Biomedical subjects

F Marotta

Publications and source records attributed to F Marotta.

At least 19 recordsLinked to original sources

PubMed2000

Flow cytometric and functional characterization of AC133+ cells from human umbilical cord blood.

AC133+ cells may represent an alternative source of transplantable haemopoietic progenitor cells to CD34+ cells. Here, we have addressed the characterization of umbilical cord blood (UCB) AC133+ cells and compared their immunophenotypic and functional features with those of UCB CD34+ cells. UCB AC133+ and CD34+ cell fractions were purified by magnetic cell sorting, analysed by flow cytometry, tested for their content in blast cell colony-forming units (CFU-Bl), erythroid and granulocyte-macrophage colony-forming units before and after expansion in the presence of various haemopoietic growth factor combinations. Median AC133+ cell yield was 62.3%, and median AC133+ population purity was 97.9%. AC133+ cells were found to contain significantly more CFU-Bl than CD34+ cells; furthermore, the replating efficiency, i.e. the number of CFU-Bl capable of generating secondary colonies, was higher in the former than in the latter cells. Both AC133+ and CD34+ cells displayed an increased ability to give rise to committed progenitors after 7-day expansion in liquid cultures. These data suggest that the AC133+ cell subset is a heterogeneous pool of immature and more differentiated cells that can be maintained and expanded in well-defined culture conditions. In comparison with CD34+ cells, UCB AC133+ cells appear to contain a higher number of early haemopoietic progenitors.

AC133 Antigen↗

PubMed1998

Detection of megakaryocyte colonies in plasma clot cultures by immunoenzymatic staining.

In vitro induced megakaryocytic differentiation/maturation of megakayocyte (meg) progenitors represents an important tool for investigating cytokine-induced in vitro thrombocytopoiesis. We have developed an assay which allows the in situ study of human meg progenitor-derived colonies, cultured on a plasma clot in the presence of cytokines. Plates were immunostained by using an anti-alpha IIb beta 3 monoclonal antibody and an alkaline phosphatase-labeled secondary antibody. alpha IIb beta 3-bearing cells were stained an intense red and were clearly differentiated from the negative cells. Processed plates were stable for some weeks at 4 degrees C. The described procedure is easy to perform and allowed us to enumerate the meg colonies and assess colony morphology and cell ploidy.

Colony-Forming Units Assay↗

PubMed1998

Growth factors increase retroviral transduction but decrease clonogenic potential of umbilical cord blood CD34+ cells.

BACKGROUND AND OBJECTIVE: The feasibility of gene marking or gene therapy protocols making use of purified CD34+ cells greatly depends on the efficiency of their stable transduction. The great potential of umbilical cord blood as a source of CD34+ cells combined with the availability of advanced cell purification procedures prompted us to evaluate whether incubation with growth factors might influence the type of cells effectively transduced by retroviral vectors. DESIGN AND METHODS: Isolated, at least 95% pure, CD34+ cells were infected with the LXSN murine retrovirus carrying the neomycin-resistance gene. Different schedules of CD34+ cell infection were performed with or without incubation for different times in the presence of Interleukin-3 (IL-3), Interleukin-6 (IL-6) and stem cell factor (SCF). Efficiency of transduction was evaluated by clonogenic assays, semiquantitative PCR and RT-PCR analyses performed either immediately or after 7 day expansion of CD34+ cells in liquid culture in the presence of erythropoietin (EPO), IL-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF). RESULTS: The results obtained indicated that the amount of transduced cells increased with the lenght of incubation with growth factors, either before or during infections. However, different types of cells were transduced depending on the duration of stimulation and infection. Thus, following one week culture of CD34+ cells in the presence of EPO, IL-3 and GM-CSF the clonogenic potential was affected dyshomogeneously. Precisely, with a single 3-hour infection performed after 12 hours of stimulation with growth factors, the clonogenic potential of the transduced cells greatly increased after one week in culture. In contrast, with a 48 hour infection, the transduced cells completely lost their clonogenic potential after one week in culture. INTERPRETATION AND CONCLUSIONS: These results demonstrate that a reasonably high transduction efficiency of purified CD34+ cells can be achieved with short schedules of incubation/infection in the absence of stroma or extracellular matrix.

Antigens, CD34↗

PubMed1997

Pure pancreatic juice from patients with chronic pancreatitis has an impaired antibacterial activity.

CONCLUSION: These data show that pure pancreatic juice of AICP patients has a markedly defective antibacterial activity. This finding might be of potential clinical interest in the understanding of the pathophysiology of the disease. BACKGROUND: The aim of the present study was to test the antibacterial activity of pure pancreatic juice in patients with chronic pancreatitis. METHODS: The study group consisted of ten patients with ethanol-induced chronic pancreatitis (AICP) and seven control patients free of pancreatic disease. All subjects had recently undergone a secretin-pancreozymin pancreatic function test. After an overnight fast, through a side-viewing endoscope, selective pancreatic duct cannulation was performed. After secretin stimulation, pure pancreatic juice was obtained. Three fractions of different molecular weights were separated. Samples were incubated with 1-mL suspension of 10(5) Escherichia coli ATCC 25,922, and log10 of colony-forming units were counted. Experiments were repeated by grading pancreatic juice concentration, pH of the medium, and inoculum size. RESULTS: No significant change of pH of pure pancreatic juice appeared between AICP and controls. Starting from 6-h observation, pure pancreatic juice of AICP patients showed a significant bacterial colonization vs controls (p < 0.01). A direct correlation appeared between bacterial colonization and either pH and dilution of pancreatic juice (p < 0.001). Antibacterial activity was independent of inoculum size, enzymatic activation or inhibition, and heat treatment. The fraction with 1000-10,000 molecular weight was the one endowed with antibacterial activity.

Adult↗

PubMed1995

Shark fin enriched diet prevents mucosal lipid abnormalities in experimental acute colitis.

Marine lipids contain eicosapentaenoic acid (EPA) which has anti-inflammatory effects. The aim of this research was to study, using the dextran sulfate induced acute colitis (AC) model, the effect of an EPA-rich shark fin supplemented diet on the mucosal lipid composition. The histology score increased in AC (p < 0.05), but only slightly in the EPA group. Similarly, colonic permeability to a intraluminally instilled water-soluble contrast medium significantly increased in the AC group, but not in EPA group. As compared with controls, the AC group showed lower levels of phosphatidylethanolamine, phosphatidylinositol, free fatty acid C20:5, and PL-FA C18:1 and C18:2 and higher levels of sphingomyelin, lysophosphatidylcholine, and C18:1 and free fatty acid C20:4 (p < 0.01) after 2 and 7 days. In the EPA group sphinogmyelin and lysophosphatidylcholine slightly increased and free fatty acid C20:4 decreased (p < 0.05) after 7 days, and no PL-FA change occurred. This study confirms the protective properties of EPA-rich marine food. EPA-enriched diet is protecting the colonic mucosa from the early derangements of lipid components occurring in this experimental AC model. This effect is likely to contribute to maintain an effective mucosal lining barrier.

Acute Disease↗

PubMed1994

Fat-soluble vitamin concentration in chronic alcohol-induced pancreatitis. Relationship with steatorrhea.

In order to study the fat-soluble vitamin concentration of patients with chronic alcohol-induced pancreatitis (CAIP) we measured vitamins A and E, total lipids, and retinol-binding protein (RBP) in the plasma of 44 patients with CAIP and 83 controls (44 healthy controls; 39 Crohn's disease patients). Mean plasma vitamin E and vitamin E/total lipid ratio were significantly lower in CAIP when compared with either control or Crohn's disease groups. A low vitamin E/total lipid ratio was found in 75% of CAIP patients (91% with steatorrhea) and a ratio less than 1.0 was virtually 100% predictive of steatorrhea. The mean plasma vitamin A level for the CAIP group was significantly lower (overall 16%, 38% with steatorrhea) than in controls. Patients with CAIP show subnormal plasma levels vitamin E more often as compared to vitamin A. Further, the plasma vitamin E/total lipids ratio may be a sensitive and practical means in the detection and follow-up of steatorrhea in these patients.

Adolescent↗

PubMed1994

Nafamostat mesilate on the course of acute pancreatitis. Protective effect on peritoneal permeability and relation with supervening pulmonary distress.

Three hundred sixty Sprague-Dawley rats were allocated into four groups, according to different content of a 24-h i.v. infusion performed 1 h after intrabiliary injection of enterokinase/sodium taurocholate to induce acute pancreatitis (AP): (1) Saline; (2) 5 micrograms/kg/h nafamostat mesilate (FUT-175); (3) 10 micrograms/kg/h FUT-175; and (4) 25 micrograms/kg/h FUT-175. Peritoneal fluid was removed and exchanged with 1 mL 3.33 M fluorescein-isothiocyanate-conjugated (FITC) dextrans of 4000-40,000 Dalton. Serial blood samples were withdrawn and examined for FITC-dextrans, phospholipase A2 (PLA2), blood gases, amylase, and lipase. As compared to control (55%), FUT-175 brought about a lower (5 micrograms/kg/h: 25%) or no mortality (10 and 25 micrograms/kg/h), and a milder histological and biochemical evidence of AP. Untreated animals with PLA2 values over two times the standard deviation showed a respiratory distress. Further, unlike group 1, FUT-175 doses as low as 5 micrograms/kg prevented the increase in peritoneal permeability to small-size molecules (up to 20,000 Dalton). In a second experiment under the same drug protocol, 1000 U/mL of PLA2 and 2 mL of pancreatitis ascites were instilled ip. Peritoneal permeability to FITC-dextrans up to 30,000 Dalton and to PLA2 significantly increased in the saline group and in the 5 micrograms/kg FUT-175 group. However, 10 micrograms/kg and 25 micrograms/kg FUT-175 doses prevented such phenomenon. In conclusion, FUT-175 proves to be a potent antiprotease molecule with a biochemical activity also against PLA2 in vivo and prevents significant transperitoneal-blood access of pancreatic enzymes.

Acute Disease↗

PubMed1993

The ventilatory response to carbon dioxide in high risk infants.

PURPOSE: To examine the ventilatory response to inspired carbon dioxide in infants considered to be at risk for sudden infant death syndrome or apnea. DESIGN: Clinical data measurement. SETTING: Infant apnea evaluation program of a university division of neonatology. PATIENTS: Fifty nine infants were full term characterized by the following diagnoses; siblings of infants who had died from sudden infant death syndrome (SIDS) (n = 7), apparent life threatening event (ALTE) (n = 24), apnea/cyanosis in the newborn nursery (n = 21), and controls. Sixty-nine infants were preterm and consisted of patients suffering from idiopathic apnea (n = 61), and bronchopulmonary dysplasia (n = 8). MEASUREMENTS: The ventilatory response to carbon dioxide was measured with a computerized waveform analyzer. MAIN RESULTS: Among full term infants no significant differences in the ventilatory slope in response to CO2 was seen. The range of mean slope was 19.4 +/- 7.6 in siblings of SIDS and 36 +/- 17 in control infants. Greater number of sibling of SIDS had slopes less than 20 ml/kg/min/mmHg in comparison to control infants. Sibling of SIDS had less increase in minute ventilation and inspiratory flow in response to CO2 administration in comparison to control infants. Preterm infants had similar slopes with a mean of 33 ml/kg/min/mmHg in infants with idiopathic apnea and 28 ml/kg/min/mmHg in infants with bronchopulmonary dysplasia. CONCLUSIONS: The large intragroup variability in the ventilatory response to inspired CO2, confirming previously reported data, comprises the benefit of this test. Thus, ventilatory response to CO2 administration is not useful in unselected patients at risk of SIDS or apnea.

Apnea↗

PubMed1993

Antibodies developing against a single recombinant interferon protein may neutralize many other interferon-alpha subtypes.

Three hepatitis C patients out of 15 who received recombinant interferon-alpha 2a (rIFN-alpha 2a) therapy developed high levels of neutralizing antibody coincident with clinical relapse. On analysis, antibodies in their sera were found to also produce some neutralization of lymphoblastoid IFN. Purified lymphoblastoid IFN was separated chromatographically into 10 fractions, each containing one or two IFN-alpha subtypes, and these were used individually to examine the subtype specificity of the neutralizing antibodies in the patients' sera. All three sera neutralized all the subtypes present in the 10 lymphoblastoid IFN fractions. These findings may explain why some patients who develop antibody-mediated resistance to treatment with rIFN-alpha 2 preparations are unable to respond again when treated instead with a human cell derived preparation containing many IFN-alpha subtypes.

Antibody Specificity↗

PubMed1993

Duodenal bicarbonate secretion induced by human epidermal growth factor in rats is partially mediated by prostaglandins.

In the present study, the effect of graded intravenous infusions of human epidermal growth factor (hEGF) 0.005, 0.05 and 0.25 micrograms/ml, with or without 4 mg/kg i.p. indomethacin pretreatment, on rat duodenal bicarbonate secretion was investigated. Perfused duodenal loops were prepared in rats which were given intravenous infusions of hEGF with or without indomethacin. Duodenal pH and pCO2 were measured at 5-min intervals for 45 min, and bicarbonate secretion was calculated. Compared to control, each dose of hEGF caused a significant dose/response rise of duodenal bicarbonate secretion. Prostaglandin release was abolished by indomethacin pretreatment. Indomethacin-pretreated rats had a significant reduction of bicarbonate secretion which was still higher than in controls. These results provide evidence that duodenal bicarbonate secretion induced by hEGF is only partly accounted for by a prostaglandin-dependent mechanism.

Animals↗

PubMed1992

Early pathological changes of endothelia in a model using LDL perfusion at physiological LDL-cholesterol concentration.

The aim of the present research was to provide further insight into the debated problem of the existence of modified LDL in vivo. For this purpose a novel model was devised for studying LDL injurious effect on endothelial cells (EC) by infusing native cholesterol rich LDL, diluted to physiological LDL cholesterol concentration. Normal rabbits were infused with LDL separated from rabbits previously fed either with standard food (I-LDL Group), 1% cholesterol (II-LDL Group) or 1% cholesterol plus probucol (IV-LDL Group). Cu++ modified II-LDL was infused as well (III-LDL Group). After dilution as above, lipid oxide (LP) significantly increased in III- and II-LDL media, as compared to I- and IV-LDL media. EC of III- and II-LDL Groups showed irregular shape and surface pattern. Further, they showed adhering clusters of monocytes, platelets and erythrocytes. Endocytic vesicles and ruthenium red-positive particles increased too. EC of IV-LDL Group were only slightly affected as compared to I-LDL Group. These data suggest that native LDL from hypercholesterolemic rabbits contain an oxidized form which is noxious to EC even when LDL is infused at physiological LDL-cholesterol concentration. This early injury is in part LP-associated and actively involves platelets and monocytes.

Animals↗

PubMed1991

Serum alpha-L-fucosidase. A more sensitive marker for hepatocellular carcinoma?

Forty-nine liver disease patients (7 chronic persistent hepatitis, CPH; 10 chronic active hepatitis, CAH; 13 liver cirrhosis, LC; 9 primary hepatocellular carcinoma, PHC, without LC; and 10 PHC with associated LC) and 20 controls were assessed for their serum alpha-L-fucosidase (ALF) and alpha-fetoprotein (AFP) levels and several routine liver injury parameters. Tumor diameter in those with hepatic cancer was assessed by angio-CT. Only ALF and AFP were significantly greater in patients with PHC and PHC + LC patients as compared to patients with LC alone. At an accepted cutoff level of 500 ng/ml, the AFP level provided 43% false negative tests. On the other hand, an ALF level exceeding 740 mumol/hr/ml provided a sensitivity of 84% with a specificity of 94%. No relationship between the ALF level and Child's criteria or with any liver injury parameter was evident. Considering all individual values, the ALF, rather than the AFP, correlated with tumor size. This finding suggests the ALF level may be of value in the early detection of PHC as well as in the follow-up of patients treated for PHC.

Adult↗

PubMed1991

Cholesterol-rich LDL perfused at physiological LDL-cholesterol concentration induces platelet aggregation and PAF-acetylhydrolase activation.

The aim of this research was to perform an in vivo study on the relationships between lipid oxide (LP), platelet aggregation and PAF-acetylhydrolase in a model using perfusion of cholesterol-rich LDL media diluted to physiological LDL-cholesterol concentration. Normal rabbits were infused with LDL (d 1.025-1.063 g/ml) extracted from rabbits previously fed either with standard food (I-LDL group), 1% cholesterol food (II-LDL group) or 1% cholesterol plus probucol (IV-LDL group). CU2+ modified II-LDL was also infused (III-LDL group). After dilution as above, LP increased significantly in III- and II-LDL media. After perfusion, LP significantly increased in III- and II-LDL groups as compared to baseline values and to control. Compared to the I-LDL group, PAF-acetylhydrolase and platelet aggregation significantly increased in III- and II-LDL groups. These data indicate the property of cholesterol- rich LDL to activate PAF-acetylhydrolase and enhance platelet aggregation, even when perfused through a medium containing a physiological LDL-cholesterol concentration.

1-Alkyl-2-acetylglycerophosphocholine Esterase↗

PubMed1991

A nine year retrospective analysis of resectable pancreatic cancer at the National Cancer Center Hospital in Tokyo: clues to diagnosis and diagnostic assessment.

A nine year retrospective survey was carried out at the National Cancer Center Hospital in order to define the diagnostic clues and most suitable diagnostic assessment in resectable pancreatic cancer patients. Forty six cases were detected (27 pancreatic head cancers, 19 pancreatic body and tail cancers). There were 8, 26, 3 and 9 cases of t1, t2, t3 and t4 tumour size cancers, respectively. Abdominal pain and/or discomfort and back pain were the most common initial symptoms and chief complaints. Jaundice was present only in pancreatic head cancer cases. Abnormal GTT and CA 19-9 were the biochemical tests most commonly found abnormal, irrespective of tumour size. ERCP followed by US and CT were the most accurate technical tests. The best care toward the awareness of the initial symptoms needs to be followed, as a first choice, by a proper biochemical (CA 19-9, GTT) and technical (US, ERCP, CT) assessment in the hope of identifying those patients whose prognosis might be improved by an early operation.

Antigens, Tumor-Associated, Carbohydrate↗

PubMed1990

Relationship between gastrin cell number, serum, antral mucosa and luminal gastrin concentration and gastric acidity in antral atrophic gastritis.

The aim of our study was to investigate the relationship between gastrin producing cell density with antral mucosa, luminal and serum gastrin concentration in antral atrophic gastritis. Our study group consisted of 17 patients: six with mild atrophic gastritis, seven with moderate atrophic gastritis and four with severe atrophic gastritis. None of the patients had type-A atrophic gastritis but the body mucosa was affected by superficial gastritis at various extent in some. A group of 15 healthy subjects served as control. All subjects underwent gastroscopic examination with multiple bioptic sampling. Radioimmunoassay was used for gastrin determination and photomicroscopy for gastrin producing cell density assessment. Electron microscopy was used to assess the gastrin producing granule density index. Patients with moderate and severe atrophic gastritis showed a lower gastric acidity and acid output as compared to control. Serum gastrin did not show significant differences among the groups. In moderate and severe atrophic gastritis, gastrin producing cell granule density index, gastrin producing cell density and antral mucosa gastrin concentration were significantly lower when compared with control and decreased with advancing of the severity of atrophic gastritis. In atrophic gastritis, however, the latter two measurements were not correlated. In moderate and severe atrophic gastritis luminal gastrin concentration significantly increased, compared with control, after the severity of atrophic gastritis. Gastrin producing cell granule density index and luminal gastrin concentration showed a significant correlation with gastric pH. These data suggest that in antral atrophic gastritis with reduced gastric acidity, the decrement of gastrin producing cells is followed by gastrin producing cell hyperfunction with increased luminal release of gastrin.

Adult↗

PubMed1990

Effect of graded intravenous doses of urogastrone on duodenal bicarbonate secretion in conscious rats: evidence of a dose-response pattern.

In the present study, the effect of graded intravenous bolus injections of urogastrone, 0.5, 5, 25 and 50 micrograms/kg, on rat duodenal bicarbonate secretion was investigated. Perfused duodenal loops were prepared in rats in a strictly controlled fashion. After full recovery, different groups of rats were injected with intravenous graded bolus doses of urogastrone. During the following 45-min study period, duodenal pH and pCO2 were measured at 5-min intervals and bicarbonate secretion was calculated accordingly. Compared to controls, each dose of urogastrone caused a significant dose-response increase in duodenal bicarbonate secretion, measured either at each 5-min reading or as total 45-min output. These results provide the evidence that urogastrone may play a role in the humoral control of duodenal alkaline secretion.

Animals↗

PubMed1990

[Oral galactose tolerance test in the study of glucose metabolism disorders in patients with liver disease].

Patients with chronic liver disease (CLD) may show an abnormal glucose tolerance test (GTT). On the other hand, it is debated whether diabetics may undergo abnormalities of liver function. The aim of our study was to see whether galactose tolerance test (GaTT) is a suitable complementary diagnostic tool in defining the glucose metabolism abnormalities characteristic of the two above mentioned clinical entities. Thirty-one patients with CLD, 16 adult diabetics and 12 healthy volunteers were considered. GTT with radioimmunoassay of plasmatic insulin (IRI) and oral GaTT were performed. CLD patients were divided into two groups: with IRI/glycemia greater than 0.4 or less than 0.4. On GaTT, galactosemia was significantly raised in CLD compared to diabetics and control. Further, a statistically significant difference was shown in the glycemic variations during GaTT between diabetics and both CLD groups. A significantly inverse ratio galactosemia/glycemia occurred during GaTT and showed a different pattern between diabetics and CLD patients with impaired insulin secretion. A careful study of galactosemia and glycemia variations during GaTT reliably defines the different patterns of glucose metabolism derangement in CLD with abnormal GTT and in diabetics.

Blood Glucose↗

PubMed1990

[Application of high resolution liquid chromatography to the test of bile acid tolerance].

In the present paper we have evaluated the accuracy of a fully automated liquid-chromatographic method to study the variations of serum bile acid concentration after oral administration of 500 mg of chenodeoxycholic as a bile acid tolerance test. The study population consisted of 11 subjects with liver cirrhosis (L.C.), 6 with chronic active hepatitis (C.A.H.) and of 15 healthy volunteers, as a control. A clear linear correlation was observed between the integrated peak area and the concentration of each bile acid. Bile acids were detected at a minimum concentration of 5 ng. Intra-assay variation, based on 10 consecutive determinations, was limited to a range of 0.42% and 3.23%. Compared to control group, L.C. and C.A.H. patients showed significantly higher levels of total bile acids and of CDCA. Significative was also the increase of glycine- and taurine-conjugates as well as the decrease of the ratio between the two. The present method, fully automated and using a low cost enzymatic reagent, has yielded an accurate analysis of bile acid fractions on a minute volume of serum in a short examination time.

Aged↗