Farmacovigilancia de las vacunas frente a la COVID-19 en farmacias comunitarias I. Protocolo / Pharmacovigilance of COVID-19 vaccines in community pharmacies I. Protocol

INTRODUCCIÓN: la aprobación en tiempo récord de las vacunas frente a la COVID-19 hace necesario implementar acciones de farmacovigilancia, tanto de reacciones infrecuentes, como a medio o largo plazo, con el fin de completar su perfil de seguridad.OBJETIVOS: colaborar en la evaluación de la seguridad de las vacunas, mediante la detección y seguimiento de sospechas de reacciones adversas en los usuarios de las farmacias.MATERIAL Y MÉTODOS. Diseño: estudio observacional prospectivo, en farmacias comunitarias de Ourense y Pontevedra, a partir de la administración de las vacunas frente al SARS-CoV-2 en febrero de 2021.Sujetos: usuarios de la farmacia que hayan recibido la vacuna, mayores de edad, y que firmen el consentimiento informado. Variables principales: número y porcentaje de sujetos que presentan al menos una reacción adversa en relación al número de pacientes incorporados al estudio; su número, tipo y frecuencia. Repercusión en su vida diaria. Procedimiento: oferta de participación en el estudio, entrevista en la zona de atención personalizada. Recogida de datos en un cuaderno de registro cumplimentado por el farmacéutico junto con el paciente. Seguimiento decenal presencial o telefónico después de la primera y segunda dosis de vacuna recibidas y mensual a partir del segundo mes de la segunda dosis. A los pacientes vacunados con una reacción considerada leve tras su evaluación, se recomendarán medidas no farmacológicas convenientes y si es necesario, se les indicará un medicamento sin receta adecuado para la resolución o alivio del problema de salud originado por la reacción. Tamaño muestral: para una precisión del 5,0 % en la estimación de una proporción mediante un intervalo de confianza al 95 % bilateral, asumiendo que el porcentaje de sujetos con al menos una reacción adversa sería del 20,0 %, resultó necesario incluir 246 participantes. (AU)

Farmacovigilancia de las vacunas frente a la COVID-19 en farmacias comunitarias II. Reacciones adversas en farmacéuticos y personal auxiliar / Pharmacovigilance of COVID-19 vaccines in community pharmacies II. Adverse reactions in pharmacists and auxiliary personnel

OBJETIVO: conocer tipo y frecuencia de las sospechas de reacciones adversas (RA) experimentadas entre los farmacéuticos comunitarios (FC) y personal auxiliar de las farmacias de Pontevedra con la primera dosis de la vacuna y su repercusión laboral y personal.MÉTODOS . Diseño: estudio observacional transversal, febrero-abril de 2021.Sujetos: farmacéuticos, técnicos y auxiliares de la provincia de Pontevedra que ejercen en contacto con los pacientes, vacunados con la primera dosis frente al SARS-CoV-2, que dieron su consentimiento explícito. Variables: número, tipo y frecuencia de RA, repercusión. Procedimiento: se habilitó un formulario en la web del Colegio de Farmacéuticos de Pontevedra. Se anunció a los FC colegiados de la provincia su existencia y la conveniencia de cumplimentarlo. Tamaño muestral: teniendo en cuenta la metodología de incorporación al estudio, mediante cumplimentación voluntaria del cuestionario, no se consideró procedente. Análisis de resultados: test de chi- cuadrado para el análisis de variables cualitativas, t de Student y Mann-Whitney para cuantitativas. La normalidad comprobada mediante Kolmogorov-Smirnov con corrección de Lilliefors. Significación estadística p<0,05.Resultados 153 participantes manifestaron síntomas compatibles con reactividad a la vacuna, 122 (93,1 %) mujeres y 31 (86,1 %) hombres. 146 (95,4 %) que recibieron Vaxzevria® (VZ), 116 (79,4 %) mujeres y 30 (20,6 %) hombres, y 7 (4,6 %) Comirnaty® Pfizer-BioNtech (CO), 6 (85,7 %) mujeres y 1 (14,3 %) hombres. Se comunicaron 823 RA, 811 (98,5 %) con VZ y 12 (1,5 %) con CO. Reacciones más frecuentes: dolor en punto de inyección, 128 (87,7 %); escalofríos, 107 (73,3 %); dolor muscular, 106 (72,6 %). El número máximo de RA manifestadas por un participante fue 12. (AU)

Farmacovigilancia de las vacunas frente a la COVID-19 en farmacias comunitarias VI. Elección de la segunda dosis / Pharmacovigilance of COVID-19 vaccines in community pharmacies VI. Choice of second dose

INTRODUCCIÓN: en Galicia se administró Vaxzevria® (VZ) como primera dosis de la vacuna frente a Covid-19 a sanitarios y trabajadores esenciales menores de 60 años. Tras la alarma social por problemas de trombosis con trombopenia detectados, la administración sanitaria gallega permitió elegir la vacuna que recibirían como segunda dosis, repitiendo con Vaxzevria® o cambiar a Comirnaty® (CO).OBJETIVOS: conocer el número de vacunados que eligen una 2ª dosis diferente, las razones por la que eligieron una u otra y las fuentes de información utilizadas para tomar la decisión.MÉTODOS: Diseño: estudio observacional transversal, en una farmacia tras la administración de la 2ª dosis de vacuna Covid-19 en mayo/junio de 2021.Sujetos: trabajadores esenciales, vacunados con Vaxzevria®, mayores de edad, que firmaron consentimiento informado. Variables: demográficas, número de vacunados que eligen una vacuna diferente como 2ª dosis, razones para ello, fuentes de información. Procedimiento: las preguntas correspondientes a este estudio estaban incluidas en el cuaderno de recogida de datos utilizado en el proyecto FARMACOVIGILANCIA DE LAS VACUNAS COVID-19. Consideraciones éticas: el estudio del que forma parte recibió informe favorable del Comité Ético de Investigación con Medicamentos de Galicia. Resultados Se entrevistó a 28 personas, 17 (60,7 %) mujeres, 11 (39,3 %) hombres; 19 (67,9 %) sanitarios. 3 (10,7 %) eligieron como 2ª dosis Comirnaty®, 2 mujeres y 1 hombre y 1 hombre no recibió la 2ª dosis por contagio tras la 1ª.Razones para repetir vacuna: recomendación de autoridades sanitarias/instituciones científicas, 13 (54,2 %); “por no cambiar”, 14 (58,3 %); evidencia científica, 8 (33,3 %). 10 (41,7 %) manifestaron más de un motivo. (AU)

Farmacovigilancia de las vacunas frente a la COVID-19 en farmacias comunitarias III. Primera dosis / Pharmacovigilance of COVID-19 vaccines in community pharmacies III. First dose

OBJETIVO: conocer tipo y frecuencia de sospechas de reacciones adversas (RA) experimentadas por la población general tras la primera dosis de la vacuna frente a COVID-19 y la repercusión sobre su actividad diaria.MÉTODOS: se describen detalladamente en el protocolo del Proyecto. Procedimiento: en un cuaderno de recogida de datos ad hoc se registraron datos demográficos, sospechas de RA y demás circunstancias referidas por los encuestados.RESULTADOS: colaboraron 10 farmacias de la provincia de Pontevedra y 2 de Ourense que incorporaron 781 casos válidos, 488 (62,5 %) mujeres y 293 (37,5 %) hombres. Media de edad 56,8 (DE=17,9) (rango=18-97) años. 389 (49,8 %) estaban incluidos en grupos de riesgo: edad ≥65 años 271 (34,7 %), 219 hipertensión arterial, 148 dislipemias, 105 neuropsiquiátricas, 88 cardiológicas, 75 diabetes, 62 EPOC/problemas respiratorios. 119 (15,2 %) anticoagulados. 64 (8,2 %) habían pasado la enfermedad.445 (57,0 %) recibieron Comirnaty® Pfizer-BioNtech (CO), 190 (24,3 %) Vaxzevria® (VZ), 104 (13,3 %) Spikevax® Moderna (SP) y 42 (5,4 %) COVID-19 Vaccine Janssen (JA). La mayoría, 491 (62,9 %) en los “vacunódromos”. 204 (26,1 %) utilizaron medicamentos como profilaxis de posibles RA. 165 (80,1 %) paracetamol.495 (63,4 %) vacunados, 321 mujeres (65,8 %) y 174 (59,4 %) hombres (diferencia no significativa), refirieron al menos una RA: 236 (53,0 %) CO, 157 (82,6 %) VZ, 69 (66,3 %) SP y 33 (80,5 %) JA. El número de RA manifestadas por los encuestados fue 1.419, con máximo de 13 por persona, resultando las más prevalentes, afectando a >10 % de los vacunados: dolor en el punto de inyección 375 (48,0 %), cansancio/fatiga 170 (21,8 %), escalofríos 118 (15,1 %), cefalea 117 (15,0 %), dolor muscular 112 (14,3 %) y fiebre 98 (12,5 %). (AU)

Farmacovigilancia de las vacunas frente a la COVID-19 en farmacias comunitarias V. Comparativa entre dosis / Pharmacovigilance of COVID-19 vaccines in community pharmacies V. Dose comparisons

OBJETIVO: conocer si hubo diferencias entre las dos dosis de la vacuna frente a COVID-19 en cuanto a tipo, frecuencia de sospechas de reacciones adversas (RA) y repercusión sobre la actividad diaria.MÉTODOS: se describen detalladamente en el protocolo del Proyecto. En esta comunicación se presentan las comparaciones estadísticas entre dosis y las relaciones con datos sociodemográficos de los encuestados.RESULTADOS: 10 farmacias de la provincia de Pontevedra y 2 de Ourense incorporaron 781 sujetos, que se redujeron a 693 tras la 2ª dosis. El número de vacunados que refirieron al menos una RA fue 495 (63,4 %) con la 1ª dosis y 312 (45,0 %) con la 2ª, p<0,05. El número de RA disminuyó con la segunda dosis, de 1.419 (1,8 DE=2,2 por vacunado), a 971 (1,2 DE=2,1 por vacunado), p<0,05. 227 encuestados sufrieron RA con las dos dosis, 266 que tuvieron RA con la 1ª no tuvieron con la 2ª, y 85 sin RA con la 1ª, sí tuvieron con la 2ª. Sexo y número de vacunados con RA: sin diferencias significativas entre sexos con la 1ª dosis. Sí con la 2ª, 218 (43,9 %) mujeres y 94 (37,1 %) hombres, p<0,05.No se encontraron diferencias significativas entre sexos con ninguna de las dos dosis en cuanto a la necesidad de atención profesional a causa de las RA ni en su repercusión sobre la actividad diaria de los vacunados. Edad y número de vacunados con RA: se encontró relación significativa inversa. 1ª dosis, media de edad de vacunados con RA 52,3 (DE=17,3) años; sin RA 64,5 (DE=16,5) años. 2ª dosis, media de edad vacunados con RA 51,1 (DE=17,5); sin RA 63,6 (DE=16,5), p<0,05. (AU)

Detección de personas en riesgo de padecer diabetes, en farmacias comunitarias españolas 2021 / Screening of people at risk of diabetes in Spanish community pharmacies 2021

JUSTIFICACIÓN: según el estudio Di@betes, estudio de prevalencia de la diabetes, se encontró que la prevalencia global de diabetes mellitus era del 13,8%, de los cuales aproximadamente la mitad (6%) no estaban diagnosticados. Las personas con diabetes tipo dos no diagnosticada presentan un alto riesgo de padecer otras enfermedades como cardíacas, dislipemias, etc. Por esta razón, la detección precoz y el tratamiento inmediato reducen la gravedad de la enfermedad, así como las futuras complicaciones e ingresos hospitalarios.OBJETIVO: conocer la prevalencia y sus características de personas con riesgo alto y/o muy alto de padecer diabetes utilizando el cuestionario de Findrisc en farmacia comunitaria, con el fin de colaborar en el diagnóstico precoz de la enfermedad.MATERIAL Y MÉTODOS: estudio observacional transversal del 15 al 30 de noviembre de 2021 en las farmacias comunitarias españolas con socios de SEFAC. Inclusión: Usuarios de la farmacia, de ≥45 años, no diagnosticados con autonomía y que consientan participar. Estudio aprobado por el Comité de Ética de la Investigación (CEICA) de Zaragoza. Variable principal: la puntuación del test de Findrisc (media±DS) y (N+%) en intervalos de riesgo. Se calculan las frecuencias relativas para las demás variables categóricas del cuestionario y también media±DS para las cuantitativas. Recogida de datos: a través SEFAC e_XPERT. Procedimiento: formación farmacéuticos participantes, captación en mostrador, realización medidas antropométricas, cumplimentación test Findrisc, información del resultado e intervención farmacéutica: -Si F<15, educación sanitaria, repetir test en 1 o 5 años en función riesgo. -Si F≥15, determinación glucemia basal y/o HbA1c. Glucemia≥110mg/dl y/o HbA1c≥5,7% derivación al médico. (AU)

Frailty and Risk of Falls in Community-Dwelling Older Adults Living in a Rural Setting. The Atahualpa Project.

BACKGROUND: Data supporting a link between frailty and risk of falls is mostly confined to individuals living in urban centers, where risk factors and lifestyles are different from that of rural settings. OBJECTIVE: To assess the association between frailty and risk of falls in older adults living in rural Ecuador. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Community-dwellers aged ≥60 years living in a rural Ecuadorian village, in whom frail status and risk of falls were assessed. MEASUREMENTS: Frailty was evaluated by the Edmonton Frailty Scale (EFS) and risk of falls by the Downton Fall Risk Index (DFRI). Multivariate models were fitted to evaluate whether frailty was associated with risk of falls (dependent variable), after adjusting for demographics, alcohol intake, cardiovascular risk factors, sleep quality, symptoms of depression, and history of an overt stroke. Correlation coefficients were constructed to assess confounders modifying this association. RESULTS: A total of 324 participants (mean age: 70.5±8 years) were included. The mean EFS score was 4.4±2.5 points, with 180 (56%) participants classified as robust, 76 (23%) as pre-frail and 68 (21%) as frail. The DFRI was positive in 87 (27%) participants. In univariate analysis, the EFS score was higher among participants with a positive DFRI (p<0.001). The number of frail individuals was higher (p<0.001), while that of robust individuals was lower (p<0.001) among those with a positive DFRI. Adjusted logistic regression models showed no association between frailty and the DFRI. Correlation coefficients showed that age, high glucose levels, and history of an overt stroke tempered the association between frailty and the risk of falls found in univariate analyses. CONCLUSIONS: Frailty is not independently associated with risk of falls in older adults living in a remote rural setting. Further studies are needed to assess the impact of frailty on the risk of falls in these populations.

Determinación de pulso irregular en farmacias comunitarias españolas. "Resultados del proyecto 'Know your pulse' 2016 y 2017" / No disponible

Objetivos: evaluar la efectividad del cribado de pulso irregular por el farmacéutico comunitario mediante el pilotaje de una metodología y procedimiento para una campaña de detección de pulso irregular en las farmacias comunitarias españolas. Métodos: estudio descriptivo transversal realizado por farmacéuticos comunitarios socios de SEFAC entre los días 24 a 29 de noviembre de 2016 y los días 5 a 11 de junio de 2017. Criterios de inclusión: usuarios ≥40 años, sin FA o con FA sin tratamiento anticoagulante. Variables principales: pulso (latidos/minuto) y CHA2DS2 vascular. Resultados: se realizaron 789 mediciones de pulso en ambas campañas, de las cuales se encontró que 41 (5,2 %) participantes presentaban un pulso irregular. 35 (4,3 %) fueron derivados a sus médicos de familia, confirmándose 12 (1,5%) diagnósticos nuevos de FA e iniciando 7 (0,9 %) de ellos tratamiento anticoagulante. Más del 80 % de los participantes no presentaba ningún síntoma clínico de FA y un 41,9 % padecía HTA. El número de pacientes con CHA2DS2 vascular ≥ 2 y pulso irregular fue de 17 (3,9 %), de los cuales 10 (58,8 %) estaban sin tratamiento; 5 (29,4 %) tenían tratamiento antiagregante y 2 (11,8 %) tratamiento anticoagulante. Conclusiones: tanto el elevado número de pacientes con FA diagnosticada y sin tratamiento como el número apreciable de pacientes con pulso irregular y sospecha de FA detectados demuestran la capacidad y eficacia de la farmacia comunitaria en la realización del cribado del pulso irregular contribuyendo así a la detección de nuevos casos de FA

No disponible

First year of pre-exposure prophylaxis implementation in France with daily or on-demand tenofovir disoproxil fumarate/emtricitabine.

BACKGROUND: In January 2016, the French Medicine Agency initiated a Temporary Recommendation for Use (TRU) to allow the use of oral intake of tenofovir disoproxil fumarate and emtricitabine for pre-exposure prophylaxis (PrEP) in adults at high risk of HIV. We report the results of the first year of PrEP implementation in France. METHODS: Data were collected by physicians using a secured web subject-monitoring interface, with two forms: an initiation form, with patients' baseline characteristics, and an HIV seroconversion form. Univariate and adjusted multivariate analysis using a logistic regression model were performed to identify baseline factors associated with on-demand PrEP regimen prescription. RESULTS: From 4 January 2016 to 28 February 2017, 3405 subjects were enrolled, with 2774 initiation forms completed; 98.1% were male and 96.9% were MSM. An on-demand regimen was prescribed to 57% of subjects. Older age (OR for participants older than 50 years = 1.76, 95% CI 1.35-2.3, P < 0.001) and site of prescription (OR of former IPERGAY sites = 2.28, 95% CI 1.84-2.83, P < 0.001) were associated with on-demand prescription. Those reporting sexually transmitted infection (STI) and condomless anal sex with at least two different partners were less likely to receive on-demand PrEP (OR = 0.68, 95% CI 0.57-0.82 and 0.75, 95% CI 0.57-0.98, respectively; P < 0.05 for all). Four breakthrough HIV infections were reported during the study, in the context of PrEP interruption or acute infection at the time of PrEP initiation. CONCLUSIONS: In a real-life setting in France, PrEP was used, either daily or on-demand, mostly by MSM, with breakthrough infections being rare.

Effects of mutations in the post-translational modification sites on the trafficking of hyaluronan synthase 2 (HAS2).

Vesicular trafficking of hyaluronan synthases (HAS1-3) from endoplasmic reticulum (ER) through Golgi to plasma membrane (PM), and either back to endosomes and lysosomes, or out into extracellular vesicles, is important for their activities. We studied how post-translational modifications affect the trafficking of HAS2 by mutagenesis of the sites of ubiquitination (K190R), phosphorylation (T110A) and O-GlcNAcylation (S221A), using Dendra2- and EGFP-HAS2 transfected into COS1 cells. Confocal microscopy showed HAS2 wild type (wt) and its K190R and S221A mutants in ER, Golgi and extracellular vesicles, while the T110A mutant remained mostly in the ER. HA synthesis was reduced by S221A, while completely blocked by K190R and T110A. Cell-surface biotinylation indicated that T110A was absent from PM, while S221A was close to the level of wt, and K190R was increased in PM. TIRF microscopy analysis gave similar results. Rab10 silencing increased HA secretion by HAS2, likely by inhibiting endocytosis of the enzyme from PM, as reported before for HAS3. Green-to-red photo-conversion of Dendra2-HAS2 constructs suggested slower decay of K190R and S221A than HAS2 wt, while T110A was barely degraded at all. S221D and S221E, the phosphomimetic mutants of this site, decayed faster and blocked hyaluronan synthesis, suggesting alternative O-GlcNAc/-PO4 substitution to regulate the stability of the enzyme. Probing the role of dynamic O-GlcNAcylation at S221 by adding glucosamine increased the half-life of only HAS2 wt. The Dendra2·HAS2 disappearance from Golgi was slower for K190R. Of the two inactive constructs, K190R co-transfected with HAS2 wt suppressed, whereas T110A had no effect on HA synthesis. Interestingly, the HAS2-stimulated shedding of extracellular vesicles was dependent on HAS residence in PM but independent of HA synthesis. The results indicate that post-translational modifications control the trafficking of HAS2, and that trafficking is an integral part of the post-translational regulation of HAS2 activity.

Epigenetic silencing of miR-124 prevents spermine oxidase regulation: implications for Helicobacter pylori-induced gastric cancer.

Chronic inflammation contributes to the development of various forms of cancer. The polyamine catabolic enzyme spermine oxidase (SMOX) is induced in chronic inflammatory conditions, including Helicobacter pylori-associated gastritis, where its production of hydrogen peroxide contributes to DNA damage and subsequent tumorigenesis. MicroRNA expression levels are also altered in inflammatory conditions; specifically, the tumor suppressor miR-124 becomes silenced by DNA methylation. We sought to determine if this repression of miR-124 is associated with elevated SMOX activity and concluded that miR-124 is indeed a negative regulator of SMOX. In gastric adenocarcinoma cells harboring highly methylated and silenced mir-124 gene loci, 5-azacytidine treatment allowed miR-124 re-expression and decreased SMOX expression. Overexpression of an exogenous miR-124-3p mimic repressed SMOX mRNA and protein expression as well as H2O2 production by >50% within 24 h. Reporter assays indicated that direct interaction of miR-124 with the 3'-untranslated region of SMOX mRNA contributes to this negative regulation. Importantly, overexpression of miR-124 before infection with H. pylori prevented the induction of SMOX believed to contribute to inflammation-associated tumorigenesis. Compelling human in vivo data from H. pylori-positive gastritis tissues indicated that the mir-124 gene loci are more heavily methylated in a Colombian population characterized by elevated SMOX expression and a high risk for gastric cancer. Furthermore, the degree of mir-124 methylation significantly correlated with SMOX expression throughout the population. These results indicate a protective role for miR-124 through the inhibition of SMOX-mediated DNA damage in the etiology of H. pylori-associated gastric cancer.

CagA C-terminal variations in Helicobacter pylori strains from Colombian patients with gastric precancerous lesions.

The C-terminus of the Helicobacter pylori CagA protein is polymorphic, bearing different EPIYA sequences (EPIYA-A, B, C or D), and one or more CagA multimerization (CM) motifs. The number of EPIYA-C motifs is associated with precancerous lesions and gastric cancer (GC). The relationship between EPIYA, CM motifs and gastric lesions was examined in H. pylori-infected Colombian patients from areas of high and low risk for GC. Genomic DNA was extracted from H. pylori strains cultured from gastric biopsies from 80 adults with dyspeptic symptoms. Sixty-seven (83.8%) of 80 strains were cagA positive. The 3' region of cagA was sequenced, and EPIYA and CM motifs were identified. CagA proteins contained one (64.2%), two (34.3%) or three EPIYA-C motifs (1.5%), all with Western type CagA-specific sequences. Strains with one EPIYA-C motif were associated with less severe gastric lesions (non-atrophic and multifocal atrophic gastritis), whereas strains with multiple EPIYA-C motifs were associated with more severe lesions (intestinal metaplasia and dysplasia) (p <0.001). In 54 strains, the CM motifs were identical to those common in Western strains. Thirteen strains from the low-risk area contained two different CM motifs: one of Western type located within the EPIYA-C segment and another following the EPIYA-C segment and resembling the CM motif found in East Asian strains. These strains induced significantly shorter projections in AGS cells and an attenuated reduction in levels of CagA upon immunodepletion of SHP-2 than strains possessing Western/Western motifs. This novel finding may partially explain the difference in GC incidence in these populations.

[Is good old minocycline a new neuroprotective drug?]. / Es la vieja minociclina un nuevo fármaco neuroprotector?

INTRODUCTION: During the last decade, the neuroprotective effects of minocycline have been a matter of an intense debate. A broad amount of contradictory studies can be found in the scientific literature, going from neuroprotection to the exacerbation of toxicity in diverse experimental models. Such differences could be the result of minocycline acting on multiple pharmacological targets. DEVELOPMENT: In the present review we will go over these pharmacological targets and the effects derived from their modulation by minocycline. Among others, its antioxidant activity derived from its chemical structure or its modulator effect on several enzymes such as nitric oxide synthase will be reviewed. Furthermore, the effects of minocycline on the intracellular pathways implicated in neurodegenerative processes including apoptosis stages, activation decision and execution will be addressed. CONCLUSIONS: All the mechanisms described herein have not escaped to a scientific community needed of new therapeutic drugs for the treatment of neurodegenerative conditions. However, the sparse clinical trials carried out so far are mainly aimed at assessing its tolerability and safety or are still in progress. We believe that more studies, both clinical and pre-clinical, should be carried out in order to ascertain the therapeutic window and the neurodegenerative disorders in which minocycline could be useful.

Prognostic value of (13)C-phenylalanine breath test on predicting survival in patients with chronic liver failure.

AIM: To evaluate the prognostic value of percentage of (13)C-phenylalanine oxidation ((13)C-PheOx) obtained by (13)C-phenylalanine breath test ((13)C-PheBT) on the survival of patients with chronic liver failure. METHODS: The hepatic function was determined by standard liver blood tests and the percentage of (13)C-PheOx in 118 chronic liver failure patients. The follow-up period was of 64 mo. Survival analysis was performed by the Kaplan-Meier method and variables that were significant (P < 0.10) in univariate analysis and subsequently introduced in a multivariate analysis according to the hazard model proposed by Cox. RESULTS: Forty-one patients died due to progressive liver failure during the follow-up period. The probability of survival at 12, 24, 36, 48 and 64 mo was 0.88, 0.78, 0.66, 0.57 and 0.19, respectively. Multivariate analysis demonstrated that Child-Pugh classes, age, creatinine and the percentage of (13)C-PheOx (HR 0.338, 95% CI: 0.150-0.762, P = 0.009) were independent predictors of survival. When Child-Pugh classes were replaced by all the parameters of the score, only albumin, bilirubin, creatinine, age and the percentage of (13)C-PheOx (HR 0.449, 95% CI: 0.206-0.979, P = 0.034) were found to be independent predictors of survival. CONCLUSION: Percentage of (13)C-PheOx obtained by (13)C-PheBT is a strong predictor of survival in patients with chronic liver disease.

Reactive oxygen species and p38 mitogen-activated protein kinase activate Bax to induce mitochondrial cytochrome c release and apoptosis in response to malonate.

Malonate, an inhibitor of mitochondrial complex II, is a widely used toxin to study neurodegeneration in Huntington's disease and ischemic stroke. We have shown previously that malonate increased reactive oxygen species (ROS) production in human SH-SY5Y neuroblastoma cells, leading to oxidative stress, cytochrome c release, and apoptotic cell death. Expression of a green fluorescent protein-Bax fusion protein in SH-SY5Y neuroblastoma cells demonstrated a Bax redistribution from the cytosol to mitochondria after 12 to 24 h of malonate treatment that coincided with mitochondrial potential collapse and chromatin condensation. Inhibition of Bax translocation using furosemide, as well as Bax gene deletion, afforded significant protection against malonate-induced apoptosis. Further experiments revealed that malonate induced a prominent increase in the level of activated p38 mitogen-activated protein (MAP) kinase and that treatment with the p38 MAP kinase inhibitor SKF86002 potently blocked malonate-induced Bax translocation and apoptosis. Treatment with vitamin E diminished ROS production, reduced the activation status of p38 MAP kinase, inhibited Bax translocation, and protected against malonate-induced apoptosis. Our data suggest that malonate-induced ROS production and subsequent p38 MAP kinase activation mediates the activation of the pro-apoptotic Bax protein to induce mitochondrial membrane permeabilization and neuronal apoptosis.

Pyruvate protects cerebellar granular cells from 6-hydroxydopamine-induced cytotoxicity by activating the Akt signaling pathway and increasing glutathione peroxidase expression.

Parkinson disease (PD) is the second-most common age-related neurodegenerative disease and is characterized by the selective destruction of dopaminergic neurons. Increasing evidence indicates that oxidative stress plays a crucial role in the pathogenesis of idiopathic PD. Anti-oxidant agents including catalase, manganese porphyrin and pyruvate confer cytoprotection to different cell cultures when challenged with 6-hydroxydopamine (6-OHDA). Herein we used rat cerebellar granular cell cultures to ascertain the plausible cellular pathways involved in pyruvate-induced cytoprotection against 0.1 mM 6-OHDA. Pyruvate provided cytoprotection in a concentration-dependent manner (2-10 mM). Consistent with its well-established anti-oxidant capacity, pyruvate (10 mM) prevented 6-OHDA-induced lipid peroxidation by blocking the rise in intracellular peroxides and maintaining the intracellular reduced glutathione (GSH) levels. Further experiments revealed that pyruvate increased Akt, but not extracellular signal-regulated kinase phosphorylation. Moreover, phosphatidylinositol 3-kinase (PI3K) inhibitors attenuated pyruvate-induced cytoprotection indicating that PI3K-mediated Akt activation is necessary for pyruvate to induce cytoprotection. On the other hand, pyruvate also up-regulated glutathione peroxidase mRNA levels, but not those of the anti-oxidant enzymes superoxide dismutase-1 and -2, catalase or the anti-apoptotic oncogenes Bcl-2 or Bcl-xL. In summary, our results strongly suggest that pyruvate, besides the anti-oxidant properties related to its structure, exerts cytoprotective actions by activating different anti-apoptotic routes that include gene regulation and Akt pathway activation.

Long term follow up of patients treated for Helicobacter pylori infection.

BACKGROUND: Helicobacter pylori infection induces progressive inflammatory changes in the gastric mucosa that may lead to gastric cancer. Understanding long term effects resulting from the cure of this infection is needed to design cancer prevention strategies. METHODS: A cohort of 795 adults with preneoplastic gastric lesions was randomised to receive anti-H pylori treatment and/or antioxidants. At the end of six years of intervention, those who did not receive anti-H pylori treatment were offered it. Gastric biopsies were obtained at baseline, and at 3, 6, and 12 years. A histopathology score was utilised to document changes in gastric lesions. Non-linear mixed models were used to estimate the cumulative effect of H pylori clearance on histopathology scores adjusted for follow up time, interventions, and confounders. RESULTS: Ninety seven per cent of subjects were H pylori positive at baseline, and 53% were positive at 12 years. Subjects accumulated 1703 person years free of infection. A multivariate model showed a significant regression in histopathology score as a function of the square of H pylori negative time. Subjects who were H pylori negative had 14.8% more regression and 13.7% less progression than patients who were positive at 12 years (p = 0.001). The rate of healing of gastric lesions occurred more rapidly as years free of infection accumulated, and was more pronounced in less advanced lesions. CONCLUSIONS: Preneoplastic gastric lesions regress at a rate equal to the square of time in patients rendered free of H pylori infection. Our findings suggest that patients with preneoplastic gastric lesions should be treated and cured of their H pylori infection.