosis. In contrast, for Lam-P1 values, a correlation with the rating scale of necrosis and fibrosis was found. CONCLUSION: Our findings confirmed the increase of PIIIP and Lamp-P1 in chronic viral liver disease, but because of the frequent overlap values, they cannot be used as substitutes for liver biopsy for diagnosis. The correlations with the histological findings indicate that these two markers can be used during the follow-up of patients receiving anti-inflammatory or anti-fibrotic treatment.

Adult↗

Immunolocalization of the costameres in human skeletal muscle fibers: confocal scanning laser microscope investigations.

BACKGROUND: The costameres in skeletal muscle fibers were first described by Pardo et al. (1983a) and have been defined as transverse circumferential elements of the cytoskeleton associated to the sarcolemma. Specific immunostaining for vinculin shows that the costameres overlie I bands. However, an exact correlation between the costameres and the Z line is uncertain, although approximately 10 proteins so far have been localized in the costameres. To define the exact localization of costameres in human skeletal muscle fibers, we carried out an immunofluorescence study using confocal scanning laser microscopy on the fascia lata muscle of adult males. METHODS: Samples were fixed in 3% paraformaldehyde; frozen sections were treated with antivinculin, antitalin, antidesmin, and anti-alpha-actinin, then immunostained with TRITC. For double localization, the TRITC-streptavidin, as a marker for vinculin and FITC-streptavidin a marker for desmin, were used. RESULTS: The distance between two subsequent transverse lines of actininf indicated that muscle fibers were well stretched. Processing, with different software functions of the images obtained using CLSM, shows that vinculin and talin are only present in the sarcolemmal lattice. Immunostaining for vinculin and double immunostaining for vinculin and desmin demonstrate that costameres superimpose underlying I bands without interruption at the Z line. Immunostaining for talin showed that the protein is located in correspondence with the I band and M line. CONCLUSIONS: We believe that costameres are "proteic machinery." The findings of the present study suggest that it is possible to determine the width and the period of each proteic component. In addition, we indicate that costameres are present in correspondence with M line.

Actinin↗

[Enamel mineralization in rats subjected to an excess fluoride diet: SEM study].

Aim of our present work is to investigate with the SEM the process of enamel mineralization in the lower incisors of albino rats submitted for 21 days (a single amelogenetic cycle) to an hyperfluoric diet (five folds more than the normal). Our observations were performed on specimens fractured 8, 12 and 16 mm from the cervical loop transversally along the major axis of the incisor. It was demonstrated that the three different phases of enamel maturation were slower, so that, when the incisor erupted, mineralization was not completed and localized areas of demineralization were present. The authors are of the opinion that all morphological changes are dependent on the effect of fluoride on ameloblasts, either during their secretory or modulatory phases. On the basis of our results attention is pointed on the possible lesions of the enamel dependent from an unwary fluoride administration, particularly when decidual teeth are still present.

Animals↗

Scanning electron microscopy of histological relapse after gluten-challenge in coeliac disease.

The steps of the morphological relapse occurring during gluten-challenge in coeliac children were for the first time investigated by scanning electron microscopy and compared with the morphological changes observed in untreated and treated coeliac patients, in pathological and normal controls. Some peculiar morphological changes, not reported up to now, were observed in treated and relapsed coeliac patients. No relationship was found among the degrees of mucosal atrophy observed by SEM and the duration of the challenge performed by an uncontrolled ingestion of gluten.

Celiac Disease↗

[Computerized reconstruction of the spatial organization of enamel rods].

Morphometric reliefs on the enamel of the lower incisor of albino rats in correspondence with the prism compaction area during the first third of the ameloblastic modulation phase have been carried out. The reliefs were done on the longitudinal and median fracture plane and on the transversal fracture plane 8 mm from the cervical ansa. Measurements of the diameters of the honeycomb pits were carried out during the deposition phase, following removal of the ameloblastic coat. The data obtained were fed into an Apple Macintosh 512K/800 computer to obtain the geometric reconstruction of an isolated prism, the relations between each portion of the prism and the respective pits and a graphic example of the architectural organization of the enamel. It was shown that each prism consists of three distinct portion, just as there are three types of honeycomb pit. The external orifice of each pit is complementary to the interdigitant portion of the Tomes process, so the study of the pits may be used to evaluate the spatial orientation of the rods and not the morphology of their section. Cross-over between rows of contiguous rods occurs throughout the thickness of the internal enamel layer. The enamel possesses a particularly suitable structure for the transformation of surface type tangential forces into stresses or prevalently compression type.

Amelogenesis↗

The biological evolution of the Leeds-Keio ligament in the human knee. A histological and ultrastructural study.

The authors report their experience in the reconstruction of the anterior cruciate ligament using the Leeds-Keio ligament in 12 patients, 7 by open arthrotomy and 5 by an arthroscopic technique. The biological evolution of the new ligament was studied macroscopically and histologically in the first 6 cases treated by open arthrotomy. Arthroscopic monitoring and biopsy was carried out 4 to 10 months after operation. In 5 cases the new ligament appeared to be intact and histologically vital, with proliferation of new tissue along the new ligament. There were broken filaments of dacron hanging from the intercondylar cavity in only one case. The first macroscopic, histological and ultrastructural results show that despite a few limitations the Leeds-Keio ligament has the features required for replacement of the anterior cruciate ligament, although it will be necessary to wait 4 or 5 years before fully evaluating the durability and function of the ligament and the best operative method.

Adolescent↗

New muscle fiber production during compensatory hypertrophy.

Morphological changes exhibited by satellite cells during compensatory hypertrophy have been observed through a scanning electron microscope on m. plantaris of young, adult rats. Compensatory hypertrophy was induced by ablation of the synergistic m. gastrocnemius. Muscles were observed 15, 30, and 60 days post-operative. A consistent increase in wet-weight of m. plantaris (60.3,77.2, and 93.7% more than the contralateral control muscle) indicated the degree of developing hypertrophy. The satellite cells exhibited the following successive changes: (1) cells enlarged and became freed of connective network sheath, (2) cell distance from the associated fibers increased though still attached to the latter, (3) subsequent cell division, giving rise to rows of cells, that with time-lapse formed elongated structures with a common sheath, (4) elongated structures developed into new muscle fibers.

Animals↗