Acute symptomatic seizures and epilepsy after mechanical thrombectomy. A prospective long-term follow-up study.

INTRODUCTION: New treatments for acute ischaemic stroke, such as mechanical thrombectomy, can achieve reperfusion of large ischaemic tissue. Some studies have suggested that reperfusion therapies can increase the risk of suffering acute symptomatic seizure (ASS) and poststroke epilepsy (PSE). The aim of the study was to determine the incidence of ASS and PSE in patients undergoing thrombectomy, and related factors. PATIENTS AND METHODS: This was a retrospective single-centre study including patients with ischaemic stroke and NIHSS> 8 treated with thrombectomy with a follow-up ≥5 years. We evaluated several epidemiological, radiological, clinical and electroencephalographic variables. RESULTS: Of the 344 included patients, 21 (6.1%) presented ASS, 53 (15.40%) died in the acute phase, and 13 (4.46%) died during the first year. The degree of reperfusion (p 0.029), advanced age (p 0.035), and haemorrhagic transformation (p 0.038) increased the risk of suffering ASS, with degree of reperfusion being an independent factor, OR 2.02 (1.21-4.64). The incidence of PSE was 4.12% in the first year, 3.72% in the second, and 1.61% in the fifth. The accumulated incidence at 5 years was 8.93%. Related risk factor for suffering PSE was ASS (p < 0.001), yielding an OR value of 2.00 (1.28-3.145). CONCLUSIONS: Thrombectomy doesn´t increase the risk of ASS. A higher percentage of reperfusion, advanced age, and haemorrhagic transformation are associated with an increased risk of ASS. ASS is a risk factor for suffering PSE. In terms of mortality, having suffered ASS and/or PSE does not increase acute or long-term mortality.

Aplicabilidad del índice SPAN-100 en una cohorte prospectiva y contemporánea de pacientes tratados con rtPA por vía intravenosa en Cataluña / Applicability of the SPAN-100 index in a prospective and contemporary cohort of patients treated with intravenous rtPA in Catalonia

Introducción: Las escalas pronósticas pueden ayudar a seleccionar pacientes para tratamientos de reperfusión. Objetivo: aplicar el índice SPAN-100 en una cohorte de pacientes tratados con rtPA por vía intravenosa y evaluar su capacidad pronóstica. Métodos: Se utilizaron datos del registro prospectivo de reperfusión de Cataluña y se seleccionaron casos consecutivos que recibieron rtPA por vía intravenosa aislado en 2011-2012. A partir del sumatorio de edad y NIHSS se categorizó la cohorte en SPAN-100 positivos [≥ 100] y SPAN-100 negativos [< 100 puntos]. Se determinaron las tasas crudas y ajustadas de hemorragia sintomática (HICS), muerte e independencia funcional (ERm 0-2) a partir del índice SPAN-100 y se calculó la curva ROC para la predicción de las principales medidas de resultado. Resultados: De los 1.685 casos incluidos, 1.405 (83%) eran SPAN-100 negativos. La tasa de HICS ajustada por sexo, ERm preictus, hipertensión, diabetes, dislipemia, cardiopatía isquémica, insuficiencia cardíaca, fibrilación auricular, ictus/AIT previos y tiempo hasta la trombólisis no fue diferente según las dos categorías pero la probabilidad de tener una ERm 0-2 al 3.er mes fue hasta casi 8 veces mayor entre los SPAN-100 negativos. El riesgo de muerte al 3.er mes fue 5 veces superior en los SPAN-100 positivos. El análisis ROC mostró especificidades altas tanto en la predicción de independencia funcional como mortalidad al 3.er mes cuando el punto de corte era de 100. Conclusiones: El índice SPAN-100 es un índice sencillo y de fácil aplicación que puede guiar la selección de pacientes para trombólisis cuando existen dudas razonables y asesorar al paciente/familia acerca de los resultados esperables

Background: Prognostic scales can be helpful for selecting patients for reperfusion treatment. This study aims to assess the prognostic ability of the recently published SPAN-100 index in a large cohort of stroke patients treated with intravenous thrombolysis (IV rtPA). Methods: Using data from the prospective registery of all reperfusion treatments administered in Catalonia, we selected patients treated with IV rtPA alone between 2011 and 2012. The SPAN-100 index was calculated as the sum of age (years) and NIHSS score, and patients in the cohort were classified as SPAN-100 positive [≥ 100] or SPAN-100 negative [< 100]. We measured raw and adjusted rates of symptomatic intracerebral haemorrhage (SICH), mortality, and 3-month functional outcome (mRS 0-2) for each SPAN-100 category. Area under the ROC curve was calculated to predict the main outcome measures. Results: We studied 1685 rtPA-treated patients, of whom 1405 (83%) were SPAN-100 negative. The SICH rates adjusted for sex, pre-stroke mRS, hypertension, diabetes, dyslipidaemia, ischaemic heart disease, heart failure, atrial fibrillation, prior TIA/stroke and time to thrombolysis did not differ between groups, but likelihood of functional independence (mRS 0-2) at 3 months was nearly 8 times higher in the SPAN-100 negative group than in the positive group. Furthermore, the 3-month mortality rate was 5 times higher in the SPAN-100 positive group. ROC curve analysis showed high specificities for predicting both functional independence and 3-month mortality for a cut-off point of 100. Conclusion: The SPAN-100 index is a simple and straightforward method that may be useful for selecting candidates for rtPA treatment in doubtful cases, and for informing patients and their relatives about likely outcomes

Ictus isquémico en pacientes en tratamiento anticoagulante por vía oral / Ischaemic stroke in patients treated with oral anticoagulants

Introducción: Los ictus cardioembólicos tienen peor pronóstico. Su prevención se basa en el tratamiento anticoagulante por vía oral (ACO), cuya principal complicación es el sangrado y, en ocasiones, su ineficacia. Pacientes y métodos: Se ha revisado retrospectivamente a 1.014 pacientes consecutivos que presentaron ictus isquémico entre 2011 y 2013. De ellos, hemos analizado a los 107 pacientes en tratamiento con ACO en el momento del ictus, con especial atención a su etiología, evolución y valor del INR en la fase aguda. Resultados: La edad media ± desviación estándar fue de 71,9 ± 10 años. Tomaba ACO desde hacía 5,9 ± 5,5 años el 98,1% por cardiopatía. Setenta y siete pacientes (72%) tenían INR < 2 y 30 (28%) un INR ≥ 2. Se encontraban en rango terapéutico 9 pacientes (8,4%). La etiología según TOAST fue cardioembólica en el 88,8% y aterotrombótica en el 1,9%. Se suspendió la anticoagulación en 48 pacientes (44,9%): 24 por transformación hemorrágica, 23 por infarto extenso y uno por endarterectomía. Se reintrodujo en 24 de ellos (50%), a los 36 días de media; en los restantes, no fue posible por fallecimiento o secuelas severas. En 9 pacientes (18,7% de potenciales candidatos) se iniciaron nuevos ACO (NACO). La evolución a los 3 meses fue mejor si el INR en la fase aguda > 1,7 respecto al INR ≤ 1,7 (mRS 0-2: 62% vs. 30,8% y fallecimiento 10% vs. 38.4%; p = 0,0004). Conclusiones: Algunos pacientes con ACO presentan ictus isquémicos, en general cardioembólicos, especialmente si el INR es infraterapéutico. Se pueden reintroducir ACO sin complicaciones, siendo el uso de NACO todavía escaso. A pesar de su ineficacia, la evolución es mejor si INR > 1,7 al inicio del ictus

Introduction: Cardioembolic stroke is associated with poorer outcomes. Prevention is based on oral anticoagulant (OAC) therapy. Haemorrhage is the main complication of OACs, which are sometimes ineffective. Patients and methods: We retrospectively reviewed 1014 consecutive patients who suffered an ischaemic stroke between 2011 and 2013, analysing those who were receiving OAC treatment at stroke onset (107 patients in total) with special attention to aetiology, outcomes, and INR value in the acute phase. Results: The mean age (SD) was 71.9 (10) years. Patients had been treated with OACs for 5.9 (5.5) years; 98.1% of them were being treated for heart disease. INR was <2 in 77 patients (72%), and 30 patients (28%) had an INR≥2. Nine patients (8.4%) had INR values within the therapeutic range. According to TOAST classification criteria, 88.8% of strokes were cardioembolic and 1.9% were atherothrombotic. Anticoagulation therapy was discontinued in 48 patients (44.9%) due to haemorrhagic transformation (24 patients), extensive infarction (23), or endarterectomy (1). Therapy was resumed in 24 patients (50%) after a mean lapse of 36 days. This was not possible in the remaining patients because of death or severe sequelae. New OACs (NOACs) were prescribed to 9 patients (18.7% of all potential candidates). At 3 months, patients with INR>1.7 in the acute phase exhibited better outcomes than patients with INR≤1.7 (mRS 0-2 in 62% vs 30.8%; death in 10% vs 38.4%; P=.0004). Conclusions: Some patients taking OACs suffer ischaemic strokes that are usually cardioembolic, especially if INR is below the therapeutic range. OACs can be resumed without complications, and NOACs are still underused. Despite cases in which treatment is ineffective, outcomes are better when INR is above 1.7 at stroke onset

Ischaemic stroke in patients treated with oral anticoagulants. / Ictus isquémico en pacientes en tratamiento anticoagulante por vía oral.

INTRODUCTION: Cardioembolic stroke is associated with poorer outcomes. Prevention is based on oral anticoagulant (OAC) therapy. Haemorrhage is the main complication of OACs, which are sometimes ineffective. PATIENTS AND METHODS: We retrospectively reviewed 1014 consecutive patients who suffered an ischaemic stroke between 2011 and 2013, analysing those who were receiving OAC treatment at stroke onset (107 patients in total) with special attention to aetiology, outcomes, and INR value in the acute phase. RESULTS: The mean age (SD) was 71.9 (10) years. Patients had been treated with OACs for 5.9 (5.5) years; 98.1% of them were being treated for heart disease. INR was <2 in 77 patients (72%), and 30 patients (28%) had an INR≥2. Nine patients (8.4%) had INR values within the therapeutic range. According to TOAST classification criteria, 88.8% of strokes were cardioembolic and 1.9% were atherothrombotic. Anticoagulation therapy was discontinued in 48 patients (44.9%) due to haemorrhagic transformation (24 patients), extensive infarction (23), or endarterectomy (1). Therapy was resumed in 24 patients (50%) after a mean lapse of 36 days. This was not possible in the remaining patients because of death or severe sequelae. New OACs (NOACs) were prescribed to 9 patients (18.7% of all potential candidates). At 3 months, patients with INR>1.7 in the acute phase exhibited better outcomes than patients with INR≤1.7 (mRS 0-2 in 62% vs 30.8%; death in 10% vs 38.4%; P=.0004). CONCLUSIONS: Some patients taking OACs suffer ischaemic strokes that are usually cardioembolic, especially if INR is below the therapeutic range. OACs can be resumed without complications, and NOACs are still underused. Despite cases in which treatment is ineffective, outcomes are better when INR is above 1.7 at stroke onset.

Applicability of the SPAN-100 index in a prospective and contemporary cohort of patients treated with intravenous rtPA in Catalonia. / Aplicabilidad del índice SPAN-100 en una cohorte prospectiva y contemporánea de pacientes tratados con rtPA por vía intravenosa en Cataluña.

BACKGROUND: Prognostic scales can be helpful for selecting patients for reperfusion treatment. This study aims to assess the prognostic ability of the recently published SPAN-100 index in a large cohort of stroke patients treated with intravenous thrombolysis (IV rtPA). METHODS: Using data from the prospective registery of all reperfusion treatments administered in Catalonia, we selected patients treated with IV rtPA alone between 2011 and 2012. The SPAN-100 index was calculated as the sum of age (years) and NIHSS score, and patients in the cohort were classified as SPAN-100 positive [≥ 100] or SPAN-100 negative [< 100]. We measured raw and adjusted rates of symptomatic intracerebral haemorrhage (SICH), mortality, and 3-month functional outcome (mRS 0-2) for each SPAN-100 category. Area under the ROC curve was calculated to predict the main outcome measures. RESULTS: We studied 1685 rtPA-treated patients, of whom 1405 (83%) were SPAN-100 negative. The SICH rates adjusted for sex, pre-stroke mRS, hypertension, diabetes, dyslipidaemia, ischaemic heart disease, heart failure, atrial fibrillation, prior TIA/stroke and time to thrombolysis did not differ between groups, but likelihood of functional independence (mRS 0-2) at 3 months was nearly 8 times higher in the SPAN-100 negative group than in the positive group. Furthermore, the 3-month mortality rate was 5 times higher in the SPAN-100 positive group. ROC curve analysis showed high specificities for predicting both functional independence and 3-month mortality for a cut-off point of 100. CONCLUSION: The SPAN-100 index is a simple and straightforward method that may be useful for selecting candidates for rtPA treatment in doubtful cases, and for informing patients and their relatives about likely outcomes.

Infarto cerebeloso: pronóstico y complicaciones de sus territorios vasculares / Cerebellar infarction: prognosis and complications of vascular territories

Introducción: Los infartos cerebelosos (IC) son infrecuentes pero pueden presentar complicaciones graves. Nuestro objetivo ha sido estudiar las características de los pacientes con IC, así como su evolución, en función del territorio afectado. Pacientes y métodos: Se han recogido datos de 124 pacientes ingresados en nuestro servicio durante un periodo de 5 años, con diagnóstico radiológico de IC, con y sin afectación de otras regiones cerebrales. Resultados: La edad media de nuestra serie es de 65,2 años, con predominio masculino (68,5%). El territorio más afectado fue la arteria cerebelosa posteroinferior (PICA) en el 49,2%, seguido de la arteria cerebelosa superior (ACS) en el 17,7% y cerebelosa anteroinferior (AICA) en el 10,5%. Se afectaron territorios supra-infratentoriales en el 13,7% y dos/tres territorios cerebelosos en el 8,9%. La etiología aterotrombótica fue más prevalente en PICA (p=0,02) y la cardioembólica en la afectación de múltiples territorios (p = 0,04), siendo similares en ACS y AICA. Se produjo transformación hemorrágica en 29 pacientes (23,4%), sobretodo en la afectación de múltiples territorios y en PICA. Se asoció hidrocefalia en 15 pacientes (12,1%, 12 de ellos PICA; p = 0,02), apareciendo de media a los 2,9±1,5 días del inicio del ictus. Al alta, la dependencia funcional (Rankin≥3) era mayor si la afectación territorial era múltiple (64% vs 31-36%; p = 0,05). Se contabilizaron 4 defunciones (3,2%).Conclusiones: Los IC tienen gran heterogeneidad. Sin embargo, cabe destacar que los infartos de PICA son los más prevalentes, su etiología suele ser aterotrombótica y son los más asociados a complicaciones graves, que ocurren durante la primera semana del ictus (AU)

Introduction: Cerebellar infarction (CI) is uncommon, but may result in severe complications. The aim of our study was to determine the characteristics of patients with CI, as well as their outcomes as regards the territories affected. Patients and methods:Data were collected from 124 patients admitted to our department during a five-year period, with a radiological diagnosis of CI, and with or without involvement of other brain territories. Results: The mean age in our series was 65.2 years, with most being male (68.5%). The posterior inferior cerebellar artery (PICA) was the most commonly affected territory at 49.2%, followed by superior cerebellar artery (SCA) at 17.7%, and anterior inferior cerebellar artery (AICA) at 10.5%. There was simultaneous supratentorial involvement in 13.7%, and two or three cerebellar arteries involved in 8.9%. The main aetiology in PICA was atherothrombosis (P=.02). On the other hand, cardio-embolism was the main origin in cases with more than one affected territory (P=.04). No particular aetiology could be found in SCA and AICA. There was haemorrhagic transformation in 29 patients (23.4%), particularly in the PICA and when other territories were involved. There was hydrocephalus in 15 patients (12.1%, 12 of them PICA; P=.02) in 2.9±1.5 days from stroke onset. At discharge, the degree of disability was worse if more than one arterial territory was involved (Rankin≥3, 64% versus 31-36%; P=.05). Four (3.2%) patients died. Conclusions: CI is very heterogeneous. Nevertheless, it is noteworthy that PICA infarction is the most frequent type and its aetiology is usually atherothrombotic. Moreover, it is the territory most frequently associated with severe complications, which take place during the first week of the stroke (AU)

[Cerebellar infarction: prognosis and complications of vascular territories]. / Infarto cerebeloso: pronóstico y complicaciones de sus territorios vasculares.

INTRODUCTION: Cerebellar infarction (CI) is uncommon, but may result in severe complications. The aim of our study was to determine the characteristics of patients with CI, as well as their outcomes as regards the territories affected. PATIENTS AND METHODS: Data were collected from 124 patients admitted to our department during a five-year period, with a radiological diagnosis of CI, and with or without involvement of other brain territories. RESULTS: The mean age in our series was 65.2 years, with most being male (68.5%). The posterior inferior cerebellar artery (PICA) was the most commonly affected territory at 49.2%, followed by superior cerebellar artery (SCA) at 17.7%, and anterior inferior cerebellar artery (AICA) at 10.5%. There was simultaneous supratentorial involvement in 13.7%, and two or three cerebellar arteries involved in 8.9%. The main aetiology in PICA was atherothrombosis (P=.02). On the other hand, cardio-embolism was the main origin in cases with more than one affected territory (P=.04). No particular aetiology could be found in SCA and AICA. There was haemorrhagic transformation in 29 patients (23.4%), particularly in the PICA and when other territories were involved. There was hydrocephalus in 15 patients (12.1%, 12 of them PICA; P=.02) in 2.9 ± 1.5 days from stroke onset. At discharge, the degree of disability was worse if more than one arterial territory was involved (Rankin ≥ 3, 64% versus 31-36%; P=.05). Four (3.2%) patients died. CONCLUSIONS: CI is very heterogeneous. Nevertheless, it is noteworthy that PICA infarction is the most frequent type and its aetiology is usually atherothrombotic. Moreover, it is the territory most frequently associated with severe complications, which take place during the first week of the stroke.

Prediction of early stroke recurrence in transient ischemic attack patients from the PROMAPA study: a comparison of prognostic risk scores.

BACKGROUND: Several clinical scales have been developed for predicting stroke recurrence. These clinical scores could be extremely useful to guide triage decisions. Our goal was to compare the very early predictive accuracy of the most relevant clinical scores [age, blood pressure, clinical features and duration of symptoms (ABCD) score, ABCD and diabetes (ABCD2) score, ABCD and brain infarction on imaging score, ABCD2 and brain infarction on imaging score, ABCD and prior TIA within 1 week of the index event (ABCD3) score, California Risk Score, Essen Stroke Risk Score and Stroke Prognosis Instrument II] in consecutive transient ischemic attack (TIA) patients. METHODS: Between April 2008 and December 2009, we included 1,255 consecutive TIA patients from 30 Spanish stroke centers (PROMAPA study). A neurologist treated all patients within the first 48 h after symptom onset. The duration and typology of clinical symptoms, vascular risk factors and etiological work-ups were prospectively recorded in a case report form in order to calculate established prognostic scores. We determined the early short-term risk of stroke (at 7 and 90 days). To evaluate the performance of each model, we calculated the area under the receiver operating characteristic curve. Cox proportional hazards multivariate analyses determining independent predictors of stroke recurrence using the different components of all clinical scores were calculated. RESULTS: We calculated clinical scales for 1,137 patients (90.6%). Seven-day and 90-day stroke risks were 2.6 and 3.8%, respectively. Large-artery atherosclerosis (LAA) was observed in 190 patients (16.7%). We could confirm the predictive value of the ABCD3 score for stroke recurrence at the 7-day follow-up [0.66, 95% confidence interval (CI) 0.54-0.77] and 90-day follow-up (0.61, 95% CI 0.52-0.70), which improved when we added vascular imaging information and derived ABCD3V scores by assigning 2 points for at least 50% symptomatic stenosis on carotid or intracranial imaging (0.69, 95% CI 0.57-0.81, and 0.63, 95% CI 0.51-0.69, respectively). When we evaluated each component of all clinical scores using Cox regression analyses, we observed that prior TIA and LAA were independent predictors of stroke recurrence at the 7-day follow-up [hazard ratio (HR) 3.97, 95% CI 1.91-8.26, p < 0.001, and HR 3.11, 95% CI 1.47-6.58, p = 0.003, respectively] and 90-day follow-up (HR 2.35, 95% CI 1.28-4.31, p = 0.006, and HR 2.20, 95% CI 1.15-4.21, p = 0.018, respectively). CONCLUSION: All published scores that do not take into account vascular imaging or prior TIA when identifying stroke risk after TIA failed to predict risk when applied by neurologists. Clinical scores were not able to replace extensive emergent diagnostic evaluations such as vascular imaging, and they should take into account unstable patients with recent prior transient episodes.

AFLPMax: a user-friendly application for computing the optimal number of amplified fragment length polymorphism markers needed in phylogenetic reconstruction.

Amplified fragment length polymorphisms (AFLPs) are widely used for phylogenetic inference especially in non-model species. Frequently, trees obtained with other nuclear or mitochondrial markers or with morphological information need additional resolution, increased branch support, or independent data sources (i.e. unlinked loci). In such cases, the use of AFLPs is a quick and cheap option. Computer simulation has shown that dominant AFLP markers lead to less accurate tree topologies than bi-allelic codominant markers such as SNPs, but this difference becomes negligible for shallow trees when using AFLP data sets that include a sufficiently large number of characters. Thus, determining how many AFLP characters are required to recover a given phylogeny is a key issue regarding the appropriateness of AFLPs for phylogenetic reconstruction. Here, we present a user-friendly, java-based graphical interface, AFLPMax, which executes an automatic pipeline of different programs providing the user with the optimal number of AFLP characters needed to recover a given phylogeny with high accuracy and support. Executables for Windows, linux and MacOS X operating systems, source code and user manual are available from: http://webs.uvigo.es/acraaj/AFLPMax.htm.

Factores clínicos y demográficos predictores de éxito del misoprostol en el tratamiento médico del aborto espontáneo / Predictors of success of misoprostol in the treatment of first trimester spontaneous abortion

Objetivos: Determinar posibles predictores de éxito del misoprostol en el tratamiento del aborto espontáneo del primer trimestre. Método: Estudio observacional descriptivo y prospectivo, realizado entre febrero de 2009 y febrero de 2010. Inclusión consecutiva de 248 mujeres con diagnostico ecográfico de aborto espontáneo del primer trimestre con tratamiento médico o quirúrgico, siendo las pacientes las que eligieron la opción terapéutica de acuerdo a los criterios de inclusión para el manejo con misoprostol. En el grupo tratamiento médico se aplicó 800 mcg de misoprostol vaginal/24horas/2 dosis, considerándose como criterio de éxito un endometrio homogéneo con grosor <15 mm en la ecografía realizada al 8° día del tratamiento. Resultados: Influyen en la tasa de éxito del misoprostol la edad de las pacientes (mejor resultado cuanto más joven, p=0,025), número de embarazos (responden mejor las primigestas, p=0,024), existencia o no de abortos (p=0,05) o legrados previos (p=0,028) (la tasa de éxito del misoprostol es mayor en las mujeres que no tienen ningún aborto o legrado previo), y tipo de sangrado vaginal que aparece como efecto secundario del misoprostol (mejorando el pronóstico cuando dicho sangrado es igual o mayor que menstruación, p=0,041). Conclusiones: Hubo predictores de éxito del misoprostol que pueden orientar el manejo, sabiendo que hubo mejor resultado en pacientes jóvenes, primigestas, sin abortos ni legrados previos y con un sangrado vaginal igual o mayor que menstruación.

Objectives: To determine possible predictors of success of misoprostol in the treatment of first trimester spontaneous abortion. Methods: Descriptive observational study and prospectively from February 2009 to February 2010. It were included 248 women which were diagnosed by ultrasound of spontaneous abortion in the first trimester and received medical or surgical treatment, depending on the patient's own choice, provided that the established clinical conditions were present. The protocol applied in the medical treatment group was 800 mcg of vaginal misoprostol/24h/2 dose. It was considered as criteria of success, the presence of a homogeneous endometrium with a thickness <15 mm in the ultrasound examination performed on the 8 th day of treatment. Results: The following variables influence the success rate of misoprostol: patient age (the younger the better outcome, p = 0.025), number of pregnancies (primiparous respond better, p = 0.024), presence or absence of abortions ( p = 0.05) or previous curettage (p = 0.028) (the success rate of misoprostol is higher in women who have no previous abortion or curettage), and type of vaginal bleeding that occurs as a side effect of misoprostol (improving prognosis when bleeding is equal to or greater than the rule, p = 0.041). Conclusions: We found predictors of success of misoprostol, which can guide the management knowing that better results can get obtained in younger patients, primigravida, no previous abortions or curettage and with a vaginal bleeding equal to or greater than the rule.

The relative contribution of band number to phylogenetic accuracy in AFLP data sets.

We examined the effect of increasing the number of sampled amplified fragment length polymorphism (AFLP) bands to reconstruct an accurate and well-supported AFLP-based phylogeny. In silico AFLP was performed using simulated DNA sequences evolving along balanced and unbalanced model trees with recent, uniform and ancient radiations and average branch lengths (from the most internal node to the tip) ranging from 0.02 to 0.05 substitutions per site. Trees were estimated by minimum evolution (ME) and maximum parsimony (MP) methods from both DNA sequences and virtual AFLP fingerprints. The comparison of the true tree with the estimated AFLP trees suggests that moderate numbers of AFLP bands are necessary to recover the correct topology with high bootstrap support values (i.e. >70%). Fewer numbers of bands are necessary for shorter tree lengths and for balanced than for unbalanced tree topologies. However, branch length estimation was rather unreliable and did not improve substantially after a certain number of bands were sampled. These results hold for different levels of genome coverage and number of taxa analysed. In silico AFLP using bacterial genomic DNA sequences recovered a well-supported tree topology that mirrored an empirical phylogeny based on a set of 31 orthologous gene sequences when as few as 263 AFLP bands were scored. These results suggest that AFLPs may be an efficient alternative to traditional DNA sequencing for accurate topology reconstruction of shallow trees when not very short ancestral branches exist.

Acute administration of grape seed proanthocyanidin extract modulates energetic metabolism in skeletal muscle and BAT mitochondria.

Proanthocyanidin consumption might reduce the risk of developing several pathologies, such as inflammation, oxidative stress and cardiovascular diseases. The beneficial effects of proanthocyanidins are attributed to their antioxidant properties, although they also can modulate gene expression at the transcriptional level. Little is known about the effect of proanthocyanidins on mitochondrial function and energy metabolism. In this context, the objective of this study was to determine the effect of an acute administration of grape seed proanthocyanidin extract (GSPE) on mitochondrial function and energy metabolism. To examine this effect, male Wistar rats fasted for fourteen hours, and then they were orally administered lard oil containing GSPE or were administered lard oil only. Liver, muscle and brown adipose tissue (BAT) were used to study enzymatic activity and gene expression of proteins related to energetic metabolism. Moreover, the gastrocnemius muscle and BAT mitochondria were used to perform high-resolution respirometry. The results showed that, after 5 h, the GSPE administration significantly lowers plasma triglycerides, free fatty acids, glycerol and urea concentrations. In skeletal muscle, GSPE lowers FATP1 mRNA levels and increases mitochondrial oxygen consumption, using pyruvate as the substrate, suggesting a promotion of glycosidic metabolism. Furthermore, GSPE increased the genetic expression of key genes in energy metabolism such as peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC1α), and modulated the enzyme activity of proteins, which are involved in the citric acid cycle and electron transport chain (ETC) in BAT. In conclusion, GSPE affects mainly the skeletal muscle and BAT mitochondria, increasing their oxidative capacity rapidly after acute supplementation.

An experimental evaluation with Drosophila melanogaster of a novel dynamic system for the management of subdivided populations in conservation programs.

A dynamic method (DM) recently proposed for the management of captive subdivided populations was evaluated using the pilot species Drosophila melanogaster. By accounting for the particular genetic population structure, the DM determines the optimal mating pairs, their contributions to progeny and the migration pattern that minimize the overall co-ancestry in the population with a control of inbreeding levels. After a pre-management period such that one of the four subpopulations had higher inbreeding and differentiation than the others, three management methods were compared for 10 generations over three replicates: (1) isolated subpopulations (IS), (2) one-migrant-per-generation rule (OMPG), (3) DM aimed to produce the same or lower inbreeding coefficient than OMPG. The DM produced the lowest co-ancestry and equal or lower inbreeding than the OMPG method throughout the experiment. The initially lower fitness and lower variation for nine microsatellite loci of the highly inbred subpopulation were restored more quickly with the DM than with the OMPG method. We provide, therefore, an empirical illustration of the usefulness of the DM as a conservation protocol for captive subdivided populations when pedigree information is available (or can be deduced) and manipulation of breeding pairs is possible.

Semi-quantitative differences in gene transcription profiles between sexes of a marine snail by a new variant of cDNA-AFLP analysis.

A variant of the cDNA-AFLP method coupled to an automated sequencer was used to quantify transcripts differentially expressed between sexes of the marine snail Littorina saxatilis. First, we conducted a validation study of the technique using known concentrations of a commercial marker. Second, we analysed six replicates of males and females from a population showing no apparent sexual dimorphism. The results confirm that the method can be properly used within the range of DNA concentrations utilized. In addition, we detected a small percentage of spots (1.8%) differentially expressed between sexes, as expected from a low to moderately sexual dimorphic species.

Grape seed proanthocyanidins correct dyslipidemia associated with a high-fat diet in rats and repress genes controlling lipogenesis and VLDL assembling in liver.

OBJECTIVE: To determine whether proanthocyanidins can protect against dyslipidemia induced by a high-fat diet (HFD) and to address the mechanisms that underlie this hypolipidemic effect. DESIGN AND MEASUREMENTS: Female Wistar rats were fed on a HFD for 13 weeks. They were divided into two groups, one of which was treated with a grape seed proanthocyanidin extract (25 mg kg(-1) of body weight) for 10 days. Plasma and liver lipids were measured by colorimetric and gravimetric analysis. Liver, muscle and adipose tissue were used to study the expression of genes involved in the synthesis and oxidation of fatty acids and lipoprotein homeostasis by real-time RT-PCR. RESULTS: The administration of proanthocyanidins normalized plasma triglyceride and LDL-cholesterol (both parameters significantly increased with the HFD) but tended to decrease hypercholesterolemia and fatty liver. Gene expression analyses revealed that proanthocyanidins repressed both the expression of hepatic key regulators of lipogenesis and very low density lipoprotein (VLDL) assembling such as SREBP1, MTP and DGAT2, all of which were overexpressed by the HFD. CONCLUSION: These findings indicate that natural proanthocyanidins improve dyslipidemia associated with HFDs, mainly by repressing lipogenesis and VLDL assembly in the liver, and support the idea that they are powerful agents for preventing and treating lipid altered metabolic states.

Evolutionary implications of discordant clines across the Baltic Mytilus hybrid zone (Mytilus edulis and Mytilus trossulus).

It is well established that reproductive isolation often arises from genome incompatibilities and that genes encoding reproductive traits are less prone to introgression. Hybrid zones of Mytilus trossulus and Mytilus edulis provide an intriguing model to assess reproductive isolation. Although gene flow is restricted in North America, introgression is pervasive in the Baltic. This study aimed at analyzing the shape of multilocus clines across the Baltic contact zone between M. edulis and M. trossulus to infer mechanisms of restriction to gene flow. We use maximum likelihood methods to construct the best fitting individual clines for five markers located on biparentally inherited autosomes and paternally and maternally inherited mitochondrial DNA (mtDNA). Strong cline shape differences among markers suggest that reproductive isolation arising from genome-wide incompatibilities is weak, and that these discrepancies possibly result from genetic drift, hybrid zone movement or marker-specific selection. However, the finding of a common cline center for M7 lysin (involved in fertilization) and paternally transmitted mtDNA (causing nuclear-mitochondrial incompatibilities in hybrids) suggest that these loci may play a role in incomplete reproductive isolation.

Interspecies transfer of female mitochondrial DNA is coupled with role-reversals and departure from neutrality in the mussel Mytilus trossulus.

Mussels of the genus Mytilus have distinct and highly diverged male and female mitochondrial DNA (mtDNA) genomes with separate routes of inheritance. Previous studies of European populations of Mytilus trossulus demonstrated that 33% of males are heteroplasmic for a second mtDNA genome of increased length and that hybridization with Mytilus edulis does not block mtDNA introgression, in contrast to reports for American populations. Here, we demonstrate that the female mtDNA type of M. edulis has replaced the resident female mtDNA type of European M. trossulus. This is supported by COIII sequence data indicating that the female mtDNA of European M. trossulus is very similar to that of M. edulis and that in phylogenetic trees, the mtDNAs of these two species cluster together but separately from American M. trossulus sequences, the latter not being disturbed by introgressive hybridization. We also provide evidence that the mtDNA genome of increased length found in heteroplasmic males of European M. trossulus derives from a recent partition of an introgressed M. edulis female type into the male route of transmission. Neutrality tests reveal that European populations of M. trossulus display an excess of replacement polymorphism within the female mtDNA type with respect to conspecific American populations, as well as a significant excess of rare variants, of a similar magnitude to those previously reported for the invading European M. edulis mtDNA. Results are consistent with a nearly neutral model of molecular evolution and suggest that selection acting on European M. trossulus mtDNA is largely independent of the nuclear genetic background.

Nonneutral evolution and differential mutation rate of gender-associated mitochondrial DNA lineages in the marine mussel Mytilus.

Mussels have two types of mitochondrial DNA (mtDNA). The M type is transmitted paternally, and the F type is transmitted maternally. To test hypotheses of the molecular evolution of both mtDNA genomes, 50 nucleotide sequences were obtained for 396 bp of the COIII gene of European populations of Mytilus edulis and the Atlantic and Mediterranean forms of M. galloprovincialis. Analysis based on the proportion of synonymous and nonsynonymous substitutions indicate that mtDNA is evolving in a non-neutral and complex fashion. Previous studies on American mussels demonstrated that the F genome experiences a higher purifying selection and that the M genome evolves faster. Here we show that these patterns also hold in European populations. However, in contrast to American populations, where an excess of replacement substitution between F and M lineages has been reported, a significant excess of replacement polymorphism within mtDNA lineages is observed in European populations of M. galloprovincialis. European populations also show an excess of replacement polymorphism within the F but not within the M genome with respect to American M. trossulus, as well as a consistent pattern of excess of rare variants in both F and M genomes. These results are consistent with a nearly neutral model of molecular evolution and a recent relaxation of selective constraints on European mtDNA. Levels of diversity are significantly higher for the M than F genome, and the M genome also accumulates synonymous and nonsynonymous substitutions at a higher rate, in contrast with earlier reports where no difference for the synonymous rate was observed. It is suggested that a subtle balance between relaxed selection and a higher mutation rate explains the faster evolutionary rate of the M lineage.

Sex-biased heteroplasmy and mitochondrial DNA inheritance in the mussel Mytilus galloprovincialis Lmk.

An exceptional mode of mtDNA inheritance involving separate maternal and paternal transmission routes has been reported recently in the mussel Mytilus edulis. This mode of inheritance provides an explanation for the high levels of heteroplasmy for two highly diverged genomes observed in males of this species. Here we provide evidence for a similar pattern of heteroplasmy in Atlantic and Mediterranean forms of the related mussel M. galloprovincialis. The results support the hypothesis that this mode of mtDNA inheritance has an ancient origin. In addition, the detection of some heteroplasmic females suggests preferential, rather than exclusive, transmission within male and female lines of descent. We also present evidence that the two highly diverged genomes display a parallel split between the Atlantic and Mediterranean forms, consistent with neutral evolution.