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Biomedical subjects

D Ricci

Publications and source records attributed to D Ricci.

At least 19 recordsLinked to original sources
PubMed2003

Modulation of pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) gene expression in isolated porcine hepatocytes perfused within a radial-flow bioreactor after low-temperature storing.

Due to the scarcity of available human livers, porcine hepatocytes are currently being evaluated as a xenogeneic cell source for extracorporeal bioartificial liver (BAL). Hypothermic storage of isolated porcine hepatocytes could support stocking of cell-loaded bioreactors for BAL use and may provide bioreactors ready to be used at the patient's bedside. For the development of this technology, it is of utmost importance to ensure cell viability and differentiated functions after low-temperature storage and following warm reperfusion. We compared cell viability, functional activity and apoptosis in isolated porcine hepatocytes which were perfused within a radial-flow bioreactor (RFB), stored at 4 degrees C and then reperfused at 37 degrees C. RFBs were loaded with 8 x 10(9), > or = 90% viable hepatocytes at 37 degrees C for 3 h. RFBs were then flushed with 4 degrees C University of Wisconsin solution (UW) and subsequently stored for 24 h or 48 h. RFBs were then reperfused for 8 h with recirculating medium plus serum at 37 degrees C . Cytochrome P450 (CYP) activity was studied before and after cold storage by means of monoethylglycinexylide (MEGX) detection in the effluent medium, after repeated lidocaine injections. After reperfusion experiments, hepatocytes were harvested for total RNA isolation. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used in order to amplify specific mRNAs for Bcl-2 and Bax genes, by using appropriate primers; beta-actin primers were used as control. Total RNA was extracted by northern blotting analysis and for Bcl-2, Bax and beta-actin RNA messenger detection, RT-PCR amplification was used. Freshly isolated hepatocytes perfused into the RFB showed a progressive increase of MEGX while a loss in Bax expression was paralleled by an increase in Bcl-2 expression, in comparison to starting hepatocytes. After 4 degrees C storage and warm reperfusion, MEGX production was preserved in 24 h- and 48 h-stored bioreactors as well as a sharp increase of Bcl-2 and a decrease of Bax mRNAs. Our study suggests that refrigeration of hepatocyte-bioreactors is a suitable strategy to maintain both viability and function of isolated hepatocytes, for up to 48 h a time-length that is compatible with long-distance delivery of ready-to-use bioreactors.

Animals↗
PubMed2002

Daptomycin morphostructural damage in Bacillus cereus visualized by atomic force microscopy.

Daptomycin is a novel, rapidly bactericidal in vitro antibiotic that is under investigation for the treatment of serious Gram-positive infections. Although daptomycin appears to disrupt membrane function, the precise mechanism of action has not been fully elucidated. Atomic force microscopy (AFM) is an innovative technique that allows high-resolution visualization and digital image manipulation of cell surface structures in 3 dimensions without the use of photons and electrons. The aim of this study was to use AFM to investigate the morphostructural changes in Bacillus cereus that occur upon daptomycin administration. The effects of daptomycin at 4x and 8x the minimal inhibitory concentration were visualized during an 8-hour incubation period. Atomic force microscopy images showed aberrant bacterial surface formations, including flattening and shrinking of cells and leakage of cytoplasm through the membrane. In addition to structural changes, the destabilization of flagella was also observed. These results support previous data suggesting that daptomycin disrupts membrane function.

Anti-Bacterial Agents↗
PubMed2001

Long-term expression of highly differentiated functions by isolated porcine hepatocytes perfused in a radial-flow bioreactor.

To overcome the limitations of standard hollow-fiber module in ensuring efficient cell perfusion and long-term expression of highly differentiated hepatocyte functions, we developed a novel bioreactor in which a three-dimensional hepatocyte culture system was perfused in radial-flow geometry. Isolated porcine hepatocytes were cultured for 2 weeks in recirculating serum-free tissue culture medium, in which NH4Cl and lidocaine were repeatedly added, and ammonia removal, urea synthesis, monoethylglycinexylide (MEGX) production, albumin secretion, Po2, Pco2, O2 consumption, and pH were measured thereafter. During the whole duration of the study, ammonia removal was paralleled by urea production, while MEGX concentration was constantly increased. Our results indicated that hepatocytes remained differentiated and metabolically active throughout the duration of the study. The radial-flow bioreactor allowed physiological contact between recirculating fluid and cells by equalizing the concentration of the perfusing components, including O2, throughout the module, suggesting a potential use of this configuration for extracorporeal liver support.

Albumins↗
PubMed2001

Spontaneous echo-contrast as an in vivo indicator of rheological imbalance in dilatative cardiomyopathy.

The aim of this study was to evaluate coagulative and hemorheologic assessment in patients with dilatative cardiomyopathy with or without spontaneous echo contrast (SEC). We studied 45 patients, 35 males and 10 females (mean age 72.1 +/- 9.2). We measured whole blood viscosity, plasmatic fibrinogen, prothrombin time (PT), activated partial thromboplastin time (aPTT), D-dimer and red cell morphology with Zipursky-Forconi method. Transthoracic and transesophageal echocardiography was performed in all patients to evaluate the presence of SEC in left atrium. We divided all the patients into two groups: the 1st group of 20 patients with SEC and Atrial Fibrillation (AF) in 80% of cases, and the 2nd group of 25 patients without SEC and AF in 31%. Our results show that in patients with SEC there is a statistically significant increase of whole blood viscosity and plasma fibrinogen in comparison with patients without SEC. Red cell morphology in all patients demonstrates a reversed EMI. D-Dimer, was out of the normal range in about 1/3 of the patients in both groups. An analysis of our results points out that in patients with SEC and AF, with a major risk factor for cardioembolic stroke, we have alterations of hemorheologic assessment with an increase of whole blood viscosity and fibrinogen that seems to be caused by an increase of red cells aggregability favoured by fibrinogen. Our conclusions are that SEC in patients with dilatative cardiomyopathy and AF is an important in vivo indicator of hemorheologic imbalance and an important marker for cardioembolic risk stroke evaluation.

Aged↗
PubMed2000

BeppoSAX and Chandra Observations of SAX J0103.2-7209 = 2E 0101.5-7225: A New Persistent 345 Second X-Ray Pulsar in the Small Magellanic Cloud.

We report the results of a 1998 July BeppoSAX observation of a field in the Small Magellanic Cloud which led to the discovery of approximately 345 s pulsations in the X-ray flux of SAX J0103.2-7209. The BeppoSAX X-ray spectrum is well fitted by an absorbed power law with a photon index of approximately 1.0 plus a blackbody component with kT=0.11 keV. The unabsorbed luminosity in the 2-10 keV energy range is approximately 1.2x1036 ergs s-1. In a very recent Chandra observation, the 345 s pulsations are also detected. The available period measurements provide a constant period derivative of -1.7 s yr-1 over the last 3 years, making SAX J0103.2-7209 one of the most rapidly spinning up X-ray pulsars known. The BeppoSAX position (30&arcsec; uncertainty radius) is consistent with that of the Einstein source 2E 0101.5-7225 and the ROSAT source RX J0103.2-7209. This source was detected at a luminosity level of a few times 1035-1036 ergs s-1 in all data sets of past X-ray missions since 1979. The ROSAT HRI and Chandra positions are consistent with that of a mV=14.8 Be spectral-type star already proposed as the likely optical counterpart of 2E 0101.5-7225. We briefly report and discuss photometric and spectroscopic data carried out at the ESO telescopes 2 days before the BeppoSAX observation. We conclude that SAX J0103.2-7209 and 2E 0101.5-7225 are the same source: a relatively young and persistent X-ray pulsar in the SMC.

Journal Article↗
PubMed2000

Transesophageal echocardiography for identifying potential cardiac sources of embolism in patients with stroke.

Pathologies of cerebral circulation are one of the most frequent causes of mortality and morbidity among the populations of Western countries. The incidence of ischemic events presumed to have a cardioembolic origin varies from 13% to 34% in the largest international multi-center studies. However, some authors have reported an incidence of general cardiogenic stroke ranging from 23% to 36% in younger patients. Transesophageal echocardiography (TEE) is a useful investigation for identifying cardiac sources of embolism in patients without vascular alterations involving the carotid and vertebral districts. This study comprised 73 patients with unexplained stroke who were investigated by both transthoracic echocardiography (TTE) and TEE. Vascular echocolor sonography had been performed in all of them with negative results. TEE demonstrated: atrial or ventricular thrombosis in 6 patients (8%), intracavitary neoplasm in 2 (2%), spontaneous echocontrast in 11 (13%), valve strands in 6 (7%), complex aortic plaque in 10 (11%), patency of the foramen ovale in 17 (19%), atrial septal aneurysm in 9 (11%), dystrophy and mitral calcifications in 9 (11%). In the other 18 patients, TEE did not reveal any anomalies. Definite sources of stroke (clots and tumors) were identified in 14% of all the lesions; however, the majority (86%) were potential sources. A comparison of the two echocardiographic techniques demonstrated a greater sensitivity and specificity of TEE. Although TEE was superior to TTE for identifying and evaluating potential embolic sources, we found both methods to be useful and would advise performing TTE before TEE. While TTE is not capable of identifying the majority of anomalies, it does provide useful information to guide subsequent transesophageal investigations.

Echocardiography↗
PubMed2000

Fractional allelic loss in non-end-stage cirrhosis: correlations with hepatocellular carcinoma development during follow-up.

Hepatocellular carcinoma (HCC) is usually preceded by cirrhosis whose genetic background is still poorly understood. The aim of this study was to evaluate, in non-end-stage cirrhosis, the fractional allelic loss (FAL) at loci mostly reported to be altered in HCC and the microsatellite instability (MSI). Twenty cases of cirrhosis were retrospectively selected. Eleven had developed an HCC during the follow-up (HCC-prone group), while 9 remained HCC-free (HCC-free group). Microdissected hepatocellular cirrhotic nodules from basal liver biopsies, were studied at 20 loci (on the chromosomal arms 1p and 1q, 3p, 4q, 6q, 7q, 8p, 13q, and 18q) and with the mononucleotide repeats BAT26 and TGFbIIR. Genetic changes were detected in both groups. Overall, the FAL index was statistically increased in the HCC-prone group (0.213) as compared to the HCC-free group (0.094; P =.044). Allelic loss at chromosomal arms 1p, 4q, 13q, 18q, and concurrent losses at more than 3 loci were confined to the HCC-prone group. In both groups, MSI was never ascertained using BAT26 and TGFbIIR. In conclusion, an increased FAL index and the lack of MSI characterize the non-end-stage cirrhosis of patients undergoing HCC during follow-up. These data emphasize the role of early clonal changes in chronic liver disease, and their potential predictive significance for clinical use.

Biopsy↗
PubMed2000

Periodic lateralized epileptiform discharges (PLEDs) as early indicator of stroke in full-term newborns.

We report on periodic lateralized epileptiform discharges (PLEDs) on EEG in two infants with neonatal convulsions. In both neonates, the EEG abnormalities were seen soon after the onset of seizures, at a time when cranial ultrasound scans were thought to be normal. Subsequent Magnetic Resonance Imaging scans demonstrated cerebral infarction in both patients. In one case, the localisation of the lesion on MRI was concordant with that of the EEG abnormalities, as usually observed in adults with focal lesions. The other infant showed similar EEG abnormalities, but her MRI showed a localised lesion in the basal ganglia, which has also been reported to be involved in the genesis of these abnormalities.

Arm↗
PubMed2000

Head growth in infants with hypoxic-ischemic encephalopathy: correlation with neonatal magnetic resonance imaging.

OBJECTIVES: The aims of the study were to establish the relationship between head growth in the first year of life with the pattern on injury on neonatal magnetic resonance imaging (MRI) in infants with hypoxic-ischemic encephalopathy (HIE) and to relate these to the neurodevelopmental outcome. METHODS: Fifty-two term infants who presented at birth with a neonatal encephalopathy consistent with HIE and who had neonatal brain MRI were entered into the study. Head circumference charts were evaluated retrospectively and the head growth over the first year of life compared with the pattern of brain lesions on MRI and with the neurodevelopmental outcome at 1 year of age. Suboptimal head growth was classified as a drop of >2 standard deviations across the percentiles with or without the development of microcephaly, which was classified as a head circumference below the third percentile. RESULTS: There was no statistical difference between the neonatal head circumferences of the infants presenting with HIE and control infants. At 12 months, microcephaly was present in 48% of the infants with HIE, compared with 3% of the controls. Suboptimal head growth was documented in 53% of the infants with HIE, compared with 3% of the controls. Suboptimal head growth was significantly associated with the pattern of brain lesions, in particular to involvement of severe white matter and to severe basal ganglia and thalamic lesions. Suboptimal head growth predicted abnormal neurodevelopmental outcome with a sensitivity of 79% and a specificity of 78%, compared with the presence of microcephaly at 1 year of age, which had a sensitivity of only 65% and a specificity of 73%. The exceptions were explained by infants with only moderate white matter abnormalities who had suboptimal head growth but normal outcome at 1 year of age and by infants with moderate basal ganglia and thalamic lesions only who had normal head growth but significant motor abnormality.

Apgar Score↗
PubMed2000

[From the great clinical studies to the management of the single patient. Medical therapy in the pre- and post-fibrinolytic era].

Medical therapy of myocardial infarction has changed over the last 30 years. The "lag phenomenon", i.e. the time lapsing from the appraisal of a new, relevant scientific evidence and its practical adoption, has been variable, and different from one treatment to another. It has been very short for GP IIb/IIIa receptor inhibitors in candidates for urgent coronary angioplasty after myocardial infarction, short for ACE inhibitors, and it has been also short for the decrease in the prescription of calcium channel inhibitors after controversial scientific evidence. This time lapse has been long for beta-blockers, that only now are used quite extensively in Italy: from less than 10% of hospitalized patients during the early '80s, to over 60% nowadays. This evolution of medical therapy has been progressive and continuous, has not been divided into two eras, and thrombolysis seems to have little to do with it. The extensive introduction of thrombolysis has simply divided patients with acute myocardial infarction into two subgroups, very different for clinical characteristics: candidates for this therapy, and "others". Candidates for thrombolysis have a much better prognosis, but for a "paradox effect" they have been studied much more than the others. Now, the bulk of publications concerning patients undergoing thrombolysis carries the risk of spreading the concept that myocardial infarction has changed. We do not think that this corresponds to reality. Probably, the concepts that have guided medical therapy have been very similar and unchanged over the years, and both medical and surgical plus coronary angioplasty therapies are today much more appropriately used than yesterday.

Adrenergic beta-Antagonists↗
PubMed2000

[Minimally invasive treatment of intraprostatic cysts].

Prostatic cysts are rare; they are to be considered in the evaluation of obstructive azoospermia. A correct diagnosis is based on transrectal ultrasound of the prostate. Herein we describe our experience with perineal ultrasound cyst aspiration. During the last 10 years, 21 infertile patients underwent ultrasound puncture of an intraprostatic cyst. Patients were placed in the lithotomy position, a 16 G needle was adopted; by means of ultrasound was possible to follow the needle entering the cyst. The content was aspirated and, if no spermatozoa were detected in the cystic liquid, sclerosing agents were introduced. The technique was performed successfully in all patients with symptomatic cyst within the prostate. Follow-up prostatic ultrasound and semen analysis were performed in 17 patients. There was 1 recurrence that was aspirated again. In 12 cases an increase in sperms number and motility was obtained, in 3 cases oligoastenospermia remained unchanged and 2 patients are still azoospermic. In 3 cases the partners become pregnant.

Cysts↗
PubMed1999

Effects of UV-C irradiation on phosphoinositide turnover in plant cells: similarities with those occurring via the formation of reactive oxygen intermediates in animal cells.

With the aim of examining the response of plant cells to UV-C irradiation, we investigated the behaviour of the phosphatidylinositol 4,5 bisphosphate (PtdIns 4,5-P2) molecule (the precursor of the phosphoinositide signal transduction cascade) by exposing callus cells from Peucedanum verticillare to UV-C (130 J m-2) and by examining the level and the fatty acid composition of PtdIns 4,5-P2 at different times after irradiation. We show that a pathway for the UV-C response includes transient PtdIns 4,5-P2 breakdown. The effect of ultraviolet rays is mimicked by H2O2 suggesting that in this plant it may be brought about by reactive oxygen intermediates (ROI), as already underlined in experimental animal models.

Animals↗
PubMed1999

Adrenoceptor involvement in the cardiovascular responses to B-HT 920 in sinoaortic denervated rats.

1. A study was made relating the involvement of alpha-adrenoceptors in the cardiovascular responses to intracerebroventricular (i.c.v.) injection of B-HT 920, a clonidine-type drug, in conscious sham-operated and sinoaortic-denervated rats. 2. Wistar rats were used, 7 days after the sham operation or sinoaortic denervation. For i.c.v. injection of drugs, a guide cannula had been previously implanted in the left lateral ventricle. 3. In sham-operated rats, cardiovascular responses to B-HT 920 (10-60 microg) were increased blood pressure and bradycardia; but, in sinoaortic-denervated rats, after the pressor response, a decrease in blood pressure also was seen. The responses to this agent were greater in sinoaortic-denervated rats than in sham-operated animals. Treatment with the alpha2-adrenoceptor antagonist yohimbine (30 microg), the imidazoline receptor antagonist idazoxan (15 microg) and the alpha1A-adrenoceptor antagonist 5-methylurapidil (15 microg) blocked the responses to B-HT 920 (30 microg). The alpha1-adrenoceptor antagonist prazosin (15 microg) and the alpha1B-adrenoceptor antagonist chloroethylclonidine (100 microg) did not modify the responses to agonist. 4. Sinoaortic denervation enhances the cardiovascular responses to B-HT 920. Moreover, the effects of i.c.v. administration of B-HT 920 could be mediated by several types of brain receptors: imidazoline receptors and alpha1A- and alpha2-adrenoceptors.

Adrenergic alpha-Agonists↗
PubMed1999

Langmuir-Blodgett films of antibodies as mediators of endothelial cell adhesion on polyurethanes.

The effect of endothelial cell adhesion on polyurethanes coated with Langmuir-Blodgett antibody films has been examined. The films were cross-linked with glutaraldehyde with the aim of providing a densely packed and covalently linked two-dimensional antibody network on the polyurethane surfaces. Our results demonstrate that although neither of the two polyurethanes examined were entirely suited to cellular adhesion, Langmuir-Blodgett antibody films, cross-linked with small concentrations of glutaraldehyde, are more suitable for endothelial cell adhesion than surfaces free of antibody.

Antibodies, Monoclonal↗
PubMed1999

Central benzodiazepine involvement in clonidine cardiovascular actions.

It is well known that the GABAergic and noradrenergic systems play an important role in blood pressure and heart rate regulation. Benzodiazepines and beta-carbolines, respectively, increase or decrease the probability of chloride-channel opening induced by GABA. The aim of this study was to determine, in conscious rats, the interaction existing between the central alpha2-adrenoceptor stimulation induced by clonidine and the facilitation or impairment of benzodiazepine receptor activity through the administration of either diazepam, a benzodiazepine receptor agonist, or methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM), an inverse benzodiazepine agonist. Clonidine (5-10 microg, intracerebroventricularly) reduced heart rate and increased mean blood pressure by activation of central alpha2-adrenoceptors. Diazepam (2 mg/kg, intravenously (i.v.)) induced an increase in heart rate, while DMCM (0.3 mg/kg, i.v.) elicited a bradycardic effect. The bradycardic effects induced by both clonidine and DMCM were antagonized by the prior administration of methylatropine (1.5 mg/kg, i.v.). DMCM (0.3 mg/kg, i.v.) prevented the clonidine effects on heart rate and mean blood pressure, while diazepam (2 mg/kg, i.v.) failed to modify these effects. Our results suggest that the bradycardic effects of clonidine are mediated by a vagal stimulation and are related to the activation of a GABAergic pathway.

Adrenergic alpha-2 Receptor Agonists↗
PubMed1999

Morphological and functional evaluation of isolated rat hepatocytes in three dimensional culture systems.

Various three-dimensional configurations, such as polyester tissue and woven-nonwoven, hydrophilic polyester fabric, either collagen-coated or uncoated, were investigated as potential scaffold for hepatocyte culture, in view of their use in bioreactors for hybrid liver support systems. Attachment, morphology and ultrastructure of primary adult rat hepatocytes were evaluated, as well as urea production and ammonium detoxification during a 24h incubation period in serum-free tissue culture medium. As control, hepatocytes were also plated onto collagen-coated dextran microcarriers and on plastic petri dishes, either collagen-coated or uncoated. In all the three-dimensional cultures, hepatocytes appeared morphologically intact without any statistically significant difference in metabolic activity. Collagen-coating did not influence cell attachment to polyester substrates, whereas woven-nonwoven hydrophilic polyester fabric may offer some potential advantages as three-dimensional system for hepatocyte culture in hybrid liver support systems.

Animals↗