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Biomedical subjects

M A Rossi

Publications and source records attributed to M A Rossi.

At least 19 recordsLinked to original sources
PubMed2005

Spatial memory in aged rats is related to PKCgamma-dependent G-protein coupling of the M1 receptor.

In the present study, individual differences in spatial memory in aged Fischer 344 (F344) rats were associated with the extent of G-protein coupling of the M1 muscarinic receptor and the dendritic-to-somal ratio of hippocampal PKCgamma (d/sPKCgamma) immunogenicity. Following testing in the eight-arm radial maze task, 7 young and 13 aged rat brains were sectioned through the dorsal hippocampal formation (HF). G-protein coupling of the M1 receptor was assessed autoradiographically using competition binding studies in the presence and absence of a G-protein uncoupler to determine high (K(H)) and low (K(L)) affinity states for agonist in the HF, neocortex, and amygdala. In aged animals, a relationship between choice accuracy in the maze and K(H), a measure of M1 receptor-G-protein coupling was seen in the dentate gyrus, CA3, CA1, and neocortex. Furthermore, choice accuracy and d/sPKCgamma immunogenicity showed a significant relationship in CA1. Lastly, a correlation was seen in the CA1 of aged animals between K(H) and d/sPKCgamma. These relationships did not hold for the amygdala. Thus, individual differences in a naturally occurring age-dependent disruption of cholinergic-PKCgamma signal transduction is associated with spatial memory dysfunction.

Age Factors↗
PubMed2005

Mitochondrial damage as an early event of monensin-induced cell injury in cultured fibroblasts L929.

The present study was designed to identify, submicroscopically, the primary organelle or target structure for monensin in cultured murine fibroblasts L929. In addition, the effect of the drug on cell size and surface membranes of the cells were analysed; cellular proliferation, collagen secretion, and necrosis and apoptosis were re-evaluated. At the lowest concentration of monensin the foremost ultrastructural alteration occurred in the mitochondria, characterized by increased matrix density with disorganized and less distinct crystae. Incubation with monensin at higher concentrations resulted in severe mitochondrial damage and marked dilatation of the Golgi apparatus and rough endoplasmic reticulum cisternae. Fibroblasts exposed to higher concentrations of monensin were enlarged with decreased number of filopodia and hollows in the surface membrane. Moreover, monensin inhibited the cell proliferation, increased immunohistochemical positiveness for collagen type I in a dose-dependent manner, and, at high concentrations, caused cell necrosis whereas apoptosis was not induced. Taken together, these results show that monensin induces early mitochondrial damage, possibly causing an energy deficit that led to inhibition of fibroblasts proliferation and accumulation of collagen causing dilatation of Golgi apparatus and rough endoplasmic reticulum. Moreover, the mitochondrial damage would also explain the monensin-induced necrosis.

Animals↗
PubMed2003

Characterization of a bovine collagen-hydroxyapatite composite scaffold for bone tissue engineering.

Different biomaterials have been used as scaffolds for bone tissue engineering. Here we characterize a biomaterial composed of sintered (1100 degrees C) and powdered hydroxyapatite (HA) and type I collagen (Coll), both of bovine origin, designed for osteoconductive and osteoinductive scaffolds. Coll/HA proportions were 1/2.6 and 1/1 (wet weight), and particles sizes varied from 200 to 400 microm. Vv (volume density) and Sv (surface to volume density) for the HA particles in the composite ranged from 0.48 +/- 0.06 to 0.55 +/- 0.02 and 5.090 +/- 0.545 to 6.366 +/- 0.289 microm(-1), respectively. Due to the relatively small changes in Vv and Sv, a macroporosity could be characterized for the biocomposite. X-ray diffraction and infrared spectroscopy showed that the sintered bone was composed essentially of HA with minimum additional groups such as surface calcium hydroxide, surface and crystal water, free carbon dioxide and possibly brushite. Mass spectrometry detected carbonates at A and B sites of HA, and weakly bound to the structure. Human osteoblasts adhered and spread on both the HA particle surface and the collagen fibers, which seemed to guide cells between adjacent particles. The biocomposite studied has several characteristics considered as ideal for its use as a scaffold for osteoconduction and osteoinduction.

Animals↗
PubMed2003

Scanning electron microscopic study of the cleaning ability of chlorhexidine as a root-canal irrigant.

AIM: To evaluate in vitro the cleaning of root-canal walls after irrigation with different irrigants. METHODOLOGY: A total of 36 recently extracted human teeth were divided into four experimental groups according to the irrigating solution used: saline; 2% chlorhexidine; 2.5% sodium hypochlorite; and 2.5% sodium hypochlorite + EDTA. The cleaning of the apical, middle and coronal thirds of the root canals was evaluated by scanning electron microscope examination using a 4-point scoring system. RESULTS: The best cleaning was obtained using 2.5% sodium hypochlorite and EDTA, followed by 2.5% sodium hypochlorite only (P < 0.05), whose cleaning was similar to chlorhexidine only in the cervical third. Cleaning by saline and 2% chlorhexidine was worse than the other two groups and was similar in all thirds. Better cleaning was found in the cervical and middle thirds for all groups with the worst results in the apical third. CONCLUSIONS: The apical third of the root canals was not cleaned as well as the middle and coronal thirds. Cleaning by chlorhexidine and saline was inferior compared to the cleaning by sodium hypochlorite with and without EDTA.

Analysis of Variance↗
PubMed2001

Schirrous invasive ductal carcinoma of the breast overexpress p53 oncoprotein.

CONTEXT: Breast cancer is the most important neoplasm in adult women, and its worldwide incidence is growing. The tumoral stroma is very important for modulating the growth and invasion of the tumor itself. The relationship between these two components is not completely understood. Schirrous carcinoma is a variant of ductal invasive carcinoma in which the stroma is very desmoplastic, and the importance of this finding still a motive for debate in the literature. OBJECTIVE: To compare the desmoplastic reactions against biological markers, such as estrogen and progesterone receptors, oncoprotein c-erbB-2 and oncoprotein p53, with the objective of studying the relationship between the tumoral stroma and epithelial cancer cells. TYPE OF STUDY: Retrospective study. SETTING: Cancer Hospital A C Camargo and Faculty of Medicine of Ribeirão Preto, University of São Paulo, São Paulo, Brazil. SAMPLE: 107 adult women operated because of ductal invasive carcinoma. The cases were separated into 4 groups according to the desmoplastic reaction - less than 15 per cent, between 15-50 per cent, 51-85 per cent, and more than 85 per cent fibrosis. The grade of fibrosis was determined by picrus-sirius staining and quantified by using a microscope with a stereo-imaging grid. Immunohistochemical methods were used to determine the expression of the hormonal receptors and c-erbB-2/p53 oncoprotein. MAIN MEASUREMENTS: Extent of desmoplastic reaction versus expression of estrogen and progesterone receptors, oncoprotein c-erbB-2, and oncoprotein p53. RESULTS: The results showed that schirrous carcinoma expresses oncoprotein p53 more frequently than other carcinomas with less extensive desmoplastic reaction. There were no differences between the grade of fibrosis and the other biological markers. CONCLUSION: The intense stromal reaction in invasive ductal carcinoma may modulate the expression of p53. Further investigations should be made with the aim of understanding how this expression determines the proliferative activity in schirrous carcinoma, and whether this overexpression is secondary to mutation of the p53 gene or due to modulation of other molecules of the stroma.

Adult↗
PubMed2001

Chronic inhibition of NO synthesis per se promotes structural intimal remodeling of the rat aorta.

OBJECTIVE: We characterized, using histomorphometry and transmission and scanning electron microscopy, the intimal remodeling of the thoracic aorta of normocholesterolemic young rats chronically-treated with N(omega)-nitro-L-arginine methylester (L-NAME) and examined the question whether these changes were caused by the lack of NO per se or by the hypertension which L-NAME administration induces. METHODS: Male Wistar rats were divided randomly into three sets: control group, standard diet/L-NAME-treated group, and standard diet/L-NAME + captopril-treated group. RESULTS: The treatment of rats with L-NAME for 4 weeks resulted in increased blood pressure (by 32% at the end of the treatment) as compared with the control value and intimal remodeling comprising a continuous layer of enlarged endothelial cells with irregular nuclear and cytoplasmic contours, lying over a thickened layer of fibrocollagenous support tissue focally expanded with lymphomononuclear cells and mainly diffuse foci of smooth muscle cells. In addition, the NO synthase inhibition caused a marked thickened tunica intima (150% thicker than the control value) and a significantly augmented intima : media ratio (126% higher than the control value). On the other hand, captopril prevented hypertension in rats simultaneously treated with L-NAME as compared with controls, and induced intimal remodeling comprising the same qualitative changes as those observed in L-NAME-treated rats. The tunica intima of l-NAME + captopril-treated rats was moderately thickened (60% increase in comparison with that of controls and 65% thinner as compared with L-NAME-treated rats). In the same way, the mean intima : media ratio of rats concomitantly treated with L-NAME and captopril was moderately increased (45% more) as compared with controls and significantly lower in comparison with rats administered L-NAME alone (36% less). CONCLUSIONS: Chronic inhibition of NO synthesis per se promotes structural intimal remodeling of the rat aorta, which is potentiated by L-NAME-induced hypertension. Most important, the present findings favor the idea that blockade of NO synthesis by causing intimal remodeling might be a primary cause, as individual biologic phenomenon, in the development of an atherosclerotic plaque.

Animals↗
PubMed2001

Experimental studies on the mechanism of phospholipase C activation by the lipid peroxidation products 4-hydroxynonenal and 2-nonenal.

The effects of three lipid peroxidation end-products, 4-hydroxynonenal (HNE), 2-nonenal (NE) and nonanal, on phosphoinositide-specific phospholipase C (PL-C) activity were studied in HL-60 cells. Enzymatic activity was determined by measuring the amounts of inositol-P3 (Ins-P3) produced by the cells incubated at 37 degrees C in the presence of the various compounds. HNE was shown to activate PL-C at concentrations of between 10(-8) and 10(-6) M; 10(-9) and 10(-8) M of NE also strongly stimulated PL-C. In contrast, nonanal failed to modify enzymatic activity. The concentrations of HNE and NE active on PL-C showed good correspondence with those that have been reported to be chemotactic towards rat neutrophils. The pretreatment of cells with 1 microM pertussis toxin completely prevented the increase of Ins-P3 production induced by HNE and NE. Maximal PL-C stimulation was produced by 10 nM NE; the degree of inositol-P3 production induced by the simultaneous addition of an equimolar dose of HNE was not significantly different from the activity value induced by NE alone, suggesting a possible competition between the two compounds. The data indicate that both HNE and NE share a common mechanism of action which, as with other better-known chemoattractants, involves PL-C activation through a G regulatory protein.

Aldehydes↗
PubMed2000

Action of 2-nonenal and 4-hydroxynonenal on phosphoinositide-specific phosopholipase C in undifferentiated and DMSO-differentiated HL-60 cells.

The promyelocytic cell line HL-60 has been used as an in vitro model to study the mechanism of action of two chemotactic aldehydes, 2-nonenal and 4-hydroxynonenal. Increasing aldehyde concentrations have been added to undifferentiated and DMSO-differentiated cells incubated at 37 degrees C and their effect on phosphoinositide-specific phospholipase C has been analysed by using a specific inositol-1,4,5-tris-phosphate assay system. Concentrations of 2-nonenal between 10(-9) and 10(-7) M significantly increased the enzymatic-activity in DMSO-differentiated HL-60 cells, while 10(-9) and 10(-8) M concentrations were active in the undifferentiated cells. 4-Hydroxynonenal was able to activate phospholipase C both in undifferentiated and DMSO-differentiated cells at concentrations ranging from 10(-8) to 10(-6) M. The concentrations of both compounds active on phospholipase C displayed a good correspondence with those which had been reported to be chemotactic towards rat neutrophils. In the case of 4-hydroxynonenal, the present results confirm its ability to activate phospholipase C, which we had previously shown in isolated neutrophil plasma membranes. The comparison of the effects of 2-nonenal and 4-hydroxynonenal on chemotaxis and phospholipase C activation suggests a common mechanism of action for both aldehydes, for which the presence of the double bond seems to be required.

Acid Phosphatase↗
PubMed2000

Interferon-gamma and tumor necrosis factor-alpha determine resistance to Paracoccidioides brasiliensis infection in mice.

To investigate the role of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) in the resistance to Paracoccidioides brasiliensis (Pb) infection, mice with homologous disruption of the IFN-gamma (GKO) or TNF-alpha receptor p55 (p55KO) were infected with the parasite. GKO and p55KO, but not wild-type (WT) mice, were unable to control the growth of yeast cells and the mice succumbed to infection by days 16 and 90 after infection, respectively. Typical inflammatory granulomas were found only in WT mice. In contrast, knockout mice presented an inflammatory infiltrate composed of a few neutrophils, mononuclear, epithelioid, and multinuclear giant cells forming incipient granulomas in GKO mice and without granuloma formation in p55KO mice. Besides, both groups of knockout mice exhibited elevated numbers of yeast forms in agreement with colony-forming unit counts in organs. Compared with WT, splenocytes from infected GKO mice cultured with the Pb F1 fraction produced lower TNF-alpha levels, whereas leukocytes from infected p55KO mice produced similar amounts of TNF-alpha but higher levels of IFN-gamma. Moreover, splenocytes from infected WT mice produced higher levels of nitric oxide (NO) resulting in a lower T-cell proliferative response to Con A than uninfected WT, or infected p55KO and GKO mice. On the contrary, the addition of IFN-gamma to splenocytes from infected GKO mice resulted in higher NO production and lower T cell proliferation. Taken together, these findings suggests that endogenous TNF-alpha, acting through the p55 receptor, and IFN-gamma mediate resistance to Pb infection and induce NO production that determines marked T cell unresponsiveness.

Animals↗
PubMed2000

Long-term survivors of heart transplantation: the Hartford Hospital experience.

Heart transplantation has been a clinical program at Hartford Hospital for the past 15 years, resulting in 206 transplants. The five-year survival rate is 69% and is 43% at 10 years. The first recipient is surviving and has had full rehabilitation. Thirteen patients have survived 10 years or more. Advances in immunosuppression are ongoing and will result in further long-term survivors. Graft vasculopathy and lack of organ donation are current problems.

Adult↗
PubMed1999

Is apoptosis a mechanism of cell death of cardiomyocytes in chronic chagasic myocarditis?

The micropathology of chronic Chagas' myocarditis reveals foci of myocardial cell loss associated with an inflammatory infiltrate composed predominantly of lymphomononuclear cells and interstitial fibrosis. The loss of myocardial cells, a devastating phenomenon in this cardiopathy, has been classically attributed to necrosis. In the present study we examined whether the loss of myocardial cells in human chronic Chagas' heart disease could result from cell death by apoptosis. A total of 11 cases of chronic chagasic myocarditis were studied: four hearts were obtained at autopsy within 8 h after death and seven endomyocardial biopsies were taken from chagasic patients with an arrhythmogenic form of the disease. The coronary arteries of all chagasic cases showed no obstructive lesions. The diagnosis of Chagas' disease was based on previously established criteria. Five cases were selected as controls: three hearts were obtained at autopsy within 8 h after death and two endomyocardial biopsies were taken from nonchagasic patients with normal myocardium morphology. As positive controls we used cardiac muscles of myocardial infarction and rat mammary glands on the fourth day after weaning. The TUNEL method was used to identify apoptotic cells in the myocardium. The expression of p53 protein, which directly or indirectly triggers apoptosis, was evaluated using immunohistochemical technique. A few apoptotic cells were stained in chronic chagasic hearts, both biopsy and autopsy cases. However, the stained nuclei were restricted to the mononuclear infiltrate accounting for about 0.5% of the mononuclear cells in the infiltrate. In contrast, the nuclei of cardiomyocytes in both regions bordering on and distant from the microfoci of myocardial cell loss were not stained by the TUNEL method. Moreover, the expression of the protein p53 in cardiomyocytes in chagasic hearts was absent. The results of the present study demonstrating negative in situ labeling of fragmented DNA associated with absence of expression of p53 provide support to the hypothesis that apoptosis is not the mechanism of cell death in chronic chagasic myocarditis. This reinforces the general opinion that the loss of cardiac muscle fibers in chagasic cardiopathy is produced by necrosis. On the other hand, the present results give support to the concept that apoptosis probably play a role in the clearing of lymphomononuclear cells in the inflammatory infiltrate in chronic chagasic myocarditis.

Adult↗
PubMed1998

Fibrosis and inflammatory cells in human chronic chagasic myocarditis: scanning electron microscopy and immunohistochemical observations.

The present study deals with both pathologic fibrosis and matrix connective tissue in chronic chagasic myocarditis. A total of 12 hearts were obtained at autopsy. Eight cases of chronic chagasic myocarditis were selected. Four cases without evidence of cardiac disease were used as controls. The diagnosis of chronic Chagas' heart disease was based on previously established criteria. A cell-maceration method was utilized to evaluate the spatial organization of the fibrillar collagen accumulation after removal of the myocardial tissue non-fibrous elements. The relationship between inflammatory cells identified by monoclonal antibodies and interstitial fibrosis stained with picrosirius red was assessed. Striking structural alterations of the collagen matrix in the perimysium were detected: increase in number and thickness of tendon-like structures, and markedly thickened and aggregated collagen strands. Besides, a diffuse increase in the thickness of collagen fibers surrounding individual myocytes, consisting of the endomysial matrix, mainly adjacent to the perimysium, could be observed. The dense-weave endomysial meshwork was composed of fine collagen fibrils, and it was continuous with those of adjacent myocytes, obscuring the lateral struts. Sometimes, thicker struts tethering myocytes to myocytes could be seen. These changes were associated with scattered dense scar-like foci, probably reflecting repair fibrosis associated with myocyte necrosis. Furthermore, the present results clearly showed the colocalization of foci of myocyte necrosis and degeneration and associated fibrosed areas and fibroblasts with T lymphocytes and macrophages. The accumulation of interstitial collagen fibers in chronic chagasic myocarditis may be expected to decrease myocardial compliance and disrupt synchronous contraction of the ventricles during systole, contributing to a spectrum of ventricular dysfunction that involve either the diastolic or systolic phase of the cardiac cycle or both. Myocardial fibrosis can be also implicated in the genesis of malignant ventricular tachyarrhythmias, major causes of sudden death among chronic chagasic patients. The increase in myocardial fibrosis could be directly related to an inflammatory reaction mainly composed of T lymphocytes and macrophages.

Adult↗
PubMed1998

CD40 ligand is not essential for the development of cell-mediated immunity and resistance to Mycobacterium tuberculosis.

It has been proposed that the induction of cellular immunity and resistance to intracellular pathogens is dependent upon CD40 ligand (CD40L). In the present study we show that this proposal is not ubiquitously supported. Mice genetically deficient in CD40L (CD40LKO) were resistant to i.v. infection with Mycobacterium tuberculosis when assessed by survival and bacteriologic burden in the spleen, liver, and lungs. Infected CD40LKO mice developed granulomas that lacked epithelioid cells and were less numerous and markedly smaller than those observed in control mice. Upon stimulation with purified protein derivative of M. tuberculosis, CD4+ T cells from infected CD40LKO mice proliferated and produced high levels of IFN-gamma but not IL-4. Finally, spleen cells from CD40LKO mice stimulated with M. tuberculosis produced IL-12, TNF, and nitric oxide levels comparable to those produced by control cells. In contrast to original proposals, these data clearly show that protective Thl immunity can be achieved against intracellular pathogens (e.g., Mycobacterium) independently of CD40L.

Animals↗
PubMed1998

Pathologic fibrosis and connective tissue matrix in left ventricular hypertrophy due to chronic arterial hypertension in humans.

OBJECTIVE: To investigate pathologic fibrosis and connective tissue matrix in left ventricular hypertrophy due to chronic arterial hypertension in humans. DESIGN AND METHODS: Seventeen human hearts were studied. Group 1 consisted of control hearts (four hearts, weighing 280 +/- 40 g each), from subjects who had had no evidence of heart disease and for whom the diagnoses of death were noncardiac. Groups 2 (five hearts, weighing 440 +/- 50 g each), 3 (five hearts, weighing 560 +/- 50 g each), and 4 (three hearts, weighing 680 +/- 60 g each) consisted of hearts from subjects who had had a history of systemic hypertension. All hearts had no valvular deformities and no evidence of ischemic disease at the postmortem examination. A cell-maceration method was employed to evaluate the myocardial connective matrix after removal of the nonfibrous elements of myocardial tissue, leaving behind a noncollapsed matrix, thus allowing a better three-dimensional view. Myocardial tissue was also processed for conventional light microscopic and morphometric studies. RESULTS: The minor transverse diameter of myocytes from hearts in groups 1-4 hearts were 13.7 +/- 7.8, 23.7 +/- 3.4, 26.6 +/- 3.7, and 32.8 +/- 5.8 microm, respectively. The volume fraction of fibrosis of the controls was 6.5%, whereas the volume fractions in hypertensive hearts increased progressively according to heart weight: 15.4, 22.9, and 31.1% for hearts in groups 2, 3, and 4, respectively. The most striking feature was the diffuse marked increase in amount of pericellular collagen weave fibers (endomysial matrix), parallel to the increase of heart weight. The hypertrophied myocytes were encased in a dense weave of collagen fibrils continuous with those of adjacent myocytes. The muscle fibers in hypertrophied hearts were markedly larger than normal, although this was extremely variable from an area to another. Besides, a diffuse increase in the number of thick collagen fibers constituting broad bands and sheets of collagen surrounding disorganized muscle bundles (perimysial matrix) was observed. Scattered dense scar-like foci, apparently replacing areas of myocyte loss, could be seen, mainly on the periphery of muscle bundles. This latter finding was more commonly observed among hypertrophied hearts from group 3 and, mainly, among hypertrophied hearts of group 4. Importantly, a progressive disarray of the connective tissue skeleton of the myocardium could be seen in parallel to the progressive increase of cardiac hypertrophy. CONCLUSIONS: The progressive accumulation of interstitial collagen fibers in left ventricular hypertrophy, in parallel to an increase in heart weight, can be expected to contribute to a spectrum of ventricular dysfunction involving either the diastolic or systolic phase of the cardiac cycle, or both, that is associated with the greater than normal arrhythmogenic risk for a hypertensive heart. Moreover, the methodology used is useful for studying the spatial organization of the collagen fibrils of the myocardium under normal and pathologic conditions.

Adult↗
PubMed1998

[Diagnostic imaging in Cushing's disease and its correlation with postsurgical clinical course].

UNLABELLED: The purpose of this paper was to assess the accuracy of computed tomography (CT) and magnetic resonance (MR) in the preoperative identification of corticotropin-secreting pituitary adenomas and to evaluate the concordance with the postoperative outcome. A total of 44 images of patients with Cushing's disease were retrospectively taken into consideration; 23 CT and 29 MR were evaluated. Patients were subdivided into remission or persistence, following the postoperative outcome and biochemical tests. In 13 out of 23 CT performed, an adenoma was found, the remaining 10 being negative. TC sensitivity was 56.5%. Twenty five MR were positive and 4 were negative. The overall sensitivity of MR in detecting ACTH-secreting adenoma was greater (86.0%, P: 0.0373, after Yates correction). After operation, 31 patients (70.5%, Cl 0.70 +/- 0.13) were considered to be in remission (27 total remission and 4 partial remission); 23 had had preoperative visualization of the pituitary tumor and 8 negative neuroradiological evaluation. Thirteen patients had persistence of disease as shown by sustained hypercotisolism in biochemical tests. In 11 of these patients, an adenoma had been preoperatively identified whereas in the remaining 2 no evidence of tumor lesion had been detected. An histologically proven tumor was found in 33 patients but findings were negative in the remaining 11 cases. Lack of histological evidence of an ACTH-secreting adenoma was more frequent in patients with persistent hypercortisolism (P < 0.0078). IN CONCLUSION: 1) MR seems to be more sensitive than CT for detecting ACTH-secreting adenomas (P: 0.037, after Yates correction); 2) no relationship was disclosed between the absence of imaging signs of adenoma and the postsurgical outcome; 3) the remission rate was significantly higher in patients with positive than in those with negative histological findings.

Adenoma↗