Modulation of growth performance, gut microflora, non-specific immunity and gene expression of proinflammatory cytokines in shabout (Tor grypus) upon dietary prebiotic supplementation.

In this study, the effect of dietary supplementation of mannan oligosaccharide (MOS) + ß-glucan (Immunogen®) was investigated on growth performance, body composition, gut microflora, innate immune responses and gene expression of some proinflammatory cytokines in shabout (Tor grypus). Shabout fingerlings (35 ± 1.2 g) were fed with basal diet (control) or basal diet supplemented with Immunogen® at 0.5, 1 and 1.5% of feed for 90 days. According to the results, growth parameters were significantly improved in fish fed with prebiotic (1 and 1.5%) for 90 days (p < 0.05). The carcass protein content was significantly higher in fish nourished by prebiotic at 1.5% of feed for 90 days compared to fish received the basal diet (p < 0.05). Feeding with various levels of Immunogen® resulted in the significant promotion of the population of intestinal Lactobacillus spp. in the prebiotic-treated groups relative to the control group (p < 0.05). Serum total globulin was significantly higher in all prebiotic groups relative to the control group at day 60. Serum bactericidal and lysozyme activities were significantly (p < 0.05) elevated after feeding with dietary prebiotic at all intervals (days 30, 60 and 90). However, the highest serum bactericidal activities were recorded in fish fed with Immunogen® at 1.5% of diet (p < 0.05). The transcription levels of interleukin 1 beta (IL-1ß), interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-α) were significantly increased in the head kidney of fish treated with dietary prebiotic at all intervals. The results show that dietary supplementation with Immunogen®, particularly at the level of 1.5%, can positively alter growth parameters, carcass protein, intestinal microflora and immune responses of shabout.

Quorum Quenching Properties and Probiotic Potentials of Intestinal Associated Bacteria in Asian Sea Bass Lates calcarifer.

Quorum quenching (QQ), the enzymatic degradation of N-acyl homoserine lactones (AHLs), has been suggested as a promising strategy to control bacterial diseases. In this study, 10 AHL-degrading bacteria isolated from the intestine of barramundi were identified by 16S rDNA sequencing. They were able to degrade both short and long-chain AHLs associated with several pathogenic Vibrio species (spp.) in fish, including N-[(RS)-3-Hydroxybutyryl]-l-homoserine lactone (3-oh-C4-HSL), N-Hexanoyl-l-homoserine lactone (C6-HSL), N-(ß-Ketocaproyl)-l-homoserine lactone (3-oxo-C6-HSL), N-(3-Oxodecanoyl)-l-homoserine lactone (3-oxo-C10-HSL), N-(3-Oxotetradecanoyl)-l-homoserine lactone (3-oxo-C14-HSL). Five QQ isolates (QQIs) belonging to the Bacillus and Shewanella genera, showed high capacity to degrade both synthetic AHLs as well as natural AHLs produced by Vibrio harveyi and Vibrio alginolyticus using the well-diffusion method and thin-layer chromatography (TLC). The genes responsible for QQ activity, including aiiA, ytnP, and aaC were also detected. Analysis of the amino acid sequences from the predicted lactonases revealed the presence of the conserved motif HxHxDH. The selected isolates were further characterized in terms of their probiotic potentials in vitro. Based on our scoring system, Bacillus thuringiensis QQ1 and Bacillus cereus QQ2 exhibited suitable probiotic characteristics, including the production of spore and exoenzymes, resistance to bile salts and pH, high potential to adhere on mucus, appropriate growth abilities, safety to barramundi, and sensitivity to antibiotics. These isolates, therefore, constitute new QQ probiotics that could be used to control vibriosis in Lates calcalifer.

Diagnostic investigations of DKK-1 and PDCD5 expression levels as independent prognostic markers of human chondrosarcoma.

In this study, we investigated the expression levels of Dickkopf-1 (DKK-1) and programmed cell death 5 (PDCD5) by using quantitative real-time PCR and immunohistochemistry in patients with chondrosarcoma. The DKK-1 mRNA levels were significantly higher in chondrosarcoma when compared with the corresponding nontumor tissues (mean ± SD: 4.23 ± 1.54; 1.54 ± 0.87; P = 0.001). PDCD5 mRNA levels were remarkably deceased in tumor tissues when compared with corresponding nontumor tissues (mean ± SD: 1.94 ± 0.73; 5.42 ± 1.73; P = 0.001). The high and moderate DKK-1 expressions were observed for 60% of chondrosarcoma samples in comparison with 27.5% of corresponding nontumor tissues (P  =  0.001). Moreover, low expression of PDCD5 was found in 67.5% of the tumor tissues when compared with the nontumor tissues (32.5%; P = 0.002). The results of this study showed that high DKK-1 expression levels were strongly related to MSTS stage (P = 0.011) and the advancement of histological grade (P < 0.001). Furthermore, the PDCD5 expression levels were correlated with histological grade (P < 0.001), MSTS stage (P = 0.016), and distant metastasis (P = 0.001). Kaplan-Meier survival and log-rank survival showed that patients with high DKK-1 levels and low PDCD5 levels were correlated with shorter overall survival (log-rank test P < 0.001). PDCD5 levels, histological grade, and tumor stage were independent predictors of overall survival. In conclusion, DKK-1 and PDCD5 can be independent predictors of overall survival in patients suffering from chondrosarcoma. © 2016 IUBMB Life, 68(7):597-601, 2016.

Downregulation of miR-148b as biomarker for early detection of hepatocellular carcinoma and may serve as a prognostic marker.

MicroRNAs (miRNAs) have a large number of various target genes in different cancer types, which may result in many biological functions. Thus, identifying the molecular mechanisms of miRNAs may effect on the complexity of cancer progression via regulation of gene. In the current study, we utilized real-time PCR to quantify the diction of miR-148b in trail hepatocellular carcinoma (HCC) specimen and normal tissues. Furthermore, we evaluated the relationship of miR-148b and clinicopathological features with survival of HCC patients. Therefore, we evaluated the level of miR-148b expression in 101 HCC patients and also in 40 normal control cases. The result suggested lower expression in tumor tissues than normal control tissues (0.96 ± 0.14; 1.84 ± 0.20, P < 0.05). Our findings suggest that the declined expression of miR-148b can considerably be linked to tumor node metastasis (TNM) stage (stages III and IV; P = 0.021) and vein invasion (P = 0.029). Nevertheless, miR-148b expression was not related to sex (P = 0.674), age (P = 0. 523), size of tumor (P = 0.507), liver cirrhosis, and histologic grade (P = 0.734). Survival analysis showed that low expression was remarkably related to overall survival (P = 0.012). Furthermore, multivariate survival test suggested that decline of miR-148b diction was linked to poor survival in HCC patients. Our results suggested that miR-148b is decreased in HCC. Therefore, we concluded that miR-148b may play its role in the prognosis of HCC.

Investigations of renal function using the level of neutrophil gelatinase-associated lipocalin associated with single-dose of cisplatin during chemotherapy.

BACKGROUND: Neutrophil gelatinase-associated lipocalin (NGAL) is known as a primary indicator of acute and chronic renal and can be effective in chronic kidney injury (CKI) with kidney tumors cisplatin (CP) chemotherapy. The aim of this study was to evaluate serum and urinary biomarker including NGAL (sNGAL and uNGAL) in canine with solid renal tumors who suffered from cisplatin after short and long-term chemotherapy. METHODS: In this study, in treatment and control groups, canine (n = 10 and n = 5) were administered cisplatin at 1.2 mg/kg/day (i.v.) for five consecutive days with CKI and without CKI, respectively. Serum and urine NGAL levels (ng/mL) were evaluated at 0, 1, 5, 9, 13, 17, 21, 25 and 29 days after drug injection versus baseline in treated and control groups. RESULTS: Canine in treatment group had shown symptoms of toxicity of cisplatin. The results indicated the higher concentrations of serum, sNGAL and uNGAL (P = 0.024; P = 0.011) compared with control group (P = 0.701, P = 0.612), (Table 2, Figs. 1 and 2). Indeed, our results showed that canine with CKI were associated with higher levels of sNGAL and uNGAL compared with control group without CKI. Moreover, the highest level of uNGAL was seen in comparison with sNGAL, after a high dose (1.2 mg/kg) administration of CP. CONCLUSION: Our data suggested that U-NGAL may be useful for monitoring of renal injury in CKI patients that exposed with cisplatin. Furthermore, a primary elevate in urinary NGAL expulsion may help in identifying cases at danger of cisplatin-induced CKI that might profit from innovative remedies to prevent cisplatin nephrotoxicity.

MiR-15/16 complex targets p70S6 kinase 1 and controls cell proliferation in MDA-MB-231 breast cancer cells.

BACKGROUND: MicroRNAs are small non-coding RNAs that regulate post-transcriptional mRNA expression by binding to 3' untranslated region (3'-UTR) of the complementary mRNA sequence resulting in translational repression and gene silencing. They act as negative regulators of gene expression and play a pivotal role in regulating apoptosis and cell proliferation. Studies have shown that miRNAs interact with p53 by regulating the activity and function of p53 through direct repression or its regulators. Mammalian target of rapamycin (mTOR) is an evolutionary conserved check point protein kinase that plays a major effect in the control of cell division via protein synthesis regulation. mTOR regulates protein synthesis through phosphorylation and inactivation of 4E-BP1 and through phosphorylation and activation of S6 kinase 1 (S6K1). These two downstream effectors of mTOR control cell growth and metabolism. In mammals, mTOR protein kinase is the central node in the nutrient and growth factor signaling and p53 plays a critical role in sensing genotoxic stress. Activation of p53 inhibits mTOR activity, which in turn regulates its downstream targets providing a cross talk among both the signaling machinery. MicroRNA-15 and 16 belong to a common precursor family and are highly conserved. Deletion or downregulation of these two microRNAs has been shown to accelerate cell division by modulating the expression of the genes involved in controlling cell cycle progression. These microRNAs may function as tumor suppressors and act on the downstream targets of p53 signaling pathway. To have a better insight of the role of miR-15/16 in regulating the cross talk of p53 and mTOR, we performed an in depth study in MDA-MB-231 breast cancer cells by performing a gain-of-function analysis with lentiviral plasmids expressing microRNA-15 and 16. METHODS: The effect of individual microRNAs on RPS6KB1 was examined by using 3'-UTR clones via luciferase based assays. The cell cycle effects were observed by flow-cytometric analysis. Reverse transcription PCR was used to explore the expression of mTOR and RPS6KB1 in cells transfected with miR-15/16. RESULTS: Overexpression of miR-15/16 led to inhibition of cell proliferation causing G1 cell cycle arrest as well as caspase-3 dependent apoptosis. Forced expression of miR-15/16 might lead to decrease in mRNA level of RPS6KB1, mTOR. The effect was a complete reversal after treatment with anti-miRs against miR-15/16 proving the specificity of the expression. In addition, the dual luciferase reporter assays indicated a clear decrease in luciferase gene expression in cells transfected with lentiviral based miR-15 and 16 plasmids indicating that miR-15/16 directly targets RPS6KB1 through its 3'-UTR binding. Further, these microRNAs also inhibit epithelial to mesenchymal transition (EMT) by targeting key proteins such as Twist1 and EZH2 clearly demonstrating its crucial role in controlling cell proliferation. CONCLUSION: This study suggests that exogenous microRNA-15/16 can target RPS6KB1, control cell proliferation and cause apoptosis in caspase-dependent manner even in the absence of functional p53.