RESUMEN
BACKGROUND: An incomplete separation of disialotransferrin (CDT) and trisialotransferrin (a non-CDT isoform) may cause false-positive CDT results in alcohol abuse testing. We describe a currently unknown disialotransferrin-trisialotransferrin-bridging phenomenon (di-tri-bridge) appearing with high prevalence in serum from liver cirrhosis patients. METHODS: Twenty one consecutive serum samples with a di-tri-bridge encountered in routine CDT HPLC (Clin-Rep(R)-CDT-on-line, Recipe) were investigated by a candidate reference CDT HPLC method, by capillary electrophoresis (Capillarys-CDT, Sebia) and by transferrin genotyping. Patients clinical background was assessed by telephone interview. RESULTS: Out of 21 consecutive serum samples showing a di-tri-bridge (and increased trisialotransferrin fractions) in HPLC as well as in CE analysis, 19 were from patients with a liver cirrhosis history. Genotyping (where applicable by the availability of DNA: n=12) yielded most frequently homozygous transferrin C1 (6x), proving that the di-tri-bridge cannot be explained by genetic transferrin variants in these samples. Other genotypes found were C2 (1x), C1C2 (4x), C1C3 (1x). CONCLUSION: The frequently seen di-tri-bridging phenomenon in transferrin HPLC analysis for patients with liver cirrhosis is not explained by genetic transferrin variants or by an increased trisialotransferrin fraction. Although further studies are needed to assess the relationship between this phenomenon and liver cirrhosis, our observation could be helpful in development of a biomarker for liver cirrhosis.
Asunto(s)
Cirrosis Hepática/sangre , Cirrosis Hepática/genética , Transferrina/metabolismo , Cromatografía Líquida de Alta Presión , Electroforesis Capilar , Femenino , Genotipo , Humanos , Masculino , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Transferrina/genéticaRESUMEN
BACKGROUND: alpha(1)-Antitrypsin (alpha(1)AT) deficiency predisposes individuals to chronic obstructive pulmonary disease (COPD) and/or liver disease. Phenotyping of the protein by isoelectric focusing is often used to characterize alpha(1)AT deficiency, but this method may lead to misdiagnosis (e.g., by missing null alleles). We evaluated a workup that included direct sequencing of the relevant parts of the gene encoding alpha(1)AT, SERPINA1 [serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 1], for patients with alpha(1)AT concentrations < or =1.0 g/L. METHODS: During a 5-year period, we identified 66 patients with alpha(1)AT concentrations < or =1.0 g/L and amplified and sequenced exons 2, 3, and 5 of the alpha(1)AT gene in these patients. To ensure that no relevant genotypes were missed, we sequenced the same exons in 48 individuals with alpha(1)AT concentrations between 1.0 and 1.5 g/L. RESULTS: Sequence analysis revealed 18 patients with combinations of disease-associated alpha(1)AT alleles: 8 homozygous for the deficient Z allele and 10 compound heterozygotes for various deficient or null alleles. We identified and named 2 new null alleles, Q0(soest) (Thr(102)-->delA, which produces a TGA stop signal at codon 112) and Q0(amersfoort) (Tyr(160)-->stop). No relevant disease-associated allele combinations were missed at a 1.0-g/L threshold. CONCLUSIONS: Up to 22% of the alleles in disease-associated alpha(1)AT allele combinations may be missed by conventional methods. Genotyping by direct sequencing of samples from patients with alpha(1)AT concentrations < or =1.0 g/L detected these alleles and identified 2 new null alleles.
Asunto(s)
Enfermedad Pulmonar Obstructiva Crónica/genética , Deficiencia de alfa 1-Antitripsina/genética , alfa 1-Antitripsina/genética , Alelos , Niño , Preescolar , Femenino , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Mutación , Linaje , Análisis de Secuencia de ProteínaRESUMEN
We assessed whether large-scale expression profiling of leukocytes of patients with essential hypertension reflects characteristics of systemic disease and whether such changes are responsive to antihypertensive therapy. Total RNA from leukocytes were obtained from untreated (n=6) and treated (n=6) hypertensive patients without apparent end-organ damage and from normotensive controls (n=9). RNA was reverse-transcribed and labeled and gene expression analyzed using a 19-K oligonucleotide microarray using dye swaps. Samples of untreated and of treated patients were pooled for each sex and compared with age- and sex-matched controls. In untreated patients, 680 genes were differentially regulated (314 up and 366 down). In the treated patients, these changes were virtually absent (4 genes up, 3 genes down). A myriad of changes was observed in pathways involved in inflammation. Inflammation-dampening interleukin receptors were decreased in expression. Intriguingly, inhibitors of cytokine signaling (the PIAS family of proteins) were differentially expressed. The expression of several genes that are involved in regulation of blood pressure were also differentially expressed: angiotensin II type 1 receptor, ANP-A receptor, endothelin-2, and 3 of the serotonin receptors were increased, whereas endothelin-converting enzyme-1 was decreased. Strikingly, virtually no changes in gene expression could be detected in hypertensive patients who had become normotensive with treatment. This observation substantiates the long-standing idea that hypertension is associated with a complex systemic response involving inflammation-related genes. Furthermore, leukocytes display differential gene expression that is of importance in blood pressure control. Importantly, treatment of blood pressure to normal values can virtually correct such disturbances.