Influence of surgical complications on kidney graft survival in recipients of simultaneous pancreas kidney transplantation.

PURPOSE: Simultaneous pancreas-kidney transplantation is the gold standard treatment for patients with end-stage renal failure secondary to insulin-dependent diabetes mellitus. This kind of transplantation is a complex operation associated with a high incidence of surgical complications and mortality risk which could influence graft survival. The aim of this study was to establish the influence of different grades of postoperative complications, classified according to Clavien-Dindo, on the rate of kidney graft loss. METHODS: We performed an observational retrospective review of all simultaneous transplantations performed between February 1989 and May 2012. Factors examined were related to recipient and donor characteristics, surgical procedures, and postoperative outcomes. For this purpose, Kaplan-Meier analyses and Cox-Regression tests are used. RESULTS: One hundred thirty-nine transplantations were performed. Complications grades I, II, and IIIa were experienced in 81 (58.3%) patients, and grades IIIb and IVa-b in 55 (39.6%). Multivariate analysis showed an influence of panel reactive antibody (hazard ratio [HR]: 10.79; P = .003), incidence of acute rejection (HR: 2.55; P = .03), and complications grouped into grades IIIb and IVa-b (HR: 3.63; P = .02). Kaplan Meier analysis showed worse kidney graft survival rate in groups grades IIIb and IVa-b compared to grades I, II, and IIIa (86.6% vs 98.7% at 1 year and 81.8% vs 97.3% at 5 years; P = .001). CONCLUSIONS: Despite being the gold standard treatment for these patients, pancreas and kidney transplantations have numerous complications which could influence the prognosis of graft kidney survival.

Identification of novel SLC3A1 gene mutations in Spanish cystinuria families and association with clinical phenotypes.

Cystinuria is an inherited metabolic disease characterized by an abnormal urinary excretion of cystine and dibasic amino acids, leading to kidney stone formation. Incidence of cystinuria in the Mediterranean Spanish population is one of the highest in the world. In view of the low prevalence of previously reported mutations in the SLC3A1 gene, analyses to identify novel variants were carried out on 20 cystinuria families. Additionally, we investigated the possible association between these molecular variants and clinical phenotypes. Genomic DNA from 48 cystinuria patients, 44 healthy relatives and 81 unrelated controls from the East Mediterranean coast of Spain was screened by conformation sensitive gel electrophoresis. Abnormal patterns were confirmed by nucleotide sequence determination and by further restriction fragment-length polymorphism. We only found 11 genetic variants within the SLC3A1 gene: five known polymorphisms (114C > A, 231T > A, 1136 + 3delT, 1332 + 7T > C and 1338G > A), four point mutations (M467T, R452W, I105R and Y461X), one single base pair deletion (1767delA) and one 2-bp insertion (1670insAT). Two of these genetic variants (I105R and 1670insAT) were described for the first time. All mutations but one were detected in families classified as Type I cystinuria due to the transmission pattern of the disease. Association analyses revealed that 231T > A (M467T), 1136 + 3delT and 1332 + 7T > C genetic variants were statistically related with urinary amino acid excretion in cystinuria patients. Although some molecular variants within the SLC3A1 gene were associated with clinical traits in cystinuria patients, the low detection rate of mutations in this gene strongly suggests that variation of the SLC3A1 is not the major genetic factor contributing to cystinuria in this Mediterranean population.

Evaluación de la salud oral en una población de drogodependientes / Evaluation of the oral heath in a drogodependientes population

Introducción: El objetivo del presente trabajo es analizar la situación de salud oral en una población adulta drogodependiente para instaurar un programa de educación sanitaria. Pacientes y métodos: Estudio descriptivo transversal. Se han evaluado un total de 53 sujetos comprendidos entre los 24 y 44 años de edad pertenecientes a un centro de ayuda al drogodependiente de la Comunidad de Madrid. La exploración se realizó de forma estandarizada en el propio centro, siguiendo los criterios de diagnostico y la ficha de soporte de la OMS. El análisis estadístico fue analizado mediante el sistema SPSS, registrando frecuencias, medias y desviación típica. Resultados: Los resultados obtenidos muestran un consumo de heroína en un 83 por ciento (n=44) de los sujetos, siendo portadores de VIH un 45,3 por ciento (n=24). La media del CAOD fue de 13.6 (+/-6.02) no encontrándose diferencias estadísticamente significativas entre los sujetos con VIH (CAOD=14.4+/-5.2) y sujetos sin VIIH (CAOD=13.03+/-6.62). El estado de la mucosa oral revela la candidiasis oral como la patología mas prevalente con un 13.2 por ciento (n=7) de los sujetos, siendo los chasquidos la patología de ATM más registrada (64.2 por ciento, n=34). Conclusiones: La alta prevalencia de caries en estos sujetos demuestran que son población de alto riesgo de caries siendo necesaria una estrecha colaboración entre los sectores sociales y sanitarios (AU)

[Sensitivity, specificity and predictive value of the genetic analysis of SLC3A1 gene variants used for the diagnosis of cystinuria among the spanish population]. / Sensibilidad, especificidad y valor predictivo del análisis genético de variantes en el gen SLC3A1 aplicado al diagnóstico de cistinuria en población española.

OBJECTIVE: To test the clinical usefulness of the analysis of point mutations R452W, M467T, 114C > A, 231T > A, 1136 + 3delT and 1332 + 7T > C in the gene SLC3A1 as well as their possible haplotypes used for the diagnosis of cystinuria in the mediterranean spanish population. MATERIAL AND METHODS: A total of 48 patients with cystinuria, 44 relatives without cystinuria, and 81 healthy controls were studied. A genetic analysis was conducted in order to identify variants in the gene SLC3A1. The sensitivity, specificity, and predictive value for each genetic variant and for the possible haplotypes were calculated. RESULTS: The specificity of mutations M467T, R452W, and 231T > A used for the diagnosis of cystinuria in the general population or for the different subtypes of cystinuria in involved families, was higher than 90%; nevertheless, none of the analysed variants reached a sensitivity higher than 80%. In the study of haplotypes, the highest sensitivity was obtained with the haplotype CTTT (83.8%); however, its specificity and predictive value were low (20.6% and 53.4%, respectively). CONCLUSIONS: The studied genetic variants did not show enough clinical usefulness.

Sensibilidad, especificidad y valor predictivo del análisis genético de variantes en el gen SLC3A1 aplicado al diagnóstico de cistinuria en población española / Sensitivity, specificity and predictive value of the genetic analysis of SLC3A1 gene variants used for the diagnosis of cystinuria among the spanish population

Objetivo. Estimar la validez clínica del análisis de las mutaciones puntuales R452W, M467T, 114C > A, 231T > A, 1136 + 3delT y 1332 + 7T > C en el gen SLC3A1, así como de sus posibles haplotipos aplicados al diagnóstico de cistinuria en población mediterránea española. Material y métodos. Se han estudiado 48 pacientes con cistinuria, 44 familiares sin cistinuria y 81 controles sanos. Se realizó un análisis genético para la identificación de variantes en el gen SLC3A1. Se calculó la sensibilidad, especificidad y valor predictivo para cada variante genética y para los posibles haplotipos. Resultados. La especificidad de las mutaciones M467T, R452W y 231T > A aplicadas al diagnóstico de cistinuria en población general o para los diferentes subtipos de cistinuria en familias afectadas fue superior al 90 por ciento; sin embargo, ninguna variante analizada alcanzó una sensibilidad superior al 80 por ciento. En el estudio de haplotipos, la sensibilidad más alta se obtuvo con el haplotipo CTTT (83,8 por ciento); sin embargo, su especificidad y valor predictivo fueron bajos (20,6 por ciento y 53,4 por ciento, respectivamente).Conclusiones. Las variantes genéticas estudiadas no mostraron suficiente validez clínica (AU)

Association between M467T and 114 C-->A variants within the SLC3A1 gene and some phenotypical traits in cystinuria patients from Spain.

Cystinuria is an inherited metabolic disease characterized by an abnormal urinary excretion of cystine and dibasic amino acids. Formation of renal calculi, recurrent infections and renal failure are the main complications of this disease. The SLC3A1 gene, which codes for a dibasic amino acid transporter protein, is involved in the pathogenesis of cystinuria. We investigated the possible association between molecular variants (M467T, E483X, T216 M and 114 C-->A) within the SLC3A1 gene and some phenotypical traits in a Spanish area. The study population consisted of 45 cystinuria patients, 42 cystinuria relatives and 81 healthy control subjects. Only the M467T mutation was found in chromosomes of cystinuria patients and relatives. However, the 114 C-->A polymorphism was detected in cystinuria patients, in relatives and in control subjects but with different prevalences. Moreover, a statistically significant association between this polymorphism and urinary amino acid levels was found in cystinuria patients (P<0.05). Subjects with the C/C genotype showed significantly higher urinary levels of cystine, arginine and their sum as compared with carriers of the A allele (P<0.05). When multiple linear regression analysis was performed in cystinuria patients, the 114 C-->A polymorphism remained significantly associated (P=0.047) with cystine levels even after controlling for age, gender and the M467T mutation. Furthermore, we also found a statistically significant interaction term (P=0.028) between M467T and 114 C-->A in determining urinary cystine levels. According to our results, the 114 C-->A polymorphism might be a marker of a functional variant in the SLC3A1 gene or in other genes related to urinary amino acid excretion in cystinuria patients.

[Genetic and molecular study of cystinuria in the Valencian community]. / Estudio genético y molecular de la cistinuria en la Comunidad Valenciana.

BACKGROUND: The aim of this study has been to know the prevalence of the four most common mutations reported in SLC3A1 gene involved in human cystinuria, and to estimate the association with different phenotypic manifestations of this disease in population of the Valencian Community, Spain. PATIENTS AND METHODS: We have carried out a cross-sectional study with a control group in 16 families with one or more members diagnosed as cystinuric patients. 149 subjects (38 cystinuric patients, 39 relatives and 72 controls) were studied. Genetic analyses were carried out using PCR (polymerase chain reaction), RFLPs (restriction fragment length polymorphisms) and sequencing of PCR products. Parametric and non parametric tests were applied in the statistical analyses. RESULTS: None of the four mutations studied were found in the control group. M467T mutation was the most prevalent in cystinuric patients as well as in relatives, showing and allelic frequency of 0.06 and 0.03 respectively. Regarding to the association analysis between genotype and phenotype, we found that, in cystinuric subjects, urinary excretion of some dibasic amino acid was higher in those who carried M467T mutation (p < 0.05). The formation of cystine crystals was also higher in cystinuric patients who carried this mutation (p < 0.05). CONCLUSIONS: Prevalences of recurrent mutations reported up to now in SLC3A1 gene are very low in cystinuric subjects of the Valencian Community, Spain. Only mutation M467T has been found, without differences between cystinuric and their relatives, although in the first ones was associated with a more accurate phenotype manifestation of the disease.

Genetic and phenotypic aspects of phenylalanine hydroxylase deficiency in Spain: molecular survey by regions.

We present an extensive study of the genetic diversity of phenylalanine hydroxylase deficiency in the Spanish phenylketonuria population. We have analysed 195 PKU patients by DGGE analysis identifying 67 different mutations which represent 89% of the total mutant chromosomes. Seventeen mutations first described in Spain have not yet been detected elsewhere; ten of these are reported here for the first time. The clinical significance of this high genetic heterogeneity was examined by analysing the genotype-phenotype correlations, mainly focusing on the mild hyperphenylalaninaemia (MHP) phenotype. The genotypes found in a group of 93 MHP patients, the largest analysed so far, are described in detail, as well as the relative frequencies of the MHP mutations identified. From the total pool of mutations, 27 can be considered severe, 18 can be defined as mild and 13 as associated with MHP. The prevalent mutations correspond to one severe mutation (IVS10nt-11), one MHP mutation (A403V) and two mild mutations (165T and V388M). The high frequency of mutations with a low degree of severity can explain the relatively higher prevalence of MHP and mild PKU phenotypes in Spain compared with NOrthern European populations. We have looked at the geographical distribution in Spain of the more common mutations, finding evidence of local mutation clustering, which could be the result of differences in the ethnic background and/or of genetic drift within each region.

Reference values of urinary excretion of cystine and dibasic aminoacids: classification of patients with cystinuria in the Valencian Community, Spain.

OBJECTIVE: Cystinuria is an autosomal-recessive disorder of the kidneys and small intestine affecting a luminal transport mechanism shared by cystine, ornithine, arginine, and lysine. Three different types of cystinuria can be distinguished according to the excretion of these amino acids in urine samples. We propose cutoff values from our population as references and we present a classification of cystinuric patients using quantitative amino acid chromatography in first morning urine samples. DESIGN AND METHODS: A random sample of forty healthy subjects belonging to general population of the Valencian Community were selected as control subjects. Cystine, lysine, arginine, and ornithine were quantified by reverse-phase HPLC. Seventy-two subjects, diagnosed previously as cystinuric by the cyanide-nitroprusside test were classified. Probands excreting more than 113.12 micromol cystine per mmol of creatinine (i.e., 1,000 micromol cystine per gram of creatinine) were classified as homozygotes. Parents of homozygotes in whom excretion of amino acids were normal were classified as heterozygotes type I. Those probands showing the excretion of at least one amino acid and the sum of urinary cystine plus the basic amino acids higher than the corresponding references ranges in our population were classified as heterozygotes type II or type III (heterozygotes non-type 1). RESULTS: We identified 24 homozygotes, 39 non-type I heterozygotes and 3 type I heterozygotes. The remaining 6 probands could not be classified. Means for cystine, lysine, arginine ornithine and their sum in homozygotes and heterozygotes non-type I were significantly (p < 0.001) in excess of the respective reference ranges. Moreover, means values in homozygotes were statistically different (p < 0.001) from heterozygotes non-type I. CONCLUSION: Urinary excretion of cystine per mmol creatinine allow us to distinguish heterozygotes from homozygotes. However, the best discriminator to distinguish non-type I heterozygotes from normal population might be the excretion of lysine per mmol creatinine. Additional studies including characterization of appropriate haplotypes should be carried out for a more precise identification of types of cystinuria.

Left anterior descending coronary artery bypass grafting through minimal thoracotomy.

BACKGROUND: In recent years, minimally invasive direct coronary artery bypass grafting has emerged as a valid tool for revascularization in a select group of patients with severe lesions of the left anterior descending coronary artery. Here we report the clinical results using two devices designed by us to facilitate the harvesting of the left internal mammary artery up to its origin and to occlude and stabilize the left anterior descending coronary artery while placing the anastomosis. METHODS: From January 1996 to January 1998, 122 patients underwent minimally invasive direct coronary artery bypass grafting in the Department of Cardiac Surgery, Favaloro Foundation. One hundred twelve patients received a single left internal mammary artery-left anterior descending coronary artery bypass graft, and in 10 patients, an additional bypass graft was performed. RESULTS: Most patients were discharged on day 2 or 3 after the procedure. Three patients (2.5%) had a perioperative myocardial infarction. The overall hospital mortality rate was 3.3% (4 patients). CONCLUSIONS: The combination of team experience, more careful dissection of the left internal mammary artery up to its origin, and use of the stabilizer-occluder and interrupted suture technique for the anastomosis has markedly improved our results.

Anonymous testing of newborn infants for HIV antibodies as a basis for estimating prevalence of HIV in childbearing women: the 1991-1994 study in Spain.

During 1991-1994, anonymous screening of newborn infants for maternal antibody to human immunodeficiency virus (HIV) was carried out in three regions of Spain: Valencia, Galicia and Sevilla. The newborn infants whose heel-stick blood eluates were satisfactory for HIV antibody tests were a consecutive series of 104 876, representing 99.3% of all newborn infants undergoing routine metabolic screening and estimated as comprising at least 98% of all births in the three regions. Enzyme immunoassay (EIA) positives were confirmed by immunoblot, yielding 246 confirmations: a rate of 2.3 per 1000. Seropositivity rates ranged from 1.4 per 1000 in Galicia to 2.1 in Sevilla and 3.1 in Valencia, and remained relatively stable in each region during the years of the study. Within socioeconomically defined subgroups of birth hospitals in Valencia and Galicia, all subgroups contained seropositives, even though there was a twofold to fivefold over-representation in the "inner city" public hospitals. To estimate the proportion of HIV-1-seropositive newborn infants who were positive for HIV-1 DNA, polymerase chain reaction (PCR) assays were performed on 165 dried blood spots that had been retained following positive immunoblot assays. Fifteen (9%) were PCR positive, and when this proportion is adjusted for the age-specific sensitivity of the method, it translates into an estimated HIV-1 transmission rate of 24% (range 18-36%). For 94,906 of the 104,876 newborn infants screened, the EIA used could detect antibodies that react with epitopes of HIV-1 and HIV-2. There were 30 newborn infants whose blood eluate was positive by this combined HIV-1/HIV-2 antibody screen and whose secondary screening with monovalent HIV-2 and HIV-1 EIA indicated that the HIV-2 reactivity was above the cut-off whereas the HIV-1 was not. Ranking these 30 results according to absolute HIV-2 reactivity and relative reactivity with respect to HIV-1 indicated that four infants were probable true HIV-2 seropositives and a total of 12 were possible HIV-2 seropositives, a prevalence of the order of 1:10000 to 1:20000 newborn infants. These anonymous population-based serological studies provide "leading-indicator" data to complement traditional AIDS surveillance for epidemiological and planning purposes.

Improved identification of heterozygotes for phenylketonuria using blood neopterin and biopterin.

A novel approach that combines information provided by the metabolism of pteridines and that of phenylalanine has been applied to the detection of heterozygotes for phenylketonuria. Phenylalanine, tyrosine, biopterin and neopterin have been measured in serum from normal controls and heterozygotes for classical phenylketonuria, before and after a phenylalanine oral load. Significant differences in neopterin and biopterin mean values in fasting serum and in the mean increase of biopterin induced by the phenylalanine load were found between groups. Inclusion of pteridine data in the discriminant analysis significantly improved the resolution of the classical phenylalanine loading test for the detection of heterozygotes for phenylketonuria.

Separation and identification of sugars and maltodextrines by thin layer chromatography: Application to biological fluids and human milk.

A monodimensional thin layer chromatography method to separate several sugars of clinical interest is described. The separation and identification of 14 sugars (L-fucose, D-galactose, D-glucose, lactose, N-acetylglucosamine, D-maltose, D-manose, L-sorbase, fructose, D-xylose, glucuronic acid, N-acetyllactosamine, 3' and 6' sialyllactose) and maltodextrines (G(2)-G(8)) is possible by using two different eluents mixtures, as well as two different detection reagents. The method has been applied to separate sugars, maltodextrines and oligosaccharides in several biological fluids (blood, urine and faeces), in an infant milk and in human milk. It is a very simple technique (with a high sensitivity) that can be used in any lab.