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Summary: Dickeya dadantii is a plant pathogen that causes soft rot disease on vegetable and potato crops. To successfully cause infection, this pathogen needs to coordinately modulate the expression of genes encoding several virulence determinants, including plant cell wall degrading enzymes (PCWDEs), type III secretion system (T3SS) and flagellar motility. Here, we uncover a novel feed‐forward signalling circuit for controlling virulence. Global RNA chaperone Hfq interacts with an Hfq‐dependent sRNA ArcZ and represses the translation of pecT, encoding a LysR‐type transcriptional regulator. We demonstrate that the ability of ArcZ to be processed to a 50 nt 3′‐ end fragment is essential for its regulation of pecT. PecT down‐regulates PCWDE and the T3SS by repressing the expression of a global post‐transcriptional regulator‐ (RsmA‐) associated sRNA encoding gene rsmB. In addition, we show that the protein levels of two cyclic di‐GMP (c‐di‐GMP) diguanylate cyclases (DGCs), GcpA and GcpL, are repressed by Hfq. Further studies show that both DGCs are essential for the Hfq‐mediated post‐transcriptional regulation on RsmB. Overall, our report provides new insights into the interplays between ubiquitous signalling transduction systems that were most studied independently and sheds light on multitiered regulatory mechanisms for a precise disease regulation in bacteria. [ABSTRACT FROM AUTHOR]

Ghrelin and obestatin are two gastrointestinal peptides obtained by post-translational processing of a common precursor, preproghrelin. The effect of obestatin on food intake is still controversial. The aim of the present study was to investigate the effects of ghrelin and obestatin on food intake in grass carp, Ctenopharyngodon idellus . Fish received intraperitoneal (IP) injection of saline, ghrelin (100 ng g − 1 BW), obestatin-like (25 ng g − 1 BW) and ghrelin in combination with obestatin-like. Ghrelin stimulation of food intake varied considerably among individual fish with 70.8% eliciting a robust response. In these high-responders, food intake was significantly increased by IP ghrelin within 2 h. Co-administration of ghrelin and obestatin-like resulted in a decrease in food intake, indicating that obestatin was able to antagonize the effect of ghrelin. However, IP obestatin-like alone could not regulate food intake in grass carp. RT-PCR analysis demonstrated that IP ghrelin peptide led to a significant increase in mRNA abundance of NPY, Y8a and Y8b genes compared to saline injected fish, while in combination with obestatin-like peptide decreased ghrelin-induced gene expressions of these three genes. IP sole obestatin-like peptide did not modify the expression levels of NPY, Y8a, Y8b, CART and POMC compared to the control group. Therefore, IP administration of obestatin-like peptide, partially blocking the ghrelin-induced appetite, investigated the possible involvement of obestatin as a mediator of the ghrelin stimulatory action on food intake, at least in “high-responders” grass carp. [ABSTRACT FROM AUTHOR]

Abstract: The R/L type pyruvate kinase (PKLR) gene was cloned from liver of grass carp (Ctenopharyngodon idella) by degenerate oligonucleotide primed PCR, including a complete open reading frame (ORF) of 1617bp. Homological protein analysis showed that the PKLR in grass carp shared a high degree of sequence identity with other known PK sequences. In addition, tissue distribution analysis showed that the highest expression of the PKLR was detected in liver. Then triplicate groups of grass carp (9 tanks) were fed twice daily to satiation with 91.8, 192.9, and 402.6gkg−1 carbohydrate diets for 8weeks, respectively. Dietary 402.6gkg−1 carbohydrate decreased growth performance, feed intake and feed utilization. With increasing dietary carbohydrates, the hepatic PK activity and mRNA expression of grass carp were up-regulated significantly. The results indicated that the PKLR play a modulation role in the adaptation of hepatic glycolysis to dietary high carbohydrates in grass carp. [Copyright &y& Elsevier]

As typical herbivorous fish, grass carp possesses greater capability to utilize carbohydrate than carnivorous and omnivorous fish. However, mechanism of carbohydrate utilization in herbivorous fish was unclear. This study is intended to investigate response of grass carp to insufficient or excessive dietary carbohydrate. Grass carp accepted isonitrogenous diets with different carbohydrate levels (15.90, 21.81 and 45.45 g 100 g −1 ) were cultured. After 8 weeks, weight gain and feed intake were highest in fish fed with medium-carbohydrate diet compared with others. Gene expression data indicated that expressions of glucose-6-phosphatase (G6Pase) were down-regulated and gene expressions of glycogen synthase (GS) were up-regulated in fish accepted lower-carbohydrate diet. In high-carbohydrate group, glucose catabolism was increased by enhanced expressions of glucokinase genes (GK). Meanwhile, fatty acid synthase (FAS) and acetyl coenzyme-A carboxylase (ACC1) genes were significantly up-regulated and lipoprotein lipase and carnitine palmitoyltransferase 1 (CPT1) genes expressions were down-regulate, inducing more lipid deposition. Subsequently, serum cholesterol and total lipid contents were significantly increased, and then induced appetite suppression through up-regulation of leptin and anorexigenic genes expressions. Furthermore, depressed food intake caused lower growth rate in grass carp with excessive carbohydrate intake. [ABSTRACT FROM AUTHOR]

Necrotrophic plant pathogens acquire nutrients from dead plant cells, which requires the disintegration of the plant cell wall and tissue structures by the pathogen. Infected plants lose tissue integrity and functional immunity as a result, exposing the nutrient rich, decayed tissues to the environment. One challenge for the necrotrophs to successfully cause secondary infection (infection spread from an initially infected plant to the nearby uninfected plants) is to effectively utilize nutrients released from hosts towards building up a large population before other saprophytes come. In this study, we observed that the necrotrophic pathogen Dickeya dadantii exhibited heterogeneity in bacterial cell length in an isogenic population during infection of potato tuber. While some cells were regular rod-shape (<10μm), the rest elongated into filamentous cells (>10μm). Short cells tended to occur at the interface of healthy and diseased tissues, during the early stage of infection when active attacking and killing is occurring, while filamentous cells tended to form at a later stage of infection. Short cells expressed all necessary virulence factors and motility, whereas filamentous cells did not engage in virulence, were non-mobile and more sensitive to environmental stress. However, compared to the short cells, the filamentous cells displayed upregulated metabolic genes and increased growth, which may benefit the pathogens to build up a large population necessary for the secondary infection. The segregation of the two subpopulations was dependent on differential production of the alarmone guanosine tetraphosphate (ppGpp). When exposed to fresh tuber tissues or freestanding water, filamentous cells quickly transformed to short virulent cells. The pathogen adaptation of cell length heterogeneity identified in this study presents a model for how some necrotrophs balance virulence and vegetative growth to maximize fitness during infection. [ABSTRACT FROM AUTHOR]

A study was conducted to determine the effects of dietary non-protein energy sources on growth, tissue lipid accumulation and lipid metabolism-related genes expression of grass carp. Triplicate groups of fish were fed for 9 weeks on four isonitrogenous (300 g kg−1) experimental diets with four levels of non-protein energy (6.52 kJ g−1 control diet, 5.32 kJ g−1 high- CEL diet, 8.46 kJ g−1 high- CHO diet and 8.53 kJ g−1 high- LIP diet respectively). Increasing dietary non-protein energy source levels did not improve the growth, and the high- CEL diet reduced the growth of grass carp. The high- CHO diet tended to induce high hepatosomatic index, with high fat and glycogen content of liver. However, the high- LIP diet caused the high mesenteric fat index, but did not increase liver fat. The m RNA abundance and activities of hepatic lipogenic enzymes were significantly increased in the high- CHO diet group, whereas the opposite tendencies were observed in the high- LIP diet group. Peroxisome proliferator-actived receptor-α ( PPARα) in liver and PPARγ in mesenteric adipose tissue were up-regulated in the high- CEL diet group. Lipoprotein lipase ( LPL) gene expression was significantly increased both in liver and mesenteric adipose tissue of fish fed the high- LIP diet, while the LPL gene expression was up-regulated in liver but down-regulated in mesenteric adipose tissue of fish fed the high- CEL diet. These findings suggest that an increase in dietary non-protein energy sources alters the genes expression of lipid metabolism and increased lipid deposition. [ABSTRACT FROM AUTHOR]

Matrix metalloproteinases (MMPs) have been involved in inflammatory and degradative processes in pathologic conditions. The purpose of this study was to investigate the protective effect of melatonin in human umbilical vein endothelial cell (HUVEC) monolayer permeability and the regulation of MMP9 induced by interleukin 1&bgr; (IL1&bgr; (IL1B)) in HUVECs. Protection studies were carried out with melatonin, a well-known antioxidant and antiinflammatory molecule. MMP9 expression was increased with IL1&bgr; induction in HUVECs. Melatonin showed a barrier-protective role by downregulation of MMP9 and upregulation of tissue inhibitor of metalloproteinase-1 expression in HUVECs. Meanwhile, melatonin also decreased sodium fluorescein permeability and counteracted the downregulation of vascular endothelial cadherin and occludin expression in HUVECs. During inflammatory stimulus, nuclear factor-kB (NF-kB) plays a significant role in regulating MMP genes expression, thus the function of NF-kB in HUVECs' barrier disruption was investigated. IL1&bgr; induced nuclear translocation of NF-kB in HUVECs and regulated MMP9 expression. However, NF-kB translocation into the nucleus was inhibited significantly by melatonin. Our results show that melatonin decreases the permeability of monolayer endothelial cell induced by IL1&bgr;. At the same time, melatonin decreased the expression and activity of MMP9 by a NF-kB-dependent pathway in HUVECs induced by IL1&bgr;. [ABSTRACT FROM AUTHOR]