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Biomedical subjects

A Marchese

Publications and source records attributed to A Marchese.

At least 73 records · Page 4Linked to original sources
PubMed1996

Detection of SHV-5 extended-spectrum beta-lactamase in Klebsiella pneumoniae strains isolated in Italy.

Thirty-five Klebsiella pneumoniae strains isolated during 1993-1994 in intensive care units of a large Italian hospital were examined for the presence of extended-spectrum beta-lactamases. Five strains showed a high level of simultaneous resistance to beta-lactam agents, including ceftazidime and aztreonam, conferred by a large (130 kb) self-transferable plasmid (in 4 of 5 strains). Isoelectrofocusing and hybridisation studies suggest that these enzymes can be identified as SHV-5 extended-spectrum beta-lactamases. Pulsed-field get electrophoresis analysis showed three different genomic fingerprinting profiles, while plasmid restriction enzyme digestion revealed three different patterns, demonstrating that the diffusion of SHV-5 beta-lactamase is not the result of a single strain or plasmid dissemination.

Conjugation, Genetic↗
PubMed1996

Myocardial contrast echocardiography in acute myocardial infarction. Pathophysiological background and clinical applications.

Myocardial contrast echocardiography is a technique used in experimental and clinical settings in order to visualize the pattern of intramyocardial perfusion. In the acute phase of myocardial infarction, regional absence of flow during myocardial contrast echocardiography delineates the area at risk of necrosis, while the definitive non-perfused area expresses infarct size. Reopening the infarct-related artery, which may be achieved spontaneously by thrombolysis or percutaneous transluminal coronary angioplasty, is not a reliable indicator of intramyocardial reperfusion. If myocardial ischaemia due to coronary occlusion has been sufficiently prolonged and severe, not only myocyte viability, but also microvascular integrity is lost. Myocardial contrast echocardiography, using intracoronary injection of sonicated contrast medium, gives information about microvascular integrity and the effective presence of intramyocardial reflow. Anatomical integrity of microvasculature does not necessarily imply preserved function, and thus the microvessel vasodilating reserve may also be impaired. Myocardial contrast echocardiography has the potential to assess alterations in microvascular function, showing, in the myocardial area with reduced coronary reserve, a relatively reduced increase in echocontrast signal intensity when an intravenous vasodilator agent is administered.

Animals↗
PubMed1996

Brodimoprim: effects of subminimal inhibitory concentrations on virulence traits of respiratory and urinary tract pathogens, and on plasmid transfer and stability.

The effects of brodimoprim, a new trimethoprim analogue, on several virulence traits of respiratory and urinary tract pathogens exposed to sub-lethal levels of the drug was studied. Adherence to tracheal epithelial cells was inhibited by brodimoprim in Klebsiella pneumoniae (41-67% reduction), Moraxella catarrhalis (87-90%) and Haemophilus influenzae (0-53%), while in Streptococcus pneumoniae binding was unaffected. With buccal epithelial cells the comparison between treated and control bacteria indicated statistically significant reduction in adherence with both S.pneumoniae and H.influenzae, (P < 0.015). With M.catarrhalis and Streptococcus pyogenes only marginal changes were detected (P > 0.05). Exoenzyme and capsule production were assessed in at least three isolates of diverse respiratory pathogens grown in the presence of sub-lethal levels of the new agent. The drug affected protease and beta-hemolysin (alpha-toxin) production in both oxacillin-susceptible and -resistant S.aureus. On the contrary, synthesis of lipase, DNase, coagulase, and beta-lactamase (S.aureus), pneumolysin (S.pneumoniae), streptolysin S, DNase, and protease (S.pyogenes), capsule (K.pneumoniae, H.influenzae and S.pneumoniae), and beta-lactamase (K.pneumoniae, H.influenzae and M.catarrhalis) were not inhibited by subminimal inhibitory concentrations (sub-MICs) of the drug. Finally, motility was blocked in urinary pathogens E.coli, P.mirabilis and P.aeruginosa, while in this latter microorganism pigment production was also affected. High molecular weight low-copy F'lac, and low molecular weight high-copy pHSG298 plasmids were eliminated from E.coli treated with sub-MIC concentrations of brodimoprim. The incidence and cured cells ranged from 9% for F'lac to 23% for pHSG298. F'lac transfer was also inhibited by the drug. When conjugation was carried out with bacteria exposed to brodimoprim (5XMIC), a reduction (50%) in the number of recombinants was noted in comparison to the control. The fact that brodimoprim interferes with the expression of some virulence traits, in particular with adherence, at sub-MIC levels may assist the drug in eradicating respiratory pathogens from the epithelial lining, thus diminishing the probability of reinfection.

Bacterial Adhesion↗
PubMed1995

Cloning, expression, and chromosomal localization of the human uridine nucleotide receptor gene.

Extracellular ATP and ADP mediate diverse physiological responses in mammalian cells, in part through the activation of G protein-coupled P2 purinoceptors. The cloning and expression of cDNAs encoding several P2 purinoceptor subtypes have enabled rapid advances in our understanding of the structural and functional properties of these receptors. The current report describes the isolation of a gene from a human genomic library that encodes a protein with the greatest similarity to the human P2U purinoceptor, a subtype that is distinguished by its ability to be activated by uridine nucleotides as well as adenine nucleotides. When expressed in a mammalian cell line, this novel receptor is activated specifically by UTP and UDP but not by ATP and ADP. Activation of this uridine nucleotide receptor resulted in increased inositol phosphate formation and calcium mobilization. Fluorescence in situ hybridization revealed that the gene encoding the uridine nucleotide receptor is located in region q13 of the X chromosome. Dendrogram analysis of the G protein-coupled P2 purinoceptors and the uridine nucleotide receptor indicates that these receptors belong to a family that may be more aptly named nucleotide receptors.

Amino Acid Sequence↗
PubMed1995

Cloning and chromosomal mapping of three novel genes, GPR9, GPR10, and GPR14, encoding receptors related to interleukin 8, neuropeptide Y, and somatostatin receptors.

We employed the polymerase chain reaction and genomic DNA library screening to clone novel human genes, GPR9 and GPR10, and a rat gene, GPR14. GPR9, GPR10, and GPR14 each encode G protein-coupled receptors. GPR10 and GPR14 are intronless within their coding regions, while GPR9 contains at least one intron. The receptor encoded by GPR9 shares the highest identity with human IL-8 receptor type B (38% overall and 53% in the transmembrane regions), followed by IL-8 receptor type A (36% overall and 51% in the transmembrane domains). GPR10 encodes a receptor that shares highest identity with the neuropeptide Y receptor (31% overall and 46% in the transmembrane domains). The receptor encoded by GPR14 shares highest identity with the somatostatin receptor SSTR 4 (27% overall and 41% in the transmembrane domains). Fluorescence in situ hybridization analysis localized GPR9 to chromosome 8p11.2-p12 and GPR10 to chromosome 10q25.3-q26.

Amino Acid Sequence↗
PubMed1995

The cloning and chromosomal mapping of two novel human opioid-somatostatin-like receptor genes, GPR7 and GPR8, expressed in discrete areas of the brain.

Following the cloning of the opioid receptors mu, kappa, and delta, we conducted a search for related receptors. Using oligonucleotides based on the opioid and also the structurally related somatostatin receptors, we amplified genomic DNA using the polymerase chain reaction and isolated fragments of novel G protein-coupled receptor genes. Two of these gene fragments designated clones 12 and 11 were used to isolate the full-length genes. The intronless coding sequences of these genes, named GPR7 and GPR8, shared 70% identity with each other, and each shared significant similarity with the sequences encoding transmembrane regions of the opioid and somatostatin receptors. GPR7 was mapped to chromosome 10q11.2-q21.1 and GPR8 to chromosome 20q13.3. Northern blot analysis using human mRNA demonstrated expression of GPR7 mainly in cerebellum and frontal cortex, while GPR8 was located mainly in the frontal cortex. In situ hybridization revealed expression of GPR7 in the human pituitary. A partial sequence of the mouse orthologue of GPR7 was obtained, and in situ hybridization demonstrated expression in discrete nuclei of brain, namely suprachiasmatic, arcuate, and ventromedial nuclei of hypothalamus. A stable cell line expressing the GPR7 gene was created, but expression levels of the receptor were low. The available pharmacology indicated binding to several opioid drugs such as bremazocine, levorphanol, and beta-FNA, but not to the opioid receptor subtype-selective mu, delta, or kappa agonists.

Amino Acid Sequence↗
PubMed1995

The human dopamine D5 receptor gene: cloning and characterization of the 5'-flanking and promoter region.

Genomic and overlapping cDNA clones encompassing the entire 5'-untranslated region of the human D5 receptor gene were cloned and sequenced. Comparison of these human D5 receptor genomic and cDNA clones revealed the presence of two exons separated by a small and variably sized intron (of either 179 or 155 bp). We have determined that the major site of transcription initiation of the D5 gene is 2125 bp upstream from the translational initiation start site. The region 5' to the transcription initiation site lacked conventional TATA and CAAT sequences, but contained several putative binding sites for transcription factors, such as Sp1 and Ap1. Luciferase reporter gene constructs containing D5 gene sequence information up to 500 bp 5' of the transcription initiation site were able to stimulate transcription only in SK-N-SH cells but not in COS-7, CHO, P19EC, NB41A3, and SK-N-MC cell lines. Promoter deletion analysis indicated that the D5 gene promoter contained a positive modulator at 119-182 and a negative modulator 251-500 bases upstream from the site of transcription initiation. In addition, in order to detect the expression of functional D5 receptor mRNAs and not those of its expressed pseudogenes, in situ hybridization analysis of monkey and human brain using a 5' D5-specific riboprobe revealed that D5 receptor mRNA was most abundant in discrete cortical areas (layers II, IV, and VI), the dentate gyrus, and hippocampal subfields with very little message detected in the striatum. Unexpectedly, D5 mRNA antisense riboprobes labeled discrete cell bodies in the pars compacta of the substantia nigra. The characterization of the genomic organization of the D5 receptor gene and of those factors involved in its transcriptional regulation may aid in our understanding of the role this gene product plays in the generation and maintenance of dopamine D1-like receptor-mediated events.

Base Sequence↗
PubMed1995

Two gene duplication events in the human and primate dopamine D5 receptor gene family.

The human dopamine D5 receptor (DRD5) gene family consists of the DRD5-encoding gene (DRD5) and the pseudogenes psi DRD5-1 and psi DRD5-2. Analysis of the 5' UTR of DRD5 and homologous regions in the pseudogenes revealed that the nucleotide identity (approx. 95%) extended for 1.9 kb and terminated at a monomeric Alu sequence in each of the pseudogenes. The presence of Alu sequences in the pseudogenes, at this point of divergence with DRD5, suggests that Alu sequences were involved in the evolution of the DRD5 family. This report is the first to describe a possible mechanism involved in the duplication of genes in the G-protein-coupled receptor (GPCR) family. The pseudogenes continue to share identity (approx. 98%) beyond this 5' UTR point of divergence with DRD5 for at least another 6 kb. Analysis of the 3' UTR of DRD5 and homologous regions in the pseudogenes revealed that the identity (approx. 95%) extends at least 14 kb, and the identity between the pseudogenes (approx. 98%) extends for at least 18 kb. Thus, the duplication unit that produced the first pseudogene was at least 16 kb, whereas the second pseudogene was at least 28 kb. We have also located two DRD5 pseudogenes in gorilla demonstrating that these closely related pseudogenes were present in a common ancestor of human and gorilla.

Animals↗
PubMed1995

Isolation of three novel human genes encoding G protein-coupled receptors.

We have cloned and mapped the chromosomal location of three novel human genes encoding G protein-coupled receptors that we have named GPR6, GPR5, and GPR4. The entire coding region for each of these genes was contained on single exons. Gene GPR6 encoded a receptor that shared closest identity (71% in the transmembrane regions) with the human orphan receptor GPR3 and was localized to chromosome 6 (q21-q22.1). Northern blot analysis revealed that GPR6 transcripts were abundant in the human putamen and to a lesser extent in the frontal cortex, hippocampus, and hypothalamus. Gene GPR5 encoded a receptor that most closely resembled the orphan receptor RBS11 (48% in the transmembrane regions) and the MIP 1 alpha/RANTES receptor (45% in the transmembrane regions) and was localized to chromosome 3 (p21.3-p21.1). Gene GPR4 shared identity (40% in the transmembrane regions) with the human platelet-activating factor receptor and was localized to chromosome 19 (q13.2-q13.3).

Amino Acid Sequence↗
PubMed1995

Antistaphylococcal activity of cefdinir, a new oral third-generation cephalosporin, alone and in combination with other antibiotics, at supra- and sub-MIC levels.

Cefdinir is one of the few oral third generation cephalosporins that shows useful activity against nosocomial Gram-positive pathogens. For this reason the anti-staphylococcal potency of the new drug, alone or in combination with other drugs was further characterized. Against penicillin-resistant, oxacillin-susceptible Staphylococcus isolates, cefdinir demonstrated useful in-vitro activity. MIC90 values (in mg/L) were 0.25 for Staphylococcus aureus (30 strains), 0.06 for Staphylococcus epidermidis (24), 0.125 for Staphylococcus hominis (10), 0.5 for both Staphylococcus xylosus (15) and Staphylococcus capitis (11) and 4 for Staphylococcus saprophyticus (10), while Staphylococcus haemolyticus (12) was less susceptible with a MIC90 value of 32. Cefdinir activity was not adversely affected by several variables such as pH, inoculum size or the presence of serum or urine. The new cephem induced a PAE on all isolates studied: with S. aureus the extent of regrowth suppression ranged from 0.8 to 1 h, and with the other species from 0.5 (S. epidermidis) to 4.1 h (S. haemolyticus). Development of resistant strains was rare. At the highest level used (10 x MIC) mutants arose with a frequency of 6 x 10(-8) with S. haemolyticus and 2 x 10(-9) with S. epidermidis. The absence of a paradoxical effect of increasing concentrations of cefdinir on its bactericidal activity was confirmed up to a value of 500-fold the MICs. When cefdinir activity was assessed in association with ciprofloxacin, netilmicin, clarithromycin, fosfomycin, rifampicin, teicoplanin and vancomycin using the chequerboard and time-kill techniques, indifference predominated with all strains and in all combinations. Synergism was detected only in 11 out of a total of 175 tests performed by the chequerboard method. Using the time-kill technique cefdinir reacted synergically in 25 of 126 tests. Antagonism was never observed. S. aureus exposed to sub-inhibitory concentrations of cefdinir failed to grow on mannitol-salt agar and to produce haemolysins, but retained coagulase activity. Penicillinase production was also lost in about 17% of the survivors. Hydrophobicity changes were detected in all species tested with the exception of S. saprophyticus.

Anti-Bacterial Agents↗
PubMed1995

Susceptibility of Streptococcus pneumoniae strains isolated in Italy to penicillin and ten other antibiotics.

The susceptibility of 312 Streptococcus pneumoniae strains isolated during 1993 in Italy has been studied. Overall resistance to penicillin, erythromycin and co-trimoxazole was 5.5, 6 and 18% respectively. The penicillin resistant isolates (nine displaying low- and eight high-level resistance) were further tested against cefotaxime, ceftriaxone, cefepime, cefodizime, ceftazidime, imipenem, meropenem and rifampicin. The low-level penicillin resistant strains were inhibited by all these drugs. Among the eight high-level penicillin resistant pneumococci, four were resistant to one or more cephalosporins. Imipenem and rifampicin remained effective against all isolates. In comparison with other European Mediterranean countries, penicillin resistance is a minor threat at present in Italy.

Anti-Bacterial Agents↗
PubMed1995

[In vitro antibacterial activity of meropenem, a new carbapenem: European data].

Meropenem is a new DHP-I stable carbapenem with a very promising microbiological, pharmacokinetic and clinical profile. The antibacterial activity of this new agent has been assessed in vitro against 8741 aerobic and 854 anaerobic strains reflecting current incidence and epidemiology in Italy, France, Germany, Spain, Switzerland and United Kingdom. Comparator agents were imipenem, ceftriaxone, vancomycin, ciprofloxacin gentamicin and amikacin. The results of this study show that meropenem has a spectrum of antibacterial activity which embraces the vast majority of clinically significant Gram-positive and Gram-negative aerobes and anaerobes. This is due in part to excellent stability to chromosomal or plasmid mediated beta-lactamases including those which hydrolyse current cephalosporins. Data from the Italian study identified meropenem as the most potent agent against all Enterobacteriaceae, with the exception of Proteus species with were most susceptible to ciprofloxacin. Moreover, meropenem was 10 times more active than the other drugs against Haemophilus and Neisseria and was active against all the anaerobic strains. Conversely, staphylococci and enterococci were more susceptible to imipenem. Overall, these European data showed that meropenem was the most powerful drug against Enterobacteriaceae and it also was the most effective drug tested against the Italian and French Pseudomonas aeruginosa strains. Meropenem was less effective than imipenem or vancomycin against Enterococcus stains but had similar activity to imipen against anaerobes. Based on this microbiological profile, the use of meropenem is appropriate in the empirical treatment of serious infections, including those caused by multiple pathogens.

Anti-Bacterial Agents↗
PubMed1995

The beta-lactamase problem: new therapeutic options.

As a consequence of their successful use in prophylaxis and therapy, bacterial resistance mediated by beta-lactamases is now widely diffused among beta-lactam antibiotics. Several effective strategies have been suggested in order to overcome this problem. One interesting option is offered by the development of a series of new beta-lactam compounds that possess a very high intrinsic stability to the hydrolytic action of the most common beta-lactamases. Among these molecules the oral third generation cephalosporins represent a significant breakthrough. Cefetamet pivoxil, because of its broad coverage of most gram-negative and gram-positive community acquired pathogens, rightly belongs to these new agents. The activity of cefetamet has been confirmed in a survey in Italy involving 4191 isolates. on this collection of strains cefetamet emerged as the most active in vitro compound, followed by cefixime, with all other comparative agents (cefuroxime, cefaclor, cephalexin, cefradoxil, ampicillin, amoxicillin-clavulanate, ampicillin-sulbactam, doxycycline, erythromycin and clindamycin) displaying lower eradication rates. According to the data gathered in the Italian survey, cefetamet can be considered the only compound, among those taken into consideration, that might be selected as the drug of choice in the empiric therapy of respiratory and urinary community-acquired infections. In fact, the prevalence of resistance to cefetamet in the most prevalent pathogens occurring in this setting is, at present, sufficiently low to render therapeutic failures, based on this parameter, highly improbable.

Anti-Bacterial Agents↗
PubMed1994

Mapping studies of two G protein-coupled receptor genes: an amino acid difference may confer a functional variation between a human and rodent receptor.

We recently isolated two orphan human G protein-coupled receptor genes designated GPR1 and GPR6. The gene GPR1 was shown to be transcribed abundantly but only in the hippocampus. Here we report the cloning of the rat GPR1 gene and report the absence of expression in hippocampus, demonstrating a functional variation for this receptor in these two species. The evolutionary history of an important sequence difference in the gene GPR1 in primate and rodent species has been examined. In contrast extensive mapping of gene GPR6 mRNA in rat brain was in keeping with the described distribution in human brain.

Amino Acid Sequence↗
PubMed1994

Cloning of human genes encoding novel G protein-coupled receptors.

We report the isolation and characterization of several novel human genes encoding G protein-coupled receptors. Each of the receptors contained the familiar seven transmembrane topography and most closely resembled peptide binding receptors. Gene GPR1 encoded a receptor protein that is intronless in the coding region and that shared identity (43% in the transmembrane regions) with the opioid receptors. Northern blot analysis revealed that GPR1 transcripts were expressed in the human hippocampus, and the gene was localized to chromosome 15q21.6. Gene GPR2 encoded a protein that most closely resembled an interleukin-8 receptor (51% in the transmembrane regions), and this gene, not expressed in the six brain regions examined, was localized to chromosome 17q21.1-q21.3. A third gene, GPR3, showed identity (56% in the transmembrane regions) with a previously characterized cDNA clone from rat and was localized to chromosome 1p35-p36.1.

Amino Acid Sequence↗
PubMed1994

[Assessment of viable myocardium after infarction with transesophageal echocardiography and myocardial echocontrastography].

Left ventricular recovery of dysfunctioning but viable myocardium can occur only in tissue in which both myocardial contractile reserve and microvascular integrity are preserved. Recent studies have demonstrated that both inotropic stimulating echo tests and myocardial contrast echocardiography can be used to assess myocardial viability in post-myocardial infarction patients. Therefore we performed a transesophageal and myocardial contrast echocardiographic evaluation of post-myocardial infarction patients to assess: the respective accuracy of post-extrasystolic potentiation and low-dose dobutamine (5 and 10 mcg) during transesophageal echocardiography in eliciting contractile reserve, and the potential of myocardial contrast echocardiography in predicting later functional recovery. Results of our studies showed comparable effects of low-dose dobutamine (5 mcg) and post-extrasystolic potentiation in increasing myocardial thickening while low-dose dobutamine (10 mcg) had a greater potential in eliciting residual contractility. Lastly, myocardial contrast echocardiography provided a clear spectrum of intramyocardial perfusion patterns in dysfunctioning areas but did not accurately correlate with later functional recovery as better predicted by low-dose dobutamine in the same segments. In conclusion, these methods represent the preferred choice of studying the perfusion-contraction match in viable myocardium thus playing an important role in prognostic and therapeutic strategies in myocardial infarction patients.

Echocardiography↗
PubMed1994

[The echocardiographic assessment of the functional variations in the left ventricle induced by isometric stress in subjects with primary cardiomyopathy in the pre-dilated phase].

The aim of this study was to assess whether a sudden increase in left ventricular (LV) afterload in a pre-dilated condition of idiopathic cardiomyopathy, in associated with some non-invasively detectable impairment of LV function. Ten subjects (Group A, 7 males, 3 females, mean age 36.7 +/- 11.4 years), submitted to cardiac catheterization because of severe symptoms related to ventricular arrhythmias, were found to have normal coronary arteries and left ventricular function, but histological features suggestive of congestive cardiomyopathy at endomyocardial biopsy. Echocardiography was performed during handgrip for 5 min at 30% of the maximal effort, in order to obtain a sharp increase in systolic LV pressure of more than 40 mm Hg. Ten healthy subjects were used as controls (Group B). Results in Group A showed a significant decrease in LV ejection fraction (from 62.4 +/- 6.9 to 57 +/- 7; p < 0.005) and LV shortening fraction (from 34.2 +/- 5.1 to 31 +/- 4.3; p < 0.01); a significant increase in LV systolic volume (from 48.2 +/- 9.1 to 60.0 +/- 12.0; p < 0.005) and systolic stress (from 114.6 +/- 33.2 to 178.8 +/- 58.2; p < 0.001). At the first minute of recovery LV function normalized quickly while LV systolic volume remained moderately increased (52.5 +/- 10.2). In conclusion, in pre-congestive dilated cardiomyopathy a latent impairment in LV function can be detected during a sharp increase of LV afterload. Isometric stress is particularly useful in eliciting these transient changes, as the method allows a reliable echocardiographic examination during maximal stress and not only during the recovery phase, as in dynamic stress.

Adult↗