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Biomedical subjects

J Caro

Publications and source records attributed to J Caro.

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

Fourth branchial arch sinus: clinical presentation, diagnostic workup, and surgical treatment.

OBJECTIVES/HYPOTHESIS: Abnormalities of the fourth branchial arch are much less common than those of the second arch and present in a different manner. The authors report their experience with five cases of fourth arch sinuses. METHODS: Retrospective chart review of patients at a tertiary care center. RESULTS: All patients presented in the first or second decade of life, and all but one had left-sided neck involvement. Four patients presented with recurrent low neck inflammatory episodes, and one with respiratory compromise. Diagnostic studies performed included ultrasound, computed tomography (CT) scan, barium swallow, magnetic resonance imaging, and fine-needle aspiration. Barium contrast studies and CT scan were the most useful in demonstrating a fourth arch sinus tract preoperatively. Surgical treatment with an emphasis on complete exposure of the recurrent laryngeal nerve and exposure of the lateral piriform sinus to facilitate complete sinus tract excision was successful in all patients. CONCLUSION: The clinical history of recurrent low neck inflammatory episodes in young patients, especially on the left side, should raise the suspicion of this entity. Investigation using barium swallow in combination with CT scanning is useful. Excision of the sinus tract taking care not to injure the recurrent laryngeal nerve is curative.

Adolescent↗
PubMed1998

Hypoxia-inducible factor 1alpha (HIF-1alpha) is a non-heme iron protein. Implications for oxygen sensing.

The hypoxia-inducible factor 1 complex (HIF-1) is involved in the transcriptional activation of several genes, like erythropoietin and vascular endothelial growth factor, that are responsive to the lack of oxygen. The HIF-1 complex is composed of two b-HLH proteins: HIF-1beta that is constitutively expressed, and HIF-1alpha, that is present only in hypoxic cells. The HIF-1alpha subunit is continuously synthesized and degraded by the ubiquitin-proteasome under oxic conditions. Hypoxia, transition metals, iron chelators, and several antioxidants stabilize the HIF-1alpha protein, allowing the formation of the transcriptionally active HIF-1 complex. The mechanisms of oxygen sensing and the pathways leading to HIF-1alpha stabilization are unclear. Because the involvement of a heme protein oxygen sensor has been postulated, we tested the heme sensor hypothesis by using a luciferase-expressing cell line (B-1), that is highly responsive to hypoxia. Exposure of B-1 cells to carbon monoxide and heme synthesis inhibitors failed to show any effect on the hypoxia responsiveness of these cells, suggesting that heme proteins are not involved in hypoxia sensing. Measurement of iron in recombinantly expressed HIF-1alpha protein revealed that this protein binds iron in vivo. Iron binding was localized to a 129-amino acid peptide between sequences 529 and 658 of the HIF-1alpha protein. Although the exact structure of the iron center has not been yet defined, a 2:1 metal/protein molar ratio suggests a di-iron center, probably similar to the one found in hemerythrin. This finding is compatible with a model where redox reaction may occur directly in the iron center of the HIF-1alpha subunit, affecting its survival in oxic conditions.

Carbon Monoxide↗
PubMed1997

The West of Scotland coronary prevention study: economic benefit analysis of primary prevention with pravastatin.

OBJECTIVE: To estimate the economic efficiency of using pravastatin to prevent the transition from health to cardiovascular disease in men with hypercholesterolaemia. DESIGN: Economic benefit analysis based on data from the West of Scotland coronary prevention study. Treatment specific hazards of developing cardiovascular disease according to various definitions were estimated. Scottish record linkage data provided disease specific survival. Cost estimates were based on extracontractual tariffs and event specific average lengths of stay calculated from the West of Scotland coronary prevention study. SUBJECTS: Men with hypercholesterolaemia similar to the subjects in the West of Scotland coronary prevention study. MAIN OUTCOME: Cost consequences, the number of transitions from health to cardiovascular disease prevented, the number needed to start treatment, and cost per life year gained. RESULTS: If 10,000 of these men started taking pravastatin, 318 of them would not make the transition from health to cardiovascular disease (number needed to treat, 31.4), at a net discounted cost of 20m Pounds over 5 years. These benefits imply an undiscounted gain of 2,460 years of life, and thus 8121 Pounds per life year gained, or 20,375 Pounds per life year gained if benefits are discounted. Restriction to the 40% of men at highest risk reduces the number needed to treat to 22.5 (5601 Pounds per life year gained (undiscounted) and 13,995 Pounds per life year gained (discounted)). CONCLUSIONS: In subjects without evidence of prior myocardial infarction but who have hypercholesterolaemia, the use of pravastatin yields substantial health benefits at a cost that is not prohibitive overall and can be quite efficient in selected high risk subgroups.

Anticholesteremic Agents↗
PubMed1997

Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes.

The hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) is involved in the activation of the erythropoietin and several other hypoxia-responsive genes. The HIF-1 complex is composed of two protein subunits: HIF-1beta/ARNT (aryl hydrocarbon receptor nuclear translocator), which is constitutively expressed, and HIF-1alpha, which is not present in normal cells but induced under hypoxic conditions. The HIF-1alpha subunit is continuously synthesized and degraded under normoxic conditions, while it accumulates rapidly following exposure to low oxygen tensions. The involvement of the ubiquitin-proteasome system in the proteolytic destruction of HIF-1 in normoxia was studied by the use of specific inhibitors of the proteasome system. Lactacystin and MG-132 were found to protect the degradation of the HIF-1 complex in cells transferred from hypoxia to normoxia. The same inhibitors were able to induce HIF-1 complex formation when added to normoxic cells. Final confirmation of the involvement of the ubiquitin-proteasome system in the regulated degradation of HIF-1alpha was obtained by the use of ts20TGR cells, which contain a temperature-sensitive mutant of E1, the ubiquitin-activating enzyme. Exposure of ts20 cells, under normoxic conditions, to the non-permissive temperature induced a rapid and progressive accumulation of HIF-1. The effect of proteasome inhibitors on the normoxic induction of HIF-1 binding activity was mimicked by the thiol reducing agent N-(2-mercaptopropionyl)-glycine and by the oxygen radical scavenger 2-acetamidoacrylic acid. Furthermore, N-(2-mercaptopropionyl)-glycine induced gene expression as measured by the stimulation of a HIF-1-luciferase expression vector and by the induction of erythropoietin mRNA in normoxic Hep 3B cells. These last findings strongly suggest that the hypoxia induced changes in HIF-1alpha stability and subsequent gene activation are mediated by redox-induced changes.

Cell Line↗
PubMed1997

Costs and effectiveness of extracorporeal gallbladder stone shock wave lithotripsy versus laparoscopic cholecystectomy. A randomized clinical trial. McGill Gallstone Treatment Group.

Thirty-five patients were randomized to extracorporeal shock-wave lithotripsy (ESWL) and 25 to laparoscopic cholecystectomy (LC). Stone disappearance occurred in only 12 of 32 ESWL patients [38% (95% CI: 21-56%)] during a 15-month follow-up. Greater incremental gains in quality of life after 6 months were observed among LC patients (p < .01). Total duration of disability was 6.8 +/- 8.5 days for ESWL, and 22.7 +/- 16.6 days for LC (p < .01). Nine (28%) patients crossed over electively to the LC group, but only 44% of these underwent LC within the next 3 years. ESWL cost Can $58.9/ day of disability saved. ESWL is limited by its selective applicability and modest stone disappearance rate. Its cost-effectiveness is largely dependent on patient acceptance of recurrent episodes of biliary colic due to the persistence of stone fragments.

Adolescent↗
PubMed1997

Hypoxia stimulates human preproendothelin-1 promoter activity in transgenic mice.

Significant elevations in endothelin (ET)-1 levels accompany many diseases, but the underlying regulatory mechanisms are unclear. To investigate the in vivo regulation of human preproendothelin-1 (PPET-1), we examined the activity of the PPET-1 promoter in transgenic mice exposed to hypoxia. Mice expressing one of three PPET-1 promoter-luciferase (PPET-1/LUC) reporter transgenes (approximately 2.5 kb, 138 bp, or none of the 5'-flanking sequences of the PPET-1 gene) were generated. LUC expression was reduced in mice with a truncated 138-bp PPET-1 promoter. Exposure of mice bearing the 2.5-kb PPET-1/LUC transgene to hypoxia (10% O2 for 24 h) increased LUC expression sixfold in pulmonary tissue but only twofold in other tissues. In situ hybridization revealed the strongest transgene expression in the pulmonary vasculature and bronchiolar epithelium. These data are consistent with the hypothesis that hypoxic induction of the PPET-1 gene leads to increased pulmonary production of ET-1 in diseases associated with low O2 tension.

Animals↗
PubMed1997

Strategies for the management of hypercholesterolaemia: a systematic review of the cost-effectiveness literature.

OBJECTIVE: To review research addressing the management of cholesterol in the prevention of coronary heart disease in order to assess the cost-effectiveness of such interventions. METHODS: A systematic review of economic evaluations identified through searches of MEDLINE and the Social Sciences Citation Index revealed 38 studies addressing the cost-effectiveness of cholesterol management. They were distinguished according to screening approaches, dietary advice and drug treatment. Most studies were not associated directly with clinical trial results, but adopted economic modelling approaches. RESULTS: Whilst there is general agreement among the majority of analyses, studies of cholesterol management concerned with screening strategies were extremely sensitive to changes in their assumptions; so much so that only a limited emphasis may be placed on specific cost-effectiveness ratios and the conclusions drawn from them. All studies considered direct costs, though many were limited to drug costs. The cost-effectiveness of primary prevention by cholesterol-lowering drugs is highly variable, depending on age at initiation of treatment and cardiovascular risk profile. Pharmacological intervention is least cost-effective in the young and the elderly. The cost-effectiveness of cholesterol-reducing agents improves when they are targeted at those at high risk. HMG-CoA reductase inhibitors are generally more effective and more cost-effective at reducing cholesterol-related coronary events than other medications. CONCLUSION: The methods and economic data upon which these studies are based need to be improved if robust policy conclusions are to be formulated.

Anticholesteremic Agents↗
PubMed1996

Absolute requirement of aryl hydrocarbon receptor nuclear translocator protein for gene activation by hypoxia.

Hypoxia-inducible factor 1 (HIF-1) is a DNA-binding heterodimeric protein complex originally described in the transcriptional activation of the erythropoietin gene by hypoxia. This protein complex is composed of two subunits, HIF-1alpha and -1beta (aryl hydrocarbon receptor nuclear translocator, ARNT). In this study, we used ARNT-deficient cells, derived from the mouse hepatoma cell line Hepa1c1c7, to further characterize HIF-1 complex formation and its relationship with gene activation by hypoxia and desferrioxamine (Df). Gel shift assays revealed that ARNT is absolutely required for the formation of the HIF-1 DNA-binding complex. Results from RNase protection assays and Northern blots showed that the lack of functional HIF-1 complex completely abrogated the response to hypoxia of vascular endothelial growth factor (VEGF) and the glycolytic enzymes aldolase A (ALDA) and phosphoglycerate kinase 1 (PGK-1), genes known to be upregulated by low oxygen tension. Desferrioxamine induction of VEGF and PGK-1 genes was reduced in the ARNT-deficient cells, but at difference with hypoxia, it was not completely suppressed. These results suggest that Df is able to activate gene transcription through HIF-1-independent mechanisms. Exposure to hypoxia or Df did not induce any changes in HIF-1alpha and -1beta mRNA levels, suggesting that posttranscriptional mechanisms are involved in HIF-1 complex activation.

Actins↗
PubMed1994

Hypoxia regulatory elements of the human vascular endothelial growth factor gene.

Vascular endothelial growth factor (VEGF) expression is highly stimulated by hypoxia, both in vitro and in vivo. Recent findings suggest that the VEGF gene utilizes an oxygen sensing mechanism similar to the one used by the erythropoietin gene. The genomic sequences that control the VEGF response to hypoxia are, however, largely unknown. In utilizing transient transfection assays in HeLa cells we determined that hypoxia/cobalt responsive enhancer elements are present at the 5' and 3' flanking regions of the human VEGF gene. The 3' enhancer is contained in a 160 bp fragment located about 60 bp downstream of the polyadenylation site. It contains a sequence stretch of about 12 bp which are highly homologous to sequences in the erythropoietin hypoxia-responsive enhancer. The 5' flanking enhancer is contained in a 100 bp fragment located about 800 bp upstream of the start site. This fragment does not contain significant homologies with the erythropoietin enhancer. Thus, it appears that the response to hypoxia of the VEGF gene is controlled by two regulatory elements; one which may be related to the erythropoietin enhancer and a second, which appears to be a completely unrelated sequence.

Base Sequence↗
PubMed1994

Hypoxic stimulation of vascular endothelial growth factor expression in vitro and in vivo.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a specific endothelial cell mitogen with potent angiogenic properties. In tumors, VEGF has been localized to the most necrotic and ischemic areas of the tissues, suggesting that local hypoxia is a potent inducer of VEGF production. Initial experiments in vitro confirmed the stimulatory effect of hypoxia on VEGF expression. The extent of this response and the mechanisms involved in oxygen sensing are poorly characterized. EXPERIMENTAL DESIGN: Confluent monolayers of malignant cell lines or primary cultures of fibroblast or endothelial cells were exposed to hypoxia or incubated with either cobalt chloride, a stimulator of erythropoietin gene expression, or sodium azide, an inhibitor of oxydative phosphorylation. VEGF expression was analyzed by Northern blot or RNase protection assays. The expression VEGF in vivo was studied in animals subjected to hypobaric hypoxia or functional anemia. RESULTS: Hypoxia greatly stimulated VEGF expression in tumor cell lines and primary fibroblast cultures. Endothelial cells, that expressed very low constitutive levels of VEGF, were resistant to hypoxic stimulation. RNase protection analysis showed that hypoxia primarily stimulated the induction of smaller and medium VEGF isoforms, i.e., the same ones expressed under normal conditions. The stimulatory effect of hypoxia on VEGF could be reproduced in vitro by cobalt chloride but not with sodium azide. In vivo, both hypoxia and anemia were found to be potent inducers of VEGF expression in several organs including heart, brain, liver, kidney, and muscle. As in vitro, cobalt was also found to be a potent stimulator of VEGF in vivo. CONCLUSIONS: Hypoxia is a potent inducer of VEGF expression in malignant as well as normal cultured cells. It is also a stimulator of VEGF expression in vivo. The VEGF gene appears to respond to hypoxia like the erythropoietin gene, and the mechanism of oxygen sensing probably is mediated by a heme-containing protein.

Anemia↗
PubMed1993

Characterization of hypoxia-responsive enhancer in the human erythropoietin gene shows presence of hypoxia-inducible 120-Kd nuclear DNA-binding protein in erythropoietin-producing and nonproducing cells.

Erythropoietin (Epo) production in response to hypoxia or cobalt is primarily mediated by activation of transcription of the Epo gene. Recently an hypoxia responsive enhancer was identified in the 3' flanking region of the mouse and human Epo genes. Using functional analysis in Hep 3B cells we define here the minimal enhancer element as a 29-bp segment starting at the Apa1 site in the 3' flanking region of the human Epo gene. Mutagenesis studies of the minimal element identified three different areas that are necessary for full enhancer activity. Electrophoretic mobility shift assays show the presence of hypoxia- and/or cobalt-inducible nuclear DNA-binding proteins that bind to one of the active sites of the enhancer. Induction of hypoxia-binding activity was abolished by Anisomycin, a potent protein synthesis inhibitor, suggesting that de novo protein synthesis is necessary for the activation process. Further characterization of DNA-binding proteins by use of UV light crosslinking identified a protein of molecular weight of approximately 120-Kd that was present only in hypoxic extracts. This protein was found to be present in hypoxic nuclear extracts from both Epo-producing and non-Epo-producing cells, suggesting that it may be involved in a more generalized mechanism of cellular response to hypoxia.

Base Sequence↗
PubMed1993

Impaired response of polycythemic mice to erythropoietin induced by protein starvation imposed after hormone administration.

The present study was performed to determine the stage of the erythropoietic pathway which is affected by starvation or protein deprivation and whose manifestation is a depressed response to exogenous erythropoietin (EPO). The response to recombinant human EPO was measured in post-hypoxic polycythemic mice by determination of 59Fe uptake into red cells, spleen and femur and/or erythroid colony forming units (CFU-E) and erythroid precursor cell concentrations in femoral marrow. Experimental mice were either starved or fed one of seven different diets whose protein (casein) content ranged from 0 to 20%. All diets were isocaloric. The response of mice maintained on the standard diet (Purina Lab chow) was taken as the normal one. Starvation during the 48-hour period immediately before EPO injection had no effect on the response to the hormone. Starvation, and protein deprivation to a lesser extent, during the 48-hour period following EPO, on the other hand, significantly reduced the response. There was a progressive increase in the response as the casein content of the diet was increased. A normal response was observed when dietary casein concentration was 10%. These findings indicate that nutritional deprivation or dietary protein alterations during the period immediately following EPO injection in polycythemic mice can have detrimental effects on the erythroid response in a model in which nutritional deprivation was relatively short and acute. They also indicate that the subnormal response is not due to a decreased size of the erythroid progenitor pool available for differentiation but to deficient rates of differentiation of erythropoietic units.

Animals↗