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Biomedical subjects

D Hernandez

Publications and source records attributed to D Hernandez.

At least 37 records · Page 2Linked to original sources
PubMed1999

Recipient factors as determinants of mortality after adult liver transplantation.

The factors that can influence the outcome of orthotopic liver transplantation (OLT) are numerous. The purpose of this study was to determine the effects of recipient preoperative factors on patient mortality. Between April 1986 and April 1998 a total of 600 OLTs were performed in our institution. We retrospectively reviewed our first 203 consecutive primary adult OLTs with at least 4 years of follow-up. A case-control comparison was performed between survivors and nonsurvivors, and differences in recipient variables were studied for their correlation with patient mortality. A logistic regression analysis was also performed. Mortality was significantly increased among those with fulminant hepatic failure (FHF) (66.6%, p = 0.003), primary cancer (63.1%, p = 0.018), females (46.1%, p = 0. 043), encephalopathy grade IV (72.7%, p = 0.012), recipients under respiratory support (69.2%, p = 0.031), and ABO-incompatible transplants (80%, p = 0.05). FHF, primary cancer, and female gender were the only variables that had a significant association with mortality in the logistic regression analysis. A higher incidence of prolonged respiratory support, bacterial and fungal infections, pneumonia, and chronic rejection contributed to the lower outcome observed in females. These results stress the need for continuous evaluation of the selection criteria of candidates for OLT suffering from primary cancer and FHF. The impact of recipient gender on mortality warrants further analysis but suggests that in the future more attention must be paid to the influence of this factor on the final outcome of OLT.

Adult↗
PubMed1999

Mouse autosomal trisomy: two's company, three's a crowd.

Autosomal trisomy causes a large proportion of all human pregnancy loss and so is a significant source of lethality in the human population. The autosomal trisomy syndromes each have a different phenotype and are probably caused by the effects of specific genes that are present in three copies, rather than the normal two. Identifying these genes will require the application of classical genetic and new genome-manipulation approaches. Recent advances in chromosome engineering are now allowing us to create precisely defined autosomal trisomies in the mouse, and so provide new routes to identifying the critical, dosage-sensitive genes that are responsible for these highly deleterious, yet very common, syndromes.

Abortion, Spontaneous↗
PubMed1999

Transchromosomal mouse embryonic stem cell lines and chimeric mice that contain freely segregating segments of human chromosome 21.

At least 8% of all human conceptions have major chromosome abnormalities and the frequency of chromosomal syndromes in newborns is >0.5%. Despite these disorders making a large contribution to human morbidity and mortality, we have little understanding of their aetiology and little molecular data on the importance of gene dosage to mammalian cells. Trisomy 21, which results in Down syndrome (DS), is the most frequent aneuploidy in humans (1 in 600 live births, up to 1 in 150 pregnancies world-wide) and is the most common known genetic cause of mental retardation. To investigate the molecular genetics of DS, we report here the creation of mice that carry different human chromosome 21 (Hsa21) fragments as a freely segregating extra chromosome. To produce these 'transchromosomal' animals, we placed a selectable marker into Hsa21 and transferred the chromosome from a human somatic cell line into mouse embryonic stem (ES) cells using irradiation microcell-mediated chromosome transfer (XMMCT). 'Transchromosomal' ES cells containing different Hsa21 regions ranging in size from approximately 50 to approximately 0.2 Mb have been used to create chimeric mice. These mice maintain Hsa21 sequences and express Hsa21 genes in multiple tissues. This novel use of the XMMCT protocol is applicable to investigations requiring the transfer of large chromosomal regions into ES or other cells and, in particular, the modelling of DS and other human aneuploidy syndromes.

Abnormalities, Multiple↗
PubMed1998

Crystallization and preliminary X-ray studies of Pseudomonas putida histidine ammonium-lyase.

Histidine ammonium-lyase from P. putida was expressed in Escherichia coli, purified to homogeneity, and crystallized by the vapour-diffusion method using polyethylene glycol 3350 as the precipitant. The crystals, which diffract to at least 2.5 A resolution, exhibit the symmetry of space group P212121, with unit-cell parameters a = 89.7, b = 138.2 and c = 164.8 A. The asymmetric unit contains a tetramer, and the crystals have a Vm value of 2.41 A3 Da-1.

Bacterial Proteins↗
PubMed1998

Three-dimensional vascular imaging using Doppler ultrasound.

BACKGROUND: We have evaluated the efficacy of using three-dimensional reconstruction of amplitude Doppler imaging data to quantitatively assess carotid artery bifurcation stenoses. METHODS: Sixty-four consecutive frames of amplitude (power) Doppler images are stored to be reassembled into a three-dimensional image representing the patent lumen. These images can then be rotated by any angle necessary to clearly view the vascular anatomy and to make quantitative ultrasound caliper measurements of the stenotic lumen and normal vessel caliber. RESULTS: Three-dimensional Doppler images accurately classified 53 of 61 vessels (87%) into categories of stenosis compared with angiography. All stenoses with >60% diameter reduction were detected and classified as such, for a sensitivity of 100%. CONCLUSIONS: Three-dimensional vascular imaging based on amplitude (power) Doppler data provides an accurate noninvasive technique for quantitative diagnosis of carotid bifurcation atherosclerotic disease, with selectable viewing projections that eliminate vessel overlap and other artifacts, and complements the hemodynamic data already available with two-dimensional duplex ultrasound.

Aged↗
PubMed1996

Down syndrome genetics: unravelling a multifactorial disorder.

Down syndrome is a common disorder affecting many tissues both during development and later on in adult life; the principle feature of all cases is a specific form of mental retardation, which is combined with a range of variable traits. Down syndrome is an aneuploidy syndrome that is caused by trisomy for human chromosome 21. While the phenotype is most likely due to a subtle increase in gene dosage of only a small minority of the estimated 500-800 genes that are present on this chromosome, the molecular genetics of Down syndrome remains speculative. However, recent advances on a number of fronts, including chromosome studies, gene identification and mouse modelling, are giving us the tools to dissect this multifactorial gene dosage disorder.

Animals↗
PubMed1996

Establishment of an in vitro assay to characterize hepatitis C virus NS3-4A protease trans-processing activity.

An in vitro cleavage system was established to measure HCV NS3 protease trans-processing activity. This system utilizes purified NS3-4A protein from baculovirus, purified substrates expressed by in vitro transcription and translation and defined buffer components. The 41-residue substrates, 5A/5B and 4A/4B, were processed efficiently in trans by wild-type NS3 but not by a catalytically inactive mutant protease; radiolabel sequencing confirmed that NS3-mediated cleavage occurred at the correct cysteine/serine sites, thereby authenticating this system. Two striking features of this in vitro assay are: (1) analogous 4B/5A and 3/4A substrates cannot be processed in trans under the same conditions, and (2) in vitro cleavage of the 5A/5B and 4A/4B sites is highly dependent on the presence of NS4A, which we show is not the case in vivo.

Amino Acid Sequence↗
PubMed1996

HLA-DR class II and ICAM-1 expression on tubular cells taken by fine-needle aspiration biopsy in renal allograft dysfunction.

BACKGROUND: Percutaneous biopsy is the method of choice for differential diagnosis of renal allograft dysfunction, although it is not risk-free. The use of less aggressive methods for diagnosis should limit the need for percutaneous biopsy to some specific situations. METHODS: We analysed 42 fine-needle aspiration biopsies from 36 kidney allograft recipients immunosuppressed with quadruple sequential therapy who suffered renal allograft dysfunction. Seven cases with stable renal function were used as controls and included as non-rejection cases in the analysis. In all aspirates the Corrected Increment was calculated and an immunocytochemical analysis of renal tubular cells with the monoclonal antibodies HLA-DR and ICAM-1 was performed. RESULTS: The Corrected Increment was increased in 13 out of 18 acute rejection cases and in one out of 31 non-rejection cases. HLA-DR expression was found in more than 30% of tubular cells from the aspirates in 16 out of 18 acute rejection cases and in eight out of 31 cases without rejection (P < 0.001). ICAM-1 expression was detected in more than 30% of tubular cells in 14 out of 18 cases with acute rejection, and in four out of 31 cases without acute rejection (P < 0.001). Interestingly, all acute vascular rejection cases (n = 6), and six out of 12 acute cellular rejection cases expressed both, HLA-DR and ICAM-1, in more than 30% of tubular cells. On the other hand, none of the non-rejection allograft dysfunctions or control samples showed more than 30% of tubular cells immunostained with both HLA-DR and ICAM-1 antibodies. CONCLUSIONS: The immunocytochemical analysis of HLA-DR and ICAM-1 on renal tubular cells taken by fine-needle aspiration biopsy, allows the diagnosis of acute cellular rejection and acute vascular rejection even when the Corrected Increment is not increased. Moreover, the risk of a core renal biopsy can be avoided when both tests are negative since an acute rejection is a remote possibility.

Acute Disease↗
PubMed1995

Mapping TNNC1, the gene that encodes cardiac troponin I in the human and the mouse.

We have mapped the TNNC1 gene, whose protein product is the cardiac TnI protein. TnI is one of the proteins that makes up the troponin complex, which mediates the response of muscle to calcium ions. The human TNNC1 locus had been assigned to a large region of chromosome 19, and we have refined the mapping position to the distal end of the chromosome by amplification of DNAs from a chromosome 19 mapping panel. We have also mapped the mouse Tnnc1 locus, by following the segregation of an intron sequence through DNAs from the European Interspecific Backcross. Tnnc1 maps close to the centromere on mouse chromosome 7.

Animals↗
PubMed1995

Highly polymorphic sequence at D21S1448 mapping close to D21S55, within the Down syndrome critical region.

We have isolated a highly polymorphic sequence from the Down syndrome critical region on human chromosome 21. This is a particularly useful marker because it lies adjacent to the locus D21S55, which is most closely associated with the major defects on Down syndrome. Other than this marker, few other variable sequences are known in this region (including other restriction fragment length polymorphisms or CA repeats) and therefore D21S1448 will be extremely helpful not only for people studying the inheritance of portions of chromosome 21 with respect to Down syndrome, but also for those carrying out linkage analysis of the chromosome.

Alleles↗
PubMed1995

In vitro autonomous proliferation in ANLL: clinical and biological significance.

In acute non-lymphoblastic leukemia (ANLL) progenitor cells frequently display a certain degree of autonomous growth. The aim of the present work was to analyze the autonomous proliferative capacity of leukemic progenitors in both de novo and secondary to myeloproliferative disorders (MPD) and myelodysplastic syndromes (MDS), acute myeloid leukemias and to correlate with clinical and biological characteristics of the disease. Clonogenic assays with and without leukocyte conditioned medium with PHA (LCM-PHA) were performed and the autonomous proliferation index (API) calculated in a series of 50 patients (34 de novo ANLL, eight secondary to MPD and eight secondary to MDS). Patients were divided into two groups according to their API, low (< or = 0.4) or high (> 0.4). Autonomous growth was observed in 84% of cases studied (82% in de novo ANLL, 75% secondary to MDS and 100% secondary to MPD). The group with the highest API (29 patients) had increased levels of hemoglobin (P = 0.006) and platelets (P = 0.01). A high API was also associated with an immature phenotype of blast cells (P = 0.02). Upon analyzing the de novo ANLL separately we observed that a high API correlated with high Hb values (P = 0.02), a lower rate of complete remission (42% vs 61%) and a lower survival rate (medium of 3 vs 10 months). These findings suggest that the capacity for autonomous proliferation can condition the clinical and biological profile of the disease.

Adult↗
PubMed1994

Mapping the gene that encodes phosphatidylinositol-specific phospholipase C-gamma 2 in the human and the mouse.

We have mapped the PLCG2 gene, which encodes the enzyme phosphatidyl inositol-specific phospholipase C-gamma 2. This is one of the phospholipases responsible for catalyzing the hydrolysis of phosphatidyl inositol in response to a great many mitogenic stimuli. PL C-gamma 2 is an essential component of the signal transduction pathway between tyrosine kinases and downstream events such as protein kinase C activation and intracellular calcium release. We assigned PLCG2 to human chromosome 16 by amplification within a somatic cell hybrid mapping panel. To position the locus at a much finer resolution, PLCG2 sequences were amplified from a chromosome 16-specific somatic cell hybrid panel, which placed the gene on the long arm of the chromosome in band 16q24.1, a region that has few known genes. We have hybridized a mouse Plcg2 open reading frame probe to mouse DNAs from the European Interspecific Backcross. The segregation pattern reveals the mouse Plcg2 locus maps to distal chromosome 8.

Animals↗
PubMed1994

Ser-143 is an essential active site residue in histidine ammonia-lyase of Pseudomonas putida.

Site directed mutagenesis was used to investigate the role of Ser-143 in enzyme activity and as a point for attack by cyanide or L-cysteine, two irreversible inhibitors of histidine ammonia-lyase (histidase). Two mutant proteins, a S143A substitution and an A142S-S143A transposition, were made. Both mutant histidases completely lost all enzymatic activity. Western blots with anti-histidase antibodies revealed that the mutant proteins were being expressed at a level equal to that of the wild-type protein. The purified mutant proteins could not incorporate [14C]cyanide nor could they generate the UV-absorbing species normally observed when L-cysteine modifies wild-type histidase. These results support the hypothesis that Ser-143 is the binding site for an as yet unidentified histidase cofactor.

Amino Acid Sequence↗
PubMed1994

Adynamic bone disease with negative aluminium staining in predialysis patients: prevalence and evolution after maintenance dialysis.

Aplastic bone disease (ABD) is a common form of renal osteodystrophy and is characterized by a defect in bone matrix formation and mineralization without an increase in osteoid thickness. The prevalence and pathogenesis of ABD in predialysis patients is largely unknown. We prospectively studied 92 unselected predialysis patients with a creatinine clearance < 10 ml/min/1.73 m2 and a mean age of 45 +/- 2 years (61 M, 31 F). None of the study patients had received any form of vitamin D therapy, and CaCO3 was the primary phosphate binder. Aplastic bone disease was observed in 30 (32%) patients. Stainable bone aluminium surface was < 3% in all ABD patients. Patients with ABD were older (52 +/- 3 versus 42 +/- 2 years; P < 0.01) and had reduced serum intact PTH compared to non-ABD patients (199 +/- 25 versus 561 +/- 87 pg/ml; P < 0.001). Patients with diabetes mellitus showed lower PTH values (179 +/- 31 versus 432 +/- 62 pg/ml; P < 0.001) and a lower incidence of advanced hyperparathyroidism bone lesions (16% versus 46%; P < 0.05) than non-diabetic patients. However, diabetes was not clearly associated with low bone turnover disease (56% in diabetics versus 41% in non-diabetics; P = 0.1). A second bone biopsy was obtained in eleven ABD patients after a period of 16.6 +/- 2.2 months on maintenance dialysis with a dialysate calcium of 7 mg/dl. Bone histology was unchanged in 10 patients, and one evolved to mild hyperparathyroidism. Trabecular bone volume did not change (22.7 +/- 1.7 versus 20.7 +/- 1.7%), and the stainable bone aluminium surface remained < 3%.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗