Seed Collection in Temperate Trees-Clean, Fast, and Effective Extraction of Populus Seeds for Laboratory Use and Long-term Storage.

Seeds ensure the growth of a new generation of plants and are thus central to maintaining plant populations and ecosystem processes. Nevertheless, much remains to be learned about seed biology and responses of germinated seedlings to environmental challenges. Experiments aiming to close these knowledge gaps critically depend on the availability of healthy, viable seeds. Here, we report a protocol for the collection of seeds from plants in the genus Populus. This genus comprises trees with a wide distribution in temperate forests and with economic relevance, used as scientific models for perennial plants. As seed characteristics can vary drastically between taxonomic groups, protocols need to be tailored carefully. Our protocol takes the delicate nature of Populus seeds into account. It uses P. deltoides as an example and provides a template to optimize bulk seed extraction for other Populus species and plants with similar seed characteristics. The protocol is designed to only use items available in most labs and households and that can be sterilized easily. The unique characteristics of this protocol allow for the fast and effective extraction of high-quality seeds. Here, we report on seed collection, extraction, cleaning, storage, and viability tests. Moreover, extracted seeds are well suited for tissue culture and experiments under sterile conditions. Seed material obtained with this protocol can be used to further our understanding of tree seed biology, seedling performance under climate change, or diversity of forest genetic resources. Key features • Populus species produce seeds that are small, delicate, non-dormant, with plenty of seed hair. Collection of seed material needs to be timed properly. • Processing, seed extraction, seed cleaning, and storage using simple, sterilizable laboratory and household items only. Obtained seeds are pure, high quality, close to 100% viability. • Seeds work well in tissue culture and in experiments under sterile conditions. • Extractability, speed, and seed germination were studied and confirmed for Populus deltoides as an example. • Can also serve as template for bulk seed collection from other Populus species and plant groups that produce delicate seeds (with no or little modifications). Graphical overview.

Imagen endoscópica de afectación gastrointestinal por linfoma del manto / Endoscopic image of gastrointestinal involvement by mantle cell lymphoma

No disponible

Resolución endoscópica de perforación gástrica con penetración a páncreas por espina de pescado / Endoscopic resolution of gastric perforation with pancreas penetration by a fish bone

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Transarterial chemoembolisation in intermediate-stage hepatocellular carcinoma. Survey on clinical practice in hospitals in the Madrid Region.

BACKGROUND: Transarterial chemoembolisation (TACE), having demonstrated survival benefits, is the treatmentof choice in intermediate-stage hepatocellular carcinoma, although there is great heterogeneity in its clinical application. MATERIAL AND METHODS: A survey was sent to the Madrid Regional hospitals to assess applicability, indications and treatment protocols. The assessment was made overall and according to the type of hospital (groups A vs. B and C). RESULTS: Seventeen out of 22 hospitals responded (8/8 group A, 9/ 14 group B-C). All do/indicate transarterial chemoembolisation, 13/17 at their own facilities. Eight of the 17 hospitals have multidisciplinary groups (5/8 A, 3/9 B-C). Nine hospitals perform > 20 procedures/year (7 group A), and 6 from group B-C request/perform < 10/year. It is performed on an "on-demand" basis in 12/17. In 5 hospitals, all the procedures use drug-eluting beads loaded with doxorubicin. The average number of procedures per patient is 2. The mean time from diagnosis of hepatocellular carcinoma to transarterial chemoembolisation is ≤ 2 months in 16 hospitals. In 11/17 hospitals, response is assessed by computed tomography. Radiological response is measured without specific criteria in 12/17 and the other five hospitals (4 group A) assessed using standardised criteria. CONCLUSION: Uniformity among the Madrid Regional hospitals was found in the indication and treatment regimen. The use of DEB-TACE has become the preferred form of TACE in clinical practice. The differentiating factors for the more specialised hospitals are a larger volume of procedures, decision-making by multidisciplinary committees and assessment of radiological response more likely to be standardised.

Evolución de cirrosis biliar primaria a síndrome de solapamiento con hepatitis autoinmune en paciente con hepatitis B crónica / Primary biliary cirrhosis-autoimmune hepatitis overlap syndrome in a patient with chronic hepatitis B infection

Se presenta el caso de una paciente con diagnóstico previo de cirrosis biliar primaria (CBP) y hepatitis B crónica en fase de portador inactivo que, en tratamiento con ácido ursodesoxicólico, presentó elevación de las transaminasas, sin datos de reactivación de su hepatitis B crónica. Se realizó biopsia hepática en la que se observaron cambios compatibles con hepatitis autoinmune (HAI) sobreañadida. Las alteraciones analíticas respondieron al tratamiento con budesonida. Se realiza una revisión sobre la entidad clínica del síndrome de solapamiento CBP-HAI y las características especiales de esta paciente para alcanzar el diagnóstico y su tratamiento (AU)

We describe the case of a female patient with a previous diagnosis of primary biliary cirrhosis (PBC) and chronic hepatitis B in inactive phase who developed increased transaminase levels with no evidence of hepatitis B virus reactivation while receiving ursodeoxycholic acid treatment. A liver biopsy showed changes compatible with overlapping autoimmune hepatitis (AIH). Budesonide treatment achieved normalization of transaminase levels. We provide a review of PBC and AIH overlap syndrome and discuss the particular features of this case that led us to this diagnosis, as well as the treatment provided (AU)

[Primary biliary cirrhosis-autoimmune hepatitis overlap syndrome in a patient with chronic hepatitis B infection]. / Evolución de cirrosis biliar primaria a síndrome de solapamiento con hepatitis autoinmune en paciente con hepatitis B crónica.

We describe the case of a female patient with a previous diagnosis of primary biliary cirrhosis (PBC) and chronic hepatitis B in inactive phase who developed increased transaminase levels with no evidence of hepatitis B virus reactivation while receiving ursodeoxycholic acid treatment. A liver biopsy showed changes compatible with overlapping autoimmune hepatitis (AIH). Budesonide treatment achieved normalization of transaminase levels. We provide a review of PBC and AIH overlap syndrome and discuss the particular features of this case that led us to this diagnosis, as well as the treatment provided.

Optimización del cultivo de líquido ascítico en el contexto clínico de la peritonitis bacteriana espontánea / Optimization of ascitic fluid culture in spontaneous bacterial peritonitis

Introducción y objetivosLa peritonitis bacteriana espontánea (PBE) es la infección del líquido ascítico (LA) en ausencia de foco infeccioso intraperitoneal en pacientes con enfermedad hepática e hipertensión portal. El objetivo del estudio fue la evaluación de la rentabilidad de los cultivos microbiológicos tras la optimización de la sistemática en su recogida en la práctica clínica habitual.MétodosSe analizaron 2 series, la primera retrospectiva con datos de 156 episodios de PBE consecutivos de enero de 2003 a diciembre de 2005 (grupo R), y la segunda con un grupo prospectivo desde octubre de 2007 a octubre de 2008, constituido por 79 episodios, en los que se optimizó el protocolo diagnóstico de PBE (grupo P), con la recogida sistemática de 10ml de LA por bote de hemocultivo.ResultadosLas características basales epidemiológicas y clínicas entre los episodios fueron similares, salvo mayor incidencia de insuficiencia renal, ascitis a tensión e íleo adinámico, y en la etiología de la hepatopatía, más frecuente de origen etílico, en el grupo P. Se consiguió aislamiento microbiológico en LA en el 12,2% de los episodios del grupo R, frente al 53,2% del grupo P (p=0,001) y en hemocultivos en el 8,5 y 26,6% (p=0,001), respectivamente. Se obtuvo aislamiento microbiológico en el 65,8% de los episodios del grupo P frente al 19,2% del grupo R (OR 8; IC 95%: 4,4-14,9; p=0,001). El microorganismo más frecuentemente aislado en la en el grupo P fue Escherichia coli (42,9%).ConclusionesLa optimización en la realización de la sistemática diagnóstica en la práctica asistencial del paciente cirrótico con ascitis aumenta su rendimiento microbiológico diagnóstico (AU)

Introduction and aimsSpontaneous bacterial peritonitis (SBP) is ascitic fluid (AF) infection in the absence of an intraperitoneal source of infection in patients with liver disease and portal hypertension. The aim of this study was to evaluate the yield of microbiological cultures to optimize their collection and systematic implementation in routine clinical practice.MethodsWe analyzed two cohorts: the first consisted of retrospective data from the clinical records of 156 consecutive episodes of SBP from January 2003 to December 2005 (group R), and the second was composed of data collected prospectively from October 2007 to October 2008, consisting of 79 episodes (group P), with systematic inoculation of 10 cc of AF in each blood culture bottle.ResultsNo significant differences were demonstrated in baseline epidemiological or clinical characteristics among episodes, except the more frequent presence of alcoholic liver disease and the incidence of tense ascites, paralytic ileus and kidney failure at diagnosis in group P. Microbiological isolation in AF was achieved in 12.2% of the episodes in group R compared with 53.2% in group P (p=0.001) and 8.5 and 26.6% (p=0.001) in blood culture, respectively. Microbiological isolation was achieved in 65.8% of episodes in group P versus 19.2% in group R (OR 8; 95% CI: 4.4-14.9; p=0.001). The most frequently isolated organism in AF in group P was Escherichia coli (42.9%).ConclusionsOptimizing the diagnostic procedure in cirrhotic patients with ascites significantly increases diagnostic microbiologic performance(AU)

Estudio descriptivo de la experiencia sobre carcinoma hepatocelular en hígado no cirrótico / Descriptive study of hepatocellular carcinoma in noncirrhotic liver

Introducción y objetivoEl carcinoma hepatocelular (CHC) es la sexta causa de cáncer. En el mundo occidental su incidencia está en aumento y la gran mayoría asientan sobre una cirrosis hepática. El objetivo del estudio fue evaluar las características del CHC en pacientes sin cirrosis.MétodosSe realizó una revisión retrospectiva de 469 pacientes con diagnóstico de CHC durante el periodo de enero de 2003 a diciembre de 2007. Se seleccionaron aquellos pacientes que cumplían criterios histológicos, o la conjunción de criterios clínicos, analíticos, radiológicos y/o de hemodinámica hepática, de ausencia de cirrosis hepática.ResultadosSe identificaron 29 pacientes con diagnóstico de CHC sobre hígado no cirrótico (6,2%). El 86,2% eran varones y con una mediana de edad de 61,6 (23 - 82) años. La enfermedad hepática fue hepatopatía crónica con fibrosis leve-moderada en el 68,9%. El 62,1% de los pacientes se encontraban asintomáticos al diagnóstico. El CHC era un nódulo único en el 86,2%, y la mediana del nódulo mayor fue 46 (20 - 150) mm. Se evidenció invasión vascular macroscópica en dos pacientes y metástasis óseas en otros dos. Se realizó tratamiento activo en 25 pacientes: 15 resección quirúrgica, 6 radiofrecuencia y 4 quimioembolización transarterial. La supervivencia acumulada a 1, 2 y 3 años, fue del 84,3; 67,2 y 50,1%, respectivamente.ConclusionesEl CHC sobre hígado no cirrótico es una patología poco frecuente, siendo tumores de gran tamaño al diagnóstico. A pesar de ello, se pueden aplicar tratamientos con intención curativa y alcanzar una supervivencia superior a la esperable en pacientes con cirrosis(AU)

Introduction and objectiveHepatocellular carcinoma (HCC) is the sixth leading cause of cancer. In western countries its impact is steadily growing and most of these tumors arise in cirrhotic liver. The aim of this study was to evaluate the incidence and characteristics of HCC developing in noncirrhotic patients. Methods: We conducted a retrospective review of 469 patients diagnosed with HCC between January 2003 and December 2007. Patients who met the histological criteria, or a combination of clinical, laboratory, imaging and hemodynamic criteria for the absence of cirrhosis were included.ResultsWe identified 29 patients with a diagnosis of HCC in non-cirrhotic liver (6.2%). Most (86.2%) were men and the median age was 61.6 (23 - 82) years. The most frequent histology of the liver was mild-moderate liver fibrosis (68.9%) and 62.1% of the patients were asymptomatic at diagnosis. HCC was a solitary nodule in 86.2%, and the median size of the main nodule was 46 (20 - 150) mm. Macroscopic vascular invasion was demonstrated in two patients and bone metastases in a further two patients. Active treatment was provided in 25 patients, consisting of tumoral resection in 15, radiofrequency ablation in six and transarterial chemoembolization in four. The overall cumulative survival at 1, 2 and 3years was 84.3%, 67.2% and 50.1%, respectively.ConclusionHCC arising in non-cirrhotic liver is uncommon and there is wide tumor extension at diagnosis. Nevertheless, in most patients, treatment with curative intent can be applied, achieving better survival than that expected patients with cirrhosis(AU)

[Descriptive study of hepatocellular carcinoma in noncirrhotic liver]. / Estudio descriptivo de la experiencia sobre carcinoma hepatocelular en hígado no cirrótico.

INTRODUCTION AND OBJECTIVE: Hepatocellular carcinoma (HCC) is the sixth leading cause of cancer. In western countries its impact is steadily growing and most of these tumors arise in cirrhotic liver. The aim of this study was to evaluate the incidence and characteristics of HCC developing in noncirrhotic patients. METHODS: We conducted a retrospective review of 469 patients diagnosed with HCC between January 2003 and December 2007. Patients who met the histological criteria, or a combination of clinical, laboratory, imaging and hemodynamic criteria for the absence of cirrhosis were included. RESULTS: We identified 29 patients with a diagnosis of HCC in non-cirrhotic liver (6.2%). Most (86.2%) were men and the median age was 61.6 (23 - 82) years. The most frequent histology of the liver was mild-moderate liver fibrosis (68.9%) and 62.1% of the patients were asymptomatic at diagnosis. HCC was a solitary nodule in 86.2%, and the median size of the main nodule was 46 (20 - 150) mm. Macroscopic vascular invasion was demonstrated in two patients and bone metastases in a further two patients. Active treatment was provided in 25 patients, consisting of tumoral resection in 15, radiofrequency ablation in six and transarterial chemoembolization in four. The overall cumulative survival at 1, 2 and 3 years was 84.3%, 67.2% and 50.1%, respectively. CONCLUSION: HCC arising in non-cirrhotic liver is uncommon and there is wide tumor extension at diagnosis. Nevertheless, in most patients, treatment with curative intent can be applied, achieving better survival than that expected patients with cirrhosis.

[Optimization of ascitic fluid culture in spontaneous bacterial peritonitis]. / Optimización del cultivo de líquido ascítico en el contexto clínico de la peritonitis bacteriana espontánea.

INTRODUCTION AND AIMS: Spontaneous bacterial peritonitis (SBP) is ascitic fluid (AF) infection in the absence of an intraperitoneal source of infection in patients with liver disease and portal hypertension. The aim of this study was to evaluate the yield of microbiological cultures to optimize their collection and systematic implementation in routine clinical practice. METHODS: We analyzed two cohorts: the first consisted of retrospective data from the clinical records of 156 consecutive episodes of SBP from January 2003 to December 2005 (group R), and the second was composed of data collected prospectively from October 2007 to October 2008, consisting of 79 episodes (group P), with systematic inoculation of 10 cc of AF in each blood culture bottle. RESULTS: No significant differences were demonstrated in baseline epidemiological or clinical characteristics among episodes, except the more frequent presence of alcoholic liver disease and the incidence of tense ascites, paralytic ileus and kidney failure at diagnosis in group P. Microbiological isolation in AF was achieved in 12.2% of the episodes in group R compared with 53.2% in group P (p = 0.001) and 8.5 and 26.6% (p = 0.001) in blood culture, respectively. Microbiological isolation was achieved in 65.8% of episodes in group P versus 19.2% in group R (OR 8; 95% CI: 4.4-14.9; p = 0.001). The most frequently isolated organism in AF in group P was Escherichia coli (42.9%). CONCLUSIONS: Optimizing the diagnostic procedure in cirrhotic patients with ascites significantly increases diagnostic microbiologic performance.

Portal hypertensive colopathy is associated with portal hypertension severity in cirrhotic patients.

AIM: To assess the prevalence of portal hypertension (PH) related colorectal lesions in liver transplant candidates, and to evaluate its association with the severity of PH. METHODS: Between October 2004 and December 2005, colonoscopy was performed in 92 cirrhotic liver transplant candidates. We described the lesions resulting from colorectal PH and their association with the grade of PH in 77 patients who underwent measurement of hepatic venous pressure gradient (HVPG). RESULTS: Mean age was 55 years and 80.7% of patients were men. The main etiology of cirrhosis was alcoholism (45.5%). Portal hypertensive colopathy (PHC) was found in 23.9%, colonic varices in 7.6% and polyps in 38% of patients (adenomatous type 65.2%). One asymptomatic patient had a well-differentiated adenocarcinoma. The manifestations of colorectal PH were not associated with the etiology of liver disease or with the Child-Pugh grade. Ninety percent of patients with colopathy presented with gastroesophageal varices (GEV), and 27.5% of patients with GEV presented with colopathy (P = 0.12). A relationship between higher values of HVPG and presence of colopathy was observed (19.9 +/- 6.2 mmHg vs 16.8 +/- 5.4 mmHg, P = 0.045), but not with the grade of colopathy (P = 0.13). Preneoplastic polyps and neoplasm (P = 0.02) and spontaneous bacterial peritonitis (P = 0.006) were more prevalent in patients with colopathy. We did not observe any association between previous beta-blocker therapy and the presence of colorectal portal hypertensive vasculopathy. CONCLUSION: PHC is common in cirrhotic liver transplant candidates and is associated with higher portal pressure.

Peritonitis bacteriana espontánea en pacientes con cirrosis mayores de 65 años / Spontaneous bacterial peritonitis in cirrhotic patients over 65 years

Fundamento y objetivo: Estudiar los episodios de peritonitis bacteriana espontánea (PBE) en pacientes cirróticos mayores de 65 años y comparar sus características y su evolución clínica dentro de una cohorte de pacientes cirróticos con PBE. Material y métodos: Se realizó un estudio observacional y retrospectivo sobre todas las altas hospitalarias con diagnóstico de PBE en el período de enero de 2003 a diciembre de 2005, con un total de 158 episodios. Se valoró el episodio de PBE desde su diagnóstico hasta la muerte o el fin de seguimiento por otras causas del paciente. Se dividió la cohorte en 2 grupos, según edad superior a 65 años (grupo 1: 73 episodios) o edad inferior o igual a 65 años (grupo 2: 86 episodios). Se realizó un estudio descriptivo y comparativo de ambos grupos y posteriormente se realizó un análisis de supervivencia mediante curvas de Kaplan-Meier (comparadas mediante el test log-rank) y valoración de los factores predictivos independientes mediante regresión de Cox en la cohorte global. Resultados: En el grupo 1 el 74% de los pacientes eran varones y la edad media (desviación estándar) fue de 72,8 (5,6) años, y la distribución según la clasificación de Child-Pugh fue del 8,2 (A), del 68,5 (B) y del 23,3% (C). La etiología de la cirrosis fue vírica en el 65,8% de los casos. Comparado con el grupo 2, se observó mayor frecuencia de insuficiencia renal al ingreso, prevalencia de hepatocarcinoma y etiología vírica, mientras que las cifras de bilirrubina eran superiores en el grupo 2. La mortalidad intrahospitalaria fue del 23,3%. Se observó una supervivencia a los 3 y a los 6 meses del 69,5 y del 54,8% y en el grupo 2 del 80,3 y del 78,4%, respectivamente (p=0,001). La edad superior a 65 años, la existencia de hepatocarcinoma y la presión arterial media superior a 75mmHg al ingreso fueron factores predictivos independientes de mortalidad (AU)

Aims: Our aim was to study the episodes of spontaneous bacterial peritonitis (SBP) in cirrhotic patients older than 65 years. Their clinical characteristics were compared with a cohort of cirrhotic patients with SBP. Methods: We conducted a retrospective study on all hospital discharged patients diagnosed with SBP in the period from January 2003 to December 2005, obtaining a total of 158 episodes. Two groups of patients were created: those over 65 (group 1: 73 episodes) and those aged }65 years (group 2: 86 episodes). A descriptive study, an analysis of survival by Kaplan Meier curves (compared with log-rank test) and a Cox regression analysis was conducted comparing the two groups. Results: In Group 1, males were 74%, the average age was 72.4 (SD 4.2) years, and the Child-Pugh distribution was 8.2% (A), 68.5% (B) and 23.3% (C).The most usual etiology of cirrhosis was viral (65.8%). In comparison with group 2, there was a superior frequency of renal impairment at admission, hepatocellular carcinoma and a viral etiology. Yet the bilirubin levels were higher in group 2. In-hospital mortality rate was 23.3%. The survival at 3 and 6 months was of 69.5% and 54.8%, while in group 2 it was 80.3% and 78.4% respectively (p=0001). An age> 65 years, the existence of hepatocellular carcinoma and a Mean Arterial Pressure <75mm Hg at admission, in the overall cohort, were independent predictors of mortality. Conclusion: An age above 65 years is an independent predictor of mortality following the development of an episode of spontaneous bacterial peritonitis in cirrhotic patients (AU)

[Spontaneous bacterial peritonitis in cirrhotic patients over 65 years]. / Peritonitis bacteriana espontánea en pacientes con cirrosis mayores de 65 años.

AIMS: Our aim was to study the episodes of spontaneous bacterial peritonitis (SBP) in cirrhotic patients older than 65 years. Their clinical characteristics were compared with a cohort of cirrhotic patients with SBP. METHODS: We conducted a retrospective study on all hospital discharged patients diagnosed with SBP in the period from January 2003 to December 2005, obtaining a total of 158 episodes. Two groups of patients were created: those over 65 (group 1: 73 episodes) and those aged 65 years (group 2: 86 episodes). A descriptive study, an analysis of survival by Kaplan Meier curves (compared with log-rank test) and a Cox regression analysis was conducted comparing the two groups. RESULTS: In Group 1, males were 74%, the average age was 72.4 (SD 4.2) years, and the Child-Pugh distribution was 8.2% (A), 68.5% (B) and 23.3% (C).The most usual etiology of cirrhosis was viral (65.8%). In comparison with group 2, there was a superior frequency of renal impairment at admission, hepatocellular carcinoma and a viral etiology. Yet the bilirubin levels were higher in group 2. In-hospital mortality rate was 23.3%. The survival at 3 and 6 months was of 69.5% and 54.8%, while in group 2 it was 80.3% and 78.4% respectively (p=0001). An age> 65 years, the existence of hepatocellular carcinoma and a Mean Arterial Pressure <75 mm Hg at admission, in the overall cohort, were independent predictors of mortality. CONCLUSION: An age above 65 years is an independent predictor of mortality following the development of an episode of spontaneous bacterial peritonitis in cirrhotic patients.

[Results of colonoscopic examination in the evaluation of cirrhotic candidates for liver transplantation]. / Resultados de la realización de colonoscopia en la evaluación de pacientes cirróticos candidatos a trasplante hepático.

AIM: To assess the presence of preneoplastic and neoplastic colonic lesions, as well as those related to portal hypertensive vasculopathy, and their association with liver disease in cirrhotic patients who are candidates for orthotopic liver transplantation (LT). METHODS: Between October 2004 and December 2005, colonoscopy was performed in 92 patients who were LT candidates, aged >50 years old or aged <50 years old but with clinical indications. RESULTS: Eighty-eight percent of the patients were > 50 years old, the mean age was 55.3 years (29-69) and 81.5% were males. The main etiology of cirrhosis was alcoholic (46.7%), and 21% were Child-Pugh class A. No abnormalities were detected in 20.7%. Polyps were discovered in 38% (35/92) of patients (adenomatous 65.2%; tubular type 86.7%). Six patients with adenomatous polyps had mild dysplasia, and one asymptomatic patient had a well-differentiated adenocarcinoma. An association was found between polyps and male sex (44% males vs 17.6% females; p=0.044) and Child-Pugh grade (63.2% Child A vs 32.9% Child B/C, p=0.016) but not with serum levels of carcinoembryonic antigen (CEA), age or etiology of liver disease. Portal hypertensive colopathy was found in 23.9%, rectal varices in 7.6% and internal or mixed hemorrhoids in 52.3%. CONCLUSION: The prevalence of preneoplastic and neoplastic colonic lesions may support the use of colonoscopy in LT candidates aged >50 years-old or with a history suggesting lower gastrointestinal bleeding or other abnormalities.