Molecular analysis of endometrial inflammation in preterm birth.

Spontaneous preterm birth (sPTB) represents the 35%-45% of all preterm birth (PTB) cases and its etiology is unknown. We investigated if the expression level of endometrial cytokines and angiogenetic factors is related to the onset of sPTB.Endometrial tissues from non-pregnant women who experienced sPTB and from non-pregnant women who did not experience sPTB were collected and examined for their expression profile. With this aim, the PCR Array analysis was performed and data were confirmed by Real-Time PCR. Differential gene expression measurements (pathological vs control tissues) showed a significant up-regulation for genes codifying for two angiogenetic factors known as connective tissue growth factor (CTGF), and coagulation factor III (F3). An increased level of expression was detected both for tyrosine kinase endothelial (TEK) and for transforming growth factor beta 2 (TGF-ß2) genes but without reaching the statistical significance. The expression level of interleukin 10 receptor alpha (IL10RA) gene was slightly decreased in pathological group compared to control one but, as well as forTEK and TGF-ß2 measurements, without reaching the statistical significance. Our work is the first to correlate the imbalance in endometrial district of non -pregnant women with sPTB. These data could suggest a new point of view whence to read sPTB. We need additional clinical and biological studies to clarify sPTB pathogenesis.

Validation of the Photography Method for Nutritional Intake Assessment in Hospitalized Elderly Subjects.

OBJECTIVE: The aim of the present study was to validate the photographic indirect method as an accurate and specific tool to assess nutritional intake in a cohort of elderly hospitalized patients. DESIGN: this is a prospective observational study. SETTING: hospital (geriatric acute ward and transitional care of IRCCSS AUO San Martino Hospital, Genoa, Italy). PARTICIPANTS: 255 consecutive elderly hospitalized patients. MEASUREMENTS: assessment of malnutrition by: Mini nutritional assessment (MNA) and abbreviated Comprehensive geriatric assessment (CIRS; Barthel index, SPMSE). The direct method (Gold standard): food dish weight (before lunch) and residual (after lunch) food dish weight and estimation of the percentage of eaten food and of residual food for each dish. The percentages of food intake and residual food were calculated according to the following formula: intake %= initial weight of the dishes- residual food weight)/ initial weight dish x100. The unit of variable was the percentage. The indirect photographic method with extrapolation of the lunch food intake by photographic method confronting initial meal and residual meal (25% quartile food dish estimation). RESULTS: The results showed a significant correlation between the direct method (weighing residual food) and the indirect photographic method(n=255; r=0.9735; p<0.001) as well as a significant positive correlation between the indirect photographic method and the food caloric estimation calculated by the direct method (n=255; r= 0.6489, p<0.001). Intraclass coefficient (ICC), showed a highly significant degree of agreement between the gold standard and the indirect photographic method (ICC: 0.69; p<0.0001). Additionally, the results showed a good inter rater agreement of the indirect photographic method (kappa-statistic measure of interrater agreement: (Z=13.04; p<0.001); agreement 70.29% e Kappa=0.5965) and a good specificity of the indirect method as it was independent on the single food item. CONCLUSIONS: The study originally provided the validation of the indirect photographic method for the assessment of nutritional intake in a vast cohort of hospitalized elderly subjects. The present results moved a step forward in the appropriate assessment of nutrition intake in frail elderly, providing an easy to use tool that may be incorporate in routine clinical practice for early and targeted therapeutic interventions.

Relationship between maternal hemodynamics and plasma natriuretic peptide concentrations during pregnancy complicated by preeclampsia and fetal growth restriction.

OBJECTIVE: A proper maternal cardiovascular adaptation to the pregnancy plays a key role for promoting an adequate uteroplacental perfusion, for ensuring normal fetal development and for preventing gestational hypertensive complications such as preeclampsia. This study aims to evaluate hemodynamic measurements obtained by noninvasive methods among preclamptic women with and without fetal growth restriction (FGR) and the relationship with plasma levels of natriuretic peptides. STUDY DESIGN: The study compared 98 pregnant women (n=48 with preeclampsia; n=50 normotensive pregnant women) and 50 nonpregnant normotensive control subjects undergoing anultrasonic cardiac output monitor (USCOM) and plasma assessment of atrial N-terminal pro B-type natriuretic peptide (NT-proBNP). The statistical analysis was carried out by analysis of variance and correlation analysis. RESULTS: Preeclampsia state is associated with increased vascular resistance (mean 1587±236 vs 978±153 dyn s cm-3) and lower cardiac output (mean 5.7±1.1 vs 6.78±0.8 l) and this hemodynamic state is associated with higher levels of NT-proBNP (mean 121.2±26.3 vs 42.5±11.4 pg ml-1); furthermore, we found an inverse correlation between maternal cardiac output and plasma levels of NT-proBNP only if preeclampsia is associated with FGR. CONCLUSION: The elevated NT-proBNP in preeclampsia may reflect ventricular stress and subclinical cardiac dysfunction worsening if FGR is present. This may have implications for the acute management of the preeclampsia and FGR women and for appropriately timed therapeutic interventions later in life.

RET genetic screening of sporadic medullary thyroid cancer (MTC) allows the preclinical diagnosis of unsuspected gene carriers and the identification of a relevant percentage of hidden familial MTC (FMTC).

OBJECTIVE: This study was aimed to demonstrate the clinical benefits of rearranged during transfection (RET) genetic screening in patients with apparently sporadic medullary thyroid cancer (MTC) not only to identify the hereditary nature of the disease in the index case but also to discover family members harbouring the same germline mutations (i.e. gene carriers) who are unaware of their condition. CONTEXT: RET genetic screening allowed the identification of germline RET mutations in apparently sporadic MTC resulting in their re-classification as hereditary forms. PATIENTS AND MEASUREMENTS: RET genetic screening was performed in 729 apparently sporadic MTC patients by direct sequencing RET exons 5, 8, 10, 11 and 13-16. Clinical and biochemical evaluation of gene carriers was also performed. RESULTS: We discovered an unsuspected germline RET mutation in 47 of 729 (6·5%) apparently sporadic MTC who were re-classified as hereditary. We found 60 of 146 (41·1%) gene carriers, 35 of whom had biochemical or clinical evidence of MTC. Thirty gene carriers underwent total thyroidectomy and 27 of 30 (90%) were persistently cured after a mean follow-up of 6·0 years. As a further result of RET genetic screening, we observed a significantly higher prevalence of familial medullary thyroid cancer (FMTC) in our series with respect to the largest series of the International RET Consortium (P = 0·0002). CONCLUSIONS: RET genetic screening of patients with apparently sporadic MTC represents a major tool for the preclinical diagnosis and early treatment of unsuspected affected family members and allows the identification of a relevant percentage of hidden FMTC.

Força muscular e mortalidade na lista de espera de transplante de fígado / Muscle strength and mortality while on a liver transplant waiting list

OBJETIVO: Avaliar a força de músculos respiratórios e de mão em pacientes na lista de espera para o transplante de fígado e associá-los a mortalidade. MATERIAIS E MÉTODOS: Foram estudados retrospectivamente 132 pacientes submetidos à avaliação fisioterapêutica de rotina e que esperavam o transplante de fígado. A força dos músculos ventilatórios foi avaliada por meio das pressões inspiratória e expiratória máximas e a força do membro superior por meio de dinamometria. Os pacientes foram divididos em dois grupos: grupo A, com 51 pacientes (14 mulheres, 50,1±12,3 anos) que morreram enquanto estavam na lista de espera e grupo B, com 81 pacientes (31 mulheres, 45,0±3,8 anos) que sobreviveram até o transplante de fígado. Foi utilizado o teste de t de Student com nível de significância de 5 por cento. RESULTADOS: Os valores médios da pressão inspiratória máxima (PImax) dos grupos A e B foram 65,7±28,0 e 77,5±33,8mmHg (p=0,04), respectivamente, e as pressões expiratórias máximas foram 72,9±32,9 e 84,4±33,1mmHg (p=0,07), respectivamente. Os valores médios da força da mão esquerda dos grupos A e B foram 18,5±8,1 e 21,5±10,5kgf (p=0,08), respectivamente, e da força da mão direita foram 20,2±9,7 e 23,5±12,5kgf (p=0,10), respectivamente. CONCLUSÕES: A PImax é menor nos pacientes que morreram enquanto aguardavam o transplante. No mesmo grupo, foi observado que a pressão expiratória máxima e a força da mão direita e esquerda foram menores, apesar de não apresentarem diferenças estatisticamente significante.

OBJECTIVE: To evaluate respiratory muscle strength and hand strength in patients on a liver transplant waiting list and to associate these with mortality. METHODS: one hundred and thirty-two patients who underwent routine physical therapy evaluation while waiting for liver transplantation were studied retrospectively. Respiratory muscle strength was assessed by measuring the maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP), and upper-limb strength was evaluated by dynamometry. The patients were divided into two groups: group A, consisting of 51 patients (14 females, 50.1±12.3 years) who died while on the waiting list; and group B, consisting of 81 patients (31 females, 45.0±3.8 years) who survived until the time of liver transplant. Student's t test was used with a 5 percent significance level. RESULTS: The mean MIP values for groups A and B were 65.7±28.0 and 77.5±33.8mmHg (p=0.04), respectively, and the mean MEP values were 72.9±32.9 and 84.4±33.1mmHg (p=0.07), respectively. The mean values for left-hand strength in groups A and B were 18.5±8.1 and 21.5±10.5kgf (p=0.08), and the mean values for right-hand strength were 20.2±9.7 and 23.5±12.5kgf (p=0.10), respectively. CONCLUSIONS: MIP was lower in the patients who died while waiting for liver transplantation. In the same group, it was observed that the MEP values and right and left-hand strength were numerically lower, although they did not reach statistically significant differences.

Interacciones industria & radiología: logros innegables, conflictos de intereses posibles: los obsequios de la industria / Radiology-industry relations: conflict of interest, achievements

Los conflictos de intereses son muy frecuentes y diversos en medicina, tienen un gran impacto tanto en la información que llega a los profesionales de la salud como en la atención médica y, en su gran mayoría no son divulgados. Para lidiar con ellos, necesitamos hacer nuestros mayores esfuerzos en la atención médica para elaborar, cumplir y desarrollar recomendaciones éticas apropiadas, además de iniciativas de educación tanto en el período formativo, como durante el ejercicio de la profesión. Esto incluye a los radiólogos y su relación con la industria.

Profesionalismo: una forma de relacion entre medicos, pacientes y sociedad

Professionalism has been tied to medical practice for long time and it has been considered in many different ways by society. A few years ago, and following a hard period of criticism centred in our fails to the commitments with service to patients and the primacy of patients' interest, we are living a revival of this concept with patient-centred professionalism, reassuring our commitment with basic aims: to put patient interest first and to serve them and community. Such issues were gathered in the ®Code or Constitutional Chart for Medical Professionalism¼, an ideal for medical professionals, that it is necessary to be grounded, focusing on the relevant real issues: medical school education, practice according to standards, ethical controlled practice, medical practice peer audited, practical spreading of professionalism principles, among others. Radiologist must adhere to these principles, and they have to consider the nature of their medical attention through imaging, engage them with some ethical principles for managing the images of their patients: safety and quality, specially radioprotection, selection of adequate patient' images, periodical updates, proper images' interpretation in the context of patient condition. Adhering, respecting and practicing these principles are aims we need to achieve if we want to preserve the future of radiology and radiologist in a highly technified but humanitarian medical attention as well.

El profesionalismo, asociado a la medicina desde épocas tempranas, ha pasado por diversas etapas de consideración en la sociedad. Tras un período reciente de cuestionamientos por priorizar intereses económicos, por sobre el cumplimiento del compromiso de servicio hacia los pacientes, apareció como reacción, un renacimiento dirigido a recuperar la primacía del servicio, que fue plasmado en el ®Código o Carta Constitucional para los Médicos¼ que propone ideales a los que ®todos los profesionales de la medicina pueden y debieran aspirar¼. Se propone la práctica de acciones concretas enfocadas hacia los temas relevantes como: docencia, ejercicio, control ético efectivo de la medicina, auditorías del trabajo médico por pares, y difusión práctica de los ideales. Los radiólogos, que ejercen a través de las imágenes, deben prestar especial atención a ciertos principios éticos básicos para un mejor futuro de la medicina y la especialidad, en el contexto de una medicina altamente tecnificada, pero humanitaria.

Octreótido, tratamiento endoscópico o ambos en la hemorragia variceal activa enpacientes cirróticos: estudio multicéntrico / Octreotide administration and/or endoscopic treatment in cirrhotic patients with acute variceal bleeding: a multicentric study

Background: Acute variceal bleeding in cirrhotic patients is an emergency with a high risk of rebleeding and death. Endoscopic procedures such as sclerotherapy or banding, combined or not with drugs such as octreotide could be considered. Aim: To assess the value of octreotide in the control of acute variceal bleeding. Patients and methods: Ninety-two patients were randomized into three groups: endoscopic therapy plus octreotide 50 Ág/h bolus and continuous infusion for 5 days (n= 36); octreotide (same dosage) (n= 13) and endoscopic therapy only (banding and/or sclerotherapy) (n= 43). Results: Haemostasis at 24 hours was achieved in 97 percent of patients with combined treatment, 69 percent of patients receiving octreotide, and 93 percent of patients with endoscopic therapy (p= 0.2). Three patients with combined treatment, four patients receiving octreotide and eight patients with endoscopic therapy, rebled during the first five days (p= 0.15). The mean of blood units transfused was similar in the three groups. No differences were observed in hospital days and side effects. At 42 days of follow up, eight patients with endoscopic therapy, one patients with combined therapy and 2 patients receiving octreotide, died (p=NS). Conclusions: Octreotide is useful in the management of acute variceal bleeding. The absence of important side effects, renders it as a safe adjuvant treatment associated with endoscopic treatment (Rev MÚd Chile 2004; 132: 285-94).

Kinetics of hydrolysis of fructooligosaccharides in mineral-buffered aqueous solutions: influence of pH and temperature.

High-performance anion exchange chromatography coupled with a pulsed amperometric detection system (HPAEC-PAD) was used to evaluate the extent of chemical hydrolysis of three fructooligosaccharides (FOS) including 1-kestose (beta-D-Fru-(2-->1)(2)-alpha-D-glucopyranoside, GF2), nystose (beta-D-Fru-(2-->1)(3)-alpha-D-glucopyranoside, GF3), and fructofuranosylnystose (beta-D-Fru-(2-->1)(4)-alpha-D-glucopyranoside, GF4). A kinetic study was carried out at 80, 90, 100, 110, and 120 degrees C in aqueous solutions buffered at pH values of 4.0, 7.0, and 9.0. Under each experimental condition, the determination of the respective amounts of reactants and hydrolysis products showed that FOS hydrolysis obeyed pseudo-first-order kinetics as the extent of hydrolysis, which decreased at increasing pH values, increased with temperature. The three oligomers were found to be degraded mainly under acidic conditions, and at the highest temperature value (120 degrees C), a quick and complete acid degradation of each FOS was observed. Using the Arrhenius equation, rate constants, half-life values, and activation energies were calculated and compared with those obtained from sucrose under the same experimental conditions. It appeared that the hydrolysis of FOS took place much more easily at acidic pH than at neutral or basic pH values.

Evidence for association of a common variant of the endothelial nitric oxide synthase gene (Glu298-->Asp polymorphism) to the presence, extent, and severity of coronary artery disease.

BACKGROUND: Genetic variants of endothelial nitric oxide synthase (eNOS) could influence individual susceptibility to coronary artery disease. OBJECTIVE: To assess whether Glu298-->Asp polymorphism of the eNOS gene is associated with the occurrence and severity of angiographically defined coronary artery disease in the Italian population. METHODS: Polymerase chain reaction/restriction fragment length polymorphism analysis was done to detect the Glu298-->Asp variant of the eNOS gene in 201 patients with coronary artery disease and 114 controls. The severity of coronary artery disease was expressed by the number of affected vessels and by the Duke scoring system. RESULTS: The frequencies of the eNOS Glu/Glu, Glu/Asp, and Asp/Asp genotypes in the coronary artery disease group were significantly different from those of controls (45.3%, 38.8%, and 15.9% v 42.1%, 51.8%, and 6.1%, respectively; chi2 = 8.589, p = 0.0136). In comparison with subjects who had a Glu298 allele in the eNOS gene, the risk of coronary artery disease was increased among Asp/Asp carriers (odds ratio 2.9, 95% confidence interval 1.2 to 6.8, p = 0.01) and was independent of the other common risk factors (p = 0.04). There was a significant association between the eNOS Glu298-->Asp variant and both the number of stenosed vessels (mean (SEM), 2.3 (0.1) for Asp/Asp v 1.9 (0.1) and 1.8 (0.1) for Glu/Glu and Glu/Asp, respectively; p = 0.01) and the Duke score (56.1 (3.1) for Asp/Asp v 46.7 (2.0) and 46.1 (1.9) for Glu/Glu and Glu/Asp, respectively; p = 0.02). CONCLUSIONS: Glu298-->Asp polymorphism of the eNOS gene appears to be associated with the presence, extent, and severity of angiographically assessed coronary artery disease.

Chronic long-term nitrate therapy: possible cytogenetic effect in humans?

Nitrates act as donors of nitric oxide (NO), a molecule with a recognized potential for genotoxicity. In order to assess whether chronic long-term nitrate therapy may increase genotoxicity, we evaluated chromosomal damage in peripheral lymphocytes of 27 ischaemic patients undergoing chronic nitrate treatment for vertical line4 years (7.9 +/- 3.1, mean +/- SD) and 18 age- and sex-matched subjects without any previous nitrate treatment. At the same time, after treatment in vitro with 0-20 microM sodium nitroprusside as NO donor, micronucleus induction and cell proliferation were also evaluated using blood from six different healthy donors. The results showed that the frequency of structural chromosomal aberrations was not significantly higher in the drug-treated group than the control [2.1 +/- 1.4 versus 1.6 +/- 1.2 (mean +/- SD); P = 0.23]. The frequency of micronucleated lymphocytes was higher in the nitrate group than in the control group (6.5 +/- 4.6 versus 3.5 +/- 2.9, P=0.01). In vitro treatment indicated a dose-dependent increase in the frequency of micronucleated lymphocytes with increasing SNP concentrations. Cytotoxicity and cell cycle delay, with a statistically significant difference with respect to control culture, were also observed. Our results suggest a possible genotoxic activity of nitrate therapy. Further studies focusing on the possible link between nitrate therapy and genotoxicity are warranted at this point.

Up-regulation of 'clearance' receptors in patients with chronic heart failure: a possible explanation for the resistance to biological effects of cardiac natriuretic hormones.

BACKGROUND: Three specific receptors for the cardiac natriuretic peptide system have been identified to date. Down-regulation of the biologically active binding sites (i.e. NPR-A and NPR-B) could explain the blunted response to cardiac natriuretic hormones observed in heart failure (HF), but not the increased metabolic clearance rate. Variations in the ratio between biological and clearance (NPR-C) receptors in target tissue may explain this increase. AIM: The aim of this study was to investigate the regulation of NPR-C receptors on platelets, in patients with HF. METHODS: Eighteen patients with HF (NYHA class: I-II, n=8; III-IV, n=10) and 18 age-matched healthy subjects were studied. The affinity constant (K(d)) and density (B(max)) of binding sites were derived by saturation assays on platelet suspensions using 125I-ANP as radioligand. RESULTS: B(max) increased as a function of the severity of disease: 21.3+/-3.3 fmol/10(9) cells in class III-IV, 11.7+/-2.2 in class I-II, and 11.6+/-1.1 in controls, respectively (P=0.0179 for class III-IV vs. controls and P=0.0451 vs. NYHA I-II). CONCLUSIONS: The increase in density of 'clearance' receptors in severe HF is theoretically consistent with the reduction in cardiac natriuretic peptide biological activity, as well as the increase in metabolic clearance rate. This suggests that clearance receptor blockade may be of potential therapeutic value in HF.

Differential ability of human endothelial cells to internalize and express exogenous DNA.

Vascular endothelium gene expression regulates blood-vessel wall interactions, vascular permeability, smooth muscle cell growth and tone. The possibility to introduce exogenous DNA or RNA sequences in endothelial cells represents a novel therapeutic approach of vascular disease. The aim of the work was to investigate the ability of endothelial cells to internalize and express exogenous DNA sequences. Human umbilical vein endothelial cells (HUVEC) were transfected with either a 780 bp fluorescein-labeled DNA (FITC-DNA) or pEGFP-C1 plasmid encoding for a green fluorescent protein (GFP), using the cationic liposome DOTAP as transfection reagent. The transfected cell population was passed through a FACScan apparatus and percentage of fluorescent cells was determined using a FACScan analysis programme. The SW620 tumor-derived cell line was used as control. The percentage of FITC-DNA positive cells was 66.0% for HUVEC and 45.0% for SW620 cells. On the contrary, the percentage of GFP-positive cells was 13.8% and 43% for HUVEC and SW620, respectively. By increasing the amount of DNA as well as the protocol of administration the percentage of GFP-positive HUVEC was enhanced suggesting a rapid degradation of DNA in the HUVEC cytoplasm.

Evidence for DNA damage in patients with coronary artery disease.

According to the "monoclonal hypothesis" of atherosclerosis, several studies suggest that cancer and atherosclerosis may have several fundamental biological mechanisms in common. Therefore, an increase in the mutation rate may be involved in the pathogenesis of atherosclerotic plaques. The aim of the study was to verify the presence of chromosomal damage in peripheral blood lymphocytes in patients with coronary artery disease by using micronucleus (MN) test, a reliable biomarker in genetic and cancer risk assessment. Subjects included 53 patients with documented coronary ischemic heart disease (group I); 10 patients with valvular heart disease in absence of atherosclerotic lesions of the coronary arteries (group II) and 16 healthy subjects, age- and sex-matched (group III) were studied as controls. For each subject, two separate cultures were performed and 1000 binucleated cells were scored for the evaluation of MN frequency. The mean (+/-S.E.M.) of MN frequency were 11.9+/-1.7, 5.9+/-1.2 and 3.6+/-0.7 in groups I, II and III, respectively. The MN frequency of group I was significantly higher than that of group III (P=0.02). In group I, MN frequency increased with the number of affected vessels (6.3+/-0.7, 13.9+/-1.6, 14.9+/-5.3 for one-, two-, and three-vessel disease, respectively). Scheffe's test showed that MN frequency was significantly higher in two-vessel compared with one-vessel disease (P=0.0077). Moreover, a positive relationship was found between MN levels and the severity of the disease, calculated by the Duke scoring system (R=0.28, P=0.032), as well as the systolic blood pressure (R=0.34, P=0.009). These results suggest that coronary artery disease in humans is a condition characterized by an increase of DNA damage, positively correlated with the severity of the atherosclerotic disease.

Evaluation of fructans in various fresh and stewed fruits by high-performance anion-exchange chromatography with pulsed amperometric detection.

Fructans are food-grade non-digestible carbohydrates that exert beneficial nutritional effects. Their characterization and quantification is required for food-labeling purposes. We describe the suitability of high-performance anion-exchange chromatography coupled with pulsed amperometric detection for the identification and quantification of fructans in fresh fruits (various apple and pear cultivars, plum, banana) as well as in commercial stewed fruits obtained from a local manufacturer. After extraction with water and appropriate filtration, inulobiose [beta-D-Fru-(2-->1)-beta-D-fructofuranoside; F2], 1-kestose [beta-D-Fru-(2-->1)2-alpha-D-glucopyranoside; GF2] and nystose [beta-D-Fru-(2-->1)3-alpha-D-glucopyranoside; GF3] were completely separated in a single 36-min run using a Dionex CarboPac PA 100 column and the new quadruple-potential waveform, originally tailored for oligosaccharide separation. No measurable amounts of F3 and GF4 were detected within the group of studied fruit products. Peak identification was realized using standards. The method is easy, reproducible, and sensitive since as little as 28 microg of sugar per gram dry matter can be quantified. Banana and plum are the varieties containing the highest levels of fructans (about 6000 microg per gram dry matter). The maturity of the fruit appears to have a great influence on the level of GF2. Samples of apple-banana stewed fruits contained the highest total fructan concentration (about 700 microg per gram dry matter). Accurate quantification of fructans will allow more precise nutritional formulation and diet selection for higher fructan consumption.

Separation and identification of enzymatic sucrose hydrolysis products by high-performance anion-exchange chromatography with pulsed amperometric detection.

An accurate carbohydrate analysis method, namely high-performance anion-exchange chromatography with pulsed amperometric detection was successfully applied to the study of sucrose hydrolysis under enzymatic (baker's yeast invertase) conditions. The hydrolysis was monitored by determining sucrose degradation and the corresponding formation of D-glucose, D-fructose and five intermediate fructans using a CarboPac PA-100 (Dionex) analytical anion-exchange column. Highly reproducible results were obtained. The unknown fructans were collected from a semi-preparative CarboPac PA-100 (Dionex) column, neutralized and then desalted on a column containing mixed bed resin AG 501-X8 (D) before identification of the chemical structure. This procedure permitted us to obtain about 20 microg of pure product which is not enough for NMR analysis. Detailed GC-MS analytical data of the methylated compounds indicated that these oligosaccharides were beta-D-Fru-(2 --> 1)-beta-D-Fru-(2 --> 1)-alpha-D-glucopyranoside (1-kestose), beta-D-Fru-(2 --> 6)-alpha-D-glucopyranoside (6-beta fructofuranosylglucose), beta-D-Fru-(2 --> 1)-beta-D-fructofuranoside (inulobiose), beta-D-Fru-(2 --> 6)-beta-D-Fru-(2 --> 1)-alpha-D-glucopyranoside (6-kestose) and beta-D-Fru-(2 --> 6)-alpha-D-Glc-(1 --> 2)-beta-D-fructofuranoside (neokestose) coeluating with a disaccharide.

Endothelin-1, endothelin-1 receptors and cardiac natriuretic peptides in failing human heart.

Endothelin (ET)-1 is a potent vasoconstrictor peptide produced in the myocardium that can exert important effects on cardiac myocyte growth and phenotype; cardiac natriuretic peptides (ANP and BNP) are known to act as physiological antagonists of ET-1. In this study a comparative determination of ET-1 receptors and of the local productions of ET-1 and of ANP and BNP was made in different sites of failing and nonfailing hearts. Tissue from right and left atrium, right and left ventricle and interventricular septum from seven adult heart transplant recipients with end-stage idiopathic dilated cardiomyopathy (functional class III and IV, with ejection fraction < 35%) and from four postmortem subjects without cardiac complications was analyzed. In failing hearts we observed a tendency to increase of density of binding sites, most evident in left ventricle (62.6+/-22.6 fmol/mg protein vs. 29.0+/-3.3, mean +/- SEM, p = ns). A prevalence of ET-A subclass, observed in all samples, resulted more pronounced in failing hearts where this increase, found in all the cardiac regions, was more evident in left ventricle (p = 0.0007 vs nonfailing hearts). The local concentrations of ET-1, ANP and BNP resulted significantly increased in failing hearts with respect to controls in all sides of the heart. In failing hearts we have observed a tendency to increase in endothelin receptor density mainly due to a significant upregulation of ET-A subtype and a parallel increase of the tissue levels of ANP, BNP and ET-1 indicating an activation of these systems in heart failure.

Sequence analysis of the aminoacylase-1 family. A new proposed signature for metalloexopeptidases.

The amino acid sequence analysis of the human and porcine aminoacylases-1, the carboxypeptidase S precursor from Saccharomyces cerevisiae, the succinyl-diaminopimelate desuccinylase from Escherichia coli, Haemophilus influenzae and Corynebacterium glutamicum, the acetylornithine deacetylase from Escherichia coli and Dictyostelium discoideum and the carboxypeptidase G(2) precursor from Pseudomonas strain, using the Basic Local Alignment Search Tool (BLAST) and the Position-Specific Iterated BLAST (PSI-BLAST), allowed us to suggest that all these enzymes, which share common functional and biochemical features, belong to the same structural family. The three amino acid blocks which were found to be highly conserved, using the CLUSTAL W program, could be assigned to the catalytic active site, based on the general three-dimensional structure of the carboxypeptidase G(2) from the Pseudomonas strain precursor. Six additional proteins with the same signature have been retrieved after performing two successive PSI-BLAST iterations using the sequence of the conserved motif, namely Lactobacillus delbrueckii aminoacyl-histidine dipeptidase, Streptomyces griseus aminopeptidase, Saccharomyces cerevisiae aminopeptidase Y precursor, two Bacillus stearothermophilus N-carbamyl-L-amino acid amidohydrolases and Pseudomonas sp. hydantoin utilization protein C. The three conserved amino acid motifs corresponded to the following blocks: (i) [S, G, A]-H-x-D-x-V; (ii) G-x-x-D; and (iii) x-E-E. This new sequence signature is clearly different from that commonly reported in the literature for proteins belonging to the ArgE/DapE/CPG2/YscS family.