Okur-Chung neurodevelopmental syndrome-linked CK2α variants have reduced kinase activity.

The Okur-Chung neurodevelopmental syndrome, or OCNDS, is a newly discovered rare neurodevelopmental disorder. It is characterized by developmental delay, intellectual disability, behavioral problems (hyperactivity, repetitive movements and social interaction deficits), hypotonia, epilepsy and language/verbalization deficits. OCNDS is linked to de novo mutations in CSNK2A1, that lead to missense or deletion/truncating variants in the encoded protein, the protein kinase CK2α. Eighteen different missense CK2α mutations have been identified to date; however, no biochemical or cell biological studies have yet been performed to clarify the functional impact of such mutations. Here, we show that 15 different missense CK2α mutations lead to varying degrees of loss of kinase activity as recombinant purified proteins and when mutants are ectopically expressed in mammalian cells. We further detect changes in the phosphoproteome of three patient-derived fibroblast lines and show that the subcellular localization of CK2α is altered for some of the OCNDS-linked variants and in patient-derived fibroblasts. Our data argue that reduced kinase activity and abnormal localization of CK2α may underlie the OCNDS phenotype.

Stability studies in biological fluids during post-analysis custody. Opiate compounds derived from heroin consumption.

In Forensic Toxicology, the evidences have to be maintained under custody for, at least, one year. Depending on the conditions and duration of storage, drug concentrations might have changed considerably since the first analysis. The aim of this study is to evaluate in vitro stability of opiate compounds, derived from heroin consumption, 6-acetylmorphine (6-MAM), morphine (MOR) and codeine (COD), in blood and urine, during post-analysis custody. Parameters evaluated were: time of custody, temperature, addition of preservative (blood) and pH (urine). Blood and urine samples were spiked with the three analytes to give a final concentration of 1000 ng/mL. The prepared samples were divided into 2 groups and stored at two temperatures (4 °C and -20 °C). Each one of these groups was subsequently divided in other two groups: with and without preservative (1%NaF) for blood, and pH 4 and 8 in the case of urine. 6-MAM, MOR and COD were analyzed by GCMS after SPE and derivatization with BSTFA. Analyses were performed in triplicate every two weeks for a year. In blood samples 6-MAM is the only compound that degrades. The best storage conditions were at -20 °C with NaF, with 6-MAM recoveries, after one year of custody, of 47.1 ± 1.5%; while in the other conditions 6-MAM disappeared after 215 days (at 4 °C with NaF), 45 days (at -20 °C without NaF) and 15 days (at 4 °C without preservative). COD does not degrade, with recoveries higher than 90%, in all of the conditions. They ranged from 89.7 ± 3.6% in samples maintained at -20 °C without NaF to 95.9 ± 2.0% in those maintained at 4 °C with NaF. MOR recoveries were lower than those of COD. They ranged from 66.9 ± 3.6%, in frozen samples added with NaF, to 78.6 ± 0.5% in refrigerated samples without preservative. In urine samples the three compounds were stable in all the studied conditions, with the exception of 6-MAM in samples at pH 8 and stored at 4 °C. In these conditions, 6-MAM disappeared after 135 days of custody; while recoveries in the other conditions ranged from 93.7 ± 6.4%, at 4 °C and pH 4, to 85.1 ± 2.0% at -20 °C and pH 8. MOR and COD recoveries were similar in the four conditions. In the case of MOR, they ranged from 82.1 ± 1.2% at 4 °C and pH 4 to 89.5 ± 6.0% at -20 °C and pH 8. As far as COD is concerned, recoveries ranged from 111.6 ± 5.8% at 4 °C and pH 8 to 102.6 ± 1.2% at 4 °C and pH 4. In conclusion, the study showed that the most labile opiate compound is 6-MAM. Its stability mainly depends on urine pH or the addition of preservative, in blood samples. The best storage conditions for samples from heroin consumers are in the freezer, at -20 °C. In addition, blood samples must be added with 1%NaF and urine samples must be buffered at pH 4.

The tooth for molecular analysis and identification : a forensic approach.

The aim of this study is to optimize laboratory preparation of teeth for DNA identification. By sectioning the tooth topographically into two different radicular portions, it was analyzed whether these portions of mineralized tissue differ in the quantity and quality of DNA they contain. 25 teeth were subject to different experimental conditions and total DNA was quantified for each individual tooth's radicular portion: apical and remaining root, according to a 2003 study by Gaytemenn and Sweet. We verified, with statistically significant figures, that the apical portion of the tooth is that which contains the greatest quantity of DNA. Different analytical procedures were studied for various polymorphic markers to evaluate the quality of the DNA. We concluded that the tooth is topographically distinct in both DNA quantity and quality. The tooth's apical portion is the preferential choice in sample preparation of dental mineralized tissue for molecular analysis and identification.

Open-label trial on efficacy and security of treatment with gemcitabine and oral modulation with tegafur and levofolinic acid (GEMTG) in patients with advanced pancreatic cancer

AIM: Advanced pancreatic cancer has a bad prognosis, with a median overall survival (OS) no longer than 4-6 months. Since the end of last century, monotherapy with gemcitabine has remained the elective therapy, but new schedules are needed in order to improve these results. We aim to evaluate the efficacy of tegafur and levofolinic acid (LV) associated with gemcitabine, as well as its toxicity, progression-free survival and OS in advanced pancreatic cancer. PATIENTS AND METHODS: An open-label, multicentric, prospective, non-controlled trial was carried out on patients with advanced or disseminated pancreatic cancer. Gemcitabine 1250 mg/m² was administered on the 1st and 8th days of the cycle, tegafur 750 mg/m²/day for 21 consecutive days and LV 25 mg/day continuously, every 28 days, with a maximum of six cycles. The primary variable was tumour overall response rate (ORR). Secondarily, time to progression (TTP), OS and scheme toxicity were determined. RESULTS: Forty patients were recruited; the male/female ratio was 30:10, with a mean age of 61 years. Forty percent had a Karnofsky index of 90% or 100%. Only 11 patients (27%) completed the six cycles of treatment, but more than 50% received three or more cycles. Dose intensity was 89.56% for gemcitabine and 87.36% for tegafur. Efficacy ORR was 22.5% (CI 95%, 6-37%). TTP was 3.87 months (CI 95%, 2.1-5.6), time to treatment failure was 2.97 months (CI 95%, 2.43-4.67) and OS 6.3 months (CI 95%, 4-7). The chemotherapeutic combination was well accepted; most haematologic and non-haematologic toxicities were grade 1 or 2. The most prevalent grade 3/4 toxicities were asthenia (30%), liver biochemistry disorders (25%), diarrhoea (15%) and stomatitis (12%). CONCLUSIONS: The administration of gemcitabine, associated with oral tegafur and leucovorin, has activity against advanced pancreatic cancer, with an adequate toxicity profile (AU)

Importancia de los polimorfismos de la ciclooxigenasa-2 en la presentación clínica de la sarcoidosis / Importance of cyclooxigenase-2 polymorphismsin the clinical presentation of sarcoidosis

OBJETIVO: Estudiar la posible relación entre las manifestaciones clínicas de la sarcoidosis y los polimorfismos del gen de laciclooxigenasa-2 (COX-2).MÉTODO: Estudio multicéntrico observacional transversal en el que participaron 7 hospitales de España. Se incluyeron pacientes diagnosticados de sarcoidosis según criterios internacionales. De cada caso se recogió edad, sexo, método diagnóstico, enzima convertidora de angiotensina, pruebas de función respiratoria, estadio radiológico y clínica del paciente en el momento del diagnóstico. Los hallazgos clínicos se agruparon en respiratorios y sistémicos. Los estudios genéticos se realizaron a partir del ADN obtenido de linfocitos de sangre periférica. El ADN se amplificó mediante PCR convencional y los polimorfismos fueron analizados por sondas de hibridación fluorescentes y curvas de disociación. Se determinaron4 variantes alélicas del gen de la COX-2: COX2.5909T>G,COX2.8473T>C, COX2.926G>C y COX2.3050G>C. RESULTADOS: La muestra se compuso de 131 casos de sarcoidosis (63 hombres; edad: 47 ± 15 años), todos con diagnóstico histológico menos 5 casos. El polimorfismo COX2.3050G>C en homocigosis resultó estar significativamente presente entre los pacientes con manifestaciones sistémicas frente al resto de pacientes (4,6% vs 0%;p=0,045). La presencia de manifestaciones sistémicas de la enfermedad estuvo significativamente asociada a los pacientes portadores del alelo C de dicho polimorfismo (34,4% vs. 18,6%; p=0,031; OR:2,3; IC 95%: 1,03-5,12). El resto de polimorfismos estudiados no estuvieron relacionados con la expresión clínica de la enfermedad. CONCLUSIÓN: La presencia de manifestaciones sistémicas parece estar relacionada con los portadores del alelo C del polimorfismoCOX2.3050G>C de la COX-2 (AU)

OBJECTIVE: To study clinical manifestations of sarcoidosis according to cyclooxigenase-2 (COX-2) polymorphisms. METHOD: Observational cross-sectional multicentre trial in which 7 Spanish hospitals participated. Patients diagnosed withs arcoidosis according to international criteria were included. Age, gender, diagnostic method, angiontens in converting enzyme, pulmonary function tests, radiological stage and clinical findings at the moment of the diagnosis were recorded for each case included. Clinical findings were grouped as respiratory or systemic. Genetic studies were performed on DNA extracted from peripheral blood lymphocytes. DNA was amplified by conventional PCR and polymorphisms were studied by Fluorescent Hybridization Probe-Melting Curves. COX-2 polymorphisms genotyped were COX2.5909T>G, COX2.8473 T>C, COX2.926 G>C y COX2.3050 G>C.RESULTS: 131 sarcoidosis patients (63 males, age: 47 ± 15years) were included. All included patients had a histological diagnosis except for 5 patients. COX2.3050G>C homozygote polymorphism resulted to be significantly present in patients with a systemic manifestation of the disease as compared with the rest of the sample(4,6% vs 0%; p = 0,045). Systemic manifestations were significantly associated with allele C carriers of this polymorphism (34.4% vs.18.6%; p = 0.031; OR: 2.3; IC 95%: 1.03 – 5.12). The rest of the studied polymorphisms were not significantly related to the clinical manifestations of the disease. CONCLUSION: Our results suggest that allele C carriers ofCOX2.3050G>C polymorphism are associated with the systemic manifestations of sarcoidosis (AU)

Association of the 3050G>C polymorphism in the cyclooxygenase 2 gene with systemic sarcoidosis.

BACKGROUND: We investigated the potential association between cyclooxygenase-2 (COX-2) gene polymorphisms and clinical manifestations of sarcoidosis. METHODS: This observational cross-sectional study involved seven hospitals in Spain. We diagnosed patients with sarcoidosis according to the International Criteria. The following variables were recorded: age, gender, initial diagnostic methods, serum angiotensin-converting enzyme (ACE) levels, pulmonary function tests, radiological stage, and clinical findings at diagnosis. Manifestations of sarcoidosis were classified as systemic vs. nonsystemic. Genotyping of four COX-2 polymorphisms (COX2.5909T>G, COX2.8473T>C, COX2.926G>C, and COX2.3050G>C) was undertaken on DNA extracted from peripheral blood lymphocytes using fluorescent hybridization probes and melting curves. RESULTS: A total of 131 sarcoid patients (63 males, mean age: 47 +/- 15 years) were studied. One hundred twenty-six of these patients had one or more positive biopsies. The results demonstrated that genotype distribution for the COX2.3050G>C polymorphism was significantly different between patients with systemic sarcoidosis and those with nonsystemic forms (p = 0.046). After adjustment for age, gender, and serum ACE levels, a significant association between the carriage of at least one C allele of the COX2.3050G>C polymorphism and systemic sarcoidosis was observed (odds ratio [OR]: 2.3; 95% confidence interval [CI]: 1.03-5.12, p = 0.031). Other polymorphisms were not associated with either clinical manifestations of the disease or serum ACE levels. CONCLUSIONS: Our results indicate for the first time that the C allele of the COX2.3050G>C polymorphism is associated with systemic sarcoidosis.

[Cardiac resynchronization through a persistent left superior vena cava]. / Resincronización cardíaca a través de una vena cava izquierda persistente.

Cardiac resynchronization therapy is effective in the treatment of patients with severe heart failure and intraventricular dysynchrony. However, we are sometimes faced with the unexpected presence of a persistent left superior vena cava. We report the case of a patient with dilated cardiomyopathy and left ventricular dysynchrony in which we implanted a resynchronization pacemaker exclusively through a persistent left superior vena cava that did not communicate with the right vena cava.

Resincronización cardíaca a través de una vena cava izquierda persistente / Cardiac resynchronization through a persistent left superior vena cava

La resincronización cardíaca es eficaz en pacientes con insuficiencia cardíaca y criterios de asincronía intraventricular. Sin embargo, durante el implante podemos encontrarnos excepcionalmente con la existencia inesperada de una vena cava izquierda persistente. Presentamos un caso de miocardiopatía dilatada en el que se implantó con éxito un dispositivo de resincronización, exclusivamente a través de una vena cava izquierda persistente no comunicada con la vena cava derecha

Cardiac resynchronization therapy is effective in the treatment of patients with severe heart failure and intraventricular dysynchrony. However, we are sometimes faced with the unexpected presence of a persistent left superior vena cava. We report the case of a patient with dilated cardiomyopathy and left ventricular dysynchrony in which we implanted a resynchronization pacemaker exclusively through a persistent left superior vena cava that did not communicate with the right vena cava

Nutrición en anorexia nerviosa / Nutrition in anorexia nervosa

La anorexia nerviosa es la enfermedad psiquiátrica más frecuente entre las mujeres jóvenes, y se caracteriza por la realización de dietas estrictas con pérdida significativa de peso y un miedo desproporcionado a su ganancia. Esta enfermedad conlleva múltiples complicaciones derivadas tanto de la desnutrición que origina como de los métodos empleados para la pérdida de peso. El tratamiento de esta afección exige un abordaje multidisciplinario y especializado, que se puede efectuar en distintos niveles asistenciales tanto ambulatorio como hospitalario, dependiendo de la situación clínica de los pacientes. Durante el proceso de renutrición, en cualquiera de los niveles asistenciales, se pretende la recuperación de un peso mínimo saludable, la normalización de la conducta alimentaria así como la corrección de las secuelas físicas y psicológicas de la malnutrición. El tratamiento inicial debe enfocarse hacia la restauración del peso, y para alcanzar los objetivos propuestos se seleccionará la vía de acceso de alimentación más apropiada; la vía oral siempre será la de elección, y se optará por la nutrición artificial sólo en situaciones de falta de cooperación o de incorrecta progresión ponderal durante el tratamiento. Si es preciso puede recurrirse a la nutrición artificial, y la nutrición enteral es preferible a la parenteral. Una vez iniciada la realimentación, la progresión en la alimentación se realizará de manera individualizada, y se incrementará progresivamente el aporte calórico hasta alcanzar los objetivos de peso propuestos. Es imprescindible el estricto control hidroelectrolítico, metabólico y físico durante la fase inicial de la realimentación para evitar y diagnosticar complicaciones que pueden aparecer, como el síndrome de renutrición

Anorexia nervosa is the most frequent psychiatric disorder among young women and is characterized by strict dieting with significant weight loss accompanied by a inordinate fear of weight gain. The disorder produces multiple complications arising from both malnutrition and from the methods used to lose weight. Treatment involves a multidisciplinary and specialized approach, which can be carried out in distinct health care levels, both inpatient and outpatient, depending on the patient's clinical status. In all levels of healthcare, during the renutrition process the aim is to recover a minimum healthy weight, achieve normal eating behavior in the patient, and correct the physical and psychological sequelae of malnutrition. Initial treatment should focus on weight gain. To achieve this goal, the most appropriate route of feeding should be selected; the route of choice is always oral, while artificial nutrition is reserved when the patient is uncooperative or there is insufficient weight gain during treatment. If artificial nutrition is unavoidable, enteral nutrition should always be preferred over parenteral nutrition. Once refeeding has been initiated, feeding progression should be individualized, progressively increasing calorie intake until the target weight has been achieved. Strict monitoring of hydroelectrolyte, metabolic and physical status is essential during the initial refeeding phase to prevent or diagnose possible complications, such as refeeding syndrome

Varón de 27 años que presenta episodio de hemoptisis: análisis del manejo clínico y pruebas complementarias / No disponible

Varón de 27 años con escasos antecedentes de interés que presentó episodio de hemoptisis franca, con imagen radiológica de nódulo pulmonar en lóbulo inferior izquierdo de aspecto benigno e hipervascularizado en la endoscopia respiratoria, demostrándose su vascularización en la arteriografía bronquial. Se realizó exéresis de la lesión por lobectomía llegando al diagnóstico anotomopatológico. En el análisis del manejo se repasará la utilidad de las técnicas empleadas en los casos de hemoptisis (AU)

No disponible

[Permanent pacing of the bundle of His after radiofrequency atrioventricular node ablation in patients with suprahisian conduction disturbances]. / Estimulación permanente del haz de His tras ablación mediante radiofrecuencia del nodo auriculoventricular en pacientes con trastorno de la conducción suprahisiano.

INTRODUCTION AND OBJECTIVES: The asynchronic contraction of the left ventricle due to left bundle branch block or right ventricular pacing is inferior from a hemodynamic point of view to the synchronic contraction through the conduction system. Several authors have reported some cases of pump failure and deterioration of mitral regurgitation after AV nodal ablation. Alternative sites of pacing such as the right ventricular outflow tract pacing have been proposed in order to avoid these complications. Direct His bundle pacing might be a new alternative for permanent pacing, however, it has not been extensively evaluated in humans yet. Our aim is to prove the feasibility of permanent His pacing in terms of stability, thresholds and pump function. POPULATION: patients without structural heart disease, selected for AV nodal ablation due to uncontrolled paroxysmal atrial fibrillation, or for pacemaker implantation due to supraHis conduction disturbance, with normal conduction system. An active fixation permanent lead was placed in His position using an steering guidewire and a diagnostic catheter as an anatomical reference. We also implanted a lead in the right atrial appendage and both were connected to a DDDR generator. Pacing thresholds and ecocardiographic ventricular function parameters were evaluated (ejection fraction, cavity size, mitral regurgitation). RESULTS: 12 patients met the inclusion criteria. Successful His pacing was achieved in 8 out of 12 cases (66%) with acceptable thresholds at implantation (1.24 +/- 0.13 volts at 0.5 ms) and during follow up at 3 months (1.31 +/- 0.20 volts at 0.5 ms). Neither a significant change in the ecocardiographic parameters not a deterioration in the clinical status caused by ablation or stimulation was evidenced. CONCLUSION: The His bundle may be the site of choice for long term pacing in patients with AV block and normal infraHis conduction system.

A study on ten short tandem repeat systems: African immigrant and Spanish population data.

This work presents the results obtained from a genetic-population study for the D1S1656 system in the population of Southwest Spain (Huelva, Cádiz and Sevilla), Spaniards of Caucasian origin from North Africa (Ceuta), as well as in the black Central West African and Moroccan immigrant populations in Spain. The results of a study of the autochtonous population of the Canary Islands (n=138), and immigrant Central West African populations in Spain (n=132), obtained for nine short tandem repeat (STR) loci (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820), as well as the amelogenin locus, all contained in Profiler Plus (Perkin-Elmer) PCR amplification kits, are also presented. Except for the FGA and VWA data on immigrant Central West African populations in Spain, no deviations from the Hardy-Weinberg equilibrium were detected.

Ablación del nodo aurículo-ventricular y marcapasos definitivo en el tratamiento de taquiarritmias auriculares: resultados y evolución a medio y largo plazo / Atrioventricular node ablation and permanent pacemaker for atrial tachyarrhythmias treatment: results and evolution at medium and long-term

Objetivos. Evaluar los resultados de la ablación por radiofrecuencia del nodo aurículo-ventricular (NAV) e implante de marcapasos definitivo (MPD) en pacientes con taquiarritmias auriculares graves, refractarias a tratamiento farmacológico y no curables con ablación, y su evolución a medio-largo plazo. Métodos. Revisar retrospectivamente las características clínicas de los enfermos, eficacia de la técnica, modos de estimulación y evolución posterior en cuanto a complicaciones, mortalidad, calidad de vida, tratamiento antiarrítmico, ingresos hospitalarios y en los pacientes que padecían insuficiencia cardíaca (ICC), capacidad funcional. Resultados. Cuarenta pacientes (14 hombres, 26 mujeres) con edad media de 68,7 (10,1) años e historia de arritmias auriculares (32 paroxísticas, 8 crónicas) de 64,8 (57,3) meses de duración. El bloqueo AV se logró en todos los casos (100 por ciento).El modo de estimulación se eligió según la arritmia de base. Hubo cuatro complicaciones no mortales. Tras un seguimiento medio de 15,4 (10,3) meses, se redujo significativamente el número de visitas a urgencias por año (5,47 [5,1] frente a 0,47 [0,97]; p < 0,001), los días de ingreso hospitalario/año (29,3 [15,6] frente a 8,3 [5,2]; p < 0,001), el número de cardioversiones eléctricas por paciente/año (2 [3,1] frente a 0,32 [1,2]; p < 0,01) y el uso medio de antiarrítmicos por paciente (2,61 [1,34] frente a 0,77 [0,75]; p < 0,001). El 92,1 por ciento de los pacientes presentó mejoría de su calidad de vida y mejoró significativamente la clase funcional de la New York Heart Asociation (NYHA) en los enfermos con ICC (2,68 [0,88] frente a [1,73] [0,8]; n = 19; p < 0,01). Se registraron cinco fallecimientos hasta el final del seguimiento. Conclusiones. En nuestra experiencia, la ablación del NAV con implante de MPD constituye una alternativa terapéutica eficaz y segura para aquellos enfermos con taquiarritmias auriculares de dificil control farmacológico y no curables mediante técnicas de ablación convencionales (AU)

A study among the population of Sevilla of death due to submersion.

Death due to submersion is of great interest from the medical-legal point of view, given the increase in nautical activity among children and adults alike over the past number of years. However, the lack of reliable statistical data concerning the impact of this specific form of death in our country must be emphasized. These are the circumstances that have led us to study the incidence of this form of death in a specific area. The population analyzed lived in the city of Sevilla during the period 1967-1993.

Group-specific component (GC) polymorphism in Cádiz (southern Spain).

The genetic polymorphism of group-specific component (GC) was analysed in a sample of 443 healthy unrelated subjects of both sexes resident in the province of Cádiz (Southern Spain). Isoelectric focusing was carried out in polyacrylamide gels followed by staining with coomassie blue R 250. The estimated gene frequencies were as follows: GC*1S = 0.6185; GC*1F = 0.1162; GC*2 = 0.2652.

Polymorphism of the plasminogen (PLG) system in Cádiz Province, southern Spain.

In this work, the authors report a plasminogen (PLG) system genetic-population study in a sample of 378 healthy subjects, of both sexes and unrelated, all resident in the province of Cádiz in Southern Spain. In this study, the PLG types were determined by isoelectric focusing in polyacrylamide gels (PAGIF), followed by staining with Coomassie blue. The genic frequencies obtained were the following: PLG A = 0.833 333 3; PLG B = 0.166 666 7.

Transferrin gene frequencies in Cádiz (southern Spain).

The genetic polymorphism of transferrin (Tf) was studied in a sample of 385 healthy unrelated subjects of both sexes resident in the province of Cádiz (southern Spain). Isoelectric focusing was carried out in polyacrylamide gels, followed by staining with Coomassie Blue R250. The gene frequencies obtained were as follows: Tf C1, 0.7922; Tf C2, 0.1883; Tf C3, 0.0195.