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Biomedical subjects

C Guarnieri

Publications and source records attributed to C Guarnieri.

At least 37 records · Page 2Linked to original sources
PubMed1991

Comparison between alpha-adrenergic- and K-opioidergic-mediated inositol (1,4,5)P3/inositol (1,3,4,5) P4 formation in adult cultured rat ventricular cardiomyocytes.

In adult cultured rat ventricular cardiac myocytes, both the alpha-adrenergic agonist phenylephrine and the selective kappa opioid receptor ligand U-50, 488H affected phosphoinositide turnover. Phenylephrine, over a time course of 10 min, caused a transient increase in Ins(1,4,5)P3 which peaked at 1 min and had returned to control at 2 min. In addition, phenylephrine produced a progressive and sustained increase in the formation of Ins (1,3,4,5)P4 which achieved a plateau after 5 min of exposure to the agonist. U-50,488H induced an increase in Ins(1,4,5)P3 which peaked at 1 min at a level significantly higher than that due to phenylephrine and was still elevated after 10 min exposure to the kappa opioid receptor agonist. In addition, U-50,488H caused a sustained increase in Ins(1,3,4,5)P4 which was comparable to that due to phenylephrine. The stimulatory effects produced by phenylephrine and U-50,488H were receptor-mediated events, since they were fully antagonized by their respective antagonists, phentolamine or Mr-1452.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1991

Reduced neutrophil superoxide generation and plasma lipoperoxide levels in pigs fed with cod liver oil.

Circulating neutrophils, isolated from pigs fed for 8 weeks with a diet supplemented with CLO, had an accentuated n-3 fatty acid incorporation into the plasma membrane, as evidenced by an approximately four-fold greater n-3/n-6 ratio as compared with the control diet group. Moreover, the neutrophils of the CLO fed pigs produced less superoxide when stimulated with PMA or f-MLP, as well as showing a more prolonged latency period before O2(-)-generation. In the plasma of pigs fed with CLO there were higher levels of thiobarbituric reactive material and lipofuscin, while the content of GSSG was similar in both dietary groups. The results of this study indicate that dietary supplementation with CLO reduces the activation of circulating neutrophils and favours the presence in the plasma of lipoperoxides.

Animals↗
PubMed1991

Scavenger effect of sulfasalazine, 5-aminosalicylic acid, and olsalazine on superoxide radical generation.

The in vitro antioxidant capacity of sulfasalazine (SASP), its metabolites (SP, 5-ASA), and olsalazine (OAZ), was studied by evaluating their effects on superoxide (O2-.) production. Assay systems were the xanthine-xanthine oxidase (X/XOD) reaction and phorbol myristate acetate (PMA)-activated polymorphonuclear leukocytes (PMNs), using the cytochrome c (cyt-c) reduction assay and a luminol-dependent chemiluminescence method. 5-ASA, SASP, and OAZ showed a dose-dependent scavenger effect in both O2-. generating systems, 5-ASA being the most powerful (greater than 50% of inhibition in the PMNs system and greater than 70% in the X/XOD system at 10 microM concentration). SP had an inhibitory effect only in the PMNs system but did not modify the activity of xanthine oxidase, thus excluding a scavenger action. These data suggest that the scavenger effect of 5-ASA, SASP, and OAZ may be an important mechanism of action.

Aminosalicylic Acids↗
PubMed1991

Enhanced superoxide production with no change of the antioxidant activity in gingival fluid of patients with chronic adult periodontitis.

In the gingival crevicular fluid (GCF) of control and chronic adult periodontitis (CAP) patients there is a spontaneous release of O2- radicals from polymorphonuclear leukocytes (PMN). The addition of the exogenous stimuli phorbol myristate acetate (PMA) decreased the O2-. formation in control GCF, while in CAP patients produced a marked enhancement of O2-. generation. The circulating PMN of control subjects did not show a spontaneous O2-. formation, differently from CAP patients. On the contrary, a similar O2-. production was measured when the circulating PMN were stimulated with PMA. Moreover, the antioxidant activity measured in 10 microliters of cell free gingival supernatant (GS) of control and CAP patients had the same values by inhibiting 12.6% and 18.9% respectively of the O2- formation supported by a xanthine/xanthine oxidase system. Probably, the protective or destructive effect of PMN in GCF of CAP patients depends on the variations of the rate of O2- formation in respect to the intrinsic antioxidant property of GS.

Adult↗
PubMed1991

[The inhibitory effect of magnesium on mitochondrial calcium uptake in ischemic and reperfused rat hearts].

Several studies suggest that the protection exerted by Mg2+ on the reperfused myocardium may be mostly due to its competitive effect with respect to Ca2+. The aim of this research was to evaluate the inhibitory action of Mg2+ on mitochondrial Ca2+ uptake in the reperfused myocardium. Hearts of male Wistar rats (250-300 g) were isolated and perfused by the Langendorff technique. Aerobic control hearts (n = 6) were perfused with a constant flow of 10 ml/min/g for 65 min. In a second group (n = 6) the hearts were aerobically equilibrated for 20 min, then subjected to 30 min of ischemia (98% reduction of coronary flow) and subsequently reperfused for 15 min at the same preischemic flow. The hearts of both groups were electrically stimulated at 300 b/min. Then, the hearts were pooled in groups of 2 each and homogenized for the isolation of mitochondria. One part of mitochondrial suspension was used to evaluate the respiratory function by a polarographic technique. The remaining part was incubated with fura-2/AM for 10 min at 30 degrees C in order to determine the kinetics of Ca2+ transport within mitochondria in the presence of succinate as substrate. Ca2+ uptake was reduced in the mitochondria of reperfused hearts with respect to control, particularly in the presence of elevated extramitochondrial Ca2+ concentrations (greater than 10 nM). On the contrary the initial rate of Ca2+ uptake was increased in the reperfused mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

[The biochemical bases of cardiovascular aging].

The majority of reports concerning the performance of senescent heart in both human and animal models states that, at rest, most hemodynamic parameters are substantially unchanged during aging, with the exception of a prolongation of left ventricular ejection time and a decrease in the early diastolic filling rate. A biochemical rationale for the increased early diastolic filling time with age seems to be the enhanced stiffness of left ventricular wall due to either myocardial hypertrophy or collagen tissue accumulation. The prolonged time of contraction and the inadequate response to stress of cardiac muscle in elderly subjects may be the consequence of changes in different steps of the excitation-contraction coupling, namely altered intracellular calcium homeostasis, reduced availability of highly energetic compounds, and impaired response to adrenergic stimulation. These defects can cause major hemodynamic changes when the heart is subjected to volume or pressure overload. In fact, during exercise, elderly subjects can reach a maximal heart rate slower than that of younger people. The possibility that the toxicity of oxygen reactive forms may provoke some age-related myocardial lesions such as structural alteration of membranes and enzyme inactivation is very attractive but the linkage between the most functional perturbations of aged hearts and peroxidative stress remains to be clarified.

Aging↗
PubMed1990

Effect of ATP depletion and phenanthroline on the spermidine-mediated decay of ornithine decarboxylase in erythroleukemia cells.

Addition of spermidine to Friend erythroleukemia cells caused a rapid decay of ornithine decarboxylase (ODC) activity and the accumulation of a ODC-antizyme complex. The induction of antizyme only partially accounted for the decrease of ODC activity by a direct inhibition of the enzyme. However, the antizyme induction was accompanied by a marked reduction of the half-life of ODC. Shift of the cells to an ATP-depleting medium prevented the spermidine-elicited decay of ODC activity as well as the accumulation of ODC-antizyme complex. However, ODC appeared to be stabilized even when ATP depletion was performed 40 min after spermidine addition, in the presence of high levels of antizyme. Similar results were obtained by treating the cells with phenanthroline, a heavy metal chelator and protease inhibitor. These findings indicate that ATP and some metalloprotease(s) may be involved in the degradation pathway of ODC, even in the presence of high levels of polyamines.

Adenosine Triphosphate↗
PubMed1990

Phosphorylation of ornithine decarboxylase in intact erythroleukemia cells.

32P-labeled ornithine decarboxylase was isolated by immunoprecipitation from murine erythroleukemia cells incubated in a medium containing [32P]ortophosphoric acid. Analysis of immunoprecipitate by SDS-polyacrylamide gel electrophoresis and autoradiography revealed a radiolabeled band, which corresponded to the position of mouse ornithine decarboxylase, phosphorylated in vitro by casein kinase-2. A preparation of casein kinase-2 purified from nuclei of erythroleukemia cells could also phosphorylate mouse ornithine decarboxylase.

Animals↗
PubMed1990

Mitochondrial function and superoxide generation from submitochondrial particles of aged rat hearts.

A decrease in heart function with ageing might be related to an impairment of mitochondrial function, since these organelles produce the greatest fraction of ATP in the myocyte. Mitochondria extracted from Wistar rat hearts at 3, 14, 18 and 24 months of age were employed to evaluate the changes of the respiratory activity during lifetime. A slight decrease of the respiratory rate (QO2) was observed in the 14 month group with respect to the 3 month group when succinate was used as substrate, whereas the respiratory control index (RCI) in the presence of glutamate or succinate increased in the 24 month group. The latter result may be related to a condition of moderate hypertrophy that generally occurs in the ageing heart. Submitochondrial particles (SMP) were also prepared to study the superoxide radicals (O2-) production at the level of rotenone or antimycin-inhibited regions of the respiratory chain. A strong elevation in the O2- generation was observed in the antimycin-inhibited region at 14 months of age; on the contrary, the rate of O2- production remained unchanged in the 24 month group in comparison to the youngest group. These observations correlate well with the enhanced tissue level of oxidized glutathione that was observed at 14 and 18 months of age. The products of lipid peroxidation (TBARS) did not change in the rat heart at any of the ages measured, whereas the levels of fluorescent substances progressively increased beginning from 18 months of age, with a greater extent in the mitochondrial compartment. The present study suggests that age does not substantially affect mitochondrial respiration and energy output in the rat heart, while a greater production by cardiac mitochondria of superoxide anions in the adult rats (14 months) might accelerate the fluorescent pigment formation.

Aging↗
PubMed1990

Reduced oxidative activity of circulating neutrophils in patients after myocardial infarction.

Circulating neutrophils isolated from patients 3-4 h after a myocardial infarction produced less O2-. compared with controls, when stimulated with phorbol myristate acetate or formyl-methionine-leucine-phenylalanine. Three days after the infarction the O2-. generation elicited by both stimuli further decreased markedly. Seven and 15 days after infarction the O2-. stimulated production was only slightly lower than or similar to the control values. The neutrophils of infarcted patients showed an augmented latency period before O2-. production compared with controls in response to exogenous stimuli, particularly three days after infarction. Electron microscopy revealed that the neutrophils isolated from the infarcted patients displayed signs of cell exhaustion with few alterations of the plasma membranes when stimulated with phorbol ester. In contrast, control neutrophils displayed alterations of the plasma membranes characteristic of active neutrophils. The results of this study indicate that the circulating neutrophils appear exhausted and functionally inhibited immediately after myocardial infarction.

Aged↗
PubMed1990

Alterations in polyunsaturated fatty acid composition of cardiac membrane phospholipids and alpha 1 adrenoceptor mediated phosphatidylinositol turnover.

STUDY OBJECTIVE - The aim of the study was to investigate the steps at which polyunsaturated fatty acids are involved in alpha 1 adrenoceptor mediated phosphatidylinositol turnover. DESIGN - Phosphatidylinositol turnover rates were investigated after preincubating neonatal rat ventricular myocytes with culture media enriched with linoleic acid (18:2n-6) or eicosapentaenoic acid (20:5n-3) to change the polyunsaturated fatty acid composition of their membrane phospholipids. EXPERIMENTAL MATERIAL - Cardiomyocytes were isolated from ventricles of 2-4 d old Wistar rats by trypsinization and were then cultured. Experiments were started 48 h after seeding, when there was a confluent monolayer of beating cardiomyocytes. MEASUREMENTS and RESULTS - In 18:2n-6 treated cells the 18:2n-6 content in the total phospholipid fraction rose from 45 to 68 nmol.mg-1 protein; in 20:5n-6 treated cells the 20:5n-3 content rose from 1.5 to 12.5 nmol.mg-1 protein, and the docosapentaenoic acid (22:5n-3) content rose from 5.1 to 14.7 nmol.mg-1 protein. The major n-3 fatty acid, 22:6n-3 (11.4 nmol.mg-1 protein), did not change after 20:5n-3 treatment. Although the phosphatidylinositol fraction showed changes paralleling those in the total phospholipids, none were significant. In this fraction the major n-3 fatty acid appeared to be 22:5n-3 (0.4 nmol.mg-1 protein). The fatty acid treated cells were prelabelled with [3H]-inositol to estimate the rate of phosphatidylinositol-4,5-bisphosphate turnover. There were no differences in the rate of [3H]-inositolphosphate formation between control, 18:2n-6 treated cells, and 20:5n-3 treated cells. Prolonged alpha 1 adrenergic stimulation of control and treated cells did not change the polyunsaturated fatty acid composition of the total phospholipid and phosphatidylinositol fractions. CONCLUSIONS - The alpha 1 adrenoceptor mediated phosphatidylinositol turnover rate is not affected by changes in polyunsaturated fatty acid composition of membrane phospholipids, neither does prolonged alpha 1 adrenergic stimulation lead to significant depletion of any specific or total polyunsaturated fatty acids in the phosphatidylinositol lipids.

Animals↗
PubMed1990

Phase I clinical and pharmacologic study of intraperitoneal cisplatin and fluorouracil in patients with advanced intraabdominal cancer.

Fluorouracil (5-FU) and cisplatin display marked therapeutic synergy in preclinical models and are effective in the treatment of a number of solid tumors when combined and administered intravenously (IV). Each drug has also been administered intraperitoneally (IP) and displays a favorable pharmacologic profile and acceptable clinical toxicity. We therefore undertook a phase I study to determine the feasibility and toxicity of combination IP chemotherapy with these agents. Thirty-one patients with histologically documented malignancy confined to the peritoneal space were treated with cisplatin 90 mg/m2 mixed with 5-FU in 2 L of lactated Ringer's solution and given IP for 4 hours every 28 days. Cohorts of at least three patients received starting 5-FU concentrations ranging from 5 mmol/L (1,300 mg in 2 L) to 20 mmol/L. The dose-limiting toxicity was neutropenia with a median granulocyte nadir of 156 cells per microliter occurring at a 5-FU dose of 20 mmol/L. Intrapatient escalation of the 5-FU dose was permitted and 15 cycles of chemotherapy were delivered at 5-FU concentrations greater than 20 mmol/L, the highest concentration being 30.7 mmol/L (8 g of 5-FU in 2L). Other toxicities included mild to moderate nausea during all cycles of therapy, vomiting in 54% of cycles, and diarrhea in 15% of cycles. Abdominal pain, renal dysfunction, peripheral neuropathy, and oral mucositis occurred infrequently and were not related to the 5-FU dose. Peritoneal fluid and plasma 5-FU concentrations were measured by high-performance liquid chromatography (HPLC) in selected patients. Mean peak plasma 5-FU concentrations ranged from 6.19 mumol/L to greater than 60 mumol/L, and peritoneal fluid to plasma 5-FU area under the curve (AUC) ratios ranged from 85 to 1,150. Nine of 15 patients with nonbulky disease had resolution of malignant ascites or at least a 50% reduction of peritoneal studding by tumor at repeat laparotomy. We conclude that combination IP chemotherapy with cisplatin and 5-FU is technically feasible and has acceptable clinical toxicity and a favorable pharmacologic profile. The recommended starting 5-FU dose for phase II trials is 3,900 mg mixed with 90 mg/m2 of cisplatin in 2 L of isotonic fluid.

Adult↗
PubMed1990

Calcium stimulates opioid receptor agonism in rat cardiac sarcolemma.

We have found that calcium stimulates, in a dose-dependent manner, the binding of opioid to kappa and delta receptors in sarcolemma from rat's ventricles isolated by hypotonic lithium bromide shock. Opioid binding was measured using [3H]-diprenorphine as a radioligand. The delta-selective agonist [D-Ala2, D-Leu5]-enkephalin and the kappa-selective agonist U-50, 488H both inhibited control and Ca(2+)-stimulated [3H]-diprenorphine binding to the sarcolemma, whereas [D-Ala2,MePhe4,Gly-(ol)5]-enkephalin was ineffective. The stimulatory effect of calcium increased the maximal binding capacity without affecting the affinity of the receptor for the ligand.

Animals↗
PubMed1990

Receptors for atrial natriuretic factor in cardiomyocytes and aortic smooth muscle.

The aim of this work was to investigate whether specific receptors for atrial natriuretic factor (ANF) are present in ventricular cardiomyocytes and aortic smooth muscle membranes. 125I-ANF was employed to test the binding of the radioligand to isolated rat cardiomyocytes. Calcium-tolerant ventricular cardiomyocytes were obtained by retrograde perfusion with collagenase. 125I-ANF binding to cardiomyocytes was highly specific (70-80%) with a KD value of 72.6 pM and a Bmax of 9.37 fmol/mg protein. In other studies, 125I-ANF binding was investigated with a membrane preparation obtained from calf thoracic aorta, from which the endothelium had been previously stripped off. In this preparation too the interaction of 125I-ANF (70-80%) was highly specific, with a KD value of 70.4 pM and a Bmax of 8.78 fmol/mg protein. These results suggest that specific receptors to atrial natriuretic factor are present both in isolated rat cardiomyocytes and in the smooth muscle of calf thoracic aorta. This second observation is in agreement with the hypothesis that the vasodilator effect of atrial natriuretic factor is due to a direct interaction between this peptide and vascular smooth muscle cells.

Animals↗
PubMed1990

Changes in myocardial mitochondrial respiration after ligation of the coronary artery in pigs.

After ligation of the left coronary artery, porcine cardiac mitochondria were isolated by homogenizing the tissue and treating the myofibrillar pellet with nagarse. When compared with unligated controls, the ischemic myocardium showed decreases in phosphocreatine (to 41%), ATP (to 56%) and in the mitochondrial respiratory control index (to 69% and 78% as measured with glutamate and succinate respectively). No changes were found in the corresponding P/O ratios. Similar results were obtained upon separation of the mitochondria into two main fractions by a density gradient technique, though only one of these fractions showed a fall in succinate-supported respiration. The results suggest that ischemia decreases the NADH-dehydrogenase activity of cardiac mitochondria.

Adenosine Diphosphate↗
PubMed1990

Influence of age on oxidative damage in mitochondria of ischemic and reperfused rat hearts.

Hearts from rats aged 3 months and 24 months respectively were isolated and subjected to a brief ischemia. The extent of myocardial injury, measured by release of creatine phosphokinase into coronary effluents and by developed tension, was greater in the young rats than in the old when compared with their corresponding non-ischemic controls. The amount of peroxidation, measured in the isolated mitochondria using the malondialdehyde method, was also greater in the younger rats. In contrast, when mitochondria from non-ischemic hearts were incubated for 20 minutes in a medium containing FeCl3, NADPH and ADP, known to generate hydroxyl radicals, significant peroxidation (together with a decrease in respiratory control indices) was obtained only from mitochondria isolated from the older rats. If, as the in vitro results suggest, the mitochondria of the old rats are not less sensitive to peroxidative attack, the difference between the effects of ischemia in the two age groups may be due to a lower rate of formation of reactive species of oxygen or to a greater anti-oxidative cytosolic capacity in the hearts of older rats. Alternatively, the overall oxidative stress following ischemia may be due to the effects of different radicals which target different parts of the mitochondrial membrane.

Aging↗