Machine Learning Analysis of Individual Tumor Lesions in Four Metastatic Colorectal Cancer Clinical Studies: Linking Tumor Heterogeneity to Overall Survival

Total tumor size (TS) metrics used in TS models in oncology do not consider tumor heterogeneity, which could help to better predict drug efficacy. We analyzed individual target lesions (iTLs) of patients with metastatic colorectal carcinoma (mCRC) to determine differences in TS dynamics by using the ClassIfication Clustering of Individual Lesions (CICIL) methodology. Results from subgroup analyses comparing genetic mutations and TS metrics were assessed and applied to survival analyses. Data from four mCRC clinical studies were analyzed (1781 patients, 6369 iTLs). CICIL was used to assess differences in lesion TS dynamics within a tissue (intra-class) or across different tissues (inter-class). First, lesions were automatically classified based on their location. Cross-correlation coefficients (CCs) determined if each pair of lesions followed similar or opposite dynamics. Finally, CCs were grouped by using the K-means clustering method. Heterogeneity in tumor dynamics was lower in the intra-class analysis than in the inter-class analysis for patients receiving cetuximab. More tumor heterogeneity was found in KRAS mutated patients compared to KRAS wild-type (KRASwt) patients and when using sum of longest diameters versus sum of products of diameters. Tumor heterogeneity quantified as the median patient’s CC was found to be a predictor of overall survival (OS) (HR = 1.44, 95% CI 1.08–1.92), especially in KRASwt patients. Intra- and inter-tumor tissue heterogeneities were assessed with CICIL. Derived metrics of heterogeneity were found to be a predictor of OS time. Considering differences between lesions’ TS dynamics could improve oncology models in favor of a better prediction of OS

Unusual remodeling of the hyalinization band in vulval lichen sclerosus by type V collagen and ECM 1 protein

OBJECTIVES: The vulva is the primary site affected in lichen sclerosus, a chronic dermatosis in women that is histologically characterized by a zone of collagen remodeling in the superior dermis. The normal physiological properties of the vulva depend on the assembly of collagen types I (COLI), III (COLIII) and V (COLV), which form heterotypic fibers, and extracellular matrix protein interactions. COLV regulates the heterotypic fiber diameter, and the preservation of its properties is important for maintaining normal tissue architecture and function. In the current work, we analyzed the expression of COLV and its relationship with COLI, COLIII, elastic fibers and extracellular matrix protein 1 in vulvar biopsies from patients with lichen sclerosus. METHODS: Skin biopsies from 21 patients with lichen sclerosus, classified according to Hewitt histological criteria, were studied and compared to clinically normal vulvar tissue (N=21). Morphology, immunohistochemistry, immunofluorescence, 3D reconstruction and morphometric analysis of COLI, COLIII, COLV deposition, elastic fibers and extracellular matrix 1 expression in a zone of collagen remodeling in the superior dermis were performed. RESULTS: A significant decrease of elastic fibers and extracellular matrix 1 protein was present in the hyalinization zone of lichen sclerosus compared to healthy controls. The non-homogeneous distribution of collagen fibers visualized under immunofluorescence in the hyalinization zone of lichen sclerosus and control skin was confirmed by histomorphometry. Lichen sclerosus dermis shows a significant increase of COLI, COLIII and COLV expression compared to the healthy controls. Significant inverse associations were found between elastic fibers and COLV and between COLV and extracellular matrix 1 expression. A direct association was found between elastic fiber content and extracellular matrix 1 expression. Tridimensional reconstruction of the heterotypic fibers of the lichen sclerosus zone of collagen remodeling confirmed the presence of densely clustered COLV. CONCLUSIONS: Increased deposition of abnormal COLV and its correlation with extracellular matrix 1 and elastic fibers suggest that COLV may be a trigger in the pathogenesis of lichen sclerosus

El virus de la D.V.B. como agente contaminante en cultivo de tejidos animales

The Hovine Viral Diarrhoea Virus (BWV)belongs to the genus Pestisvirus. It is the cause or reproductive disorders in catlle and is widespread in the conuntry. There are noncytophatogenic strains of the BVDV very difficult to detect inthe primary tissue culture and cell lines, ordinarilly used in diagnostic test or research activities. Foetal calf serum (FCS) is the common source of contamulation for the animal tissue culture, because of the poor quality controll during its collection and markenting under local conditions. It is possibly to fmd two kind of situations related to BVDV and the presence of viral particles; in each case is possible to have problems with the laboratory procedures. In order to avoid BWV tissue culture contamination, in our laboratory it has been used an indirect inmunofluorescense tedinique for the detectionofthe Wusinfoetalcaüserumandtissue cultures, havingresults asfoilows: 28.5%positives from 32 FCS samples from differnt sources and 42.8% from tissue cuitures from different animal origen and several research centres. Come recomdations are made for the prevention and control ot this problem at diagnostic and researchlaboratorylevelliietheuse of BI, indirect peroxidase techniques, or inmunopresitation for the detection of the Wus, and the use of a few number of fwtuses for each lot of FCS in order to reduce the risk of contamination from this source.El Pestisvirus que produce la Diarrea Viral Bovina (V.D.V.B.), ocasiona problemas reproductivos en el ganado vacuno y se encuentra ampliamente difundido en el país. La presencia de cepas no citopatogénicas del mismo, las cuales normalmente no se evidencian en los cultivos celulares contaminados con éste virus, han constituido un Iimitante para el establecimiento y mantenimiento de cultivos primarios y líneas celulares libres del V.D.V.B., para uso rutinario en actividades diagnósticas o investigativas. El suero fetal bovino (SFB) es la fuente usual de contaminación para los cultivos de tejidos animales, dadas las escasas previsiones para el control de su calidad, ya que la recolección y mercadeo del SFB se realiza con criterios artesanales y sin los debidos controles de calidad. Específicamente en el caso del V.D.V.B., se pueden presentar dos situaciones con el SFB: que contenga partículas virales o que posea niveles de anticuerpos contra el virus; en ambos casos se darán problemas en los procedimientos realizados con dichos sueros. Con el fin de controlar las contaminaciones de los cultivos celulares con el VDVB, en el Postgrado de la Facultad de Medicina Veterinaria de la Universidad Nacional, se ha empleado la técnica de Inmunofluorescencia Indirecta (IFl) para detectarlas tanto, en el SFB como en diferentes cultivos celulares, obteniéndose para el SFB el 25.8% de positivos de un total de 32 muestras de diferentes orígenes y para los cultivos celulares el 42.8% de positivos de 28 lotes, procedentes de diferente origen animal y centros de investigación, resultados que son comparables con los obtenidos por otros laboratorios internacionales como el National Animal Diseases Laboratory (NADL) del Departamento de Agricultura de USA. Dentro de las recomendaciones para la prevención y control del problema en laboratorios de diagnóstico e investigación médica y veterinaria, que empleen tecnologías relacionadas con manejo y mantenimiento de cultivos celulares, están las de implementar técnicas para la detección del virus como la IFI, la peroxidasa o la Inmunoprecipitación, al igual que el manejo de un bajo número de fetos en la conformación de lotes de SFB

Caryocar brasiliense camb protects against genomic and oxidative damage in urethane-induced lung carcinogenesis

The antioxidant effects of Caryocar brasiliense Camb, commonly known as the pequi fruit, have not been evaluated to determine their protective effects against oxidative damage in lung carcinogenesis. In the present study, we evaluated the role of pequi fruit against urethane-induced DNA damage and oxidative stress in forty 8-12 week old male BALB/C mice. An in vivo comet assay was performed to assess DNA damage in lung tissues and changes in lipid peroxidation and redox cycle antioxidants were monitored for oxidative stress. Prior supplementation with pequi oil or its extract (15 µL, 60 days) significantly reduced urethane-induced oxidative stress. A protective effect against DNA damage was associated with the modulation of lipid peroxidation and low protein and gene expression of nitric oxide synthase. These findings suggest that the intake of pequi fruit might protect against in vivo genotoxicity and oxidative stress

Effects of Milled Maize Stalks on the Productive Response of Grazing Dairy Cows.

The productive response of grazing dairy cows was evaluated, using milled corn stalks in the diet. The study was developed in two different settings, in Ecuador (Costa and Sierra regions). On the coast farm (29.1 ha), cows grazed on Bermuda grass (Cynodon nlemfuensis) and Guinea grass (Panicum maximum) with several types of legumes (Lysicarpus, Centrosema, Desmodium, Galactia), supplemented with corn stalks cv. INIAP 125. The animals received 0.46 kg beginning at 3 kg, and milled maize stalks in 30 and 28-day periods, respectively (M-30 and M-28), and control without stalks for 36 days (M-0). The farm in the other region (14.2 ha) had 23 cows grazing on Kikuyo grass (P. clandestinum) and ryegrass-white clover (L. perenne and whole maize stalks and T. (60-70% ripe grain), at a rate of 18 kg green/cow/day for 48 days; and balanced supplement, at a rate of 0.5 kg/ milk liter, after the fourth kilogram, along with minerals. In both cases the forage had effects (P < 0.05) on cow response. In the Sierra area, the increase was 1.68 kg/cow, and in the coast, it was 1.1 and 2.5 kg/cow). Maize stalks served as a nutritional complement for poorly consumed grass areas in both regions; milk production/animal was increased, and the costs were reduced

Reclaiming Sustainable Space: A Study of Degrowth Activists

While degrowth debates typically encompass abstract ecological and economic arguments against growth, our study considers how degrowth-minded activism becomes interwoven with the production and consumption of space and place. Drawing illustrative insights from an ethnographic study in the city of Seville, our findings reveal a configuration of practices (accessibility, self-organisation, reproduction and conviviality) through which degrowthminded activism infuses urban life with non-capitalist processes and logics. Consequently, our work contributes to a paucity of studies theorizing the production/consumption of space in relation to broader processes of capitalist development. In doing so, we also promote a more humane consideration of the spatial dimensions through which more equitable ways of living are constituted

Efecto de la inclusión del forraje de maíz molido en la respuesta productiva de vacas lecheras en pastoreo.

Se evaluó la respuesta productiva de vacas lecheras en pastoreo al incluir forraje de maíz molido en su dieta. El estudio se realizó en dos ambientes diferentes de Ecuador (Región Costa y Sierra). En la costa, en una finca de 29,1 ha,las vacas pastaron asociaciones de estrella (Cynodon nlemfuensis) y saboya (Panicum máximum) con leguminosas devarios géneros como Alysicarpus, Centrosema, Desmodium, Galactia, suple-mentadas con forraje de maíz cv. INIAP 125. Los animales recibieron 0,46 kg a partir del tercer kilogramo, y forraje de maíz molido en períodos de 30 y 28 días, respectivamente (M-30 y M-28) y control sin forraje por 36 días (M-0). En la otra región, la propiedad tenía 14,2 ha y 23 vacas en ordeño; estas pastaron asociaciones de kikuyo (P. clandestinum) y ryegra-sstrébol blanco (L. perenne y T. repens) y forraje de maíz planta completa (60 a 70 % grano maduro), a razón de 18 kg verde/vaca/día durante 48 días, y balanceado a razón de 0,5 kg/l de leche después del cuarto kilogramo y minerales. El forraje influyó en ambos casos (P < 0,05) en la respuesta de las vacas; para la zona de la sierra el incremento fue de 1,68 kg/vaca de aumento en la leche y en la costa el aumento fue entre 1,1 y 2,5 kg/vaca. El forraje de maíz complementó nutri-cionalmente las asociaciones bajo pastoreo en ambas regiones, incrementó el rendimiento lechero/animal y redujo los costos.Effect of Ground CornStalks on the Productive Response of Dairy Grazing Cows ABSTRACTThe productive response of dairy grazing cows using ground cornstalks was assessed. The study was developed in two different Ecuadoran settings (Costa Region and Sierra). At the Costa, on a 29.1 ha farm, the cows grazed on star clusters (Cynodon nlemfuensis) and panic grass(Panicum máximum), with legumes of different genera, such as Alysicarpus, Centrosema, Desmodium, Galactia, supplemented with corn stalks, cv. INIAP 125. The animals received 0.46 kg from the third kg onward, and ground corn stalks in 30- and 28- day periods, respectively (M-30 and M-28), and a control group without stalks for 36 days (M-0). In another region, the estate had 14.2 ha and 23 milking cows, grazing on clusters of West African pennisetum (P. clandestinum) and ryegrass white clover (L. perenne and T. repens) and whole corn stalks (60 -70 % ripe grain), at a ratio of 18 kg green/cow/day for 48 days and balanced at a ratio of 0.5 kg/l of milk following the fourth kilogram, and minerals. In both cases, stalks effected (P < 0.05) on cow response. In the Sierra region, the milk yield increase observed was 1.68 kg/cow, and in the Costa region, it was between 1.1 and 2.5 kg/cow. Corn stalks meant nutritional supplementation to grazing clusters in both regions, increased milk/animal yields, and cost reductions

HypoxamiRs Profiling Identify miR-765 as a Regulator of the Early Stages of Vasculogenic Mimicry in SKOV3 Ovarian Cancer Cells

Vasculogenic mimicry (VM) is a novel cancer hallmark in which malignant cells develop matrix-associated 3D tubular networks with a lumen under hypoxia to supply nutrients needed for tumor growth. Recent studies showed that microRNAs (miRNAs) may have a role in VM regulation. In this study, we examined the relevance of hypoxia-regulated miRNAs (hypoxamiRs) in the early stages of VM formation. Data showed that after 48 h hypoxia and 12 h incubation on matrigel SKOV3 ovarian cancer cells undergo the formation of matrix-associated intercellular connections referred hereafter as 3D channels-like structures, which arose previous to the apparition of canonical tubular structures representative of VM. Comprehensive profiling of 754 mature miRNAs at the onset of hypoxia-induced 3D channels-like structures showed that 11 hypoxamiRs were modulated (FC>1.5; p < 0.05) in SKOV3 cells (9 downregulated and 2 upregulated). Bioinformatic analysis of the set of regulated miRNAs showed that they might impact cellular pathways related with tumorigenesis. Moreover, overall survival analysis in a cohort of ovarian cancer patients (n = 485) indicated that low miR-765, miR-193b, miR-148a and high miR-138 levels were associated with worst patients outcome. In particular, miR-765 was severely downregulated after hypoxia (FC < 32.02; p < 0.05), and predicted to target a number of protein-encoding genes involved in angiogenesis and VM. Functional assays showed that ectopic restoration of miR-765 in SKOV3 cells resulted in a significant inhibition of hypoxia-induced 3D channels-like formation that was associated with a reduced number of branch points and patterned tubular-like structures. Mechanistic studies confirmed that miR-765 decreased the levels of VEGFA, AKT1 and SRC-α transducers and exerted a negative regulation of VEGFA by specific binding to its 3‘UTR. Finally, overall survival analysis of a cohort of ovarian cancer patients (n = 1435) indicates that high levels of VEGFA, AKT1 and SRC-α and low miR-765 expression were associated with worst patients outcome. In conclusion, here we reported a novel hypoxamiRs signature which constitutes a molecular guide for further clinical and functional studies on the early stages of VM. Our data also suggested that miR-765 coordinates the formation of 3D channels-like structures through modulation of VEGFA/AKT1/SRC-α axis in SKOV3 ovarian cancer cells

Imaging tumour hypoxia with positron emission tomography.

Hypoxia, a hallmark of most solid tumours, is a negative prognostic factor due to its association with an aggressive tumour phenotype and therapeutic resistance. Given its prominent role in oncology, accurate detection of hypoxia is important, as it impacts on prognosis and could influence treatment planning. A variety of approaches have been explored over the years for detecting and monitoring changes in hypoxia in tumours, including biological markers and noninvasive imaging techniques. Positron emission tomography (PET) is the preferred method for imaging tumour hypoxia due to its high specificity and sensitivity to probe physiological processes in vivo, as well as the ability to provide information about intracellular oxygenation levels. This review provides an overview of imaging hypoxia with PET, with an emphasis on the advantages and limitations of the currently available hypoxia radiotracers.Cancer Research UK (CRUK) funded the National Cancer Research Institute (NCRI) PET Research Working party to organise a meeting to discuss imaging cancer with hypoxia tracers and Positron Emission Tomography. IF was funded by CRUK and is also supported by the Chief Scientific Office. ALH is supported by CRUK and the Breast Cancer Research Foundation. RM is funded by NIHR Cambridge Biomedical Research Centre.This is the accepted manuscript. The final version is available from Nature Publishing at http://www.nature.com/bjc/journal/vaop/ncurrent/full/bjc2014610a.html