Genetic analysis in retinoblastoma and peripheral blood correlation.

OBJECTIVE: To determine the importance of intratumoral genetic analysis in the diagnosis of germ-line mutations in patients with retinoblastoma. To underline the importance of performing these genetic tests in every case of retinoblastoma. METHOD: Intratumoral genetic analysis of RB1 mutation was performed on 17 enucleated eyes that were non-responsive to conservative treatment. Patients had no family history of retinoblastoma, and lesions were always single. The identified mutations were then also studied in peripheral blood analysis. RESULTS: There were 12 (70.6%) cases with positive results in intratumoral analysis. In 8 cases (47.1%) mutation of both RB1 alelli were detected, and in 4 (23.5%) cases only one allele was found mutated. In 5 patients (29.4%) no mutation was identified. In the first hit, mutations comprised 7 frameshift or nonsense and 2 splice, whereas in the second hit, one splice mutation, 2 nonsense and 8 loss of heterozygosity were identified. Among 6 patients where intratumoral analysis detected a single mutation associated with a loss of heterozygosity, the peripheral blood analysis was able to detect the same mutation in 3 cases (50%). CONCLUSIONS: Intratumoral genetic analysis of sporadic retinoblastoma can detect germ-line mutations. These patients are at higher risk of bilateralization and development of second tumors or trilateral retinoblastoma. Genetic screening is recommended in every patient diagnosed with retinoblastoma.

[Genetic analysis results of patients with a retinoblastoma refractory to systemic chemotherapy]. / Resultados del analisis genético de los pacientes con retinoblastoma resistente a quimioterapia sistémica.

OBJECTIVE: To analyze the genetic alterations identified in the RB1 gene in retinoblastoma patients who do not respond to systemic chemotherapy. METHODS: A genetic analysis was performed on 115 patients with retinoblastoma, 40 of whom had received systemic chemotherapy, and 29 of them had bilateral disease. Descriptive and retrospective study. Non-responders were considered as patients who are finally enucleated. RESULTS: Patients with deletion type mutations are those with less preservation of the eyeball (Pearson Chi-square, P=.055). Patients with an impaired nonsense-frameshift type are more likely to preserve the eyeball. Of the 3 patients who had undergone bilateral enucleation, 2 of them had deletions and one missense alteration. Survival analysis (Kaplan-Meier curve) shows that patients with deletion type mutations are more resistance to chemotherapy, are suffering higher rates of enucleation, and for a shorter period of time (log rank [Mantel-Cox] with a significance level of P=.053), which are also associated with increased rate of being bilateral. CONCLUSIONS: Patients with a genotype show increased resistance to chemotherapy should be evaluated more closely and treated with various therapeutic weapons early. Patients that have deletions in the RB1 gene are at increased risk of chemoresistance. It is likely that other genetic alterations other than RB1 gene may be related to tumor aggressiveness and treatment resistance.

Central obesity and altered peripheral adipose tissue gene expression characterize the NAFLD patient with insulin resistance: Role of nutrition and insulin challenge.

OBJECTIVE: Insulin resistance (IR) and white adipose tissue (WAT) dysfunction frequently are associated with nonalcoholic fatty liver disease (NAFLD); however, the pathogenic mechanisms contributing to their clustering are not well defined. The aim of this study was to define some nutritional, anthropometric, metabolic, and genetic mechanisms contributing to their clustering. METHODS: Forty-five (20 men, 25 women) patients (age 45.7 ± 11.1 y) with recent diagnosis of NAFLD were grouped according to IR state. Energy balance was assessed using a food questionnaire and indirect calorimetry, and body composition with anthropometry and dual-energy x-ray absorptiometry. Biochemical and hormonal parameters combined with adipose tissue gene expression were determined. Microarray analysis of gene expression was performed in a subset of WAT samples from IR patients (n = 9), in the fasted state, after specific test meals (monounsaturated fatty acid [MUFA], saturated fat [SAT], and carbohydrate-rich) and after being challenged with insulin. RESULTS: IR patients exhibited higher trunk fat to leg fat ratio (P < 0.05) and had a higher ratio of SAT/MUFA fat intake (P < 0.05) than insulin-sensitive (IS) individuals. Deposition of fat in the trunk but not in the leg was directly related to liver enzyme levels (P < 0.05). IR patients also had lower adiponectin serum levels and leptin (LEP) mRNA expression in WAT compared with IS patients (P < 0.01 and P < 0.05, respectively). Microarray analysis after insulin challenge confirmed that insulin treatment induces the expression of PPARG gene and LEP and decreases GCGR gene (P < 0.05 for all) in WAT. No changes in these genes were observed in the postprandial state induced after the acute effect of specific diets. CONCLUSIONS: Patients exhibiting NAFLD and IR had preferential central fat deposition directly related to their serum alanine aminotransferase levels. These patients showed peripheral adipose tissue dysfunction and exhibited inappropriately low LEP biosynthesis that could be partially restored after anabolic conditions induced by insulin signaling.

[Trilateral retinoblastoma. Correlation between the genetic anomalies of the RB1 gene and the presence of pineal gland cysts]. / Retinoblastoma trilateral. Correlación de las alteraciones genéticas del gen RB1 y la presencia de quistes de la glándula pineal.

OBJETIVE: To determine the correlation between the presence of genetic anomalies identified in the RB1 gene and the development of trilateral retinoblastoma. METHOD: No patients with primitive neuroectodermal tumour (PNET) were identified out of a total of 206 patients, but there were 17 cases of pineal cysts, of which 11 had a genetic study. RESULTS: Of the 11 patients who had a genetic study performed, the anomaly in the germinal line was identified in 8 cases, which was equivalent to 100% of the bilateral retinoblastomas, and 25% of the unilateral ones. It is more common to find a germinal mutation in patients with bilateral disease (P=.024). There are no significant differences in the type of anomaly identified, although the nonsense-frameshift type is more frequent in cases with bilateral involvement. Identification of the genetic anomaly is more frequent in patients who have pineal cysts (Fisher test; P=.490). Nine of the 17 patients received systemic chemotherapy (52.29% of the cases), which could be able to prevent the development of PNET. Although a certain trend was observed in all the mentioned parameters, there was a relationship between, the presence of pineal cysts and bilateral disease (Pearson Chi X2: P=.191), a known family history (Fisher test; P=.114) and age of early diagnosis (Fisher test; P=.114). There were no significant differences in the mutation type identified. CONCLUSIONS: Considering pineal cysts as a pre-malignant form of pinealoblastoma, we found a relationship between the germinal line mutation of the RB1 gene and the cases with bilateral or unilateral retinoblastoma.

Heart rate variability and phantom pain in male amputees: application of linear and nonlinear methods.

Phantom-limb pain (PLP) is a phenomenon that may appear among people with amputation. Some studies reveal that 70% of people with amputation experience PLP years postamputation. There is a lack of scientific evidence about the cause of PLP. It has been hypothesized that the autonomic nervous system (ANS) could be involved in the mechanism that triggers PLP, but this hypothesis remains unclear. The aim of this study was to correlate ANS function, through heart rate variability (HRV) analysis, with PLP in adult males with amputation. The study population comprised 35 subjects, with 27 reporting PLP often or always. The rest of the subjects did not report any PLP. In order to calculate linear and nonlinear parameters of HRV, all subjects underwent 10 min of resting heart rate monitoring. The study did not find correlations between HRV parameters and PLP. Most of the subjects showed decreased values in linear parameters of HRV while nonlinear values were normal. HRV is not implicated in PLP. Linear and nonlinear methods for HRV analysis might reflect different physiological phenomena; while linear values place people with amputation at cardiovascular risk, nonlinear values indicate normality.

Epidemiología de la tuberculosis / Epidemiology of tuberculosis

La tuberculosis, “vieja conocida de la Neumología”, constituye todavía una enfermedad con una alta morbimortalidad en el mundo lo que hace que, pese a estar en el siglo XXI, continúe recabando nuestra atención, se planteen grandes retos como el control de la misma y surgan nuevos problemas como la emergencia de la tuberculosis multirresistente. Así pues, constituye para la Organización Mundial de la Salud una enfermedad de gran interés que le ha llevado a desarrollar un programa específico y ambicioso que pretende su erradicación para el año 2050. En este capítulo detallamos algunos de los aspectos más interesantes de la epidemiología de esta enfermedad, con una atención especial para el problema de la tuberculosis multirresistente y se exponen datos actual

Tuberculosis, that “old acquaintance of Pneumology”, is still a disease with a high morbidity and mortality in the world. This is why, in spite of our having entered the XXI century, it continues to demand our attention, great challenges continue to be faced, such as bringing it under control, and new problems continue to arise, such as the emergence of multiresistant tuberculosis. Thus, for the World Health Organisation, it represents a disease of great importance that has led this body to develop a specific and ambitious program that aims at its eradication by the year 2050. In this article we detail some of the most interesting aspects of the epidemiology of this disease, with special attention paid to the problem of multiresistant tuberculosis, and we present up to date data on the morbidity and mortality of tuberculosis in the world

[Other obstructive diseases: byssinosis, chronic bronchitis and occupational COPD and eosinophilic bronchitis]. / Otras enfermedades obstructivas: bisinosis, bronquitis crónica y EPOC de origen laboral y bronquitis eosinofílica.

Besides occupational asthma and diseases derived from acute inhalation, other obstructive diseases also have an occupational origin. Although at present byssinosis is a rare disease in Spain, we describe its characteristics because of its historical interest amongst occupational respiratory diseases and because it is still relevant in developing countries. Chronic bronchitis can also be related to exposure at work to dust and smoke, and is often referred to as "industrial bronchitis". Historically, the relation of CPOD to occupation has been subject to controversy, but nowadays this relationship is accepted; we describe the present evidence supporting this relationship. Finally, we describe eosinophilic bronchitis without asthma, an entity that was described for the first time in 1989 and that can sometimes have an occupational origin, sharing aetiological agents with occupational asthma.

Otras enfermedades obstructivas: bisinosis, bronquitis crónica y EPOC de origen laboral y bronquitis eosinofílica / Other obstructive diseases: byssinosis, chronic bronchitis and occupational COPD and eosinophilic bronchitis

Además del asma ocupacional y las enfermedadesderivadas de la inhalación aguda, otras enfermedadesobstructivas también reconocen un origen laboral.Aunque en la actualidad la bisinosis es una enfermedadrara en España, describimos las características de lamisma por su interés histórico dentro de las enfermedadesrespiratorias de origen laboral y porque todavíasigue vigente en los países en vías de desarrollo. Labronquitis crónica también puede estar relacionadacon la exposición laboral a polvos y humos, con frecuenciadenominada “bronquitis industrial”. La relaciónocupacional de la EPOC ha sido más controvertidaa lo largo de la historia, pero en la actualidad estarelación está aceptada; describimos la evidencia actualque sustenta esta relación. En último lugar, describimosla bronquitis eosinofílica sin asma, entidad descritapor primera vez en 1989 y que en ocasiones tambiénpuede tener un origen laboral, compartiendo agentesetiológicos con el asma ocupacional

Besides occupational asthma and diseases derivedfrom acute inhalation, other obstructive diseases alsohave an occupational origin. Although at presentbyssinosis is a rare disease in Spain, we describe itscharacteristics because of its historical interestamongst occupational respiratory diseases andbecause it is still relevant in developing countries.Chronic bronchitis can also be related to exposure atwork to dust and smoke, and is often referred to as“industrial bronchitis”. Historically, the relation ofCPOD to occupation has been subject to controversy,but nowadays this relationship is accepted; wedescribe the present evidence supporting thisrelationship. Finally, we describe eosinophilicbronchitis without asthma, an entity that wasdescribed for the first time in 1989 and that cansometimes have an occupational origin, sharingaetiological agents with occupational asthma

Simultaneous determination of cefepime and grepafloxacin in human urine by high-performance liquid chromatography.

A liquid chromatographic method with UV detection for simultaneous determination of cefepime and grepafloxacin has been developed. The method uses a C18 column, equipped with a pre-column of the same material, and acetonitrile-0.1 M phosphoric acid/sodium hydroxide buffer (pH 3.0)-0.01 M n-octylamine (pH 3.0) as mobile phase in gradient mode. Mobile flow rate and sample volume injected were 1.3 mL min(-1) and 20 microL, respectively. Detection wavelengths were 259 nm for cefepime and 278 nm for grepafloxacin. The retention times were 4.03 min for cefepime and 8.85 min for grepafloxacin, with detection limits of 1.0 and 1.1 microg mL(-1), respectively. The method was applied to the determination of both antibiotics in spiked samples of human urine.

Characterization and hardening of concrete with ultrasonic testing.

In this study, we describe a technique which can be used to characterize some relevant properties of 26 cylindrical samples (15 x 30 cm2) of concrete. The characterization has been performed, according to Spanish regulations in force, by some destructive and ultrasound-based techniques using frequencies of 40 kHz. Samples were manufactured using different water/cement ratios (w/c), ranging from 0.48 to 0.80, in order to simulate different values of compressive strength at each sample. We have correlated the propagation velocity v of ultrasonic waves through the samples to compressive strength R values. As some other authors remark, there exists an exponential relationship between the two above parameters. We have found that a highly linear relationship is present between R and w/c concentration at the samples. Nevertheless, when the same linear model is adopted to describe the relationship between v and w/c, the value of r decreases significantly. Thus, we have performed a multiple regression analysis which takes into account the impact of different concrete constituents (water, cement, sand, etc.) on ultrasound propagation speed. One of the most relevant practical issues addressed in our study is the estimation of the hardening curve of concrete, which can be used to quantify the viability of applying the proposed method in a real scenario. Subsequently, we also show a detailed analysis of the temporal evolution of v and R through 61 days, beginning at the date where the samples were manufactured. After analyzing both parameters separately, a double reciprocal relationship is deduced. Using the above parameters, we develop an NDE-based model which can be used to estimate hardening time of concrete samples.

Motivation in sport: an achievement goal perspective in young Spanish recreational athletes.

This study examined the relationship between goal orientations, perceived motivational climate, enjoyment, satisfaction, perception of ability, and preference for tasks with different levels of difficulty in Spanish athletes. To measure these variables 323 recreational athletes completed the Spanish versions of the Perception of Success Questionnaire, Perception of Significant Others' Sport Success Criteria Questionnaire, Enjoyment/Boredom scale, and Satisfaction in Sport Questionnaire. To measure perception of ability, athletes responded to two items reflecting a general perception and a comparative perception of their ability. Finally, preference for a task of different difficulty was measured by two items reflecting preference for easy and for challenging tasks. Analysis showed that scores on a positive ego orientation and negative task orientation were associated with maladaptive motivational patterns in sport. Such patterns involve lower perceptual ability, preference for easy tasks, and less enjoyment and satisfaction in sport activities.

Spectrofluorimetric determination of levofloxacin in tablets, human urine and serum.

A spectrofluorimetric method to determine levofloxacin is proposed and applied to determine the substance in tablets and spiked human urine and serum. The fluorimetric method allow the determination of 20-3000 ng ml(-1) of levofloxacin in aqueous solution containing acetic acid-sodium acetate buffer (pH 4) with lambda(exc)=292 and lambda(em)=494 nm, respectively. Micelle enhanced fluorescence improves the sensibility and allows levofloxacin direct measurement in spiked Human serum (5 mug ml(-1)) and urine (420 mug ml(-1)), in 8 mM sodium dodecyl sulphate solutions at pH 5.

Spectrophotometric determination of gallium in alloys and fly-ash with pyridoxal derivatives of thiocarbohydrazide and carbohydrazide.

The symmetric derivatives of pyridoxal with thiocarbohydrazide and carbohydrazide, and the asymmetric derivatives of pyridoxal and salicylaldehyde with the same hydrazides have been synthesized and their analytical potential for spectrophotometric and kinetic fluorimetric determination of metal ions was studied. Gallium(III) and PyMAU(1,3-bis{[4-(2-methyl-3-hydroxy-5-hydroxymethyl)pyridyl]methyleneaminourea at pH = 4.2 form a complex with a single absorption maximum at 425 nm, which can be extracted into cyclohexanone in the presence of a controlled amount of sodium perchlorate. The extract has maximum absorbance at 435 nm. Both systems can be used for determining gallium. The optimal range of gallium concentration for measurement in a 1-cm cell is 0.5-1.25 gmg/ml for the procedure in homogeneous medium ((425) = 3.76 x 10(4).mole(-1).cm(-1)) and 0.25-1.25 mug/ml for the extraction procedure ((435) = 5.30 x 10(4) 1.mole(-1).cm(-1). The latter procedure has been applied to the determination of gallium in alloys and fly-ash.

Derivatives of carbohydrazide, thiocarbohydrazide and diaminoguanidine as photometric analytical reagents-I 1,3-bis[(2-pyridyl)methyleneamino]thiourea and 1,3-bis[(2-pyridyl)methyleneamino]guanidine.

1,3-Bis[(2-pyridyl)methyleneamino]thiourea (PMAT) and 1,3-bis[(2-pyridyl)methyleneamino]-guanidine (PMAG) have been prepared. They have been examined and characterized by infrared and ultraviolet spectroscopy. A spectrophotometric method has been used for determination of the protonation constants of the reagents. Finally, a spectrophotometric survey has been made of the reactions of various cations with PMAT and PMAG.