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Biomedical subjects

M Marino

Publications and source records attributed to M Marino.

At least 73 records · Page 4Linked to original sources
PubMed2000

Mitogenic signal of epidermal growth factor in chick embryo hepatocytes: role of PKCalpha.

In the present study we investigated the role of two isoforms of protein kinase C in the mitogenic signal of epidermal growth factor (EGF) in primary culture of chick embryo hepatocytes. The down-regulation of PKCalpha by long-term exposure to phorbol myristate acetate (PMA) provoked a reduced mitogenic response to EGF while the down-regulation of PKCepsilon with oligonucleotide antisense had no effect on the stimulation of DNA synthesis, assayed as thymidine incorporation. EGF enhanced H3 diacylglycerol (DAG) production by cells preincubated with H3myristic acid, but did not increase the production of inositol 1-4-5-trisphosphate (IP3). EGF produced an increase in the release of arachidonic acid and prostaglandin E2 (PGE2) in the extracellular medium. The increase was blocked by specific inhibitors (quinacrine and AACOCF3) of phospholipase A2 (PLA2) and was inhibited by down-regulation of PKCalpha, demostrating that this isoform is involved in arachidonic acid production. DAG and arachidonic acid produced an additional effect on thymidine incorporation. The treatment with PLA2 inhibitors, which block the increase in arachidonic acid, decreased the effect of EGF on DNA synthesis. These results suggest that in chick embryo hepatocytes PKCalpha is the main isoform involved in EGF-induced DNA synthesis. Its rapid activation is dependent on DAG production and induces an increased production of arachidonic acid and prostaglandin which are involved in the mitogenic activity.

Animals↗
PubMed1999

High-performance liquid chromatography separation methods for the analysis of peptide nucleic acids.

An analytical analysis of peptide nucleic acids (PNAs) was carried out by reversed-phase HPLC using a solvent system comprised of aqueous trifluoroacetic acid and acetonitrile. A regression equation was obtained which represents the relationship of the molecular mass, sequence composition and retention time. This equation can be used to estimate the retention time of a known PNA under certain HPLC conditions. In addition to this equation, new HPLC conditions were also optimized which can be used for separation of pure PNAs.

Acetonitriles↗
PubMed1999

The ins and outs of meiosis.

During oogenesis the oocyte is arrested in meiosis twice. First at prophase I, then a second time at metaphase I in many invertebrates and in metaphase II in the vast majority of vertebrates. Meiosis resumption is triggered by the sperm. This article examines mechanisms that cause oocytes' arrest in meiosis and how spermatozoa help the oocyte to get out of this cellular predicament. J. Exp. Zool. (Mol. Dev. Evol.) 285:226-236, 1999.

Animals↗
PubMed1999

Inhibition of experimental autoimmune encephalomyelitis in SJL mice by oral administration of retro-inverso derivative of encephalitogenic epitope P87-99.

Retro-inverso modification of peptides preserves parent peptide overall topology and provides at the same time stability to proteolysis, leading to derivatives with prolonged half-life in vitro and in vivo. In this study the encephalitogenic epitope P87 - 99 of myelin basic protein has been prepared in the retro-inverso form to examine its biological activity in a murine model of multiple sclerosis. Experiments of in vivo T cell tolerance induction in SJL mice revealed that the retro-inverso peptide was able to induce a selective T cell hyporesponsiveness, as measured by a reduction in the proliferative response of lymphnode T cells after antigen challenge. Oral administration of retro-inverso peptide decreased the disease severity significantly and delayed considerably the disease onset in treated mice. Enhancement of resistance to proteolysis by retro-inverso modification of encephalitogenic epitopes may increase the therapeutic value of oral tolerance induction in the treatment of multiple sclerosis and other Th1-associated inflammatory disorders.

Administration, Oral↗
PubMed1999

The blood perfused isolated heart: characterization of the model.

We have characterized the aerobic blood-perfused isolated heart model evaluating the hemodynamics and metabolism of both the blood donor animal and the isolated organ. Anaesthesia of the blood donor with sodium pentobarbital (30 mg/kg) increases arterial concentration of non esterified fatty acids (NEFA) from 80 +/- 6 to 452 +/- 70 microM; p < 0.01. Injection of 1,000 U/kg heparin causes a second significant increase from 452 +/- 70 to 1012 +/- 104 microM; p < 0.01. Insertion of the perfusion circuit, without the isolated heart, causes a reduction in blood pressure of the blood donor and a significant increase in norepinephrine from 277 +/- 44 to 634 +/- 130 pg/ml; p < 0.05. Two hours of aerobic perfusion of the isolated heart inserted in the perfusion circuit, decreases arterial pressure of the blood donor with a concomitant increase of plasma norepinephrine from 475 +/- 150 to 841 +/- 159 pg/ml; p < 0.05. Developed pressure, oxygen consumption, glucose and NEFA uptake of the isolated heart remain constant during two hours of aerobic perfusion, NEFA being the preferred substrate. Tissue content of high energy phosphates at the end of the perfusion is high and similar to that observed "in vivo". Despite this, there is a release of lactate and CPK from the isolated heart. We conclude that: 1) the model allows accurate measurement of hemodynamics and metabolism of both the isolated heart and the blood donor animal; 2) the perfusion procedure modifies the substrates concentration of the blood donor animal which, in turn, results in the preferential NEFA utilization of the isolated heart. These changes do not affect the functional parameters of the perfused heart.

Animals↗
PubMed1999

Phenotype of mice and macrophages deficient in both phagocyte oxidase and inducible nitric oxide synthase.

The two genetically established antimicrobial mechanisms of macrophages are production of reactive oxygen intermediates by phagocyte oxidase (phox) and reactive nitrogen intermediates by inducible nitric oxide synthase (NOS2). Mice doubly deficient in both enzymes (gp91(phox-/-)/NOS2(-/-)) formed massive abscesses containing commensal organisms, mostly enteric bacteria, even when reared under specific pathogen-free conditions with antibiotics. Neither parental strain showed such infections. Thus, phox and NOS2 appear to compensate for each other's deficiency in providing resistance to indigenous bacteria, and no other pathway does so fully. Macrophages from gp91(phox-/-)/NOS2(-/-) mice could not kill virulent Listeria. Their killing of S. typhimurium, E. coli, and attenuated Listeria was markedly diminished but demonstrable, establishing the existence of a mechanism of macrophage antibacterial activity independent of phox and NOS2.

Abscess↗
PubMed1999

Structure of the lnlB leucine-rich repeats, a domain that triggers host cell invasion by the bacterial pathogen L. monocytogenes.

The L. monocytogenes protein lnlB activates phosphoinositide 3-kinase and induces phagocytosis in several mammalian cell types. The 1.86 A resolution X-ray crystal structure of the leucine-rich repeat domain of lnlB that is both necessary and sufficient to induce phagocytosis is presented here. The structure supports a crucial role for calcium in host cell invasion by L. monocytogenes and supplies a rationale for its function. Calciums are bound to the protein in an unusually exposed manner that suggests that the metals may act as a bridge between lnlB and mammalian cell surface receptors. The structure also identifies surfaces on the curved and elongated molecule that may constitute additional interaction sites in forming a bacterial-mammalian signaling complex.

Amino Acid Sequence↗
PubMed1999

Nicotinamide alters the calcium release pattern and the degradation of MPF activity after fertilisation in ascidian oocytes.

Fertilisation in ascidian oocytes triggers a plasma membrane current, the release of intracellular calcium and the degradation of Maturation Promoting Factor (MPF) activity leading to the completion of meiosis and the initiation of embryo development. We have previously shown that the fertilisation current in ascidians is produced through the metabolism of nicotinamide nucleotide (NN) metabolites to ADP ribose. In this study we have used nicotinamide to test whether NN metabolism plays additional roles in fertilisation in ascidians. Nicotinamide treatment blocked calcium-induced calcium release (CICR) and arrested the cell cycle prior to the completion of meiosis I. Nicotinamide further prevented the abolition of MPF activity after fertilisation. Interestingly, nicotinamide treatment caused ascidian oocytes to form interphase-like pronuclei after fertilisation, despite the high MPF activity. The data demonstrate that NN metabolism is involved in calcium signalling through CICR and further suggest that a NN metabolite acts as a messenger connecting MPF activity to the formation of the meiotic apparatus.

Animals↗
PubMed1999

Juvenile polyposis: case report and assessment of the neoplastic risk in 271 patients reported in the literature.

A case of juvenile polyposis is reported and 271 cases are collected from the literature. The risk for neoplasia is analyzed and the endoscopic or surgical therapy is discussed. The authors conclude that juvenile polyposis should be considered as a challenge to the surgeon with regard to familial adenomatous syndromes, and strongly recommend a close follow-up of patients with juvenile polyposis.

Adenomatous Polyposis Coli↗
PubMed1999

Antimicrobial activity of the essential oils of Thymus vulgaris L. measured using a bioimpedometric method.

The essential oils obtained from Thymus vulgaris L. harvested at four ontogenetic stages were evaluated for their biological activity and chemical composition. The thyme essential oils were tested for their inhibitory effects against nine strains of gram-negative bacteria and six strains of gram-positive bacteria. The bioimpedance method was chosen for studying the antibacterial activity of the essential oils and the parameter chosen for defining and quantifying the antibacterial activity of the essential oils was the detection time. The plate counting technique was used to study the inhibitory effect by direct contact. All the thyme essential oils examined had a significant bacteriostatic activity against the microorganisms tested. This activity was more marked against the gram-positive bacteria. The oil from thyme in full flower was the most effective at stopping the growth of the microbial species examined. The oils tested were also shown to have good antibacterial activity by direct contact, which appeared to be more marked against the gram-negative bacteria. Only a few of the species were capable of recovering at least 50% of their metabolic function after contact with the inhibitor, while most of the strains were shown to have been inactivated almost completely. Escherichia coli O157:H7 was the most sensitive species, given that after contact with even the lowest concentration of oil cells could not be recovered.

Anti-Bacterial Agents↗
PubMed1999

[Dermatofibrosarcoma protuberans: authors' experience].

Dermatofibrosarcoma protuberans is an uncommon slow-growing cutaneous neoplasm. Surgical excision is an effective therapeutic approach although a significant number of local recurrences has been documented in the literature. In the present paper the Authors report 3 cases of dermatofibrosarcoma protuberans. All cases have been surgically treated with a wide local excision extended up to 3 cm from the tumor borders. No local recurrence has been reported. The Authors conclude that the surgical technique employed could reduce the local recurrence rate of dermatofibrosarcoma protuberans.

Abdomen↗
PubMed1998

Estrogens cause rapid activation of IP3-PKC-alpha signal transduction pathway in HEPG2 cells.

The mechanisms through which steroids affect target cells are not fully understood. In addition to the classic model, there is now increasing evidence that steroids can exert rapid actions. It must still be elucidated if rapid and slow estrogen actions produce co-operative and/or integrative functions. The effects of estrogen on inositol trisphosphate (IP3) production and PKC-alpha levels on membrane in the HEPG2 cell line have been investigated. Results show that estrogen addition to HEPG2 cells causes a rapid increase of IP3 production. The effect was totally inhibited by pre-incubation with tyrosine-kinase inhibitor genisteine and with the anti-estrogen ICI 182,780. An increased PKC-alpha level on the membrane fraction was present 30 min after estrogen exposure. The strong signal could elicit a variety of cellular responses such as modulation of ion channel, stimulation of cell proliferation, and phosphorylation of cytosolic ER. The ability of estrogen to trigger IP3 production in human hepatoma cells is a novel aspect of estrogen action that requires the current model of hormone stimulation target cells to be revised.

Carcinoma, Hepatocellular↗
PubMed1998

Ins and outs of meiosis in ascidians.

Ascidian oocytes are blocked in metaphase (M) of the first meiotic division. Fertilization triggers the completion of meiosis without any further arrest. In this review, we have analyzed the mechanisms that regulate the progression through meiosis in these oocytes. A primary signal from the fertilizing spermatozoon, probably soluble sperm factor(s), induces intracellular calcium release by activating the IP3 and CICR pathways and gates the fertilization current by triggering the generation of ADP ribose (ADPr). The calcium oscillations are not required for the inactivation of MPF observed at M-I release; however, ADPr may be indirectly involved in the activity of MPF associated kinase, Cdc2. MPF activity reaches a second peak at M-II followed by subsequent inactivation. Progression to M-II is dependent on the intracellular calcium oscillations. MAP kinase (MAPK) activity decreases at M-I exit and remains low during the completion of meiosis. Finally, although Cdc2, Cyclin B and MAPK-like proteins have been identified in ascidian oocytes, components of CSF still remain to be identified.

Adenosine Diphosphate Ribose↗
PubMed1998

Age-dependent accumulation of dolichol in rat liver: is tissue dolichol a biomarker of aging?

Dolichols are long hydrophobic molecules broadly distributed in all tissues and cellular membranes of eukariotic cells. Dolichol affects membrane structure and fluidity, membrane-associated protein activities, and membrane sensitivity to oxidative stress. Reports have shown that dolichols exhibit a remarkable (6- to 30-fold) age-related increase in the tissues of adult and mature rats and of old flies, mice, and humans. In our longitudinal study, the age-related accumulation of dolichol was monitored in the liver tissue of male Sprague Dawley rats fed ad libitum up to age of 27 months. In addition 24-month-old rats subjected to different regimens of anti-aging diet restriction (40% calorie restriction or every-other-day feeding ad libitum) were tested. A parallel study of the accumulation of carbonyl in liver protein (a proposed biomarker of aging) was made. In addition, the age-related decline of liver autophagy/proteolysis was studied in isolated liver cells, in view of the essential role of this function in liver membrane maintenance. Results show that an age-dependent accumulation of dolichol can be observed in the liver of the rats fed ad libitum but not in the liver of 24-month-old food-restricted rats, that accumulation of dolichol precedes the accumulation of altered liver proteins, and that dolichol accumulation is accompanied by a decline in liver autophagy. It is concluded that dolichol accumulation satisfies the proposed primary and secondary applicable criteria and the desirable features required to be qualified as a biomarker of aging.

Aging↗
PubMed1998

Ammonification in Bacillus subtilis utilizing dissimilatory nitrite reductase is dependent on resDE.

During anaerobic nitrate respiration Bacillus subtilis reduces nitrate via nitrite to ammonia. No denitrification products were observed. B. subtilis wild-type cells and a nitrate reductase mutant grew anaerobically with nitrite as an electron acceptor. Oxygen-sensitive dissimilatory nitrite reductase activity was demonstrated in cell extracts prepared from both strains with benzyl viologen as an electron donor and nitrite as an electron acceptor. The anaerobic expression of the discovered nitrite reductase activity was dependent on the regulatory system encoded by resDE. Mutation of the gene encoding the regulatory Fnr had no negative effect on dissimilatory nitrite reductase formation.

Ammonia↗