Biomedical subjects

M Fiore

Publications and source records attributed to M Fiore.

At least 37 records · Page 2Linked to original sources

PubMed1999

Dimethyl sulfoxide restores contact inhibition-induced growth arrest and inhibits cell density-dependent apoptosis in hamster cells.

Most nontransformed cell lines respond to confluence by arresting the cell cycle in a viable G(1) phase, whereas immortalized cell lines growing in monolayer do not stop cell cycle progression in response to high cell density and are subjected to density-dependent apoptosis. We have examined the effects, in terms of cell growth, apoptosis, and expression of adhesion molecules of culturing contact inhibition-deficient hamster cells in the presence of dimethyl sulfoxide (DMSO). Addition of 1.5% DMSO to the growth medium for 96 h arrested Chinese hamster ovary (CHO) cells in the G(1) phase as a confluent monolayer, associated with a remarkable increase in the expression of the cyclin-dependent kinase inhibitor p27. Cells cultured in DMSO-containing medium showed increased levels of cadherins and alpha5beta1 and beta1 integrin complexes. Cell exposure to DMSO also reduced both cell density-dependent apoptosis and necrosis and resulted in increased Bcl-2 expression. These results converge to indicate that DMSO restores contact inhibition-induced growth arrest and prevents high-density-dependent apoptosis and suggest that the effect of DMSO may be mediated by intracellular signaling triggered by cell-extracellular matrix and cell-cell interactions. Both p27 and bcl-2 appear to be involved in the resumption of growth control accompanying cell adhesion in DMSO-exposed CHO cells.

3T3 Cells↗

PubMed1999

Inducible expression of the cell surface heparan sulfate proteoglycan syndecan-2 (fibroglycan) on human activated macrophages can regulate fibroblast growth factor action.

Monocyte/macrophages play important roles in regulating tissue growth and angiogenesis through the controlled release of heparin-binding growth factors such as fibroblast growth factor (FGF), vascular endothelial growth factor, and heparin binding epidermal growth factor. The action of these potent growth mediators is known to be regulated by adsorption to heparan sulfate proteoglycans (HSPGs) on the surface and within the extracellular matrix of other neighboring cells, which respectively promote or restrict interactions with their signal-transducing receptors on target cells. Here we report on the nature of HSPGs inducibly expressed on the surface of macrophages that confer these cells with the capacity to regulate endogenous growth factor activity. We reveal that activated human macrophages express only a single major 48-kDa cell surface HSPG, syndecan-2 (fibroglycan) as the result of de novo RNA and protein synthesis. In addition, we demonstrate this macrophage HSPG selectively binds the macrophage-derived growth factors FGF-2, vascular endothelial growth factor and heparin binding EGF and can present FGF-2 in a form that transactivates receptor-bearing BaF32 cells. These results define a novel and unique proteoglycan profile for macrophages and imply a key role for syndecan-2 in the delivery of sequestered growth factors by inflammatory macrophages for productive binding to their appropriate target cells in vivo.

Angiogenesis Inducing Agents↗

PubMed1999

Overexpression of tumour necrosis factor alpha in the brain of transgenic mice differentially alters nerve growth factor levels and choline acetyltransferase activity.

Tumour necrosis factor alpha (TNF-alpha) is a pleiotrophic cytokine synthesized primarily by macrophages and monocytes, which exerts a variety of biological activities during inflammatory responses, immune reactions, and wound healing. Within the central nervous system (CNS), the basal levels of TNF-alpha are almost undetectable, but increase after neurological insults. Using transgenic mice expressing high levels of TNF-alpha in the CNS, we investigated the effect of this cytokine on the levels of brain nerve growth factor (NGF), a neurotrophin playing a crucial role in the development, maintenance and regeneration of basal forebrain cholinergic neurons. The immunoenzymatic assay and in situ hybridization revealed that the constitutive expression of NGF decreased in the hippocampus, increased in the hypothalamus, while remained unchanged in the cortex. Moreover, septal cholinergic neurons which receive trophic support from NGF produced in the hippocampus display loss of choline acetyltransferase immunoreactivity, suggesting that the reduced availability of NGF may influence negatively the synthesis of brain cholinergic neurons. These observations indicate that the basal level of brain NGF can be influenced negatively or positively by local expression of TNF-alpha and that this cytokine, through dose-dependent regulation of NGF synthesis and release, may be involved in neurodegenerative events associated with aging.

Age Factors↗

PubMed1999

Prenatal methylazoxymethanol acetate alters behavior and brain NGF levels in young rats: a possible correlation with the development of schizophrenia-like deficits.

It has been hypothesized that a deleterious key contribution to schizophrenia (SZ) development is a failure of migration and setting of young neurons into their appropriate cortical target sites, particularly in the entorhinal cortex (EC). To test this hypothesis in an animal model, we injected, in pregnant rats, on gestational day (GD) 9, or 10, or 11, or 12, the antimitotic compound methylazoxymethanolacetate (MAM) known to cause EC neuronal loss. We investigated whether or not EC disruption during prenatal development is able to affect behavior, including memory and learning, and brain nerve growth factor (NGF). Prenatally MAM treated young rats didn't display gross behavioral changes in social interaction, open-field and novel object investigation tests. By contrast, GD11 and GD12 MAM treated rats had a retardation in passive avoidance acquisition, while, in GD12 animals, pain sensitivity was reduced. GD12 animals also showed increased NGF in the EC and remaining cortex. MAM treated animals showed no changes in paw NGF or substance P levels suggesting that the altered nociceptive response is not related to local downregulation of these two molecules. The possibility that these behavioral and biochemical alterations might be associated with the onset of SZ is discussed.

Analysis of Variance↗

PubMed1999

Song behavior, NGF level and NPY distribution in the brain of adult male zebra finches.

The aim of the present study was to investigate the role of nerve growth factor (NGF) and neuropeptide Y (NPY) in the higher vocal center (HVC) on singing behavior of adult male zebra finches. The results of our studies show: (a) that NGF is present in the brain of these birds and it is higher in the HVC than in the other neostriatal tissues; (b) that exogenous administration of NGF or NGF-antibody had no discernible effect on singing behavior; and (c) that NGF enhances the NPY immunoreactivity in neurons and fibers localized in HVC and other areas of the neostriatum and hippocampus whereas anti-NGF decreased NPY stained cells in the hippocampus. These studies indicate that NGF is produced in the brain of zebra finch and that it plays a role in the regulation of NPY.

Animals↗

PubMed1999

Psychological and symptom distress in terminal cancer patients with met and unmet needs.

This study identified the needs of terminal cancer patients, investigated the factors associated with unmet needs, and assessed psychological and symptom distress associated with unsolved needs. Ninety-four patients were randomly selected from 324 patients admitted for palliative care in 13 Italian centers. Two self-administered questionnaires (the Symptom Distress Scale and the Psychological Distress Inventory) were administered to all the patients. Patients needs were identified using a semi-structured interview, aimed at exploring five areas: physiological needs, safety needs, love and belonging needs, self-esteem needs, self-fulfillment needs. A content analysis of the answers defined 11 needs, and identified patients with unmet needs. The most frequent unmet needs were symptom control (62.8%), occupational functioning (62.1%), and emotional support (51.7%). The less frequently reported needs were those related to personal care (14.6%), financial support (14.1%), and emotional closeness (13.8%). Low functional state was significantly associated with a high proportion of patients with unmet needs of personal care, information, communication, occupational functioning, and emotional closeness. Patients with unmet needs showed significantly higher psychological and symptom distress for most needs. This study provides some suggestions about the concerns that should be carefully considered during the late stage of cancer.

Aged↗

PubMed1999

Inhibitory action of melatonin on H2O2- and cyclophosphamide-induced DNA damage.

Melatonin, the pineal gland hormone known for its ability to modulate circadian rhythm, has recently been studied in its several functions. It is believed to inhibit cancer growth, to stimulate the immune system and to act as an antioxidant. In particular, this latter activity is ascribed to two different mechanisms: stimulation of radical detoxifying enzymes and scavenging of free radicals. We used this compound in mammalian cells in vitro to investigate its mechanism of action in modulating DNA damage. Cytogenetic and cytofluorimetric analyses were performed. We show that melatonin is able to modulate chromosome damage (chromosomal aberrations and sister chromatid exchanges) induced by cyclophosphamide. Conversely, its involvement in modulating oxidative processes, thereby reducing DNA damage, is less clear. In particular, melatonin is able to decrease H2O2-induced chromosomal aberrations but not sister chromatid exchanges and has been found to induce oxygen species in a cytofluorimetric test (DCFH assay).

Animals↗

PubMed1998

Infection with Schistosoma mansoni in mice induces changes in nociception and exploratory behavior.

In this study, CD-1 mice were infected percutaneously with 1600 cercariae of Schistosoma mansoni and their pain sensitivity and exploratory behavior were analyzed in well-standardized tests (hot-plate, hole-board, open-field, novel object investigation and black/white box). Schistosome infection produced body weight reduction, increased analgesia, induced changes in the number of fecal pellets emitted during the hole-board and the black/white tests, induced decreased locomotion in the open-field, decreased sniffing, rearing, wall-rearing and time spent in exploratory activity. The infection also lengthened the latency time to the first transition from the white into the black compartment in the black/white box, index of enhanced anxiety. The present findings indicate that the analgesia is one of the main effects of the disease suggesting that schistosome infection induces maladaptive response in exploratory behavior and in locomotor activity of the host associated with altered motivational and attentional levels. Furthermore, though mouse behavioral changes appear to be similar to those observed in parasite/host systems where the changes are supposed to be adaptive for the parasite, in the case of Schistosoma/mouse system, the changes in host behavior resulted to be not adaptive for the parasite.

Animals↗

PubMed1998

DNA damage and cytotoxicity induced by beta-lapachone: relation to poly(ADP-ribose) polymerase inhibition.

beta-Lapachone (3,4-dihydro-2,2-dimethyl-2H-naphtho[1,2-b]pyran-5, 6-dione) was previously shown to enhance the lethality of X-rays and radiomimetic agents and its radiosensitizing role in mammalian cells was attributed to a possible interference with topoisomerase I activity. Furthermore, beta-lapachone alone was found to induce chromosomal damage in Chinese hamster ovary (CHO) cells. The aim of the present study was to further elucidate the possible mechanisms by which beta-lapachone exerts its genotoxic action in cultured mammalian cells. Flow cytometry analysis of beta-lapachone-treated CHO cells indicated a selective cytotoxic effect upon S phase of the cell cycle. beta-lapachone produced DNA strand breaks as determined by alkaline elution assay; alkaline elution profiles from treated cells showed a bimodal dose-response pattern, with a threshold dose above which a massive dose-independent DNA degradation was observed. Furthermore, beta-lapachone increased the capacity of crude CHO cellular extracts to unwind supercoiled plasmid DNA, while significantly inhibiting in vitro poly(ADP-ribose) polymerase (PARP). These results suggest that damage induction is probably mediated by the interaction between beta-lapachone and cellular enzymatic function(s), rather than reflecting a direct action on the DNA. We suggest that the inhibition of PARP plays a central role in the complex biological effects induced by beta-lapachone in CHO cells.

Animals↗

PubMed1998

Exploratory and displacement behavior in transgenic mice expressing high levels of brain TNF-alpha.

Studies reported recently have shown that tumor necrosis factor-alpha (TNF-alpha) a cytokine released by macrophages and monocytes plays a key role in inflammatory processes and immune and neuro-endocrine regulation. TNF-alpha is also produced in the central nervous system (CNS). However, the role of this cytokine in the CNS is largely unknown, although evidence indicates that it is involved in various neurobehavioral manifestations. Using transgenic mice expressing high amounts of murine TNF-alpha transgene in the neurons of the CNS, we investigated the stereotyped, exploratory, and displacement activities in the hole-board and black/white box. Transgenic mice and their normal control littermates were hybrids of the CBA x C57BL/6 genetic backgrounds and were obtained by backcrossing the CBA x C57BL/6 founder female and her progeny with F1 hybrid mates. Transgenic mice did not show changes in the stereotyped behavior on the hole-board, but they displayed several alterations in the exploratory activities both in the hole-board and black/white box. Transgenic mice also exhibited an increase in grooming when exposed to a highly unfamiliar environmental stimuli in the black/white box. The study suggests that supranormal endogenous TNF-alpha in the brain affects the behavioral responses to stressful conditions.

Animals↗

PubMed1998

Occupancy of dipeptidyl peptidase IV activates an associated tyrosine kinase and triggers an apoptotic signal in human hepatocarcinoma cells.

Dipeptidyl peptidase IV (CD26/DPP-IV) is an ectoenzyme expressed on different cell types. Signaling properties and functional consequences of the CD26 triggering have been elucidated mostly on T cells, where the molecule delivers a costimulatory signal that potentiates T-cell activation through the T-cell receptor. We conducted studies in the human hepatocarcinoma-derived PLC/PRF/5 cell line to examine the signal transduction through CD26 and its functional properties in the absence of other T-cell-specific membrane molecules. Engagement of CD26 in PLC/PRF/5 cells through a specific antibody induces tyrosine phosphorylation of several proteins with maximal intensity 15 minutes after the stimulation. This effect was under the negative regulatory control of CD45 tyrosine phosphatase, in that the addition of orthovanadate clearly enhanced the phosphorylation events. Using in vitro kinase assays with CD26 immunoprecipitates, we observed that a protein or proteins with kinase activity are coprecipitated with the CD26 molecule. In addition, unlike Jurkat T cells, in which CD26 expression exerts a protective effect against apoptosis, in PLC/PRF/5 cells CD26 occupancy delivers a potent apoptotic signal. This effect was also observed in HepG2 cells, thus indicating that it represents a more general phenomenon occurring in different liver neoplastic cell lines.

Apoptosis↗

PubMed1998

Chronic unexplained liver disease in children with primary immunodeficiency syndromes.

Liver disease may be found in patients with primary immunodeficiency syndromes because of the high risk of infection with hepatotropic viruses related to the treatment with blood derivatives. The prevalence of liver disease in these patients and its etiology, however, is still not completely understood. We have evaluated the prevalence and the etiology of liver disease in children with different forms of primary immunodeficiencies. Thirty patients included in the study underwent molecular studies to detect common hepatotropic viruses, including hepatitis C and G viruses. Liver involvement was found in 11 of 30 (36.6%) patients. All patients with liver disease had deficiencies of specific immunity, with a prevalence in this subgroup of 47.8%. Liver disease was more severe in patients with T and B cell combined immune disorders than in those with a selective T cell immunodeficiency. Moreover, the severity of the disease correlated with an overall more rapid fatal outcome. A viral etiology was found in only six of these patients, whereas in the remaining five patients, no cause of liver injury was identified. In the virally infected patients, hepatitis C virus was the most common viral agent. In patients with immunodeficiencies, there is a high prevalence of liver disease not fully explained on the basis of the common viral infections.

Adolescent↗

PubMed1998

Apoptosis as a mechanism of peripheral blood mononuclear cell death after measles and varicella-zoster virus infections in children.

Viral infections may induce an acquired form of immunodeficiency, generally lasting a few weeks. In the more severe form, such as HIV infection, the immunodeficiency is permanent. Programmed death of T cells represents one of the mechanisms by which HIV determines the T cell functional impairment, finally resulting in the destruction of T cells. In this study, we evaluated whether an altered regulation of apoptosis was also implicated in the anergy associated with the common measles or varicella-zoster virus (VZV) infections in infancy. A spontaneous apoptosis of peripheral blood mononuclear cells was observed in children who had suffered from these infections as long as 6 mo after the acute disease. Apoptosis was demonstrated through analysis of cellular DNA content, morphologic evidence of cell nuclei shrinkage, and by analysis of DNA degradation. Stimulation of T cells through anti-CD4 MAb increased the number of apoptotic cells with a maximal effect 72 h after the stimulation. Our results suggest that apoptosis may account for the anergy that follows acute viral infections in infancy.

Apoptosis↗

PubMed1997

TNF-alpha expressed in the brain of transgenic mice lowers central tyroxine hydroxylase immunoreactivity and alters grooming behavior.

Tumor necrosis factor-alpha (TNF-alpha) is a cytokine involved in a wide range of biological effects both in physiological and non-physiological conditions. It is also produced in the central nervous system (CNS) where it has been implicated in reparative processes after traumatic injuries and in CNS demyelination, neurodegeneration and inflammation. Using transgenic mice (Tg-m) expressing TNF-alpha specifically in the CNS, we showed that the overexpression of this cytokine reduced tyroxine hydroxylase immunoreactivity (TH-ir) in the caudate-putamen and in the dorsomedial hypothalamic areas and impaired grooming behavior. We also showed that this behavior is increased following anti-nerve growth factor injection. These findings support the hypothesis, proposed by others, that TNF-alpha is involved in the degenerative processes which occur in Parkinson's disease.

Animals↗

PubMed1997

Deregulated apoptosis is a hallmark of the Fanconi anemia syndrome.

Fanconi anemia (FA) is a genetic human disorder associated with bone marrow failure and predisposition to cancer. FA cells show poor growth capacity and spontaneous chromosomal anomalies as well as cellular and chromosomal hypersensitivity to DNA cross-linking agents such as mitomycin C (MMC). Because it is likely that disruption of the apoptotic control would lead to such a phenotype, we investigated the implication of apoptosis in the FA syndrome. It is shown that, although demonstrating a high frequency of spontaneous apoptosis, FA cells from four genetic complementation groups are deficient in gamma-ray-induced apoptosis and their MMC hypersensitivity is not due to apoptosis. Fas is a cell surface receptor belonging to the tumor necrosis factor receptor family and is involved in apoptosis. We show that, independently of DNA damage, the alteration in the control of apoptosis in FA concerns also the pathway initiated by Fas activation. Finally, ectopic expression of the wild-type FAC gene corrects the MMC hypersensitivity and anomalies in apoptosis and cell cycle response in FA cells. Altogether, these findings strongly implicate the FA genes as playing a major role in the control of apoptosis. Thus, further studies with FA syndrome will be instrumental toward molecularly dissecting the apoptotic pathways.

Apoptosis↗

PubMed1997

Nerve growth factor effects on the song control system of zebra finches.

The aim of this experiment was to test whether or not nerve growth factor (NGF) is involved in cholinergic processes in the avian brain, by injecting NGF into the higher vocal center (HVC) and examining its effects on adult male zebra finch song. Since NGF has been hypothesized to protect cells after injury, some birds received both NGF and ibotenic acid (IBO) lesions of HVC, while others received either NGF or IBO or neither (SHAM). Only the IBO-treated birds showed alterations in song. Although there was no evidence of cell preservation in the immunocytochemical and morphological analysis NGF appears to prevent the IBO induced impairment in song augmenting the activity of the remaining neurons and enhancing brain repair.

Animals↗

PubMed1997

Role of TNF-alpha but not NGF in murine hyperalgesia induced by parasitic infection.

Using adult mice infected with the trematode Schistosoma mansoni, we observed that this infection induces both thermal hyperalgesia and an increase in the levels of nerve growth factor in the paws. To explore the mechanism involved in peripheral hypersensitivity during chronic infection, mice were infected with 60 cercariae of S. mansoni and injected 17 weeks later with nerve growth factor, anti-nerve growth factor or with other molecules known to be associated with hyperalgesic processes. The results of these studies showed that antibodies against tumor necrosis factor-alpha, but not against nerve growth factor, reduce thermal sensitivity in schistosome infected mice, suggesting that this cytokine but not NGF plays a crucial role in schistosome-induced thermal hyperalgesia. Treatments with anti-inflammatory drugs support this hypothesis.

Animals↗

PubMed1997

Removal of the submaxillary salivary glands and infection with the trematode Schistosoma mansoni alters exploratory behavior and pain thresholds in female mice.

In this study, CD-1 female mice, deprived of the submaxillary salivary glands, were infected with S. mansoni and their behavior was observed 15 weeks after infection, when the eggs of the parasite are present in the brain. Sialectomized infected mice showed changes in exploratory activity, sniffing, and wall-rearing in the open-field and in the black/white box, but no differences in pain sensitivity were observed on the hot plate. The present results suggest that the modifications in the behavior of sialectomized infected mice might be associated with the inability of the animals to cope with the aversive effects of the infection and, most probably, with modifications in the levels of polypeptides released into the bloodstream by the salivary glands, affecting the NGF-responsive cells of the nervous, endocrine, and immune systems.

Animals↗