Integrated Multidimensional Analysis Is Required for Accurate Prognostic Biomarkers in Colorectal Cancer

CRC cancer is one of the deadliest diseases in Western countries. In order to develop prognostic biomarkers for CRC (colorectal cancer) aggressiveness, we analyzed retrospectively 267 CRC patients via a novel, multidimensional biomarker platform. Using nanofluidic technology for qPCR analysis and quantitative fluorescent immunohistochemistry for protein analysis, we assessed 33 microRNAs, 124 mRNAs and 9 protein antigens. Analysis was conducted in each single dimension (microRNA, gene or protein) using both the multivariate Cox model and Kaplan-Meier method. Thereafter, we simplified the censored survival data into binary response data (aggressive vs. non aggressive cancer). Subsequently, we integrated the data into a diagnostic score using sliced inverse regression for sufficient dimension reduction. Accuracy was assessed using area under the receiver operating characteristic curve (AUC). Single dimension analysis led to the discovery of individual factors that were significant predictors of outcome. These included seven specific microRNAs, four genes, and one protein. When these factors were quantified individually as predictors of aggressive disease, the highest demonstrable area under the curve (AUC) was 0.68. By contrast, when all results from single dimensions were combined into integrated biomarkers, AUCs were dramatically increased with values approaching and even exceeding 0.9. Single dimension analysis generates statistically significant predictors, but their predictive strengths are suboptimal for clinical utility. A novel, multidimensional integrated approach overcomes these deficiencies. Newly derived integrated biomarkers have the potential to meaningfully guide the selection of therapeutic strategies for individual patients while elucidating molecular mechanisms driving disease progression.     

The investigation of the combined effects of gamma irradiation and environmental manipulation on mortality and sterility of Rhyzopertha dominica (Fabricius) (Coleoptera: Bostrichidae)

With respect to the limitations use of methyl bromide and phosphine, employing ionizing radiation to control stored product pests has attracted great attention. The aim of this study is the investigation of the combined effects of gamma irradiation as viable alternatives to synthetic insecticides and environmental management on mortality and sterility of Rhyzopertha dominica (Fabricius) in a wheat cultivar (Gascogen). The effect of doses range from 30 to 2000?Gy gamma irradiation in combination with manipulation of temperature (15, 21, 27 and 32?°C) and relative humidity (20%, 50%, 65% and 85%) on 5–10?days old adults of R. dominica in Gascogen cultivar of wheat were explored. The experiments were repeated three times and conducted in The Nuclear Agriculture Research School in Karaj and laboratory of shahid steki silo in shahre-kord. Probit analysis revealed that both temperature and relative humidity had combined effects when used with gamma irradiation. The lowest doses of gamma ray required to kill 25% (14.2?Gy) and 50% (610.8?Gy) of the population (LD₂₅ and LD₅₀) were recorded at 21?°C and 85% relative humidity respectively. The low dose for 99% mortality (LD₉₉; 2386.7?Gy) was recorded for beetles maintained at 21?°C and 50% relative humidity. The effect of temperature (15, 21, 27 and 32?°C) on sterility caused by gamma irradiation was also investigated. The results showed that the F₁ generation emerged only when the beetles were treated with doses of 0–100?Gy at 32?°C and 0–70?Gy at 27?°C. These results indicate that temperature and relative humidity play an important role in the susceptibility of the lesser grain beetle to gamma irradiation. The results suggest that controlling the efficiency of gamma radiation through environmental control allows the use of low doses of gamma radiation that have a less harmful effect on human health, non-target organisms and seed agronomic features.     

In vitro cardiomyogenic potential of human umbilical vein-derived mesenchymal stem cells

Cardiomyocyte loss in the ischemically injured human heart often leads to irreversible defects in cardiac function. Recently, cellular cardiomyoplasty with mesenchymal stem cells, which are multipotent cells with the ability to differentiate into specialized cells under appropriate stimuli, has emerged as a new approach for repairing damaged myocardium. In the present study, the potential of human umbilical cord-derived mesenchymal stem cells to differentiate into cells with characteristics of cardiomyocyte was investigated. Mesenchymal stem cells were isolated from endothelial/subendothelial layers of the human umbilical cords using a method similar to that of human umbilical vein endothelial cell isolation. Isolated cells were characterized by transdifferentiation ability to adipocytes and osteoblasts, and also with flow cytometry analysis. After treatment with 5-azacytidine, the human umbilical cord-derived mesenchymal stem cells were morphologically transformed into cardiomyocyte-like cells and expressed cardiac differentiation markers. During the differentiation, cells were monitored by a phase contrast microscope and their morphological changes were demonstrated. Immunostaining of the differentiated cells for sarcomeric myosin (MF20), desmin, cardiac troponin I, and sarcomeric alpha-actinin was positive. RT-PCR analysis showed that these differentiated cells express cardiac-specific genes. Transmission electron microscopy revealed a cardiomyocyte-like ultrastructure and typical sarcomers. These observations confirm that human umbilical cord-derived mesenchymal stem cells can be chemically transformed into cardiomyocytes and can be considered as a source of cells for cellular cardiomyoplasty.     

Genotyping of Acanthamoeba isolates from clinical and environmental specimens in Iran

In this study, 15 Acanthamoeba isolates from AK patients and 10 environmental samples (water, soil and animal-origin samples) were classified at the genotype level based on the sequence analysis of the Diagnostic Fragment 3 (DF3) of Acanthamoeba small subunit rRNA gene. The obtained results revealed that most of these Acanthamoeba strains belonged to genotype T4 both in clinical and environmental samples. The presence T11 genotype in clinical samples was also revealed after the genotyping analysis and to our knowledge this is the first report of T11 genotype in Iran. Moreover, the isolation of T4 genotype from cow faeces in this study highlights a possible transmission of Acanthamoeba through animal faeces in Iran.Overall, the widespread distribution of pathogenic Acanthamoeba T4 across the environmental sources and the increasing number of contact lens wearers in Iran, demands more awareness within the public and health professionals as this pathogen is emerging as a risk for human health in Iran and worldwide.     

Intralateral hypothalamic area injection of isoproterenol and propranolol affects food and water intake in broilers

The role played by adrenergic system in water intake, especially food intake, has long been known in mammals. In avian species, there have been many experiments exploring the effects of the adrenergic system in different sites in the central nervous system in meat- and egg-type poultry. This study was designed to examine the possible effects of intralateral hypothalamic area (ILHy) microinjections of a beta-adrenergic agonist, isoproterenol, and a beta-adrenoceptor blocker, propranolol, on food and water intake in 3-h food-deprived and 3-h water-deprived broiler cockerels. Our findings suggest that the β-adrenergic system directly affects food especially water intake in broilers. Although isoproterenol significantly (P ≤ 0.05) decreased food intake for the first 15 min, it reduced food intake during the experiment. Isoproterenol reduced water intake significantly (P ≤ 0.05), which was abolished by pretreatment with propranolol. It is proposed that β-adrenoceptors in LHy play a direct and indirect role in the regulation of food especially water intake in broiler cockerels.     

Hepatic mechanisms of the Walnut antidiabetic effect in mice

The present study was designed to explore the mechanism of action of walnut (the seed of Juglans regia) leaf and ridge on hepatic glucose metabolism in diabetic mice. Experimental diabetes was induced by intravenous administration of streptozotocin (60 mg/kg)and confirmed with an increase of blood glucose, 90–100% of the control, 72 hours later. Isolated extracts from walnut leaf and ridges were administered in a single effective dose of 400 mg/kg orally. Firstly, blood glucose was determined every 1 hour until 5 hours post administration of extracts. In the second experiment, the liver was surgically removed, 2 hours post treatment of diabetic animals with extracts, homogenized and used for measurement of key enzymes of glycogenolysis (glycogen phosphorylase, GP) and gluconeogenesis (phosphoenolpyruvate carboxykinase, PEPCK). Treatment by both leaf and ridge extracts decreased blood glucose and liver PEPCK activity and increased blood insulin and liver GP activity. It is concluded that walnut is able to lower blood glucose through inhibition of hepatic gluconeogenesis and secretion of pancreatic insulin.