Therapeutic Potential of Targeting the Cytochrome P450 Enzymes Using Lopinavir/Ritonavir in Colorectal Cancer: A Study in Monolayers, Spheroids and In Vivo Models.

Cytochrome P450 (CYP450) enzyme has been shown to be expressed in colorectal cancer (CRC) and its dysregulation is linked to tumor progression and a poor prognosis. Here we investigated the therapeutic potential of targeting CYP450 using lopinavir/ritonavir in CRC. The integrative systems biology method and RNAseq were utilized to investigate the differential levels of genes associated with patients with colorectal cancer. The antiproliferative activity of lopinavir/ritonavir was evaluated in both monolayer and 3-dimensional (3D) models, followed by wound-healing assays. The effectiveness of targeting CYP450 was examined in a mouse model, followed by histopathological analysis, biochemical tests (MDA, SOD, thiol, and CAT), and RT-PCR. The data of dysregulation expressed genes (DEG) revealed 1268 upregulated and 1074 down-regulated genes in CRC. Among the top-score genes and dysregulated pathways, CYPs were detected and associated with poor prognosis of patients with CRC. Inhibition of CYP450 reduced cell proliferation via modulating survivin, Chop, CYP13a, and induction of cell death, as detected by AnnexinV/PI staining. This agent suppressed the migratory behaviors of cells by induction of E-cadherin. Moreover, lopinavir/ritonavir suppressed tumor growth and fibrosis, which correlated with a reduction in SOD/thiol levels and increased MDA levels. Our findings illustrated the therapeutic potential of targeting the CYP450 using lopinavir/ritonavir in colorectal cancer, supporting future investigations on this novel therapeutic approach for the treatment of CRC.

Evaluating the effects of direct-fed microbial supplementation on the performance, milk quality and fatty acid of mid-lactating dairy cows.

BACKGROUND: The objective of the experiment was to investigate the effect of a mixture of direct-fed microbial (DFM) on feed intake, nutrient digestibility, milk yield and composition, milk fatty acid and blood parameter in crossbred mid-lactating cows. METHODS: Twenty-four crossbred Holstein cows (body weight = 650±15 kg; days in milk = 100±20; daily milk yield = 25±3 kg) were used in a completely randomized design with three treatments: (1) CON, without DFM; (2) LS, inoculation with Lactobacillus fermentum (4.5 × 108 CFU/day) plus Saccharomyces cerevisiae (1.4×1010 CFU/day); and (3) LSM, inoculation with LS plus Megasphaera elsdenii (4.5 × 108 CFU/day). All animals received the same ration with 45.7% forage and 54.3% concentrate. RESULTS: Results showed that the highest feed intake was observed in treatments LS and LSM (p = 0.02). Compared with the CON, milk production, 4% fat-corrected milk, energy-corrected milk, fat (kg/day), protein (kg/day) and lactose (kg/day), FE and percent of fat were increased (p<0.05) by LSM, but unaffected by LS. Also, compared with the CON, both LS and LSM increased antioxidant activity (p<0.05). The concentration of C18:2c n-6 increased significantly in treatment LSM compared with the CON (p = 0.003). The concentration of C20:0 increased significantly in treatment LS compared with the CON (p = 0.004). The highest concentrations of insulin, glucose, triglyceride and cholesterol were observed by LSM (p<0.05). Compared with the CON, both LS and LSM increased blood monocyte, neutrophil, eosinophil and basophil (p<0.05), and blood lymphocyte was increased (p = 0.02) only by LSM. CONCLUSIONS: The results of the research showed that the use of DFMs had no effect on the digestibility, microbial load and the major part of fatty acids in milk. However, it improved feed intake, milk yield and antioxidant activity of milk and also increased the milk concentration of C18:2 n-6.

Rosemary (Rosmarinus officinalis L.) essential oil alleviates testis failure induced by Etoposide in male rats.

Rosemary (Rosmarinus officinalis L.) is a shrub used to treat hepatic, intestinal, renal, respiratory, and reproductive failures. Etoposide a plant-based compound derived from Podophyllum pelltatum, has been used for human malignancies treatment. However, it induces testis, and hepatic failures. In the present study, impact of rosemary essential oil against testis failure, lipid parameters, and hepatic enzymes in male rats has been studied. Forty male Wistar albino rats were grouped in a completely randomized design with Etoposide injection (ETO), rosemary supplementation (ROS), with Etoposide injection and rosemary supplement (ETO+ROS), and control rats with no Etoposide injection and no rosemary (CON). The experiment lasted for seven consecutive weeks including one week as acclimatization time. At the end of the experiment, rats were sacrificed by cervical dislocation, and blood samples were analyzed for serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), low-density lipoprotein-Cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), total cholesterol (TC), total Protein (TP), glucose (GLU) and testosterone. The left testis was harvested for histological examination. Results showed that rats with Etoposide injection had higher ALT, AST, and ALP the control rats. No significant difference was found among treatments in terms of glucose concentration in blood. Rosemary supplemntaion decreased cholesterol and TG concentration and increased HDL concentration in male rats. Furthermore, administration of rosemary essential oil increased blood testosterone but decreased ALT and AST. The epithelial height of seminiferous tubules was decreased significantly in ET as compared with CON. Rosemary essential oil lessened the adverse effect of Etopside on epithelial height in rat testis as it is shown in ET+ROS. In conclusion, dietary supplementation of rosemary essential oil alleviated liver toxicity and functional testis damage induced by Etopside.

Transcriptome analysis provides new insights into plants responses under phosphate starvation in association with chilling stress.

BACKGROUND: Chilling temperature reduces the rate of photosynthesis in plants, which is more pronounced in association with phosphate (Pi) starvation. Previous studies showed that Pi resupply improves recovery of the rate of photosynthesis in plants much better under combination of dual stresses than in non-chilled samples. However, the underlying mechanism remains poorly understood. RESULTS: In this study, RNA-seq analysis showed the expression level of 41 photosynthetic genes in plant roots increased under phosphate starvation associated with 4 °C (-P 4 °C) compared to -P 23 °C. Moreover, iron uptake increased significantly in the stem cell niche (SCN) of wild type (WT) roots in -P 4 °C. In contrast, lower iron concentrations were found in SCN of aluminum activated malate transporter 1 (almt1) and its transcription factor, sensitive to protein rhizotoxicity 1 (stop1) mutants under -P 4 °C. The Fe content examined by ICP-MS analysis in -P 4 °C treated almt1 was 98.5 ng/µg, which was only 17% of that of seedlings grown under -P 23 °C. Average plastid number in almt1 root cells under -P 4 °C was less than -P 23 °C. Furthermore, stop1 and almt1 single mutants both exhibited increased primary root elongation than WT under combined stresses. In addition, dark treatment blocked the root elongation phenotype of stop1 and almt1. CONCLUSIONS: Induction of photosynthetic gene expression and increased iron accumulation in roots is required for plant adjustment to chilling in association with phosphate starvation.

Determination of intra-axial brain tumors cellularity through the analysis of T2 Relaxation time of brain tumors before surgery using MATLAB software.

INTRODUCTION: Timely diagnosis of brain tumors could considerably affect the process of patient treatment. To do so, para-clinical methods, particularly MRI, cannot be ignored. MRI has so far answered significant questions regarding tumor characteristics, as well as helping neurosurgeons. In order to detect the tumor cellularity, neuro-surgeons currently have to sample specimens by biopsy and then send them to the pathology unit. The aim of this study is to determine the tumor cellularity in the brain. METHODS: In this cross-sectional study, 32 patients (18 males and 14 females from 18-77 y/o) were admitted to the neurosurgery department of Shohada-E Tajrish Hospital in Tehran, Iran from April 2012 to February 2014. In addition to routine pulse sequences, T2W Multi echo pulse sequences were taken and the images were analyzed using the MATLAB software to determine the brain tumor cellularity, compared with the biopsy. RESULTS: These findings illustrate the need for more T2 relaxation time decreases, the higher classes of tumors will stand out in the designed table. In this study, the results show T2 relaxation time with a 85% diagnostic weight, compared with the biopsy, to determine the brain tumor cellularity (p<0.05). CONCLUSION: Our results indicate that the T2 relaxation time feature is the best method to distinguish and present the degree of intra-axial brain tumors cellularity (85% accuracy compared to biopsy). The use of more data is recommended in order to increase the percent accuracy of this techniques.

A study of the relationship between gender/age and apparent diffusion coefficient values in spleen of healthy adults using diffusion-weighted magnetic resonance imaging.

BACKGROUND: Diffusion-weighted magnetic resonance imaging (DWI) systems are very effective in detecting strokes, and they also have shown significant promise in the detection of fibrosis and cirrhosis of the liver. However, such systems have the disadvantages of poor reproducibility and noise, which can diminish the accuracy of the apparent diffusion coefficients (ADCs) provided by the DWI process. The main aim of this study was to determine the relationship between the age and gender of healthy adults in terms of the ADC values of the spleen measured by DWI. METHODS: Sixty-nine subjects selected for this study from people who were referred to the Tabesh Medical Imaging Center in Tabriz, Iran, in 2013. Each subject underwent echo-planar DWI for her or his ADC values of the spleen with b-values of 50, 400, and 800 s/mm2, and the resulting ADC values were evaluated. RESULTS: No significant differences were observed in ADC values of the spleen among the female and male participants or those from various ages (P>0.05). CONCLUSIONS: Based on the findings of this study, it was concluded that the effect of age and gender on the spleen's ADC values can be omitted from the spleen-diagnosis procedure. In other words, the spleen's ADC values are not related to the age or the gender of healthy adults.