Valoración de resultados tras 112 radioembolizaciones con 90Y-microesferas / Evaluation of results after 112 radioembolizations with 90Y-microspheres

Objetivo Para conocer los resultados de la radioembolización (transarterial radioembolization o TARE), en el tratamiento de tumores hepáticos, se realizó una valoración retrospectiva tras 112 TARE con 90Y-microesferas administradas en 82 pacientes en un único hospital, analizando la eficacia y la seguridad, tras un seguimiento mayor o igual a 1 año post-TARE en todos los pacientes, y evaluando la posible relación entre la respuesta al tratamiento y la supervivencia de los pacientes. Material y métodos Se administraron 57 TARE únicas y 55 TARE múltiples en pacientes con hepatocarcinoma (53), metástasis hepáticas (25) y colangiocarcinoma (4), con evaluación previa multidisciplinar clínica, angiográfica y gammagráfica (planar/SPECT/SPECT-TC con 99mTc-MAA), modelo multicompartimental (ecuaciones MIRD), valoración gammagráfica post-TARE (planar/SPECT/SPECT-TC), seguimiento clínico-radiológico, evaluación de respuesta tumoral (criterios mRECIST) y análisis (Kaplan Meier) de supervivencia libre de progresión (SLP) y supervivencia global (SG). Resultados La intención terapéutica fue paliativa (82%) y como puente a trasplante hepático/resección quirúrgica (17%). Se obtuvo respuesta (R), completa o parcial, en el 65,9% de los casos. Al año post-TARE estaban libres de progresión el 34,7% de los pacientes con R y 19,2% de los no R (p:0,003), con SG del 80% para los R y 37,5% para los no R (p:0,001). Las curvas de supervivencia mostraron mediana de SG de 18 meses (95% IC 15,7-20,3) para los R y 9 meses (95% IC 6,1-11,8) para los no R (p:0,03). Efectos secundarios leves (27,6%) y severos (5,3%) resueltos, sin mayor incidencia tras TARE múltiple. Conclusiones La TARE con 90Y-microesferas en pacientes adecuadamente seleccionados con tumores hepáticos, aporta eficacia terapéutica y bajo índice de toxicidad, con SLP y SG superiores en los pacientes con respuesta a la TARE respecto a los que no respondieron (AU)

Aim To determine the results of radioembolization transarterial (TARE), in the treatment of liver tumors, a retrospective evaluation was performed after 112 TARE with 90Y-microspheres administered in 82 patients in a single hospital, analyzing efficacy and safety, after a follow-up greater than or equal to 1 year post-TARE in all patients, and evaluating the possible relationship between treatment response and patient survival Material and methods We have administered 57 single TARE and 55 multiple TARE in patients with hepatocellular carcinoma (53), liver metastases (25) and cholangiocarcinoma (4), with prior multidisciplinary evaluation, clinical, angiographic and gammagraphic (planar/SPECT/SPECT-CT with 99mTc-MAA), multicompartment model (MIRD equations), post-TARE screening (planar/SPECT/SPECT-CT), clinical and radiological follow-up, tumor response evaluation (mRECIST criteria) and Kaplan–Meier analysis to determine progression-free survival (PFS) and overall survival (OS). Results Therapeutic intention was palliative (82%) and as bridge to liver transplantation/surgical resection (17%). We obtained response (R), complete or partial, in 65.9% of cases. One year after TARE 34.7% of patients with R and 19.2% of non-R were progression-free (p: 0.003), with OS of 80% for R and 37.5% for non-R (p: 0.001). Survival analysis showed median OS of 18 months (95% CI 15.7–20.3) for R and 9 months (95% CI 6.1–11.8) for non-R (p: 0.03). We found mild (27.6%) and severe (5.3%) side effects, all of them resolved, without higher incidence after multiple TARE. Conclusion TARE with 90Y-microspheres, in appropriately selected patients with liver tumors, provides therapeutic efficacy and low rate of toxicity, with higher PFS and OS in patients with TARE response compared to those who did not respond (AU)

Evaluation of results after 112 radioembolizations with 90Y-microspheres.

AIM: To determine the results of radioembolization transarterial (TARE), in the treatment of liver tumors, a retrospective evaluation was performed after 112 TARE with 90Y-microspheres administered in 82 patients in a single hospital, analyzing efficacy and safety, after a follow-up greater than or equal to 1 year post-TARE in all patients, and evaluating the possible relationship between treatment response and patient survival. MATERIAL AND METHODS: We have administered 57 single TARE and 55 multiple TARE in patients with hepatocellular carcinoma (53), liver metastases (25) and cholangiocarcinoma (4), with prior multidisciplinary evaluation, clinical, angiographic and gammagraphic (planar/SPECT/SPECT-CT with 99mTc-MAA), multicompartment model (MIRD equations), post-TARE screening (planar/SPECT/SPECT-CT), clinical and radiological follow-up, tumor response evaluation (mRECIST criteria) and Kaplan-Meier analysis to determine progression-free survival (PFS) and overall survival (OS). RESULTS: Therapeutic intention was palliative (82%) and as bridge to liver transplantation/surgical resection (17%). We obtained response (R), complete or partial, in 65.9% of cases. One year after TARE 34.7% of patients with R and 19.2% of non-R were progression-free (P: .003), with OS of 80% for R and 37.5% for non-R (P: .001). Survival analysis showed median OS of 18 months (95% CI 15.7-20.3) for R and 9 months (95% CI 6.1-11.8) for non-R (P: .03). We found mild (27.6%) and severe (5.3%) side effects, all of them resolved, without higher incidence after multiple TARE. CONCLUSION: TARE with 90Y-microspheres, in appropriately selected patients with liver tumors, provides therapeutic efficacy and low rate of toxicity, with higher PFS and OS in patients with TARE response compared to those who did not respond.

Progreso y utilidad actual de la radiómica dentro del estudio PET/TC en cáncer de mama no metastásico: una revisión sistemática / Progress and current utility of radiomics in PET/CT study of non-metastatic breast cancer: A systematic review

Objetivo Sintetizar la evidencia actual sobre la utilidad de la radiómica en el análisis de la imagen PET/TC en cáncer de mama local o localmente avanzado y evaluar la calidad metodológica de los estudios radiómicos publicados al respecto. Material y métodos Revisión sistemática de artículos en distintas bases de datos hasta 2021 utilizando los términos «PET», «radiomics», «texture», «breast». Se seleccionaron solo artículos con datos humanos y que incluyeran una imagen de PET en su análisis. Se excluyeron estudios con datos de pruebas y menos de 20 pacientes. De cada artículo se extrajo el tamaño muestral, el radiotrazador utilizado, la técnica de imagen y las características de imagen extraídas. Se determinó la calidad metodológica de los estudios mediante el instrumento QUADAS-2. Resultados Se seleccionaron 18 artículos. El diseño retrospectivo fue el más utilizado. La característica radiómica más estudiada fue el SUVmax. Diversos parámetros radiómicos se correlacionaron con la caracterización tumoral, y la heterogeneidad tumoral demostró utilidad para predecir el curso de la enfermedad y la respuesta al tratamiento. La mayoría de los artículos mostraron un alto riesgo de sesgo, derivado principalmente de la selección de pacientes. Conclusiones Se observó una alta probabilidad de sesgo en los artículos publicados. La radiómica es un campo aún en desarrollo y son necesarios más estudios para demostrar su utilidad en la práctica clínica habitual. La herramienta QUADAS-2 permite la valoración crítica de la calidad metodológica de la evidencia disponible. Pese a las limitaciones, la radiómica se muestra como una herramienta que puede ayudar a conseguir un manejo oncológico personalizado en el cáncer de mama (AU)

Aim To synthesize the current evidence of the usefulness of radiomics in PET/CT image analysis in local and locally advanced breast cancer. Also, to evaluate the methodological quality of the radiomic studies published. Methods Systematic review of articles in different databases until 2021 using the terms «PET», «radiomics», «texture», «breast». Only articles with human data and that included a PET image were included. Studies with simulated data and with less than 20 patients were excluded. The sample size, radiotracer used, imaging technique, and radiomics characteristics were extracted from each article. The methodological quality of the studies was determined using the QUADAS-2 tool. Results Eighteen articles were selected. The retrospective design was the most used. The most studied radiomic characteristic was SUVmax. Several radiomic parameters were correlated with tumor characterization, and tumor heterogeneity proved useful for predicting disease course and response to treatment. Most articles showed a high risk of bias, mainly from the patient selection. Conclusions A high probability of bias was observed in most of the published articles. Radiomics is a developing field and more studies are needed to demonstrate its usefulness in routine clinical practice. The QUADAS-2 tool allows critical assessment of the methodological quality of the available evidence. Despite its limitations, radiomics is shown to be an instrument that can help to achieve personalized oncologic management of breast cancer (AU)

Progress and current utility of radiomics in PET/CT study of non-metastatic breast cancer: A systematic review.

AIM: To synthesize the current evidence of the usefulness of radiomics in PET/CT image analysis in local and locally advanced breast cancer. Also, to evaluate the methodological quality of the radiomic studies published. METHODS: Systematic review of articles in different databases until 2021 using the terms "PET", "radiomics", "texture", "breast". Only articles with human data and that included a PET image were included. Studies with simulated data and with less than 20 patients were excluded. Were extracted sample size, radiotracer used, imaging technique, and radiomics characteristics from each article. The methodological quality of the studies was determined using the QUADAS-2 tool. RESULTS: 18 articles were selected. The retrospective design was the most used. The most studied radiomic characteristic was SUVmax. Several radiomic parameters were correlated with tumor characterization, and tumor heterogeneity proved useful for predicting disease course and response to treatment. Most articles showed a high risk of bias, mainly from the patient selection. CONCLUSIONS: A high probability of bias was observed in most of the published articles. Radiomics is a developing field and more studies are needed to demonstrate its usefulness in routine clinical practice. The QUADAS-2 tool allows critical assessment of the methodological quality of the available evidence. Despite its limitations, radiomics is shown to be an instrument that can help to achieve personalized oncologic management of breast cancer.

Periodontitis and cerebrovascular disease: a new novel in medicine.

The aim of this review is to determine if there is a relationship between periodontal disease and stroke. The included case-control and cohort studies mediate the incidence of stroke and periodontal disease by analyzing different parameters. A literature review was carried out in PubMed, Scopus and Embase databases using the key word "stroke" AND "periodontal disease". An amount of 932 articles came out from our research on these three databases. These articles were selected according to PRISMA criteria. The following inclusion criteria were established: studies conducted in humans, articles published in English and published in the last ten years. Exclusion criteria were: experimental studies on animals, articles published more than 10 years ago, non-English language articles, articles of non-indexed journals, and articles not directly related to the association between stroke and periodontitis. These criteria reduced the number of articles from 932 to 399. At the end, articles that appeared to be repeated in different databases have been eliminated: 254 articles remained. All these articles titles were reviewed by the authors, who decided whether or not to include them in the review. We selected an amount of 43 articles. These studies were reviewed by reading the titles and abstracts, and by finally selecting the ones with the same topic of this review. When titles or abstracts were not clear, the complete article was read. At the end 7 articles were selected. In addition, 2 systematic reviews and 1 article, cited in the discussion, and regarding the protocol used in patients suffering from cardiovascular diseases and periodontitis, were selected. The quality of these articles was evaluated through the JADAD system. In conclusion, patients with stroke have a higher prevalence of periodontitis.

Producción de biofilm y presencia de genes icaABCD en cepas de Staphylococcus aureus aisladas de leche cruda / Biofilm production and presence of genes icaABCD in Staphylococcus aureus strains isolated from raw milk

En la patogénesis de la mastitis estafilocócica, el biofilm se considera un marcador de virulencia que permite la adherencia del microorganismo al epitelio mamario. Con la finalidad de determinar la producción de biofilm y detectar la presencia de genes icaABCD, se analizaron 30 cepas de S. aureus aisladas de leche cruda obtenida de vacas con mastitis subclínica. El biofilm se determinó por el método cuantitativo sobre microplacas de cultivo celular y los genes icaABCD por reacción en cadena de la polimerasa. En 96,67% de las cepas de S. aureus se observó producción de biofilm. De éstas 43,34% fueron fuertes productoras, 30,00% moderadas productoras, 23,33% débiles productoras y 3,33% no productoras de biofilm. La totalidad de los genes icaADCD se obtuvo en 6,66% de los aislamientos. Todas las cepas, excepto una, presentaron icaA o icaD. En 56,66% de los aislamientos se identificaron genes icaA/icaD. En todas las cepas productoras de biofilm se identificaron genes del operón icaABCD, lo cual evidencia que poseen un factor de virulencia que facilita su persistencia en el tejido mamario. Además, el biofilm constituye una fuente de contaminación microbiana que puede conducir a fallas en los procesos de producción que generen problemas de inocuidad de los alimentos.

In the pathogenesis of staphylococcal mastitis the biofilm is considered a marker of virulence that allows the adhesion of the microorganism to the mammary epithelium. In order to determine the production of biofilm and the presence of icaABCD gene, 30 S. aureus strains isolated from raw milk obtained from cows with subclinical mastitis were analyzed. Biofilm production was determined by the quantitative method on microplates for cell culture and the presence of genes icaABCD by polymerase chain reaction. Biofilm production was observed in 96.67% of strains of S. aureus. These 43.34% were strong producers, 30.00% producing moderate, 23.33% weak producers and 3.33% non producing biofilm. All of the icaABCD genes was obtained at 6.66% of the isolates. All strains, except one, presented icaA or icaD. 56,66% of isolates identified genes icaA/icaD. The icaABCD operon genes were identified in all biofilm-producing strains, which demonstrates that they possess a virulence factor that facilitates its persistence in the breast tissue. In addition, the biofilm is a source of microbial contamination which can lead to failures in production processes that generate problems of food safety.

Rapid detection and evolutionary analysis of Legionella pneumophila serogroup 1 sequence type 47.

OBJECTIVES: Legionella pneumophila serogroup 1 (Lp1) sequence type 47 is the leading cause of legionellosis in north-western Europe, but, surprisingly, it is rarely isolated from environmental samples. Comparative genomics was applied to develop a PCR assay and to better understand the evolution of this strain. METHODS: Comparative analysis of 36 genomes representative of the Lp species was used to identify specific PCR targets, which were then evaluated in silico on 545 sequenced genomes and in vitro on 436 Legionella strains, 106 respiratory samples, and three environmental samples from proven ST47 sources. Phylogenetic analyses were performed to understand the evolution of ST47. RESULTS: The gene LPO_1073 was characterized as being 100% conserved in all 129 ST47 genomes analysed. A real-time PCR designed to detect LPO_1073 was positive for all 110 ST47 strains tested and agreed with culture and typing results previously obtained for 106 respiratory samples. The three environmental samples were also positive. Surprisingly, 26 of the 44 ST109 strains tested among 342 non-ST47 strains scored positive for LPO_1073. SNP-based phylogenetic analysis was undertaken to understand this result: the PCR-positive ST109 genomes were almost identical to ST47 genomes, with the exception of a recombined region probably acquired by ST47 from a ST62(-like) strain. CONCLUSION: The genomic analysis allowed the design of a highly specific PCR assay for rapid detection of ST47 strains. Furthermore, it allowed us to uncover the evolution of ST47 strains from ST109 by homologous recombination with ST62. We hypothesize that this recombination generated the leading cause of legionellosis in north-western Europe.

Molecular epidemiology, phylogeny and evolution of Legionella.

Legionella are opportunistic pathogens that develop in aquatic environments where they multiply in protozoa. When infected aerosols reach the human respiratory tract they may accidentally infect the alveolar macrophages leading to a severe pneumonia called Legionnaires' disease (LD). The ability of Legionella to survive within host-cells is strictly dependent on the Dot/Icm Type 4 Secretion System that translocates a large repertoire of effectors into the host cell cytosol. Although Legionella is a large genus comprising nearly 60 species that are worldwide distributed, only about half of them have been involved in LD cases. Strikingly, the species Legionella pneumophila alone is responsible for 90% of all LD cases. The present review summarizes the molecular approaches that are used for L. pneumophila genotyping with a major focus on the contribution of whole genome sequencing (WGS) to the investigation of local L. pneumophila outbreaks and global epidemiology studies. We report the newest knowledge regarding the phylogeny and the evolution of Legionella and then focus on virulence evolution of those Legionella species that are known to have the capacity to infect humans. Finally, we discuss the evolutionary forces and adaptation mechanisms acting on the Dot/Icm system itself as well as the role of mobile genetic elements (MGE) encoding T4ASSs and of gene duplications in the evolution of Legionella and its adaptation to different hosts and lifestyles.

Trasplante laparoscópico de metanefros: un paso inicial al xenotrasplante renal / Laparoscopic transplantation of metanephroi: A first step to kidney xenotransplantation

Antecedentes: Una solución novedosa a la escasez de riñones para trasplante puede ser el xenotrasplante de riñones embrionarios. Objetivo: Conocer la viabilidad del trasplante alogénico laparoscópico de metanefros (M) en conejos. Material y método: Se realizó disección microscópica para obtener metanefros en embriones de 14 días de edad (24 M), 15 (20 M) y 16 (26 M). Mediante laparoscopia abdominal de un puerto se insertó percutáneamente una aguja raquídea y por ella, mediante un catéter epidural, depositamos el metanefros cerca de un vaso sanguíneo patente en la grasa retroperitoneal. Setenta metanefros se trasplantaron a 18 conejos. Tres semanas después los animales fueron explorados por cirugía abierta. Se analizó la comparación de la madurez embrionaria, las variables morfométricas de los metanefros y la tasa de desarrollo de los metanefros trasplantados. Resultados: El límite temporal inferior para la extracción de metanefros en conejos es el día 14. Tres semanas postrasplante crecieron a una mínima expresión solo 3/24 M de 14 días (12,5%). Por el contrario, 10/20 (50%) de los de 15 días y 12/26 (46,1%) de los de 16 días de edad crecieron y se diferenciaron de tal manera que se habían desarrollado normalmente los glomérulos, túbulos proximales y distales y conductos colectores. No se detectaron cambios inmunológicos relevantes en sangre periférica. Conclusiones: Describimos, por primera vez en la literatura, el trasplante laparoscópico alogénico de metanefros de embriones como una técnica factible y no invasiva. Los receptores no necesitaron inmunosupresión

Background: Embryonic kidney xenotransplantation could represent a new solution to the scarcity of kidneys for transplantation. Objective: To determine the feasibility of allogeneic laparoscopic transplantation of metanephroi (M) in rabbits. Material and method: Microscopic dissection was conducted to obtain metanephroi from 14-day-old (24 M), 15-day-old (20 M) and 16-day-old (26 M) embryos. Using single-port abdominal laparoscopy, a spinal needle was inserted percutaneously, through which the metanephroi were deposited (using an epidural catheter) close to a patent blood vessel in the retroperitoneal fat. Seventy metanephroi were transplanted to 18 rabbits. Three weeks later, the animals were examined through open surgery. We compared the embryonic maturity, the morphometric variables of the metanephroi and the development rate of the transplanted metanephroi. Results: The lower time limit for the extraction of metanephroi from the rabbits was day 14. Three weeks after transplantation, only 3/24 14-day-old metanephroi grew at minimal expression (12.5%). In contrast, 10/20 (50%) 15-day-old and 12/26 (46.1%) 16-day-old metanephroi grew. These metanephroi had differentiated sufficiently for the glomeruli, proximal and distal tubules and collecting ducts to develop normally. We detected no relevant immunological changes in the peripheral blood. Conclusions: We have described for the first time in the literature the allogeneic laparoscopic transplantation of metanephroi from embryos as a feasible and noninvasive technique. The recipients did not require immunosuppression

Laparoscopic transplantation of metanephroi: A first step to kidney xenotransplantation.

BACKGROUND: Embryonic kidney xenotransplantation could represent a new solution to the scarcity of kidneys for transplantation. OBJECTIVE: To determine the feasibility of allogeneic laparoscopic transplantation of metanephroi (M) in rabbits. MATERIAL AND METHOD: Microscopic dissection was conducted to obtain metanephroi from 14-day-old (24M), 15-day-old (20M) and 16-day-old (26M) embryos. Using single-port abdominal laparoscopy, a spinal needle was inserted percutaneously, through which the metanephroi were deposited (using an epidural catheter) close to a patent blood vessel in the retroperitoneal fat. Seventy metanephroi were transplanted to 18 rabbits. Three weeks later, the animals were examined through open surgery. We compared the embryonic maturity, the morphometric variables of the metanephroi and the development rate of the transplanted metanephroi. RESULTS: The lower time limit for the extraction of metanephroi from the rabbits was day 14. Three weeks after transplantation, only 3/24 14-day-old metanephroi grew at minimal expression (12.5%). In contrast, 10/20 (50%) 15-day-old and 12/26 (46.1%) 16-day-old metanephroi grew. These metanephroi had differentiated sufficiently for the glomeruli, proximal and distal tubules and collecting ducts to develop normally. We detected no relevant immunological changes in the peripheral blood. CONCLUSIONS: We have described for the first time in the literature the allogeneic laparoscopic transplantation of metanephroi from embryos as a feasible and noninvasive technique. The recipients did not require immunosuppression.

Producción de Enterotoxina y Biofilm en Aislamientos Clínicos de Staphylococcus aureus Resistente a Meticilina / Enterotoxin and Biofilm Production in Clinical Isolates of Methicillin-Resistant Staphylococcus aureus

S. aureus se ha convertido en un problema de salud pública, debido a la dificultad que representa el tratamiento de las infecciones causadas por SARM. El propósito de esta investigación fue determinar la producción de enterotoxinas A, B, C y D y la producción de biofilm en aislamientos de SARM. Se estudiaron 50 cepas aisladas de diferentes tipos de muestras clínicas. La detección de enterotoxinas se realizó por la técnica de aglutinación en fase reversa y la producción de biofilm mediante: agar rojo congo y el método en microplacas de cultivos celulares. La producción de enterotoxina se observó en 9 cepas (18%), siendo la enterotoxina D (64%) la más prevalente, seguida de la B (27%) y la A (9%). Se demostró una asociación significativa entre la producción de enterotoxina y el tipo de muestra de la que provenía la cepa. La producción de biofilm se constató en 30% y 98% de las cepas por los métodos de agar rojo congo y microplacas de cultivos celulares, respectivamente; sólo en 15 cepas (30%) se observó correlación de ambos ensayos, se demostró que el método en microplacas de cultivo celular es más eficaz para detectar la producción de biofilm en S. aureus.

S. aureus has become a public health problem, due to the difficulty of treating infections caused by methicillin-resistant S. aureus (MRSA). The purpose of this research was to determine the production of enterotoxins A, B, C and D and the production of biofilm in clinical isolates of MRSA. Fifty MRSA strains isolated from different types of clinical samples were studied. Detection of enterotoxins was carried out using the technique of reversed phase agglutination, while biofilm production was studied through two tests: Congo red agar and the microplate cell culture method. Enterotoxin production was observed in 9 strains (18%); enterotoxin D (64%) was the most prevalent, followed by B (27%) and A (9%). A significant association was shown between enterotoxin production capacity and the type of sample that came from the strain. Biofilm production was found in 30% and 98% of the strains using the Congo red Agar and microplate cell culture methods, respectively. A correlation of both trials was observed in only 15 strains (30%). It was shown that the microplate cell culture method is more effective for detecting biofilm production in S. aureus strains.

Metabolic differences in hepatocytes of obese and lean pigs.

There are important differences in terms of metabolic activity, energy utilization and capacity of protein and fat deposition when Iberian and modern pigs are compared. Primary culture of hepatocytes was used to evaluate hepatic function and sensitivity to hormones between breeds without the interference of circulating blood factors. Hepatocytes were isolated from pure Iberian (n=10) and Landrace (n=8) pigs of similar BW (24.5±12.1 and 32.9±6.1 kg BW, respectively), by collagenase perfusion. Monolayers were established in medium containing fetal bovine serum for 1 day and switched to serum-free medium for the remainder of the culture period. Hepatocytes were maintained in William's E supplemented with ß-mercaptoethanol (0.1 mM), glutamine (2 mM), antibiotics (gentamicin, penicillin, streptomycin and amphotericin B), dimethyl sulfoxide (1 µg/ml), dexamethasone (10-8 M), insulin (0.173 and 17.3 nM) and glucagon (0.287, 2.87 and 28.7 nM) for 24 to 48 h. Gluconeogenesis (GNG), glycogen degradation, triglycerides (TG) content and esterification, ß-hydroxybutyrate (BHB) synthesis, IGF-1 synthesis, albumin and urea synthesis were determined. Iberian pigs had greater capacity of GNG than Landrace (24%, P<0.05), although no difference in glycogen degradation was found (P>0.10). TG content and esterification tended to be lower in hepatocytes from Iberian compared with Landrace pigs (12% and 31%, respectively; 0.100.10). IGF-1 synthesis was diminished in hepatocytes from Iberian compared with Landrace pigs (16%, P<0.05). On the contrary, rate of albumin synthesis was greater in Iberian compared with Landrace pigs (58%, P<0.05). Finally, the capacity of urea synthesis was lower in hepatocytes of Iberian compared with Landrace pigs (37%, P<0.05). When ammonia was added to the media, urea concentration increased (648%, 1108% and 2791% when 0 mM was compared with 2.5, 5 and 10 mM, respectively). Urea synthesis increased on increasing ammonia content (55% and 325% when 0 mM was compared with 5 and 10 mM, respectively; P<0.0001). In conclusion, the genetic background accounts for important differences in protein and energy metabolism pathways found in primary culture of hepatocytes from lean and obese pigs.

Proteomic analysis of the somatic and surface compartments from Dirofilaria immitis adult worms.

Dirofilaria immitis (hearthworm) is a filarial roundworm transmitted by mosquitoes to different vertebrate hosts (dogs, cats and humans, among others), causing dirofilariosis. The adult worms reside in the pulmonary arteries affecting vessels and tissues and resulting in different pathological manifestations. Worms migrate to the heart and surrounding major vessels in heavy infections. Dirofilariosis can result in serious damage to affected hosts. In the last few years, a re-emergence of the disease driven by the climate change has been pointed out. Very recently, the knowledge at molecular level of this parasite has been extended by the published studies on its genome and transcriptome. Nevertheless, studies on the expression of defined protein sets in different parasite compartments and the corresponding role of those proteins in the host-parasite relationship have been relatively scarce to date. These include the description of the adult worm secretome, and some of the proteins eliciting humoural immune responses and those related with plasminogen binding in secreted and surface extracts of the parasite. Here, we investigate by proteomics the somatic and surface compartments of the D. immitis adult worm, adding new information on protein expression and localization that would facilitate a deeper understanding of the host-parasite relationships in dirofilariosis.

Tratamientos fisioterápicos para pacientes con hombro doloroso tras un accidente cerebrovascular. revisión sistemática / Physiotherapy treatments for patients with shoulder pain after stroke. A systematic review

OBJETIVO: Determinar, sobre la base de la evidencia científica actual, los tratamientos fisioterápicos que son más eficaces en la prevención y el tratamiento del hombro doloroso hemipléjico (HDH). Estrategia de búsqueda: Se llevó a cabo una revisión sistemática de ensayos controlados en las bases de datos PubMed, PEDro, Cochrane Library Plus, SCOPUS, Web of Knowledge, y búsqueda manual. Selección de estudios: Se incluyó un total de 16 estudios, que incluían tratamientos fisioterápicos. Síntesis de resultados: Del análisis de los 16 ensayos clínicos se extrajeron como técnicas de fisioterapia eficaces en el alivio del dolor de hombro: sujeción de hombro, acupresión con aromaterapia, masaje suave de espalda, punción seca profunda, crioterapia, terapia robótica, estimulación eléctrica transcutánea del nervio (TENS), estimulación eléctrica funcional y terapia basada en el concepto Bobath. CONCLUSIÓN: Aunque existe controversia respecto al uso de la estimulación eléctrica funcional, el resultado favorable del resto de los tratamientos mencionados sugiere que el tratamiento fisioterápico es fundamental para prevenir o tratar el HDH

OBJECTIVE: To determine which physiotherapy treatments are more effective in the prevention and treatment of hemiplegic shoulder pain (HSP) based on current scientific evidence. Search strategy: A systematic review of controlled trials was conducted in the PubMed, PEDro, Cochrane Library Plus, SCOPUS, Web of Knowledge databases as well as a manual searching. Screening of studies. A total of 16 studies included physiotherapy treatments were included. Synthesis of the results. Based on the analysis of 16 studies, the following physiotherapy technique considered effective in relieving shoulder pain were found: shoulder strapping, aromatherapy acupressure, slow-stroke back massage, deep dry needling, cryotherapy, therapeutic robotic system, transcutaneous electrical nerve stimulation, functional electrical stimulation and therapy based on the Bobath concept. CONCLUSION: Although there is controversy regarding the use of functional electrical stimulation, the favorable outcome of other treatments mentioned suggests that physical therapy is essential to prevent or treat the HSP

Impact of dietary betaine and conjugated linoleic acid on insulin sensitivity, protein and fat metabolism of obese pigs.

To determine possible mechanisms of action that might explain the nutrient partitioning effect of betaine and conjugated linoleic acid (CLA) in Iberian pigs and to address potential adverse effects, twenty gilts were restrictively fed from 20 to 50 kg BW Control, 0.5% betaine, 1% CLA or 0.5% betaine + 1% CLA diets. Serum hormones and metabolites profile were determined at 30 kg BW and an oral glucose test was performed before slaughter. Pigs were slaughtered at 50 kg BW and livers were obtained for chemical and histological analysis. Decreased serum urea in pigs fed betaine and betaine + CLA diets (11%; P = 0.0001) indicated a more efficient N utilization. The increase in serum triacylglycerol (58% and 28%, respectively; P = 0.0098) indicated that CLA and betaine + CLA could have reduced adipose tissue triacylglycerol synthesis from preformed fatty acids. Serum glucose, low-density lipoprotein (LDL) cholesterol and non-esterified fatty acids were unaffected. CLA and betaine + CLA altered serum lipids profile, although liver of pigs fed CLA diet presented no histopathological changes and triglyceride content was not different from Control pigs. Compared with controls, serum growth hormone decreased (20% to 23%; P = 0.0209) for all treatments. Although serum insulin increased in CLA, and especially in betaine + CLA pigs (28% and 83%; P = 0.0001), indices of insulin resistance were unaffected. In conclusion, CLA, and especially betaine + CLA, induced changes in biochemical parameters and hormones that may partially explain a nutrient partitioning effect in young pigs. Nevertheless, they exhibited weak, although detrimental, effects on blood lipids. Moreover, although livers were chemically and histologically normal, pigs fed CLA diet challenged with a glucose load had higher serum glucose than controls.

Differences in portal appearance of lysine and methionine in Iberian and Landrace pigs.

Compared to modern breeds, Iberian pigs have lower rates of muscle protein deposition and greater viscera weight. Factors that limit growth performance of Iberian pigs are unknown. We hypothesized that differences in net portal appearance of the essential AA Lys and Met might partially explain the lower growth rate reported in Iberian pigs compared to modern breeds. Net portal appearance of AA was measured in 6 Iberian and 6 Landrace gilts (28 kg BW) fitted with chronic catheters in the portal vein, carotid artery, and mesenteric vein. Blood samples were taken for 6 h after feeding 2 isoenergetic diets (14.0 to 14.5 MJ ME/kg DM) with low CP (LCP) and high CP (HCP) (13 vs. 16%, respectively) in a crossover design after a 1-wk adaptation. Net portal appearance of Lys was greater (P < 0.05) when pigs consumed the HCP than LCP diet (23 vs. 15 µmol/min); breeds did not differ. Net portal appearance of Met tended to be higher (P = 0.076) for pigs fed HCP than LCP diet (4.4 vs. 3.6 µmol/min); breeds did not differ. In conclusion, differences in portal appearance of Lys and Met did not exist between Iberian and Landrace pigs fed barley (Hordeum vulgare)-soybean (Glycine max) diets with low and high protein concentration.

Comparative proteomic analysis of Fasciola hepatica juveniles and Schistosoma bovis schistosomula.

Protein interactions between host and parasites can influence the infection success and severity. The aim of this investigation was to identify the proteins from two trematodes potentially localized at the host-parasite interface. We performed the proteomic profiles from in vivo obtained immature lung stage Schistosoma bovis schistosomula and in vitro excysted juveniles from Fasciola hepatica, parasites of ruminants and man usually giving rise to chronic infections. Proteomes from those parasites were obtained after digestion with trypsin and the peptides generated were identified by mass spectrometry, both before and after parasites' treatment with 70% methanol. The comparison of the two proteome sets from each parasite and between them, the analysis of their relative abundance and of their potential exposure to the host from living parasites, together with the specific immunolocalization of two of the identified molecules, show that this approach could assist in the identification of parasite exposed proteins and in the definition of molecules common for the two parasites with potential interaction with the host. Further characterization of these molecules could guide to define new common anti-parasitic targets and potential vaccine candidates.

Specific real-time PCR for simultaneous detection and identification of Legionella pneumophila serogroup 1 in water and clinical samples.

Legionella pneumophila, a bacterium that replicates within aquatic amoebae and persists in the environment as a free-living microbe, is the causative agent of Legionnaires' disease. Among the many Legionella species described, L. pneumophila is associated with 90% of human disease, and within the 15 serogroups (Sg), L. pneumophila Sg1 causes more than 84% of Legionnaires' disease worldwide. Thus, rapid and specific identification of L. pneumophila Sg1 is of the utmost importance for evaluation of the contamination of collective water systems and the risk posed. Previously we had shown that about 20 kb of the 33-kb locus carrying the genes coding for the proteins involved in lipopolysaccharide biosynthesis (LPS gene cluster) by L. pneumophila was highly specific for Sg1 strains and that three genes (lpp0831, wzm, and wzt) may serve as genetic markers. Here we report the sequencing and comparative analyses of this specific region of the LPS gene cluster in L. pneumophila Sg6, -10, -12, -13, and -14. Indeed, the wzm and wzt genes were present only in the Sg1 LPS gene cluster, which showed a very specific gene content with respect to the other five serogroups investigated. Based on this observation, we designed primers and developed a classical and a real-time PCR method for the detection and simultaneous identification of L. pneumophila Sg1 in clinical and environmental isolates. Evaluation of the selected primers with 454 Legionella and 38 non-Legionella strains demonstrated 100% specificity. Sensitivity, specificity, and predictive values were further evaluated with 209 DNA extracts from water samples of hospital water supply systems and with 96 respiratory specimens. The results showed that the newly developed quantitative Sg1-specific PCR method is a highly specific and efficient tool for the surveillance and rapid detection of high-risk L. pneumophila Sg1 in water and clinical samples.

Susceptibilidad a los agentes antimicrobianos en cepas de Staphylococcus Aureus aisladas en leche de bovinos con mastitis subclínica y leche de tanque / Antimicrobial Susceptibility of Staphylococcus aureus Strains Isolated from Subclinical Bovine Mastitis Milk and Bulk Tank Milk

Staphylococcus aureus es uno de los principales patógenos bacterianos de mastitis bovina en el estado Zulia - Venezuela. El objetivo del presente estudio fue determinar la susceptibilidad a los agentes antimicrobianos de cepas de S. aureus aisladas de muestras de leche de cuartos mamarios con mastitis subclínica y leche de tanque en tres fincas del estado Zulia. Un total de 88 cepas fueron analizadas, de estos aislamientos 81 fueron aislados de leche de cuartos mamarios y 7 de leche de tanque. A las cepas se le determinó la sensibilidad a los agentes antimicrobianos por el método de difusión del disco en agar, correspondientes a: vancomicina (Va), estreptomicina (S), clindamicina (Cc), eritromicina (E), tetraciclina (Te), penicilina (Pe), ciprofloxacina (Cip), cefoxitina (Fox), cloranfenicol (C), enrofloxacina (Enr), rifampicina (Rif), trimetoprim/sulfametoxazol (SXT), oxacilina (Ox) y gentamicina (G). La concentración inhibitoria mínima (CIM) se determinó a Pe, Va, Ox, y E. Las cepas de S. aureus aisladas de leche de cuartos mamarios fueron resistentes a Pe (12,3%), Enr (8,6%), Cip, S (4,9%, respectivamente), E (3,7%), Cc, y Rif (2,5%, respectivamente). Las cepas aisladas de leche de tanque fueron resistentes a Pe y Te (28,6%) y Cc (14, 3%). Los resultados indican que los aislamientos de S. aureus provenientes de leche de cuartos mamarios con mastitis subclínica fueron más susceptibles a todos los agentes antimicrobianos (70%) en comparación con las cepas procedentes de la leche de tanque (57%).

Staphylococcus aureus is one of the main bacterial pathogens involved in bovine mastitis in Zulia State-Venezuela. The objective of this research was to determine the susceptibility to antimicrobial agents of Staphylococcus aureus isolated from the mammary quarter milk of subclinical mastitis cows and from bulk tank milk in three dairy farms of Zulia State. A total of 88 strains were analyzed, of these isolates 81 were isolated from quarter milk and 7 from bulk tank milk. The susceptibility to antimicrobial agents of those strains was determined by agar disk diffusion test. The antibiotics tested were: vancomycin (Va), streptomycin (S), clindamycin (Cc), erithromycin (E), tetracycline (Te), penicillin (Pe), ciprofloxacin (Cip), cefoxitin (Fox), chloramfenicol (C), enrofloxacin (Enr), rifampicin (Rif), trimethoprim/sulfametoxazole (SXT), oxacillin (Ox) and gentamicin. (G). Minimum Inhibitory Concentration (MIC) for Pe, Va, Ox and E was determined. S. aureus strains isolated from quarter milk were resistant to Pe (12.3%), Enr (8.6%), Cip, S (4.9%, respectively), E (3.7%), Cc, and Rif (2.5%, respectively). Strains isolated from bulk tank milk were resistant to Pe and Te (28.6%, respectively) and Cc (14.3%). The results indicate that isolates of S. aureus from mammary quarter milk of subclinical mastitis were more susceptible to all antimicrobial agents (70%) compared with strains from the bulk tank milk (57%).

The Legionella pneumophila F-box protein Lpp2082 (AnkB) modulates ubiquitination of the host protein parvin B and promotes intracellular replication.

The environmental pathogen Legionella pneumophila encodes three proteins containing F-box domains and additional protein-protein interaction domains, reminiscent of eukaryotic SCF ubiquitin-protein ligases. Here we show that the F-box proteins of L. pneumophila strain Paris are Dot/Icm effectors involved in the accumulation of ubiquitinated proteins associated with the Legionella-containing vacuole. Single, double and triple mutants of the F-box protein encoding genes were impaired in infection of Acanthamoeba castellanii, THP-1 macrophages and human lung epithelial cells. Lpp2082/AnkB was essential for infection of the lungs of A/J mice in vivo, and bound Skp1, the interaction partner of the SCF complex in mammalian cells, similar to AnkB from strain AA100/130b. Using a yeast two-hybrid screen and co-immunoprecipitation analysis we identified ParvB a protein present in focal adhesions and in lamellipodia, as a target. Immunofluorescence analysis confirmed that ectopically expressed Lpp2082/AnkB colocalized with ParvB at the periphery of lamellipodia. Unexpectedly, ubiquitination tests revealed that Lpp2082/AnkB diminishes endogenous ubiquitination of ParvB. Based on these results we propose that L. pneumophila modulates ubiquitination of ParvB by competing with eukaryotic E3 ligases for the specific protein-protein interaction site of ParvB, thereby revealing a new mechanism by which L. pneumophila may employ translocated effector proteins to promote bacterial survival.