Frequency of switch from cyclosporine to FK 506 before and after Neoral use in pediatric liver transplantation.
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Administration, Oral↗
Biomedical subjects
S Riva
Publications and source records attributed to S Riva.
Steatocystoma multiplex and leuconychia in a child with Alagille syndrome.
Alagille syndrome is a rare autosomal dominant developmental disorder, characterized by congenital paucity of interlobular bile ducts, peculiar facies, posterior embryotoxon, bone abnormalities, and peripheral pulmonary artery stenosis. Cutaneous involvement in this disorder is quite rare and the association of Alagille syndrome with multiple comedones and cysts has been described only once. Here, we report the clinical and histological findings of a 7-year-old patient affected by Alagille syndrome who presented multiple cystic lesions and comedones reminiscent of steatocystoma multiplex and a congenital total true leuconychia. Although unexplained, the association of this syndrome with a developmental disorder of the pilosebaceous unit may not be fortuitous.
Alagille Syndrome↗
No treatment for low-risk thrombocythaemia: results from a prospective study.
Essential thrombocythaemia (ET) is a chronic myeloproliferative disorder characterized by the occurrence of thromboembolic episodes, particularly in patients aged > 60 years or with a previous history of thrombosis, and/or by haemorrhages in patients with an exceedingly high platelet count. In these subgroups of patients the use of cytoreductive therapy is beneficial in terms of risk/benefit ratio. Only limited anecdotal data are available on the thrombotic or haemorrhagic risk and survival in young asymptomatic ET patients with a platelet count < 1500 x 10(9)/l. Therefore the optimal management of these patients is unknown. To assess the incidence of thrombosis and haemorrhages in this group of patients we carried out a prospective observational study in a cohort of 65 patients with ET, aged < 60 years, with no history of thrombosis or haemorrhage and platelet count < 1500 x 10(9)/l, and in 65 age- and sex-matched controls. Patients were not treated with cytoreductive therapy until the occurrence of thrombosis or haemorrhage. Arterial or venous thrombotic events were objectively documented both in cases and in controls. The median follow-up was 4.1 years, with an incidence of thrombosis in patients and controls of 1.91 and 1.50 cases/100 patient-years, respectively. The age- and sex-adjusted risk rate ratio was 1.43 (95% CI 0.37-5.4). Only three minor haemorrhagic episodes occurred in patients, with an incidence of 1.12 cases/100 patient-years. Pregnancy and surgery were not associated with thrombosis in these patients. We conclude that the thrombotic risk in young ET patients, with no thrombotic history and a platelet count < 1500 x 10(9)/l, is not increased compared to the normal population and that a conservative therapeutic approach should therefore be considered in these patients.
Adolescent↗
[The hemodynamics laboratory without cine film. A comparison between the production costs of 35-mm film and the recordable compact disk].
BACKGROUND: For more than 35 years, cine film has been used as the standard recording medium for coronary angiography. At present, the DICOM and the CD-R format have been established as the standard media for the transport of cardiac angiographic images in place of cine angiographic film. The aim of the study was to compare the media production costs between two cardiac catheterization laboratories, with and without cine film. METHODS: We collected data from two different laboratories in the same geographic area which perform about 1000 procedures/year, using a similar digital x-ray imaging system. In one lab, images are recorded on 35-mm film at 25 frames/sec. In the other one, the image support is based on a CD-R. For each laboratory we considered both direct patient and variable equipment costs. Direct patient costs in the film-lab include: cine film, processing chemicals, processing labor, chemical disposal, maintenance; in the digital lab: CD-R costs and masterization time. Equipment costs in the film-lab include: cine camera, cine film processor and cine projector; in the filmless lab a DICOM formatter and a review workstation. The equipment amortization costs are considered over a three-year period. RESULTS: Total direct patient costs are 90,000 lira for the film and 14,000 lira for the CD-R. Equipment costs are 193,000,000 lira in the film-lab and 150,000,000 lira in the filmless one. Overall cost per patient is 154,300 lira for the cine film and 64,000 lira for the CD-R. CONCLUSIONS: This study shows that the media costs per patient for a digital DICOM CD-R format system are substantially less than for 35-mm film, permitting savings of more than 90,000,000 Italian lira per year in a mid-volume cardiac catheterization laboratory.
Compact Disks↗
Growth-dependent and growth-independent translation of messengers for heterogeneous nuclear ribonucleoproteins.
The hnRNP A1 transcript has a relatively short 5'- untranslated region (UTR) starting with a pyrimidine tract similar to that of mRNAs encoded by the TOP [terminal oligo(pyrimidine)] genes in vertebrates. Such genes code for ribosomal proteins and for other proteins directly or indirectly involved in the production and function of the translation apparatus. As expected from the role of the pyrimidine tract in the translational regulation of TOP mRNAs, the A1 mRNA is more efficiently loaded onto polysomes in growing than in resting cells. On the other hand, a less stringent regulation with respect to that of other TOP mRNAs is observed, partially due to the presence of multiple transcription start sites within the pyrimidine tract, where transcripts with shorter TOP sequences are less sensitive to regulation. Thus, from the point of view of structural features and translation behaviour the A1 mRNA can be included in the class of TOP genes, suggesting a possible role of A1 in translation. Interestingly, a TOP-like behaviour was observed for hnRNP I mRNA but not for hnRNP C1/C2 and A2/B1 mRNAs, indicating the existence of two classes of hnRNPs with different translational regulation.
Base Sequence↗
A repeated element in the human lamin B2 gene covers most of an intron and reiterates the exon/intron junction.
Nuclear lamins are intermediate filament-type proteins forming a fibrillar meshwork that underlies the inner nuclear membrane. We have previously reported the identification of the human lamin B2 gene that maps to the subtelomeric band p13.3 of chromosome 19 in close proximity of a human DNA replication origin. Here we report the identification within the human lamin B2 gene of a novel repeated element (variable number of tandem repeats: VNTR) that appears to have a very recent origin, being absent in the genome of mouse and primates such as cercopitheques, lemurs and macaques. The VNTR is adjacent to exon 8 of the lamin B2 gene which, albeit encoding the nuclear localization signal of the protein, is highly divergent both at amino acid and nucleotide level among species. Moreover the VNTR, characterized by a repeated unit of about 100 bp, covers most of intron 8 of the gene and reiterates both the last 7 bp of the upstream exon and the exon/intron junction. RT-PCR experiments carried out on HeLa cell RNA suggest that none of the downstream junctions is used during the processing of the lamin B2 pre-mRNA.
Alternative Splicing↗
Sequence determinants for hnRNP I protein nuclear localization.
hnRNP I, also referred to as polypyrimidine tract binding protein, is one of the proteins associated with nascent pre-mRNA in the heterogeneous nuclear ribonucleoprotein (hnRNP) complexes. As for all karyophilic proteins, the nuclear import of hnRNP proteins requires specific sequence determinants that in many instances differ from the canonical import signal. In order to identify the sequences responsible for the nuclear localization, various hnRNP I portions were fused to a reporter protein (bacterial chloramphenicol acetyl transferase) and used in transient transfection assay. By this approach we identified a 60-amino-acid sequence located at the amino terminus of hnRNP I (designated NLD-I) that is both necessary and sufficient for nuclear localization. NLD-I represents a novel bipartite type of nuclear localization signal that bears no resemblance to other nuclear localization determinants so far identified in hnRNP proteins.
3T3 Cells↗
Analysis of mandelonitrile lyase and beta-glucosidase from sweet almonds by combined electrophoretic techniques.
Almonds are a rich source of mandelonitrile lyase (oxynitrilase) and beta-glucosidase. The isolation of these two enzymes from sweet almonds requires fractional ammonium sulfate precipitation followed by ion-exchange chromatography on diethylaminoethyl-(DEAE) and carboxymethylcellulose (CMC) columns. In the present investigation different electrophoretic techniques such as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing in immobilized pH gradients (IEF-IPG), and capillary electrophoresis were used to characterize these two enzymes. For the first time, beta-glucosidase and oxynitrilase were separated in an immobilized pH gradient of one pH unit. Capillary zone electrophoresis (CZE) was an excellent tool for analysis of the purity of enzyme preparations, achieving complete separation of various protein constituents in only 15 min. CZE showed a resolving capacity for the separation of enzyme forms comparable to that of isoelectric focusing in an immobilized pH gradient.
Aldehyde-Lyases↗
Effect of organic cosolvents on the stability and activity of the beta-1,4-galactosyltransferase from bovine colostrum.
The influence of various organic cosolvents on the stability and activity of the beta-1,4-galactosyltransferase from bovine colostrum (GalT) and of its ancillary enzyme UDP-galactose-4'-epimerase has been investigated using the glucosylated alkaloid colchicoside (1) as a model substrate. It has been found that some cosolvents, such as Me2SO and MeOH, can be used up to 20% v/v without any influence on the performance of these enzymes, while others, such as tetrahydrofuran, rapidly inactivated GalT at concentrations as low as 5% v/v. These results have been exploited for the galactosylation of the poorly water soluble coumarinic glucoside fraxin (2).
Animals↗
[Improvement of accessory pathways radiofrequency catheter ablation by arterial or venous epicardial mapping].
Failure of radiofrequency catheter ablation for atrioventricular reciprocating tachycardia may be related to imprecise location of accessory pathways. We have tested the safety and efficacy in improving successful rate of the procedure of a new technique of epicardial mapping of the atrioventricular sulcus by means of a small diameter (2.5F) 16 polar electrode catheter with a soft tip and a minor interelectrode and intercouple distance (2-6-2). The catheter was advanced via a right femoral approach into the coronary sinus or its branches, and the right coronary artery. We report 5 patients who underwent epicardial mapping-guided radiofrequency catheter ablation who had been previously treated with 1 or more (range 1-4) unsuccessful traditional mapping of the atrioventricular sulcus. Epicardial mapping was performed by means of selective catheterization of the coronary sinus in 4 cases, and of the right coronary artery in 1. The accessory pathways was precisely localized and ablated in all patients (mean 8 +/- 1.5 radiofrequency pulses, and 32 +/- 6 min fluoroscopy duration). No procedure or catheterization-related complications were observed. In conclusion, the technique of epicardial mapping used in this study proved to be safe and effective in localizing accessory pathways in selected cases, thereby enhancing radiofrequency catheter ablation success rate. The main advantage of this atraumatic catheter as compared to the traditional ones are the femoral approach and the possibility to advance the catheter to the most anterior aspect of the great cardiac vein. The epicardial mapping is thus a feasible alternative to traditional mapping, particularly in cases in which previous procedures have failed due to a complex arrhythmogenic substrate and or congenital abnormalities.
Catheter Ablation↗
Synthetic applications of NAD(P)(H)-dependent enzymes.
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Bacteria↗
Utilization of the same DNA replication origin by human cells of different derivation.
In the past, a highly sensitive and efficient method was developed to map DNA replication origins in human cells, based on quantitative PCR performed on nascent DNA samples. This method allowed the identification of a replication origin in the myeloid HL-60 cell line, located on chromosome 19 within an approximately 500 bp segment near the lamin B2 gene [Giacca et al. (1994) Proc. Natl. Acad. Sci. USA, 91, 7119]. The same procedure has now been further simplified and extended to a variety of other exponentially growing human cells of different histological derivation (three neural, one connectival and one epithelial), with a nearly diploid chromosomal content. In all the six cell lines tested, the origin activity within the lamin B2 gene domain was localized to the same region. Furthermore, the lamin B2 origin was also found to be active in stimulated, but not in quiescent, peripheral blood lymphocytes.
Bone Marrow Cells↗
hnRNP A1 selectively interacts through its Gly-rich domain with different RNA-binding proteins.
Heterogeneous nuclear ribonucleoproteins (hnRNPs) are abundant nuclear polypeptides, most likely involved in different steps of pre-mRNA processing. Protein A1 (34 kDa), a prominent member of the hnRNP family, seems to act by modulating the RNA secondary structure and by antagonizing some splicing factors (SR proteins) in splice-site selection and exon skipping/inclusion. A role of A1 in the nucleo-cytoplasmic transport of RNA has also been proposed. These activities might depend not only on the RNA-binding properties of the protein but also on specific protein-protein interactions. Here we report that A1 can indeed selectively interact, in vitro, both with itself and with other hnRNP basic "core" proteins. Such selective binding is mediated exclusively by the Gly-rich C-terminal domain, where a novel protein-binding motif constituted by hydrophobic repeats can be envisaged. The same domain is necessary and sufficient to promote specific interaction in vivo, as assayed by the yeast two-hybrid assay. Moreover, an in vitro interaction with some SR proteins was also observed. These observations suggest that diverse and specific protein-protein interactions might contribute to the different functions of the hnRNP A1 protein in mRNA maturation.
Amino Acid Sequence↗
In vivo protein-DNA interactions at human DNA replication origin.
Protein-DNA interactions were studied in vivo at the region containing a human DNA replication origin, located at the 3' end of the lamin B2 gene and partially overlapping the promoter of another gene, located downstream. DNase I treatment of nuclei isolated from both exponentially growing and nonproliferating HL-60 cells showed that this region has an altered, highly accessible, chromatin structure. High-resolution analysis of protein-DNA interactions in a 600-bp area encompassing the origin was carried out by the in vivo footprinting technique based on the ligation-mediated polymerase chain reaction. In growing HL-60 cells, footprints at sequences homologous to binding sites for known transcription factors (members of the basic-helix-loop-helix family, nuclear respiratory factor 1, transcription factor Sp1, and upstream binding factor) were detected in the region corresponding to the promoter of the downstream gene. Upon conversion of cells to a nonproliferative state, a reduction in the intensity of these footprints was observed that paralleled the diminished transcriptional activity of the genomic area. In addition to these protections, in close correspondence to the replication initiation site, a prominent footprint was detected that extended over 70 nucleotides on one strand only. This footprint was absent from nonproliferating HL-60 cells, indicating that this specific protein-DNA interaction might be involved in the process of origin activation.
Base Sequence↗
The roles of heterogeneous nuclear ribonucleoproteins (hnRNP) in RNA metabolism.
In eukaryotic cells, messenger RNAs are formed by extensive post-transcriptional processing of primary transcripts, assembled with a large number of proteins and processing factors in ribonucleoprotein complexes. The protein moiety of these complexes mainly constitutes a class of about 20 major polypeptides called heterogeneous nuclear ribonucleoproteins or hnRNPs. The function and the mechanism of action of hnRNPs is still not fully understood, but the identification of RNA binding domains and RNA binding specificities, and the development of new functional assays, has stimulated interest in them. In contrast to previous models that hypothesised a mere structural (histone-like) function, a more diversified and dynamic role for these proteins is now emerging. In fact, they can be viewed as a subset of the trans-acting pre-mRNA maturation factors. They might actively participate in post-transcriptional events such as regulated splicing and mRNA export. Moreover, recent data suggest an involvement of some of these proteins in molecular diseases. Here we present an overview of the most relevant properties of hnRNPs and discuss some emerging ideas on their roles.
Amino Acid Sequence↗
A two-step efficient chemoenzymatic synthesis of flavonoid glycoside malonates.
A simple and high-yielding protocol for the malonylation of some flavonoid glycosides is described. The two-step synthesis is based on the regioselective enzymatic introduction of a benzylmalonyl group by catalysis with the lipase from Candida antarctica, followed by Pd/C hydrogenolysis of the benzyl moiety.
Acylation↗
Persistence of single echo beat inducibility after selective ablation of the slow pathway in patients with atrioventricular nodal reentrant tachycardia: relationship to the functional properties of the atrioventricular node and clinical implications.
INTRODUCTION: Residual slow pathway conduction with or without reentrant echo beats has been reported in 25% to 30% of patients undergoing ablation for AV nodal reentrant tachycardia (AVNRT). METHODS AND RESULTS: Fifty-eight consecutive patients (aged 45 +/- 12 years) with slow-fast AVNRT underwent radiofrequency catheter ablation of the slow AV nodal pathway (SP). Residual slow-fast echo beat was documented in 21 (36%) of 58 patients (group A). The pre- and postablation AH intervals triggering the echo beats were similar (346 +/- 8 msec vs 352 +/- 6 msec, P = NS), as were the pre- and postablation echo zones (55 +/- 6 msec vs 52 +/- 5 msec, P = NS) and functional refractory period of the SP. A consistent prolongation of the AV nodal effective refractory period (AVN-ERP; from 265 +/- 28 msec to 340 +/- 50 msec, P < 0.001) and the Wenckebach cycle length (WBCL; from 298 +/- 41 msec to 438 +/- 43 msec, P < 0.001) was observed in all patients with abolition of SP conduction (group B). In group A patients, the prolongation of WBCL was less (285 +/- 33 msec preablation, and 334 +/- 41 msec postablation, P < 0.001). Additional pulses abolished the residual echo in 16 of 21 patients, and further prolongation of the AVN-ERP and WBCL comparable to those found in patients without a residual echo beat was observed. During 19 +/- 8 months follow-up, no patient had clinical recurrence of AVNRT. CONCLUSION: Residual single echo beat after SP ablation for AVNRT reflects the persistence of some portion of the SP with unchanged functional conduction properties whose prognostic significance is uncertain. A consistent increase of WBCL can be a reliable marker of complete abolition of slow pathway conduction and termination of AVNRT.
Adult↗
Bacteria and bacterial cell wall constituents induce the production of regulatory cytokines in dendritic cell clones.
The primary function of dendritic cells (DC) is the uptake, processing, and presentation of antigens to unprimed T cells, but the regulation of these functions is largely unknown. The study of the signals that maintain DC in a resting state or that drive their activation has been hampered by the difficulties in obtaining pure DC populations. The availability of immortalized DC clones from different tissues (spleen and skin) allowed us to investigate the regulation of cytokine production in response to physiological signals in the absence of contaminating cells. The DC clones exhibited the phenotypical and functional features of DC precursors and could phagocytose, albeit at a low rate, whole bacteria. Heat-inactivated bacteria and bacterial cell wall products were tested for cytokine induction. Lipopolysaccharide, lipoteichoic acid, and gram-negative bacteria were potent inducers of tumor necrosis factor alpha and interleukin 6 release, whereas gram-positive bacteria were less efficient. The results suggest that microbial infections can directly promote cytokine DC release of relevant inflammatory responses as well as in the autocrine activation of DC.
Animals↗