Agouti-related protein neuron circuits that regulate appetite

WOS: 000348622800002PubMed ID: 25402352New tools for mapping and manipulating molecularly defined neural circuits have improved the understanding of how the central nervous system regulates appetite. Studies that focused on Agouti-related protein neurons, a starvation-sensitive hypothalamic population, have identified multiple circuit elements that can elicit or suppress feeding behavior. Distinct axon projections of this neuron population point to different circuits that regulate long-term appetite, short-term feeding, or visceral malaise-mediated anorexia. Here, we review recent studies examining these neural circuits that control food intake

Interaction of RNA-binding protein HuR and miR-466i regulates GM-CSF expression.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) produced by T helper 17 (Th17) cells plays an essential role in autoimmune diseases. Transcriptional regulation of Th17 cell differentiation has been extensively studied, but post-transcriptional regulation of Th17 cell differentiation has remained less well characterized. The RNA-binding protein HuR functions to promote the stability of target mRNAs via binding the AU-rich elements of the 3\u27 untranslated region (3\u27UTR) of numerous pro-inflammatory cytokines including IL-4, IL-13, IL-17 and TNF-α. However, whether HuR regulates GM-CSF expression in Th17 cells has not been fully investigated. Here we showed that HuR conditional knockout (KO) Th17 cells have decreased GM-CSF mRNA in comparison with wild-type (WT) Th17 cells, and that HuR binds directly to GM-CSF mRNA 3\u27UTR. Interestingly, HuR deficiency increased the levels of certain microRNA expression in Th17 cells; for example, miR-466i functioned to mediate GM-CSF and IL-17 mRNA decay, which was confirmed by in vitro luciferase assay. Furthermore, we found that HuR promoted Mxi1 expression to inhibit certain miRNA expression. Taken together, these findings indicate that interaction of HuR and miR-466i orchestrates GM-CSF expression in Th17 cells

A comprehensive study of volatile fatty acids production from batch reactor to anaerobic sequencing batch reactor by using cheese processing wastewater

Volatile fatty acids (VFAs) has great potential for closed-loop production in dairy industries via resource recovery from waste-streams. In the current study, the transition of VFA production from batch reactor to anaerobic sequencing batch reactor (ASBR) by using cheese industry wastewater under alkali pH was evaluated with respect to seed sludge structure, microbial diversity and reactor type. The transition from the batch reactor to the ASBR demonstrated that the maximum VFA production yield (g COD/g SCOD) was comparable in two reactors (batch: 0.97; ASBR: 0.94), whereas, the dominant acid type was different (batch: 49% lactic acid; ASBR: 80% propionic acid). There was a significant correlation between the productions of butyric acid with Gracilibacteraceae and Desulfovibrionaceae; propionic acid with Desulfovibrionaceae and Synergistaceae; lactic acid with Pseudomonadaceae and Rhodocyclaceae. The high VFA production efficiency can be achieved by long term reactor operation, which enables the shift from industrial waste-streams to biorefineries

HuR overexpression in MB231 breast cancer cells

Abstract only availableCancer cells share acquired capabilities necessary for their malignant transformation. These "hallmarks of cancer" include increased proliferation, self-sufficiency in growth signals, insensitivity to antigrowth signals, evasion of apoptosis, angiogenesis and metastasis (Hanahan and Weinberg 2000). HuR is a RNA-binding protein which has been implicated in regulating mRNAs involved in each of these characteristics. We hypothesize that HuR maintains the growth characteristics of malignant cancer cells through the stabilization and increased translation of cancer relevant genes. If HuR does enhance malignancy then the overexpression of HuR would amplify the capabilites of malignant cancer cells and increase cell proliferation. This hypothesis was tested by creating a breast cancer cell line that stably overexpresses HuR. A vector overexpressing HuR was created by ligating a PCR amplified insert containing HuR and a HA hemagluttin tag into a Zeocin resistant episomal plasmid. Cells normally express HuR, so the tag was used to distinguish the overexpressed HuR from endogenous HuR. This plasmid was used to transfect MB-231 estrogen receptor-negative breast cancer cells. After transfection, Zeocin selected against the cells that did not incorporate the plasmid. Western Blots for the surviving cells revealed that HA HuR was expressed, implying that the cells were overexpressing HuR. Proliferation assays of heterogenous populations of both HA HuR-containing and normal MB231 cells yield no difference in cell division. Further experiments will use homogenous populations that highly overexpress HuR to see if HuR overexpression alters the proliferation and cell cycle capabilities of these cells. References: "Hallmarks of Cancer" Hanahan, Douglas and Weinberg, Robert A. Cell. Vol. 100, 57-70. 200

The role of post-transcriptional regulation in chemokine gene expression in inflammation and allergy.

The aim of this review is to discuss recent advances in the understanding of the regulation of chemokine expression occurring during chronic inflammatory conditions, such as allergic diseases. The focus will be on current data, which suggest that post-transcriptional regulation plays a larger role in chemokine gene regulation than previously recognised. In particular, a growing body of data indicates that mechanisms controlling mRNA stability may be relevant in determining, or maintaining, the increased levels of chemokine gene expression in this context. Such regulatory pathways may be important targets of novel anti-inflammatory strategies

Bioconversion of food waste to volatile fatty acids: impact of microbial community, pH and retention time

Bio-based production of materials from waste streams is a pivotal aspect in a circular economy. This study aimed to investigate the influence of inoculum (three different sludge taken from anaerobic digestors), pH (5 & 10) and retention time on production of total volatile fatty acids (VFAs), VFA composition as well as the microbial community during anaerobic digestion of food waste. The highest VFA production was ∼22000 ± 1036 mg COD/L and 12927 ± 1029 mg COD/L on day 15 using the inoculum acclimated to food waste at pH 10 and pH 5, respectively. Acetic acid was the dominant VFA in the batch reactors with initial alkaline conditions, whereas both propionic and acetic acids were the dominant products in the acidic condition. Firmicutes, Chloroflexi and Bacteroidetes had the highest relative abundance in the reactors. VFA generation was positively correlated to the relative abundance of Firmicutes

Long-term effects of neoadjuvant chemoradiotherapy followed by sphincter-preserving resection on anal sphincter function in relation to quality of life among locally advanced rectal cancer patients: a cross-sectional analysis

BACKGROUND: There is growing recognition for the consequences of rectal cancer treatment to maintain an adequate functional sphincter in the long-term rather than preserving the anal sphincter itself. This study aims to evaluate long-term effects of neoadjuvant chemoradiotherapy (nCRT) followed by sphincter-preserving resection on anal sphincter function in relation to quality of life (QoL) among locally advanced rectal cancer patients. METHODS: Twenty-nine patients treated with nCRT followed by low anterior resection surgery were included in this study. Data on patient demographics, tumor location and symptoms of urgency and fecal soiling were recorded and evaluated with respect to Wexner Fecal Incontinence Scoring Scale, European Organization for Research and Cancer (EORTC) cancer-specific (EORTC QLQ-C30) and colorectal cancer-specific (EORTC QLQ-CR38) questionnaires and anorectal manometrical findings. Correlation of manometrical findings with Wexner Scale, EORTC QLQ-CR38 scores and EORTC QLQ-C30 scores was also evaluated. RESULTS: Median follow-up was 45.6 months (ranged 7.5–98 months. Higher scores for incontinence for gas (p = 0.001), liquid (p = 0.048) and solid (p = 0.019) stool, need to wear pad (p = 0.001) and alteration in life style (p = 0.004) in Wexner scale, while lower scores for future perspective (p = 0.010) and higher scores for defecation problems (p = 0.001) in EORTC QLQ-CR38 were noted in patients with than without urgency. Manometrical findings of resting pressure (mmHg) was positively correlated with body image (r = 0.435, p = 0.030) and sexual functioning (r = 0.479, p = 0.011) items of functional scale, while rectal sensory threshold (RST) volume (mL) was positively correlated with defecation problems (r = 0.424, p = 0.031) items of symptom scale in EORTC QLQ-CR38 and negatively correlated with social function domain (r = −0.479, p = 0.024) in EORTC QLQ-C30. RST volume was also positively correlated with Wexner scores including incontinence for liquid stool (r = 0.459, p = 0.024), need to wear pad (r = 0.466, p = 0.022) and alteration in lifestyle (r = 0.425, p = 0.038). CONCLUSION: The high risk of developing functional anal impairment as well as the systematic registration of not only oncological but also functional and QoL related outcomes seem important in rectal cancer patients in the long-term disease follow-up

Transcriptomic-wide discovery of direct and indirect HuR RNA targets in activated CD4+ T cells

Due to poor correlation between steady state mRNA levels and protein product, purely transcriptomic profiling methods may miss genes posttranscriptionally regulated by RNA binding proteins (RBPs) and microRNAs (miRNAs). RNA immunoprecipitation (RIP) methods developed to identify in vivo targets of RBPs have greatly elucidated those mRNAs which may be regulated via transcript stability and translation. The RBP HuR (ELAVL1) and family members are major stabilizers of mRNA. Many labs have identified HuR mRNA targets; however, many of these analyses have been performed in cell lines and oftentimes are not independent biological replicates. Little is known about how HuR target mRNAs behave in conditional knock-out models. In the present work, we performed HuR RIP-Seq and RNA-Seq to investigate HuR direct and indirect targets using a novel conditional knock-out model of HuR genetic ablation during CD4+ T activation and Th2 differentiation. Using independent biological replicates, we generated a high coverage RIP-Seq data set (>160 million reads) that was analyzed using bioinformatics methods specifically designed to find direct mRNA targets in RIP-Seq data. Simultaneously, another set of independent biological replicates were sequenced by RNA-Seq (>425 million reads) to identify indirect HuR targets. These direct and indirect targets were combined to determine canonical pathways in CD4+ T cell activation and differentiation for which HuR plays an important role. We show that HuR may regulate genes in multiple canonical pathways involved in T cell activation especially the CD28 family signaling pathway. These data provide insights into potential HuR-regulated genes during T cell activation and immune mechanisms