Anaerobic degradation of dairy wastewater in intermittent UASB reactors: influence of effluent recirculation

This work studied the influence of effluent recirculation upon the kinetics of anaerobic degradation of dairy wastewater in the feedless phase of intermittent upflow anaerobic sludge bed (UASB) reactors. Several laboratory-scale tests were performed with different organic loads in closed circuit UASB reactors inoculated with adapted flocculent sludge. The data obtained were used for determination of specific substrate removal rates and specific methane production rates, and adjusted to kinetic models. A high initial substrate removal was observed in all tests due to adsorption of organic matter onto the anaerobic biomass which was not accompanied by biological substrate degradation as measured by methane production. Initial methane production rate was about 45% of initial soluble and colloidal substrate removal rate. This discrepancy between methane production rate and substrate removal rate was observed mainly on the first day of all experiments and was attenuated on the second day, suggesting that the feedless period of intermittent UASB reactors treating dairy wastewater should be longer than one day. Effluent recirculation expressively raised the rate of removal of soluble and colloidal substrate and methane productivity, as compared with results for similar assays in batch reactors without recirculation. The observed bed expansion was due to the biogas production and the application of effluent recirculation led to a sludge bed contraction after all the substrates were degraded. The settleability of the anaerobic sludge improved by the introduction of effluent recirculation this effect being more pronounced for the higher loads

Regulatory Architecture of the Neuronal Cacng2/Tarpγ2 Gene Promoter: Multiple Repressive Domains, a Polymorphic Regulatory Short Tandem Repeat, and Bidirectional Organization with Co-regulated lncRNAs

CACNG2 (TARPγ2, Stargazin) is a multi-functional regulator of excitatory neurotransmission and has been implicated in the pathological processes of several brain diseases. Cacng2 function is dependent upon expression level, but currently, little is known about the molecular mechanisms that control expression of this gene. To address this deficit and investigate disease-related gene variants, we have cloned and characterized the rat Cacng2 promoter and have defined three major features: (i) multiple repressive domains that include an array of RE-1 silencing transcription factor (REST) elements, and a calcium regulatory element-binding factor (CaRF) element, (ii) a (poly-GA) short tandem repeat (STR), and (iii) bidirectional organization with expressed lncRNAs. Functional activity of the promoter was demonstrated in transfected neuronal cell lines (HT22 and PC12), but although selective removal of REST and CaRF domains was shown to enhance promoter-driven transcription, the enhanced Cacng2 promoter constructs were still about fivefold weaker than a comparable rat Synapsin-1 promoter sequence. Direct evidence of REST activity at the Cacng2 promoter was obtained through co-transfection with an established dominant-negative REST (DNR) construct. Investigation of the GA-repeat STR revealed polymorphism across both animal strains and species, and size variation was also observed in absence epilepsy disease model cohorts (Genetic Absence Epilepsy Rats, Strasbourg [GAERS] and non-epileptic control [NEC] rats). These data provide evidence of a genotype (STR)-phenotype correlation that may be unique with respect to proximal gene regulatory sequence in the demonstrated absence of other promoter, or 3′ UTR variants in GAERS rats. However, although transcriptional regulatory activity of the STR was demonstrated in further transfection studies, we did not find a GAERS vs. NEC difference, indicating that this specific STR length variation may only be relevant in the context of other (Cacna1h and Kcnk9) gene variants in this disease model. Additional studies revealed further (bidirectional) complexity at the Cacng2 promoter, and we identified novel, co-regulated, antisense rat lncRNAs that are paired with Cacng2 mRNA. These studies have provided novel insights into the organization of a synaptic protein gene promoter, describing multiple repressive and modulatory domains that can mediate diverse regulatory inputs