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Abstract Immune checkpoint blockade (ICB) approaches have changed the therapeutic landscape for many tumor types. However, half of cutaneous squamous cell carcinoma (cSCC) patients remain unresponsive or develop resistance. Here, we show that, during cSCC progression in male mice, cancer cells acquire epithelial/mesenchymal plasticity and change their immune checkpoint (IC) ligand profile according to their features, dictating the IC pathways involved in immune evasion. Epithelial cancer cells, through the PD-1/PD-L1 pathway, and mesenchymal cancer cells, through the CTLA-4/CD80 and TIGIT/CD155 pathways, differentially block antitumor immune responses and determine the response to ICB therapies. Accordingly, the anti-PD-L1/TIGIT combination is the most effective strategy for blocking the growth of cSCCs that contain both epithelial and mesenchymal cancer cells. The expression of E-cadherin/Vimentin/CD80/CD155 proteins in cSCC, HNSCC and melanoma patient samples predicts response to anti-PD-1/PD-L1 therapy. Collectively, our findings indicate that the selection of ICB therapies should take into account the epithelial/mesenchymal features of cancer cells.

Background Tebentafusp, a bispecific (gp100×CD3) ImmTAC, significantly improved overall survival (OS) outcomes for HLA-A*02:01+ adult patients with untreated metastatic uveal melanoma (mUM) and showed promising survival in previously treated mUM with 1-year OS of 62% in the primary analysis of study IMCgp100-102. Here we report long-term outcomes from this phase 1/2 study in pretreated mUM.Patients and methods Patients with previously treated mUM received tebentafusp weekly intravenous at 20 µg dose 1, 30 µg dose 2 and either 54, 64, 68, or 73 µg (phase 1) or 68 µg (phase 2) dose 3+. The primary objective was overall response rate. Secondary objectives included OS and safety. OS was estimated by Kaplan-Meier methods. Association between OS and baseline covariates, on-treatment Response Evaluation Criteria in Solid Tumors (RECIST) response, baseline tumor biopsy and circulating-tumor DNA (ctDNA) changes were assessed.Results 146 patients were treated with tebentafusp: 19 in phase 1 and 127 in phase 2. With a median follow-up duration of 48.5 months, the median OS was 17.4 months (95% CI, 13.1 to 22.8), and the 1-year, 2-year, 3-year and 4-year OS rates were 62%, 40%, 23% and 14%, respectively. Improved survival was associated with lower ctDNA baseline levels and greater ctDNA reductions by week 9 on-treatment, with 100% 1-year, 73% 2-year and 45% 3-year OS rates for patients with ctDNA clearance. Baseline gp100 expression was not associated with survival, despite more RECIST responses among patients with higher expression. No new safety signals were reported with long-term dosing.Conclusions This study represents the longest follow-up of a Tcell receptor bispecific to date and confirms the durable survival benefits achieved with tebentafusp in previously treated mUM with good tolerability long-term. A role for ctDNA reduction as an early indicator of clinical benefit was again suggested for patients treated with tebentafusp.

Antecedentes: Dada su importancia para evaluar la digestibilidad de los alimentos para rumiantes, el artículo se propuso brindar información sobre algunos aspectos generales del desarrollo de la técnica de producción de gas in vitro y referirse a los usos que hoy día se dan a la misma. Desarrollo: Los orígenes de la técnica se remontan a los trabajos en 1979. Desde entonces ha habido cambios en su forma de aplicarla manteniendo la esencia de medir la producción de gas que se produce cuando se inocula la muestra con líquido ruminal, heces o enzimas en un medio mineral amortiguado por bicarbonatos que libera una cantidad de gas proporcional a los ácidos grasos volátiles producidos. La forma en que se mide la cantidad de gas ha cambiado hasta llegar a la utilización de transductores que se acoplan a computadoras. La técnica ha sido usada para múltiples estudios que se reseñan en el artículo, a la par que la referencia a algunos trabajos dedicados a su perfeccionamiento a partir de dificultades que se presentaron. Conclusiones: Se concluye que la técnica in vitro de producción de gases es probablemente la herramienta más versátil de medición de la digestibilidad; presenta procedimientos didácticos y accesibles; acarrea menos costos; además representa una alternativa más compatible para las legislaciones de bienestar animal, al usar las heces como inóculo, y que queda aún por hacer para lograr una mejor precisión y exactitud y la posible aplicación en otros campos de evaluación de los alimentos para animales. Palabras clave: digestibilidad, ética animal, evaluación de alimentos, heces depuestas (Fuente: AIMS)

Dentro de la multiplicidad de factores de la alimentación, se identifican el conocimiento o los saberes que parten de información que se adquiere a través de diferentes medios, incidiendo en la modificación del acto alimentario. El objetivo de la investi-gación fue analizar desde un enfoque de género el conocimiento sobre la sustentabilidad de la dieta de adultos mexicanos. Se aplicó una encuesta electrónica a 559 habitantes del centro del país. El instrumento incluyó 21 variables que permitieron identificar las características sociodemográficas y los saberes sobre el impacto ambiental de la alimentación. Por medio de Chi cuadrada se pudo establecer que la muestra, de la cual la mitad corresponde a mujeres jóvenes de una zona urbana, presenta diferencias estadística-mente significativas en cuanto a la disposición para adoptar una alimentación con características sustentables, teniendo la percep-ción de las frutas como un factor importante en patrones alimentarios de menor impacto ambiental; esto contrastado entre sexos. La similitud entre conocimientos de sustentabilidad de la dieta entre los grupos indica una misma disposición hacia un consumo de alimentos ambientalmente favorables. Sin embargo, los encuestados presentan una percepción de la carne como punto importante para una alimentación sostenible, que podría deberse a la falta de educación nutricional y ambiental.

Abstract Background . Up to 20% of people with HIV (PWH) who undergo virologically suppressed antiretroviral therapy (ART) fail to experience complete immune restoration. We recently reported that plasma anti-CD4 IgG (antiCD4IgG) autoantibodies from immune non-responders specifically deplete CD4 + T cells via antibody-dependent cytotoxicity. However, the mechanism of antiCD4IgG production remains unclear. Methods . Blood samples were collected from 16 healthy individuals and 25 PWH on suppressive ART. IgG subclass, plasma lipopolysaccharide (LPS), and antiCD4IgG levels were measured by ELISA. Gene profiles in B cells were analyzed by microarray and quantitative PCR. Furthermore, a patient-derived antiCD4IgG–producing B cell line was generated and stimulated with LPS in vitro. B cell IgG class switch recombination (CSR) was evaluated in response to LPS in splenic B cells from C57/B6 mice in vitro. Results . Increased plasma anti-CD4 IgGs in PWH were predominantly IgG1 and associated with increased plasma LPS levels as well as B cell expression of TLR2, TLR4, and MyD88 mRNA in vivo. Furthermore, LPS stimulation induced antiCD4IgG production in the antiCD4IgG B cell line in vitro. Finally, LPS promoted CSR in vitro. Conclusion . Our findings suggest that persistent LPS translocation may promote anti-CD4 autoreactive B cell activation and antiCD4IgG production in PWH on ART, which may contribute to gradual CD4 + T cell depletion. This study suggests that reversing a compromised mucosal barrier could improve ART outcomes in PWH who fail to experience complete immune restoration.

La dieta desequilibrada impacta en el medioambiente a través de la agricultura, como un sector de alta emisión de gases de efecto invernadero (GEI) que usa el 70% de las reservas mundiales de agua. Además, existe una desigualdad en la distribución y uso de este recurso que se acrecentó durante el confinamiento por coronavirus. El objetivo de esta investigación fue identificar los cambios en la dieta y el cuidado del agua en la zona centro de México durante la pandemia de COVID-19. Encontrándose un aumento en el consumo de verduras (71.7%) y frutas (64.9%). En el cuidado del agua en casa la atención de fugas (58.4%) y regulación del tiempo de baño en regadera (54.5%) mostraron un incremento. Así mismo, se identificaron grupos caracterizados por acciones dirigidas a una alimentación adecuada y menos contaminante, un mejor cuidado del agua y la percepción de beneficios en la participación del cuidado del medio ambiente. La emisión de CO2 semanal de la dieta permitió identificar características del consumo de alimentos durante pandemia y una participación hacia mejorar el ecosistema. Dando bases para plantear estrategias que partan de las actividades que llevan a cabo las personas o que fomenten una mayor incidencia de estas.

Herbal formulas supplemented have improved the health of ruminants. Our objectives were to determine the effects of supplemental Indian polyherbal phytogenic sources (IndPPs) on the growth performance, blood chemistry, blood cells, and antibody amounts of dairy calves. Forty Holstein calves (Initial BW 45.8 ± 7.2 kg) were randomly assigned to treatments: 0, 1, 2, and 3 gd-1 of IndPPs. The herbal supplement was characterized by GC-MS analysis. The calves treated with IndPPs additive did not affect the average productive variable involved in growth. The use of IndPPs linearly reduces the blood serum biochemistry glucose (P

Antecedentes: Los estudios del valor nutritivo de las hojas peciolos de Dichrostachys cinerea son insuficientes para definir el secado de las muestras en estudios de digestibilidad in vitro. El objetivo fue calcular .la producción de gas in vitro y parámetros de mejor ajuste a modelos como indicadores de la digestibilidad del follaje de la planta a dos alturas y con dos métodos de secado. Métodos: Las muestras se colectaron manualmente de forma aleatoria de 10 árboles diferentes (de 1 m y 2 m de altura), en época de seca, simulando el ramoneo de la planta en su totalidad. Cada muestra se dividió para ser sometida a dos tratamientos de secado (a temperatura ambiente aproximadamente por 96 h y a 550 C en estufa), hasta peso constante. Se evaluó la influencia de la altura de la planta y temperatura de secado en los volúmenes de gas in vitro producido a las 24, 48 y 72 h, utilizando como inóculo heces bovinas, mediante análisis de varianza de clasificación simple de las combinaciones de factores. Resultados: La producción de gas in vitro de hojas peciolos de plantas de 2 m, secadas a 55 °C fue significativamente menor que el resto de las combinaciones temperatura-altura evaluadas. Conclusiones: Los parámetros de mejor ajuste a las ecuaciones de producción de gas in vitro de hojas peciolo de Dichrostachys cirenea muestran la posibilidad de su uso en la alimentación de rumiantes debido a su potencial nutricional similar al de varias especies arbustivas forrajeras de la familia leguminosa. Palabras clave: valor nutritivo, leguminosas, follaje, digestibilidad in vitro (Fuente: AIMS)

Whether intermittent strategies of delivering drugs can improve cancer patients survival is still unclear. Here, the authors reports the results of a randomized phase II clinical trial aimed to compare the efficacy and safety of two dosing regimens (continuous and intermittent) of vemurafenib and cobimetinib combination as first-line treatment of patients with unresectable or metastatic advanced melanoma with BRAFV600 mutation

The triple-negative breast cancer (TNBC) subtype comprises approximately 15% of all breast cancers and is associated with poor long-term outcomes. Classical chemotherapy remains the standard of treatment, with toxicity and resistance being major limitations. TNBC is a high metabolic group, and antimetabolic drugs are effective in inhibiting TNBC cell growth. We analyzed the combined effect of chemotherapy and antimetabolic drug combinations in MDA-MB-231, MDA-MB-468 and HCC1143 human TNBC cell lines. Cells were treated with each drug or with drug combinations at a range of concentrations to establish the half-maximal inhibitory concentrations (IC50). The dose-effects of each drug or drug combination were calculated, and the synergistic or antagonistic effects of drug combinations were defined. Chemotherapy and antimetabolic drugs exhibited growth inhibitory effects on TNBC cell lines. Antimetabolic drugs targeting the glycolysis pathway had a synergistic effect with chemotherapy drugs, and antiglycolysis drug combinations also had a synergistic effect. The use of these drug combinations could lead to new therapeutic strategies that reduce chemotherapy drug doses, decreasing their toxic effect, or that maintain the doses but enhance their efficacy by their synergistic effect with other drugs.

El uso, así como el efecto productivo y económico de compuestos herbales en producción cunícola ha sido poco estudiado, por tal motivo el objetivo fue evaluar el efecto de una mezcla poliherbal rica en conjugados de colina a base de Trachyspermum ammi, Achyranthes aspera, Azadirachta indica y Citrullus colocynthis, sobre la respuesta económica y productiva de conejos para carne. Para lo cual se utilizaron 40 conejos Nueva Zelanda X California (30 días de edad), los cuales fueron aleatorizados en cinco grupos (0.0, 200, 400, 600 y 800 mg kg-1 de MS poliherbal, BioCholina®), el experimento tuvo una duración de 34 días. Se evaluó la respuesta productiva, calidad de la carne e indicadores económicos. Se utilizó un diseño completamente al azar con un arreglo de polinomios ortogonales para determinar efectos lineales y cuadráticos, con un nivel de significancia de P

Antecedentes: Los estudios del valor nutritivo de las hojas peciolos de Dichrostachys cinerea son insuficientes para definir el secado de las muestras en estudios de digestibilidad in vitro. El objetivo fue calcular .la producción de gas in vitro y parámetros de mejor ajuste a modelos como indicadores de la digestibilidad del follaje de la planta a dos alturas y con dos métodos de secado. Métodos: Las muestras se colectaron manualmente de forma aleatoria de 10 árboles diferentes (de 1 m y 2 m de altura), en época de seca, simulando el ramoneo de la planta en su totalidad. Cada muestra se dividió para ser sometida a dos tratamientos de secado (a temperatura ambiente aproximadamente por 96 h y a 550 C en estufa), hasta peso constante. Se evaluó la influencia de la altura de la planta y temperatura de secado en los volúmenes de gas in vitro producido a las 24, 48 y 72 h, utilizando como inóculo heces bovinas, mediante análisis de varianza de clasificación simple de las combinaciones de factores. Resultados: La producción de gas in vitro de hojas peciolos de plantas de 2 m, secadas a 55 °C fue significativamente menor que el resto de las combinaciones temperatura-altura evaluadas. Conclusiones: Los parámetros de mejor ajuste a las ecuaciones de producción de gas in vitro de hojas peciolo de Dichrostachys cirenea muestran la posibilidad de su uso en la alimentación de rumiantes debido a su potencial nutricional similar al de varias especies arbustivas forrajeras de la familia leguminosa.

Abstract Background A considerable portion of the HIV pandemic is composed of people under antiretroviral therapy, many of whom get a late diagnosis. Patients starting antiretroviral therapy (ART) at a very advanced stage of HIV disease attain a low recovery of CD4 T cells. Factors associated with poor recovery are incompletely described. This study aimed at finding variables associated with CD4 T cell recovery in late-presenting HIV patients. Methods We studied a cohort of HIV+ patients initiating ART with very low basal CD4 T cell counts. We defined immune recovery as the net increase in circulating CD4 T cell counts after one year on ART. We analyzed diverse routine laboratory determinations at different times using Least Absolute Shrinkage and Selection Operator (LASSO), adaptive LASSO and Conditional Inference Random Forest. Results CD4/CD8 ratio, % CD4 T cells and CD8 T cell counts at different times were the main recovery correlates, validated by all approaches. Unexpectedly, basal hematocrit was a consistent predictor. Additionally, week 24 creatinine had a high lasso coefficient, and alkaline phosphatase had a high conditional inference random forest coefficients, although neither was verified by other tests. Conclusions CD4 T cell proportions are associated with CD4 T cell recovery, independently of cell counts. Inflammation-related variables could also affect reconstitution. These accessible variables may reflect underlying mechanisms and could improve the follow up of patients starting ART with an advanced HIV infection.

Abstract Background Metabolomics has a great potential in the development of new biomarkers in cancer and it has experiment recent technical advances. Methods In this study, metabolomics and gene expression data from 67 localized (stage I to IIIB) breast cancer tumor samples were analyzed, using (1) probabilistic graphical models to define associations using quantitative data without other a priori information; and (2) Flux Balance Analysis and flux activities to characterize differences in metabolic pathways. Results On the one hand, both analyses highlighted the importance of glutamine in breast cancer. Moreover, cell experiments showed that treating breast cancer cells with drugs targeting glutamine metabolism significantly affects cell viability. On the other hand, these computational methods suggested some hypotheses and have demonstrated their utility in the analysis of metabolomics data and in associating metabolomics with patient’s clinical outcome. Conclusions Computational analyses applied to metabolomics data suggested that glutamine metabolism is a relevant process in breast cancer. Cell experiments confirmed this hypothesis. In addition, these computational analyses allow associating metabolomics data with patient prognosis.

An experiment was conducted to evaluate the effect of a polyherbal mixture based on Withania somnifera, Ocimum tenuiflorum, Tinospora cordifolia and Emblica officinalis, on lamb’s growth and blood metabolites. Thirty-two lambs (initial body weight BW 25 ± 4.1 kg) were randomly assigned to the treatments which consisted of the polyherbal ImmuPlus at dietary concentrations of 0, 0.1, 0.2 and 0.3% of dry matter (DM) for 60 days. There was a quadratic response to polyherbal in average daily gain (p < 0.01) and feed conversion (p < 0.05). The optimal concentration estimated by regression for maximum ADG was 0.14 ± 0.08% DM of ImmuPlus. Ruminal DM digestibility was not affected but pH was reduced linearly (p < 0.05). Ruminal N-NH3 (p < 0.01) and total VFA concentration showed a quadratic increment (p < 0.05) whereas butyrate proportion was increased linearly (p < 0.05). There was no effect in most blood metabolites related to energy metabolism, but uric acid was linearly reduced (p < 0.01) and bilirubin showed a quadratic decrement (p < 0.05). Mean corpuscular haemoglobin (p < 0.01) increased linearly (p < 0.10), lymphocytes and segmented neutrophils showed a quadratic response (p < 0.01) and platelets showed a linear reduction (linear p < 0.05). The inclusion of ImmuPlus improved the daily gain in finishing lambs and feed efficiency at 0.1% DM without altering blood chemistry and blood biometry.

Trastuzumab therapy has dramatically changed breast cancer prognosis. Consensus documents recommend a close monitoring during therapy, not always feasible, especially in metastatic breast cancer. The purpose of this study is to describe trastuzumab cardiotoxicity in metastatic breast cancer patients to understand how to improve cardiovascular monitoring. We retrospectively studied metastatic breast cancer patients scheduled for trastuzumab therapy (2001-2018). All patients underwent a baseline evaluation and monitoring during therapy. Cardiotoxicity was defined as symptomatic heart failure or asymptomatic decrease in left ventricular ejection fraction > 10% from baseline and < 53%. Ninety-two women were included, mean age 61 years (±14.43), median follow-up 42.5 months (IQR 26-74). Fourteen percent developed cardiotoxicity: two heart failure with preserved left ventricular ejection fraction, three heart failure with reduced left ventricular ejection fraction, and eight asymptomatic decreased in left ventricular ejection fraction. Eighty-one percent of cardiac dysfunction cases occurred within the first 4 years and on median of 31 months from trastuzumab initiation. Thus, in metastatic breast cancer patients, trastuzumab-mediated cardiotoxicity occurred more frequently during the first 4 years. These data should be considered to optimize follow-up protocols.

Immunotherapy based on anti-PD1 antibodies has improved the outcome of advanced melanoma. However, prediction of response to immunotherapy remains an unmet need in the field. Tumor PD-L1 expression, mutational burden, gene profiles and microbiome profiles have been proposed as potential markers but are not used in clinical practice. Probabilistic graphical models and classificatory algorithms were used to classify melanoma tumor samples from a TCGA cohort. A cohort of patients with advanced melanoma treated with PD-1 inhibitors was also analyzed. We established that gene expression data can be grouped in two different layers of information: immune and molecular. In the TCGA, the molecular classification provided information on processes such as epidermis development and keratinization, melanogenesis, and extracellular space and membrane. The immune layer classification was able to distinguish between responders and non-responders to immunotherapy in an independent series of patients with advanced melanoma treated with PD-1 inhibitors. We established that the immune information is independent than molecular features of the tumors in melanoma TCGA cohort, and an immune classification of these tumors was established. This immune classification was capable to determine what patients are going to respond to immunotherapy in a new cohort of patients with advanced melanoma treated with PD-1 inhibitors Therefore, this immune signature could be useful to the clinicians to identify those patients who will respond to immunotherapy.

Abstract Background Muscle-invasive bladder tumors are associated with a high risk of relapse and metastasis even after neoadjuvant chemotherapy and radical cystectomy. Therefore, further therapeutic options are needed and molecular characterization of the disease may help to identify new targets. The aim of this study was to characterize muscle-invasive bladder tumors at the molecular level using computational analyses. Methods The TCGA cohort of muscle-invasive bladder cancer patients was used to describe these tumors. Probabilistic graphical models, layer analyses based on sparse k-means coupled with Consensus Cluster, and Flux Balance Analysis were applied to characterize muscle-invasive bladder tumors at a functional level. Results Luminal and Basal groups were identified, and an immune molecular layer with independent value was also described. Luminal tumors showed decreased activity in the nodes of epidermis development and extracellular matrix, and increased activity in the node of steroid metabolism leading to a higher expression of the androgen receptor. This fact points to the androgen receptor as a therapeutic target in this group. Basal tumors were highly proliferative according to Flux Balance Analysis, which makes these tumors good candidates for neoadjuvant chemotherapy. The Immune-high group showed a higher degree of expression of immune biomarkers, suggesting that this group may benefit from immune therapy. Conclusions Our approach, based on layer analyses, established a Luminal group candidate for therapy with androgen receptor inhibitors, a proliferative Basal group which seems to be a good candidate for chemotherapy, and an immune-high group candidate for immunotherapy.

Dentro de las producciones pecuarias, las enfermedades entéricas tienen un papel importante, ya que generan severas pérdidas económicas derivadas de la mortalidad, depresión del crecimiento y disminución del índice de conversión. Rotavirus es considerado una causa importante de gastroenteritis viral aguda en humanos y animales jóvenes en todo el mundo, estando asociado a diarreas deshidratantes en granjas de producción animal. El objetivo de este estudio fue identificar rotavirus en granjas de producción cunícola ubicadas en la región sur-oriente del Estado de México a través de RT-PCR. Se recolectaron muestras de 39 pequeñas granjas de traspatio ubicadas dentro de los 13 municipios que conforman la región estudiada. Un total de 147 muestras se analizaron, 99 de conejos sanos y 48 de conejos con signología entérica, de las cuales se identificó Rotavirus en 9 de ellas (18.7 %), mientras que en conejos sanos no hubo identificación. Los resultados concuerdan con la suposición de que en conejos con signología entérica, los virus parecen tener un papel importante, pero no decisivo, y que la participación de rotavirus no es determinante para la presentación de la enfermedad. Se plantea la posibilidad de que Rotavirus no sea endémico en granjas cunícolas de la región sur-oriente del Estado de México, encontrándose ocasionalmente, sin inducir episodios entéricos graves de forma individual; no obstante, podría ser un factor importante en la presentación de brotes entéricos en asociación con otros agentes patógenos. Este es uno de los primeros reportes de la presencia de Rotavirus en conejos con signología entérica en México.

Reseña de libro: Emergencias de lo poshumano. Desafíos arquitectónicos y pedagógicos desde un margen disciplinar. Universidad de Alicante. ISBN: 978-84-9717-769-6.

Cancer immunotherapy based on the use of antibodies targeting the so-called checkpoint inhibitors, such as programmed cell death-1 receptor, its ligand, or CTLA-4, has shown durable clinical benefit and survival improvement in melanoma and other tumors. However, there are some special situations that could be a challenge for clinical management. Persons with chronic infections, such as HIV-1 or viral hepatitis, latent tuberculosis, or a history of solid organ transplantation, could be candidates for cancer immunotherapy, but their management requires a multidisciplinary approach. The Spanish Melanoma Group (GEM) panel in collaboration with experts in virology and immunology from different centers in Spain reviewed the literature and developed evidence-based guidelines for cancer immunotherapy management in patients with chronic infections and immunosuppression. These are the first clinical guidelines for cancer immunotherapy treatment in special challenging populations. Cancer immunotherapy in chronically infected or immunosuppressed patients is feasible but needs a multidisciplinary approach in order to decrease the risk of complications related to the coexistent comorbidities.

A polyherbal feed mixture containing (Achyrantes aspera, Trachyspermum ammi, Citrullus colocynthis, Andrographis paniculata, and Azadirachta indica) was evaluated in growing calves through blood chemistry, blood biometry, and gene expression during the pre-ruminant to weaning period. Forty Holstein calves (initial BW 45.6 ± 3.2 kg; 22.8 ± 0.9 days post birth) from a dairy farm were randomly assigned to the following treatments: 0, 3, 4, and 5 g/d of a polyherbal mixture, dosed in colloid gels with gelatin. Calves were housed in individual outdoor boxes with ad libitum access to a 21.5% CP calf starter and water and fed individually with a mixture of milk and a non-medicated milk replacer (22% CP). Blood samples were collected on day 59 for blood chemistry, blood biometry, and gene expression analysis in leukocyte through microarray assays. Immunoglobulins were quantified by enzyme-linked immunosorbent assay. The animals treated with the polyherbal mixture showed a quadratic effect on final body weight, daily weight gain, final hip height, and final thoracic girth. The best performance results were obtained with a treatment dose of 4 g/d. The serum IgG increased linearly with the treatment doses. Gene set enrichment analysis of upregulated genes revealed that the three biological processes with higher fold change were tight junction, mucin type O-Glycan biosynthesis, and intestinal immune network for IgA production. Also, these upregulated genes influenced arachidonic acid metabolism, and pantothenate and CoA biosynthesis. Gene ontology enrichment analysis indicated that the pathways enriched were PELP1 estrogen receptor interacting protein pathways, nuclear receptors in lipid metabolism and toxicity, tight junction, ECM-receptor interaction, thyroid hormone signaling pathways, vascular smooth muscle contraction, ribosome function, glutamatergic synapse pathway, focal adhesion, Hippo, calcium, and MAPK signaling pathways.

Renal cell carcinoma comprises a variety of entities, the most common being the clear-cell, papillary and chromophobe subtypes. These subtypes are related to different clinical evolution; however, most therapies have been developed for clear-cell carcinoma and there is not a specific treatment based on different subtypes. In this study, one hundred and sixty-four paraffin samples from primary nephrectomies for localized tumors were analyzed. MiRNAs were isolated and measured by microRNA arrays. Significance Analysis of Microarrays and Consensus Cluster algorithm were used to characterize different renal subtypes. The analyses showed that chromophobe renal tumors are a homogeneous group characterized by an overexpression of miR 1229, miR 10a, miR 182, miR 1208, miR 222, miR 221, miR 891b, miR 629-5p and miR 221-5p. On the other hand, clear cell renal carcinomas presented two different groups inside this histological subtype, with differences in miRNAs that regulate focal adhesion, transcription, apoptosis and angiogenesis processes. Specifically, one of the defined groups had an overexpression of proangiogenic microRNAs miR185, miR126 and miR130a. In conclusion, differences in miRNA expression profiles between histological renal subtypes were established. In addition, clear cell renal carcinomas had different expression of proangiogenic miRNAs. With the emergence of antiangiogenic drugs, these differences could be used as therapeutic targets in the future or as a selection method for tailoring personalized treatments.

Naringenin is flavonoid mainly found in citrus fruits which has shown several biological properties. In this work, we evaluated the therapeutic potential of the flavonoid Naringenin. Five-month-old B6.MRL-Faslpr/J lupus-prone mice were administered daily orally with Naringenin for seven months. We showed that Naringenin treatment at 50 or 100 mg/kg inhibited the splenomegaly and decreased the levels of anti-nuclear and anti-dsDNA autoantibodies. Furthermore, a reduction in serum concentration of TNF-α, IFN-γ and IL-6 was observed in the mice provided with Naringenin. Interestingly, serum levels of IL-10 increased. Naringenin decreased the frequency and absolute numbers of splenic effector memory T cells. Additionally, in order to be able to evaluate whether Naringenin prevented kidney damage, twelve-week-old MRL/MpJ-Faslpr/J mice, an accelerated lupus model, were orally administered with Naringenin at 100 mg/kg for six weeks. Surprisingly, Naringenin treatment prevented kidney damage and reduced the development of fibrosis similar to cyclophosphamide group. Moreover, Naringenin treatment increased the percentage of regulatory T cells in this aggressive model of lupus. Together, these results suggest a potential ability of Naringenin to reduce the autoimmunity in lupus-prone mice by modulation of T-cell subsets and cytokines profile that mitigate the development of important lupus clinical manifestations.

Breast cancer is a heterogeneous disease. In clinical practice, tumors are classified as hormonal receptor positive, Her2 positive and triple negative tumors. In previous works, our group defined a new hormonal receptor positive subgroup, the TN-like subtype, which had a prognosis and a molecular profile more similar to triple negative tumors. In this study, proteomics and Bayesian networks were used to characterize protein relationships in 96 breast tumor samples. Components obtained by these methods had a clear functional structure. The analysis of these components suggested differences in processes such as mitochondrial function or extracellular matrix between breast cancer subtypes, including our new defined subtype TN-like. In addition, one of the components, mainly related with extracellular matrix processes, had prognostic value in this cohort. Functional approaches allow to build hypotheses about regulatory mechanisms and to establish new relationships among proteins in the breast cancer context.

CD38 is a transmembrane glycoprotein expressed by T-cells. It has been reported that patients with systemic lupus erythematosus (SLE) showed increased CD38+CD25+ T-cells correlating with immune activation and clinical signs. Contrariwise, CD38 deficiency in murine models has shown enhanced autoimmunity development. Recent studies have suggested that CD38+ regulatory T-cells are more suppressive than CD38− regulatory T-cells. Thus, we have suggested that CD38 overexpression in SLE patients could play a role in regulating immune activation cells instead of enhancing it. This study found a correlation between CD38 with FoxP3 expression and immunosuppressive molecules (CD69, IL-10, CTLA-4, and PD-1) in T-cells from lupus-prone mice (B6.MRL-Faslpr/J). Additionally, B6.MRL-Faslpr/J mice showed a decreased proportion of CD38+ Treg cells regarding wild-type mice (WT). Furthermore, Regulatory T-Cells (Treg cells) from CD38-/- mice showed impairment in expressing immunosuppressive molecules and proliferation after stimulation through the T-cell receptor (TCR). Finally, we demonstrated an increased ratio of IFN-γ/IL-10 secretion in CD38-/- splenocytes stimulated with anti-CD3 compared with the WT. Altogether, our data suggest that CD38 represents an element in maintaining activated and proliferative Treg cells. Consequently, CD38 could have a crucial role in immune tolerance, preventing SLE development through Treg cells.

The ability of alkylseleno/alkyltelluroacetylenes such as bis(selenomethylethynyl)-perfluorobenzene (4F-Se) to act as a ditopic chalcogen bond (ChB) donor in co-crystals with ditopic Lewis bases such as 4,4′-bipyridine is extended here to the octafluorobiphenylene analog, 4,4′-bis(selenomethylethynyl)-perfluorobiphenyl (8F-Se), with the more electron-rich 4,4′-bipyridylethane (bpe), showing in the 1:1 (8F-Se)•(bpe) co-crystal a shorter and more linear C−Se•••N ChB interaction than in (4F-Se)•(bpe), with Se•••N distances down to 2.958(2) Å at 150 K, i.e., a reduction ratio of 0.85 vs. the van der Waals contact distance.

Immune reconstitution inflammatory syndrome (IRIS) is an exacerbated immune response that can occur to HIV+ patients after initiating antiretroviral therapy (ART). IRIS pathogenesis is unclear, but dysfunctional and exhausted cells have been reported in IRIS patients, and the TIM-3/Gal-9 axis has been associated with chronic phases of viral infection. This study aimed to evaluate the soluble levels of TIM-3 and Gal-9 and their relationship with IRIS development. TIM-3, Gal-9, TNF-α, IFN-γ, IL-6, TNFR1, TNFR2, E-cadherin, ADAM10, and ADAM17 were measured to search for IRIS-associated biomarkers in plasma samples from 0-, 4-, 8-, 12-, and 24-weeks after ART initiation of 61 HIV+ patients (15 patients developed IRIS, and 46 did not). We found that patients who developed IRIS had higher levels of TIM-3 [median 4806, IQR: 3206–6182] at the time of the IRIS events, compared to any other follow-up time evaluated in these patients or compared with a control group of patients who did not develop IRIS. Similarly, IRIS patients had a higher TNF-α level [median 10.89, IQR: 8.36–12.34] at IRIS events than any other follow-up time evaluated. Other molecules related to the TIM-3 and TNF-α pathway (Gal-9, IL-6, IFN-γ, TNFR1, TNFR2, ADAM-10, and ADAM-17) did not change during the IRIS events. In conclusion, our data suggest that a high level of soluble TIM-3 and TNF-α could be used as an IRIS biomarker.

Se evaluó la seropositividad a Neospora caninum mediante ensayo inmunoenzimático (ELISA) y se detectó el ADN por PCR anidada con iniciadores externos Np 21-4 e internos Np 9-10 en leucocitos de vacas del sistema de producción de leche en pequeña escala en Amecameca, Estado de México. Se estudió un hato con 34 hembras adultas realizando el seguimiento con muestreos sanguíneos durante cinco meses; también, se analizaron dos muestras de encéfalo de fetos abortados y se constató la secuencia de los amplicones en el Genbank. La concordancia entre técnicas fue comparada mediante la prueba Kappa. La seroprevalencia por ELISA fue de 85.3 %; y se detectó ADN del parásito en 89.4 % de las vacas. Una muestra de encéfalo fue positiva. La similitud de los amplicones osciló entre 91 y 96 %, el índice Kappa fue de 0.41. El empleo de leucocitos aumenta la probabilidad de amplicar ADN de N. caninum.

El objetivo fue estimar la seroprevalencia de neosporosis, diarrea viral bovina, rinotraqueítis infecciosa bovina, brucelosis y leptospirosis; así como, determinar su asociación con el antecedente de aborto e identicar los factores de riesgo. Mediante un estudio de casos y controles de 29 unidades de producción, se seleccionaron 57 vacas con antecedente de aborto y 121 controles. Se analizaron por serología y fetos abortados por PCR para N. caninum. Excepto para brucelosis, se detectaron vacas seropositivas correspondiendo la mayor seroprevalencia a N. caninum (51.7 %) y la menor (11.8 %) para Leptospirosis. 11.8 % de las vacas fueron seropositivas a N. caninum y diarrea viral bovina de manera simultánea, presentando el 100% de ellas registros de al menos un aborto. Se comprobó la presencia de ADN de N. caninum en un feto, concluyéndose que N. caninum es causa determinante en la presentación de aborto en el sistema de producción de leche en pequeña escala.

Resumen Este articulo intenta demostrar que, a diferencia del pensamiento dualista de los filósofos griegos clásicos que concebían el tiempo como la antítesis imperfecta y finita de la eternidad inrempornl, el pensamiento hebreo antiguo (representado en la Biblia) entiende que el tiempo es inseparable de la existencia de los seres y que, en consecuencia, la eternidad es el tiempo infinito de los seres inmortales. Por lo tanto, la idea de que "en la eternidad el tiempo no existe o dejará de existir" es absurda y extraña para la mente de los escritores bíblicos. Nos interesa estudiar la concepción hebrea antigua del tiempo, porque su comprensión nos permitiría interpretar la teología de la Biblia de una manera más acertada y realizar una reflexión sistemática sobre bases más auténticamente bíblicas. Summary The present paper tries co show rhat -unlike che dualicy of classic Greek philosophers' thoughc which believed in time as the imperfect and finite and antithesis of unremporal eternity- che ancient Hebrew thought (as represented in the Bible) understands rhat time is inseparable from che existence of beings and that -as a consequence- eternity is the infinice time of immoral beings. Accordingly, che idea that "in erernity, time does not exist interested in che ancient Hebrew conception of time because its understanding would allow us to make a much more accurate inrerpretation of che theology of che Bible, as well as to achieve a systematic retlection on che actually auchenric biblical grounds. Résumé Cet article tache de démontrer que, contrairement a la pensée dualistc des philosophes grecs classiques, qui corn;:urent le temps comme l'antithése imparfaitc et limité de l'éternité incorporelle, la pensée hébra'ique ancienne (qui apparair dans la Bible) a la notion de que le remps est inséparable de l'e.xistence des etres et que, par conséquent, l'éternité esr le temps infini des etres immortels. C'est pour celá que l'idée de que "dans l'éternité le remps n'éxistera plus ou arrivéra a sa fin" est absurde et étrangére á la mentalicé des écrivains bibliques. Nous nous intéressons dans l'én1de de l'ancienne conception hébraique du temps, parce que si nous arrivons a la comprendre, nous pourrons interpéter la théologie biblique d'une facon plus exacte, et réfléchir sistématiquement sur des fondements bibliques plus sürs.

Resumen La Biblia ofrece una descripción inspirada de la fe: ésta es "la certeza de lo que se espera y la convicción de lo que no se ve" (Hebreos 11: 1). Esto significa que la fe nos proyecta más allá del tiempo y del lugar en que ahora nos encontramos dentro de las coordenadas espacio-temporales. Quien tiene fe religiosa cree en la existencia de Dios. Por otra parte, quien cree en la capacidad del hombre para descubrir la verdad también experimenta un cierto tipo de fe. La fe es necesaria en la búsqueda de la verdad, sea ésta religiosa o científica. Todo el que mira hacia el futuro buscando la verdad con la esperanza de encontrarla, halla que la fe es un elemento indispensable que estimula y sostiene al espíritu inquisitivo. La fe es parte de la experiencia humana, e involucra tanto la razón como los sentimientos en la búsqueda de la verdad. Abstract The Bible offers an inspired description of faith: it is "the substance of things hoped for, the evidence of things not seen" (Hebrews 11: 1). This means faith can project us beyond our present time and place, which is found within time and place co-ordinates. A person with religious faith believes in che existence of God. On the other hand, those who believe in man's ability to find truth also experience sorne kind of faith. When looking for truth, faith is needed, be it religious or scientific. All those who look forward to a furure search for truth in the hope of finding it, also find that faith is an essential element which encourages and upholds the inquisitive spirit. Faith is part of human experience, and involves both reason and feelings in its search for truth. Résumé La Bible nous offre une description inspirée de la foi: c'est "une ferme assurance des choses qu'on espere, une démonstration de celles qu'on ne voit pas" (Hébreux 11:1). Ca veut dire que la fo¡ nous projette au-dela du temps et de la place ou nous sommes maintenant au-dedans les coordonnées de l'espace et le temps. Celui qui a une foi réhgieuse croit que Die u existe. Par contre, celui que croit en la capacité de l'homme pour découvrir la vérité, il aussi a une espece de foi. La foi est nécessaire pour la recherche de la vérité, soit elle réligieuse ou scientifique. Tout celui qui regarde l'avenir pour chercher la vérité, avec l'espérance de la trouver, constate que la foi est un instrument indispensable qui stimule et soutient l'esprit inquisitif. La fo¡ fait partie de l'expérience humaine, et implique a la fois la raison et les sentiments dans la recherche de la vérité.

Triple-negative breast cancer (TNBC) accounts for 15-20% of all breast cancers and usually requires the administration of adjuvant chemotherapy after surgery but even with this treatment many patients still suffer from a relapse. The main objective of this study was to identify proteomics-based biomarkers that predict the response to standard adjuvant chemotherapy, so that patients at are not going to benefit from it can be offered therapeutic alternatives.We analyzed the proteome of a retrospective series of formalin-fixed, paraffin-embedded TNBC tissue applying high-throughput label-free quantitative proteomics. We identified several protein signatures with predictive value, which were validated with quantitative targeted proteomics in an independent cohort of patients and further evaluated in publicly available transcriptomics data.Using univariate Cox analysis, a panel of 18 proteins was significantly associated with distant metastasis-free survival of patients (p

To compare the concordance in risk classification between the EndoPredict and the MammaPrint scores obtained for the same cancer samples on 40 estrogen-receptor positive/HER2-negative breast carcinomas.Formalin-fixed, paraffin-embedded invasive breast carcinoma tissues that were previously analyzed with MammaPrint as part of routine care of the patients, and were classified as high-risk (20 patients) and low-risk (20 patients), were selected to be analyzed by the EndoPredict assay, a second generation gene expression test that combines expression of 8 genes (EP score) with two clinicopathological variables (tumor size and nodal status, EPclin score).The EP score classified 15 patients as low-risk and 25 patients as high-risk. EPclin re-classified 5 of the 25 EP high-risk patients into low-risk, resulting in a total of 20 high-risk and 20 low-risk tumors. EP score and MammaPrint score were significantly correlated (p = 0.008). Twelve of 20 samples classified as low-risk by MammaPrint were also low-risk by EP score (60%). 17 of 20 MammaPrint high-risk tumors were also high-risk by EP score. The overall concordance between EP score and MammaPrint was 72.5% (κ = 0.45, (95% CI, 0.182 to 0.718)). EPclin score also correlated with MammaPrint results (p = 0.004). Discrepancies between both tests occurred in 10 cases: 5 MammaPrint low-risk patients were classified as EPclin high-risk and 5 high-risk MammaPrint were classified as low-risk by EPclin and overall concordance of 75% (κ = 0.5, (95% CI, 0.232 to 0.768)).This pilot study demonstrates a limited concordance between MammaPrint and EndoPredict. Differences in results could be explained by the inclusion of different gene sets in each platform, the use of different methodology, and the inclusion of clinicopathological parameters, such as tumor size and nodal status, in the EndoPredict test.

Professor Dunitz questions the usefulness of ascribing crystalline structural stability to individual atom–atom intermolecular interactions viewed as bonding (hence stabilizing) whenever linked by a bond path. An alternative view is expressed in the present essay that articulates the validity and usefulness of the bond path concept in a crystallographic and crystal engineering context.

Se determinaron algunos indicadores del valor nutritivo de Albizia lebbeck, Calliandra surinamensis, Erythrina variegata, Gliricidia sepium y Leucaena leucocephala para rumiantes y cerdos. Se emplearon análisis químicos, la técnica de producción de gas in vitro y la de la bolsa móvil de nailon. Las concentraciones de proteína bruta estuvieron entre 12,7 y 23,1 %. Existieron diferencias significativas en la producción de gases entre especies, así como en las digestibilidades en cerdos. El follaje de E. variegata fue el más aceptado por los cerdos. Los follajes de E. variegata, L. leucocephala y G. sepium tienen el mejor valor nutritivo para los rumiantes, mientras que los follajes de E. variegata, L. leucocephala y A. lebbeck tienen las mejores digestibilidades en cerdos.

Se determinaron indicadores in vitro del valor nutritivo de Moringa oleifera, durante la época de seca, para rumiantes en el municipio de Camagüey, Cuba. Esta especie florece en las zonas de Santayana, Imán y La Belén). Los valores de materia seca (20,0 y 23,2 %) y cenizas (7,8 y 10,0 %) no presentaron diferencias significativas; no sucedió así con la proteína bruta (23,6 y 26,0 %) y la fibra neutro detergente (17,0 y 25,9 %). Los mayores valores de proteína bruta y materia seca se encontraron en La Belén; los menores fueron los de Santayana. En los cultivos de las tres zonas no se detectaron saponinas ni alcaloides; sin embargo, se evidenció la presencia de taninos y taninos condensados, con mayor incidencia en Santayana. La producción de gas in vitro con heces bovinas, demostró diferencias significativas a las 12; 24 y 96 h de incubación. El valor nutritivo de la moringa en estas localidades es alto.

Anti-angiogenic therapy benefits many patients with advanced renal cell carcinoma (RCC), but there is still a need for predictive markers that help in selecting the best therapy for individual patients. MicroRNAs (miRNAs) regulate cancer cell behavior and may be attractive biomarkers for prognosis and prediction of response. Forty-four patients with RCC were recruited into this observational prospective study conducted in nine Spanish institutions. Peripheral blood samples were taken before initiation of therapy and 14 days later in patients receiving first-line therapy with sunitinib for advanced RCC. miRNA expression in peripheral blood was assessed using microarrays and L2 boosting was applied to filtered miRNA expression data. Several models predicting poor and prolonged response to sunitinib were constructed and evaluated by binary logistic regression. Blood samples from 38 patients and 287 miRNAs were evaluated. Twenty-eight miRNAs of the 287 were related to poor response and 23 of the 287 were related to prolonged response to sunitinib treatment. Predictive models identified populations with differences in the established end points. In the poor response group, median time to progression was 3.5 months and the overall survival was 8.5, whereas in the prolonged response group these values were 24 and 29.5 months, respectively. Ontology analyses pointed out to cancer-related pathways, such angiogenesis and apoptosis. miRNA expression signatures, measured in peripheral blood, may stratify patients with advanced RCC according to their response to first-line therapy with sunitinib, improving diagnostic accuracy. After proper validation, these signatures could be used to tailor therapy in this setting.

En 1982 se firmó un convenio entre el Centro Internacional de Agricultura Tropical (CIAT), con sede en Palmira, y dos instituciones oficiales panameñas de investigación agrícola: el Instituto de Investigaciones Agropecuarias de Panamá (IDlAP) y la Facultad de Ciencias Agropecuarias (FCA) de la Universidad de Panamá. Este convenio se realizó entre 1982 y 1988 con el objetivo principal de obtener germoplasma mejorado de arroz para las condiciones de riego y secano favorecido. Este trabajo analiza el desarrollo de dicho convenio y los logros alcanzados. Se identificaron sitios claves para las evaluaciones de los. materiales segregantes (FrF8) Y se estableció un flujo del material genético el cual unía los distintos sitios de selección en Panamá con los programas de arroz del ClAT y las pruebas internacionales de arroz de América Latina y el Caribe. Se evaluaron 38305 líneas segregantes, incluyendo viveros internacionales; el porcentaje promedio de selección fue del 13%. La susceptibilidad a las principales enfermedades y a la sequía fueron los factores mas importantes de selección. Once variedades liberadas para diversos ecosistemas en siete países se originaron en este proyecto. Estos resultados indican la importancia de la colaboración interinstitucional e indican que los beneficios obtenidos se extendieron mas allá de las fronteras y tiempo contemplados inicialmente en este proyecto.

Recent reports demonstrate the feasibility of quantifying gene expression by using RNA isolated from blocks of formalin-fixed, paraffin-embedded (FFPE) tumor tissue. The development of molecular tests for clinical use based on archival materials would be of great utility in the search for and validation of important genes or gene expression profiles. In this study, we compared the performance of different normalization strategies in the correlation of quantitative data between fresh frozen (FF) and FFPE samples and analyzed the parameters that characterize such correlation for each gene. Total RNA extracted from FFPE samples presented a shift in raw cycle threshold (Cq) values that can be explained by its extensive degradation. Proper normalization can compensate for the effects of RNA degradation in gene expression measurements. We show that correlation between normalized expression values is better for moderately to highly expressed genes whose expression varies significantly between samples. Nevertheless, some genes had no correlation. These genes should not be included in molecular tests for clinical use based on FFPE samples. Our results could serve as a guide when developing clinical diagnostic tests based on RT-qPCR analyses of FFPE tissues in the coming era of treatment decision-making based on gene expression profiling.

BackgroundTrastuzumab improves survival outcomes in patients with HER2+ metastatic breast cancer. The Long-Her study was designed to identify clinical and molecular markers that could differentiate long-term survivors from patients having early progression after trastuzumab treatment.MethodsData were collected from women with HER2-positive metastatic breast cancer treated with trastuzumab that experienced a response or stable disease during at least 3 years. Patients having a progression in the first year of therapy with trastuzumab were used as a control. Genes related with trastuzumab resistance were identified and investigated for network and gene functional interrelation. Models predicting poor response to trastuzumab were constructed and evaluated. Finally, a mutational status analysis of selected genes was performed in HER2 positive breast cancer samples.Results103 patients were registered in the Long-HER study, of whom 71 had obtained a durable complete response. Median age was 58 years. Metastatic disease was diagnosed after a median of 24.7 months since primary diagnosis. Metastases were present in the liver (25%), lungs (25%), bones (23%) and soft tissues (23%), with 20% of patients having multiple locations of metastases. Median duration of response was 55 months. The molecular analysis included 35 patients from the group with complete response and 18 patients in a control poor-response group. Absence of trastuzumab as part of adjuvant therapy was the only clinical factor associated with long-term survival. Gene ontology analysis demonstrated that PI3K pathway was associated with poor response to trastuzumab-based therapy: tumours in the control group usually had four or five alterations in this pathway, whereas tumours in the Long-HER group had two alterations at most.ConclusionsTrastuzumab may provide a substantial long-term survival benefit in a selected group of patients. Whole genome expression analysis comparing long-term survivors vs. a control group predicted early progression after trastuzumab-based therapy. Multiple alterations in genes related to the PI3K-mTOR pathway seem to be required to confer resistance to this therapy.

In the title compound, C26H24BrNO5·H2O, a synthetic analogue of natural flavagline, the cyclopentane ring adopts an envelope conformation (the flap atom bearing the phenyl group) and the vicinal phenyl and bromophenyl groups are slightly shifted relative to each other [CPh—C—C—CPhBr = 36.3 (2)°]. Intramolecular N—H...O and C—H...O hydrogen bonds form S(5) motifs. In the crystal, the organic and the water molecules are linked by an O—H...O hydrogen bond. Pairs of organic and water molecules, located about inversion centers, interact through O—H...O hydrogen bonds, forming R44(20) and R44(26) motifs, which together lead to C22(9) motifs. The crystal packing is also characterized by N—H...O and C—H...O hydrogen bonds between neighbouring organic molecules, forming R22(10) and R22(18) motifs, respectively.

El espacio público como meta-laboratorio e interfaz a partir del cual entender el propio espacio de la ciudad mediante el dibujo. Tres experiencias reali- zadas por PKMN en el ámbito del aprendizaje activo que alteran las relaciones entre ciudad y usuario implementando nuevas capas gráficas informativas o simbólicas en el paisaje urbano en tiempo real.

With the completion of the human genome sequence, biomedical sciences have entered in the "omics" era, mainly due to high-throughput genomics techniques and the recent application of mass spectrometry to proteomics analyses. However, there is still a time lag between these technological advances and their application in the clinical setting. Our work is designed to build bridges between high-performance proteomics and clinical routine. Protein extracts were obtained from fresh frozen normal lung and non-small cell lung cancer samples. We applied a phosphopeptide enrichment followed by LC-MS/MS. Subsequent label-free quantification and bioinformatics analyses were performed. We assessed protein patterns on these samples, showing dozens of differential markers between normal and tumor tissue. Gene ontology and interactome analyses identified signaling pathways altered on tumor tissue. We have identified two proteins, PTRF/cavin-1 and MIF, which are differentially expressed between normal lung and non-small cell lung cancer. These potential biomarkers were validated using western blot and immunohistochemistry. The application of discovery-based proteomics analyses in clinical samples allowed us to identify new potential biomarkers and therapeutic targets in non-small cell lung cancer.

BACKGROUND:Proteomics is expected to play a key role in cancer biomarker discovery. Although it has become feasible to rapidly analyze proteins from crude cell extracts using mass spectrometry, complex sample composition hampers this type of measurement. Therefore, for effective proteome analysis, it becomes critical to enrich samples for the analytes of interest. Despite that one-third of the proteins in eukaryotic cells are thought to be phosphorylated at some point in their life cycle, only a low percentage of intracellular proteins is phosphorylated at a given time. METHODOLOGY/PRINCIPAL FINDINGS:In this work, we have applied chromatographic phosphopeptide enrichment techniques to reduce the complexity of human clinical samples. A novel method for high-throughput peptide profiling of human tumor samples, using Parallel IMAC and MALDI-TOF MS, is described. We have applied this methodology to analyze human normal and cancer lung samples in the search for new biomarkers. Using a highly reproducible spectral processing algorithm to produce peptide mass profiles with minimal variability across the samples, lineal discriminant-based and decision tree-based classification models were generated. These models can distinguish normal from tumor samples, as well as differentiate the various non-small cell lung cancer histological subtypes. CONCLUSIONS/SIGNIFICANCE:A novel, optimized sample preparation method and a careful data acquisition strategy is described for high-throughput peptide profiling of small amounts of human normal lung and lung cancer samples. We show that the appropriate combination of peptide expression values is able to discriminate normal lung from non-small cell lung cancer samples and among different histological subtypes. Our study does emphasize the great potential of proteomics in the molecular characterization of cancer.

INTRODUCTION: Gene profiling may improve prognostic accuracy in patients with early breast cancer, but this technology is not widely available. We used commercial assays for qRT-PCR to assess the performance of the gene profiles included in the 70-Gene Signature, the Recurrence Score and the Two-Gene Ratio. METHODS: 153 patients with early breast cancer and a minimum follow-up of 5 years were included. All tumours were positive for hormonal receptors and 38% had positive lymph nodes; 64% of patients received adjuvant chemotherapy. RNA was extracted from formalin-fixed paraffin-embedded (FFPE) specimens using a specific kit. qRT-PCR amplifications were performed with TaqMan Gene Expression Assays products. We applied the three gene-expression-based models to our patient cohort to compare the predictions derived from these gene sets. RESULTS: After a median follow-up of 91 months, 22% of patients relapsed. The distant metastasis-free survival (DMFS) at 5 years was calculated for each profile. For the 70-Gene Signature, DMFS was 95% -good prognosis- versus 66% -poor prognosis. In the case of the Recurrence Score, DMFS was 98%, 81% and 69% for low, intermediate and high-risk groups, respectively. Finally, for the Two-Gene Ratio, DMFS was 86% versus 70%. The 70-Gene Signature and the Recurrence Score were highly informative in identifying patients with distant metastasis, even in multivariate analysis. CONCLUSION: Commercially available assays for qRT-PCR can be used to assess the prognostic utility of previously published gene expression profiles in FFPE material from patients with early breast cancer. Our results, with the use of a different platform and with different material, confirm the robustness of the 70-Gene Signature and represent an independent test for the Recurrence Score, using different primer/probe sets.