Subject(s)
Cryopyrin-Associated Periodic Syndromes/complications , Intraocular Pressure , Ocular Hypertension/etiology , Chronic Disease , Cryopyrin-Associated Periodic Syndromes/diagnosis , Gonioscopy , Humans , Male , Microscopy, Acoustic , Middle Aged , Ocular Hypertension/diagnosis , Ocular Hypertension/physiopathology , Rare DiseasesABSTRACT
The degradation rate and efficiency of digestion processes is typically measured by introducing a substrate or pollutant into a digester and then monitoring the effluents for the pollutant or substrate, a costly and slow process. A new method for rapid measurement of the rates and efficiencies of anaerobic degradation of pollutants and lignocellulose substrates from various pretreatments is described. The method uses micro-reactors (10-30mL) containing a mixed culture of anaerobic bacteria obtained from a working anaerobic digester. The rate of degradation of pollutants and metabolic heat rate are measured in parallel sets of micro-reactors. Measurement of metabolic rate and pollutant degradation simultaneously is an effective means of rapidly examining pollutant degradation on a micro-scale. Calorimetric measurements alone allow rapid, relative evaluation of various substrate pretreatment methods.
Subject(s)
Bacteria, Anaerobic/physiology , Bacterial Proteins/metabolism , Bioreactors/microbiology , Calorimetry/methods , Cell Culture Techniques/instrumentation , Energy Metabolism/physiology , Equipment Failure Analysis/methodsABSTRACT
Hepatic stellate cells (HSCs) are key cellular components of hepatic wound healing and fibrosis. There is emerging evidence that the fibrogenic function of HSCs may be influenced by neurochemical and neurotrophic factors. This study addresses the potential for the serotonin (5-HT) system to influence HSC biology. Rat and human HSCs express the 5-HT1B, 5-HT1F 5-HT2A 5-HT2B, and 5-HT7 receptors, with expression of 5-HT1B 5-HT2A and 5-HT2B being induced on HSC activation. Induction of 5-HT2A and 5-HT2B was 106+/-39- and 52+/-8.5-fold that of quiescent cells, respectively. 5-HT2B was strongly associated with fibrotic tissue in diseased rat liver. Treatment of HSCs with 5-HT2 antagonists suppressed proliferation and elevated their rate of apoptosis; by contrast 5-HT was protective against nerve growth factor-induced apoptosis. 5-HT synergized with platelet-derived growth factor to stimulate increased HSC proliferation. HSCs were shown to express a functional serotonin transporter and to participate in both active uptake and release of 5-HT. We conclude that HSCs express key regulatory components of the 5-HT system enabling them to store and release 5-HT and to respond to the neurotransmitter in a profibrogenic manner. Antagonists that selectively target the 5-HT class of receptors may be exploited as antifibrotic drugs.
Subject(s)
Liver Cirrhosis/metabolism , Liver/metabolism , Receptors, Serotonin/metabolism , Serotonin Antagonists/pharmacology , Serotonin Plasma Membrane Transport Proteins/metabolism , Serotonin/metabolism , Animals , Apoptosis/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Drug Design , Humans , Liver/injuries , Liver/pathology , Liver Cirrhosis/drug therapy , Liver Cirrhosis/pathology , Male , Nerve Growth Factor/pharmacology , Neurotransmitter Agents/pharmacology , Rats , Rats, Sprague-Dawley , Serotonin Antagonists/therapeutic useABSTRACT
Murine Nramp1 encodes a divalent cation transporter that is expressed in late endosomes/lysosomes of macrophages, and the transported cations facilitate intracellular pathogen growth control. The Nramp1 promoter is TATA box-deficient, has two initiator elements, and is repressed by c-Myc, in accordance with the notion that genes that deplete the iron content of the cell cytosol antagonize cell growth. Repression via c-Myc occurs at the initiator elements, whereas a c-Myc-interacting protein (Miz-1) stimulates transcription. Here we demonstrate that a non-canonical E box (CAACTG) inhibits basal promoter activity and activation by Miz-1. A consensus Sp1-binding site or GC box is also necessary for Miz-1-dependent transactivation, but not repression. Repression occurs by c-Myc competing with p300/CBP for binding Miz-1. Our results show that an Sp1 site mutant inhibits coactivation by p300 and that the murine Nramp1 promoter is preferentially expressed within macrophages (relative to a beta-actin control) compared with non-macrophage cells. The effect of the Sp1 site mutation on promoter function shows cell-type specificity: stimulation in COS-1 and inhibition in RAW264.7 cells. Miz-1-directed RNA interference confirms a stimulatory role for Miz-1 in Nramp1 promoter function. c-Myc, Miz-1, and Sp1 were identified as binding to the Nramp1 core promoter in control cells and following acute stimulation with interferon-gamma and lipopolysaccharide. These results provide a description of sites that modulate the activity of the initiator-binding protein Miz-1 and indicate a stimulatory role for GC box-binding factors in macrophages and a inhibitory role for E box elements in proliferating cells.
