Eficacia antiparasitaria in vitro del extracto de quebracho (Schinopsis balansae) sobre larvas infectantes de Haemonchus contortus de ovinos / In vitro antiparasitic efficacy of the quebracho extract (Schinopsis balansae) on infecting larvae of Haemonchus contortus of sheep

RESUMEN El objetivo de este trabajo fue evaluar in vitro la eficacia del extracto de quebracho (Schinopsis spp.), rico en taninos condensados, en el control de H. contortus de ovinos, ya que existen evidencias de que estos taninos pueden reducir la excreción de huevos, la fecundidad de las hembras y la carga de parásitos adultos. Para evaluar el efecto antihelmíntico in vitro sobre larvas infectantes de H. contortus susceptibles a todos los grupos químicos, se utilizó el test de inhibición de migración larval (IML) a 3 concentraciones diferentes (5 mg/ml, 15 mg/ml y 30 mg/ml). El efecto de los tratamientos fue analizado mediante un análisis de varianza y la estimación de las diferencias entre grupos se realizó por medio de la prueba LSD Fisher. Los resultados del test in vitro demostraron una reducción de la migración larval que varió entre el 74% y el 80%, a las concentraciones de entre 5 mg/ml y 30 mg/ml. Del análisis de varianza surgen diferencias significativas entre tratamientos (p = 0,0494). Al realizar la prueba de comparación de medias se evidenciaron diferencias significativas (p < 0,05) entre los promedios de migración a las diluciones de 5 mg/ml y 15 mg/ml, y de 5 mg/ml y 30 mg/ml, mientras que no se detectaron diferencias significativas entre la dilución de 15 mg/ml y 30 mg/ml. Estos resultados señalaron que el extracto de quebracho, a las diluciones evaluadas in vitro, presentó actividad antihelmíntica sobre larvas L3 susceptibles de H. contortus. Sin embargo, se requiere ampliar los estudios in vivo para demostrar un efecto antihelmíntico en ovinos.

ABSTRACT The objective of this work was to evaluate in vitro efficacy of the quebracho extract (Schinopsis spp.), rich in condensed tannins, against H. contortus in sheep, since there is evidence that this tanninsthese tannins can reduce egg excretion, fecundity of females and the burden of adult parasites. A larval migration inhibition (IML) test with 3 different concentration (5 mg/ml, 15 mg/ml, and 30 mg/ml) was used to evaluate the in vitro anthelmintic effect upon iInfective H. contortus larvae,from a susceptible strain to all chemical groupswere utilized with 3 diferentconcentration (5mg/ml, 15mg/ml, and 30mg/ml). The effect of the treatments was submitted to a variance analysis and the estimation of the differences between groups was evaluated using LSD Fisher test. Results from the in vitro test, revealed a reduction of the larval migration that varies from 74% to 80%, at the concentrations between 5 mg/ml to 30 mg/ml. From the analysis of variance, significant differences appear between treatments (p = 0,0494). After When performing the mean comparison test were performed, significant differences (p < 0,05) were found between the migration averages at dilutions of 5 mg/ml and 15 mg/ml, and between 5 mg/ml and 30 mg/ml, while were no't detected significant differences between the dilution of 15 mg/ml and 30 mg/ml. These results indicated that quebracho extract at the dilutions evaluated in vitro showed anthelmintic activity on L3 susceptible to H. contortus. However, it is necessary to conduct further studies in vivo to demonstrate an anthelmintic effect in sheep.

Halting pro-survival autophagy by TGFß inhibition in bone marrow fibroblasts overcomes bortezomib resistance in multiple myeloma patients.

Bortezomib (bort) has improved overall survival in patients with multiple myeloma (MM), but the majority of them develop drug resistance. In this study, we demonstrate that bone marrow (BM) fibroblasts (cancer-associated fibroblasts; CAFs) from bort-resistant patients are insensitive to bort and protect the RPMI8226 and patients' plasma cells against bort-induced apoptosis. Bort triggers CAFs to produce high levels of interleukin (IL)-6, IL-8, insulin-like growth factor (IGF)-1 and transforming growth factor (TGF) ß. Proteomic studies on CAFs demonstrate that bort resistance parallels activation of oxidative stress and pro-survival autophagy. Indeed, bort induces reactive oxygen species in bort-resistant CAFs and activates autophagy by increasing light chain 3 protein (LC3)-II and inhibiting p62 and phospho-mammalian target of rapamycin. The small-interfering RNA knockdown of Atg7, and treatment with 3-methyladenine, restores bort sensitivity in bort-resistant CAFs and produces cytotoxicity in plasma cells co-cultured with CAFs. In the syngeneic 5T33 MM model, bort-treatment induces the expansion of LC3-II(+) CAFs. TGFß mediates bort-induced autophagy, and its blockade by LY2109761, a selective TßRI/II inhibitor, reduces the expression of p-Smad2/3 and LC3-II and induces apoptosis in bort-resistant CAFs. A combination of bort and LY2109761 synergistically induces apoptosis of RPMI8226 co-cultured with bort-resistant CAFs. These data define a key role for CAFs in bort resistance of plasma cells and provide the basis for a novel targeted therapeutic approach.

Comparative proteomic profiling of Hodgkin lymphoma cell lines.

Classical Hodgkin lymphoma (cHL) is a malignancy with complex pathogenesis. The hallmark of the disease is the presence of large mononucleated Hodgkin and bi- or multinucleated Reed/Sternberg (H/RS) cells. The origin of HRS cells in cHL is controversial as these cells show the coexpression of markers of several lineages. Using a proteomic approach, we compared the protein expression profile of cHL models of T- and B-cell derivation to find proteins differentially expressed in these cell lines. A total of 67 proteins were found differentially expressed between the two cell lines including metabolic proteins and proteins involved in the regulation of the cytoskeleton and/or cell migration, which were further validated by western blotting. Additionally, the expression of selected B- and T-cell antigens was also assessed by flow cytometry to reveal significant differences in the expression of different surface markers. Bioinformatics analysis was then applied to our dataset to find enriched pathways and networks, and to identify possible key regulators. In the present study, a proteomic approach was used to compare the protein expression profiles of two cHL cell lines. The identified proteins and/or networks, many of which not previously related to cHL, may be important to better define the pathogenesis of the disease, to identify novel diagnostic markers, and to design new therapeutic strategies.

The impact of proteomics in the understanding of the molecular basis of Paclitaxel-resistance in ovarian tumors.

The current therapy for ovarian cancer has advanced from alkylating agents, to a combination of carboplatinum and paclitaxel offering increased survival. Although most patients respond to this first-line therapy, initially, the majority of these patients relapse within 2 years. The mechanisms responsible for acquired drug resistance in ovarian cancer have been elucidated only in part. They include i) enhanced drug export, ii) activation/inhibition of intracellular signalling pathways, iii) molecular alterations in tubulin isotype composition. A better understanding of these mechanisms is needed, in order to develop new approaches, aimed at overcoming resistance to anticancer agents, and to reveal the complexity of causes, which contribute to drug resistance. In this review we offer an updated overview of proteomic studies on the molecular mechanisms of paclitaxel resistance. These proteomic studies also identify potential targets for modulating drug resistance, that could be predictive of response to chemotherapy in ovarian carcinomas.

SU-E-J-180: A Characterization of the LAP Aquarius Phantom for External LAP Laser Alignment and MR Geometric Distortion Verification for the use of SRS Patient Simulation.

PURPOSE: To explore additional application of the new Aquarius external laser alignment verification Phantom by LAP (Aq-LAP Phantom) examining geometric accuracy of magnetic resonance images (MRI) commonly used for planning intracranial stereotactic radiation surgery (ICSRS) cases. METHODS: Newly designed external patient alignment lasers were first aligned by the Aq-LAP Phantom at a Siemens Magneton Vario 3T MR unit. The scans were then performed with the T1 Axial 3D MPRAGE protocols with 0.9 mm temporal resolution, which may be used for ICSRS. They also include FLAIR, T2 BLADE and Diffusion Axial TRACE imaging acquisitions with 1 mm temporal resolution. The MRI will be fused to 1 mm cut computerized tomography (CT) images acquired by a Siemens Somatom Sensation Open©. The geometric distortions (GD) were measured against the CT in all axial, sagital, and coronal directions at different levels. RESULTS: MR images of the Aquarius Phantom indicate a distinct similarity between the nonlinear GD along the z-axis crosshair and typical magnetic field gradient nonlinearity. There is linear correlation between MR divergence datasets of distorted crosshairs (p-values from 0.57 to 0.00), and nonlinear correlation between MR divergence datasets of the distorted crosshair with the CT divergence datasets of the cross plane (p-values from 8.45×10̂-4 to 1.38×10̂-46). The margin of error exceeded no more than 0.29 mm. GDs up to about 2 mm are observed at the distal regions of the longitudinal axis in the SRS treatment planning MR images. CONCLUSIONS: Using the Aquarius Phantom, one is able to detect GD in ICSRS planning MRI acquisitions, and align the external LAP patient alignment lasers by following the LAP QA. Based on the results, one may recommend using the Aquarius Phantom to determine if margins should be included for SRS treatment planning. The Aquarius Phantom, used for laser alignment and geometric distortion detection, was provided by LAP of America.

¿Es concordante el tamaño tumoral medido en resonancia magnética mamaria y el de la biopsia quirúrgica en pacientes con cáncer de mama, tratadas con quimioterapia neoadyuvante? / Is breast MRI concordant with pathologic tumor size in breast cancer patients treated with neo-adjuvant chemotherapy?

Neoadjuvant chemotherapy (NACT) is a treatment used in those breast cancers initially inoperable due to their size, and also in operable breast cancers where NACT could increase the rate of conservative breast surgery. To assess tumor response to treatment, clinical examination, mammography, ultrasound, and breast MRI are used, the latter being the modality that yields the best correlation with histologic tumor volume. We evaluated the correlation of tumor sizes as measured by MRI versus surgical pathological specimen in breast cancers treated with NACT. Eighteen patients underwent MRI to monitor NACT; in 15 (83 percent) of them the final biopsy was obtained. In this group a very good correlation was observed, with a mean difference between MRI and histology of 4 mm regarding tumor volumen, which has allowed an adequate management of patients in our daily practice.

La quimioterapia neoadyuvante (QTNA) es un tratamiento usado en aquellos cánceres mamarios cuyo tamaño los hace inoperables al momento del diagnóstico y en cánceres mamarios operables, pero cuyo uso podría permitir una cirugía conservadora. Para evaluar la respuesta al tratamiento, se ha utilizado el examen clínico, la mamografía, el ultrasonido y la resonancia magnética mamaria, siendo ésta última la que mejor correlación tiene con el tamaño tumoral histológico. Quisimos evaluar la concordancia del tamaño tumoral medido en resonancia magnética con el de la biopsia quirúrgica, en cánceres mamarios tratados con QTNA. Dieciocho pacientes se realizaron resonancia magnética para monitorización de QTNA, en 15 (83 por ciento) de ellas se obtuvo la biopsia definitiva. En este grupo observamos una muy buena correlación, con una diferencia promedio de 4 mm, entre el tamaño de la resonancia y el de la histología, lo que permitió en nuestro medio un adecuado manejo de las pacientes.

Utility of high-resolution computed tomography and BAL in cryptogenic organizing pneumonia.

BACKGROUND: Cryptogenic organizing pneumonia (COP) is a rare disease, and its diagnosis requires histological confirmation. The objective of our study was to describe the findings of the thoracic high-resolution computed tomography (HR-CT) and bronchoalveolar lavage (BAL) in patients with confirmed COP and evaluate the complementary diagnostic use of BAL and thoracic HR-CT. METHODS: Patients recorded in the registry of interstitial pulmonary diseases between 1991 and 2008 were located and the COP patients selected. RESULTS: We identified 21 patients with histological confirmation of COP. The median age was 58.0 ± 15.9 years, and 61.9% of patients were female. The most frequent thoracic HR-CT profile was patchy infiltrate (71.4%), followed by parenchymatous consolidation (42.9%). The most frequent BAL profile was mixed alveolitis (62%) with lymphocyte predominance, a CD4/CD8 index of 0.4 and foamy macrophages. The effectiveness of transbronchial biopsy was 66.6%. The diagnostic utility of Poletti's BAL criteria gives us a specificity of 88.8%, although the sensitivity obtained was low. The specificity of certain HR-CT profiles is 99%. In addition, we observed a complementary use of the HR-CT and the BAL. CONCLUSIONS: The imaging findings and BAL could be useful for patients with appropriate clinical presentation and for those whose transbronchial biopsy is negative or for whom a confirmatory biopsy cannot be performed.

Biomolecular pathogenesis of borderline ovarian tumors: focusing target discovery through proteogenomics.

Tumors of the epithelial surface account for about 80% of all ovarian neoplasms and exhibit a heterogeneous histological classification affecting survival. Tumors of low malignant potential, defined as borderline ovarian tumors(BOTs), have a markedly better survival and low recurrence, even if surgery still represents the common management for this type of cancer. It is still debated in the literature if BOTs can be considered as intermediate precursors in the progression to high grade ovarian tumors. Evidences now propose that high-grade serous carcinomas are not associated with a defined precursor lesion. Together with histopathological studies, mutations of KRAS, BRAF and p53 genes, microsatellite instability (MSI)and under- or over-expression of many genes and proteins have been used to address this question. Despite the large body of data summarized, a limited number of molecules proved to be useful in elucidating BOTs pathogenesis and only a few of these showed possible application in the therapy. We believe that high-throughput technologies would help to overcome these limitations offering the promise of a better understanding of BOTs classification. The aim is to guide the diagnosis and prognosis of BOTs to develop new possible therapeutic molecular targets avoiding surgery.

Enteroclisis (enterografía) por tomografía computada / Enteroclysis (enterography) by computed tomography

La enterografía con TCMC se ha transformado en el método de imagen de elección en el estudio del Intestino delgado (ID), dada su alta sensibilidad y comparativamente bajo costo. Se basa en la distensión del ID usando contraste "neutro" y adquisición volumétrica con TCMC después de la inyección de contraste yodado. La distensión del ID y el contraste yodado sumado a la capacidad multiplanar de la TCMC permite evaluar las características de la pared intestinal tales como grosor, presencia de tumores, procesos inflamatorios y compromiso de la grasa vecina. La distensión del lumen facilita la demostración de las oclusiones intestinales de bajo grado.

MDCT enterography has become the best imaging modality to study the small bowel due to its high sensibility, specificity and relatively low cost. It is based in the small bowel distention with neutral contrast and intravenous iodine contrast. These two factors with the multiplanar capability of ultrafast MDCT allows to show the characteristics of the small bowel wall such as thickness, inflammation, tumor and alterations of the surrounding fat. The stenotic areas are also nicely shown because of the distention of the lumen.

Ultrasonografía endorectal: estado actual / Endorectal ultrasonography: current status

La ultrasonografía endorectal (USER) ha sido ampliamente usada en patología recto anal debido a su bajo costo, alta disponibilidad y alto rendimiento en centros especializados. Sin embargo, su principal limitación es la dependencia en la experiencia del examinador. La técnica se caracteriza por el uso de un transductor especialmente dedicado, circular, que permite obtener imágenes axiales en 360 grados perpendiculares al eje longitudinal del recto y ano. Su principal utilidad en la actualidad se extiende al estudio de la incontinencia fecal, generalmente producida por el daño del esfinter anal por trauma obstétrico. Los desgarros propios del trauma como también la atrofia del músculo esfinteriano producto de la denervación del esfinter post parto son demostrados con alta confiabilidad. Los procesos inflamatorios pueden también ser demostrados por éste método. El trayecto fistuloso puede ser seguido en toda su extensión especialmente si es inyectado con H2O2 que lo hace más fácilmente visible. La RM sin embargo ha sido empleada con frecuencia dada la gran calidad de imágenes que pueden ser obtenidas. En la actualidad, en el cáncer de recto su rol se limita solamente a aquellas pequeñas neoplasias en estadio T1 o en algunos casos estadios T2 cuando se planea una resección local. Sin embargo, hoy en día, la Resonancia Magnética, debido a su alta resolución espacial y de contraste tisular es el método de elección en la estadificación del cáncer rectal.

Diagnóstico de infección invasora en quemaduras: comparación de tres métodos / Diagnosis of invasive infection in burns: comparison of 3 methods

En 10 pacientes con quemaduras de tercer grado de 19ñ4,5 por ciento de superficie corporal comprometida, se tomaron biopsias de la herida el día 5§, 12§ y 18§ para comparar prospectivamente el rendimiento individual de la biopsia contemporánea por congelación, biopsia diferida por inclusión y el recuento de colonias por gramo de tejido, de los pacientes con infección invasora y potencial desarrollo de sepsis secundaria. En 6 casos se produjo infección invasora clínica. De ellos, 2 pacientes presentaron sépsis. En 4 pacientes no se identificó infección invasora. No hubo mortalidad. En los pacientes con infección invasora, 15/18 muestras evidenciaron gérmenes en tejido vital en la biopsia contemporánea. En la biopsia diferida, 16/18 y 14/18 muestras mostraron gérmenes e inflamación, lo que es comparable para ambos métodos. En el grupo sin infección invasora, 2/12 muestras señalaron gérmenes con ambas técnicas. El recuento de coloniaspor gramo fue 10 5 en 14/18 muestras en el grupo con infección invasora y 10 3 en 8/12 muestras en el segundo grupo, diferencia estadísticamente significativa. Se concluye que en una experiencia inicial, los métodos biópticos y microbiológicos cuantitativos han sido efectivos para diagnosticar precozmente la infección invasora de la herida con una predicción de 78 y 83 por ciento respectivamente, lo que permitirá ajustar oportunamente el tratamiento local y sistémico en futuros casos de alto riesgo y evitar la sepsis. Se destaca la ventaja en la rapidez de información que ofrece la biopsia contemporánea por congelación

Infección en quemaduras: evaluación de antimicrobianos, escarectomía y balneoterapia / Burns infection: antimicrobial agents evaluation, scarectomy and balneotherapy

Se compara retrospectivamente la incidencia de infección local, sépsis y mortalidad de 178 pacientes quemados graves tratados entre 1975 y 1987. Hasta 1978 no hubo tratamiento local específico. Desde 1979 se regularizó el uso de antimicrobiano local. En 1982 se agregó escarectomía tangencial y en 1986, balneoterapia en solución de hipoclorito de sodio 125 ppm y asistencia nutricional intensiva. El número de pacientes, edad y superficie comprometida en los cuatro períodos son comparables (f > 0.05). La superficie corporal quemada (SCQ) media fue de 14.7% y la edad media de 42,6 años. La SCQ del grupo con sepsis fue 23,9% (f = 0,007) y su edad media fue de 46,2 años (f = 0,42). La mortalidad por sepsis fue de 17,3%, con SCQ media de 31,5% y la edad media de 62 años; estadísticamente mayores que los no fallecidos (f = 0,0001). El principal germen aislado en sepsis fue Pseudomona aeruginosa con incidencia de 43,4%. Se concluye que las mejoras en el tratamiento redujeron la incidencia de infección local y sepsis; la incidencia de sepsis crece con el aumento de la SCQ; la mortalidad no se modificó con las mejoras de tratamiento y depende como factor principal de la SCQ y edad de los pacientes