Simultaneous Heterotrophic Nitrification and Aerobic Denitrification of Water after Sludge Dewatering in Two Sequential Moving Bed Biofilm Reactors (MBBR).

Water after sludge dewatering, also known as reject water from anaerobic digestion, is recycled back to the main wastewater treatment inlet in the wastewater treatment plant Porsgrunn, Norway, causing periodic process disturbance due to high ammonium of 568 (±76.7) mg/L and total chemical oxygen demand (tCOD) of 2825 (±526) mg/L. The main aim of this study was the simultaneous treatment of reject water ammonium and COD using two pilot-scale sequential moving bed biofilm reactors (MBBR) implemented in the main wastewater treatment stream. The two pilot MBBRs each had a working volume of 67.4 L. The biofilm carriers used had a protected surface area of 650 m2/m3 with a 60% filling ratio. The results indicate that the combined ammonia removal efficiency (ARE) in both reactors was 65.9%, while the nitrite accumulation rate (NAR) and nitrate production rate (NPR) were 80.2 and 19.8%, respectively. Over 28% of the reject water's tCOD was removed in both reactors. The heterotrophic nitrification and oxygen tolerant aerobic denitrification were the key biological mechanisms found for the ammonium removal in both reactors. The dominant bacterial family in both reactors was Alcaligenaceae, capable of simultaneous heterotrophic nitrification and denitrification. Moreover, microbial families that were found with equal potential for application of simultaneous heterotrophic nitrification and aerobic denitrification including Cloacamonaceae, Alcaligenaceae, Comamonadaceae, Microbacteriaceae, and Anaerolinaceae.

Molecular and Functional Sex Differences of Noradrenergic Neurons in the Mouse Locus Coeruleus.

Preclinical work has long focused on male animals, though biological sex clearly influences risk for certain diseases, including many psychiatric disorders. Such disorders are often treated by drugs targeting the CNS norepinephrine system. Despite roles for noradrenergic neurons in behavior and neuropsychiatric disease models, their molecular characterization has lagged. We profiled mouse noradrenergic neurons in vivo, defining over 3,000 high-confidence transcripts expressed therein, including druggable receptors. We uncovered remarkable sex differences in gene expression, including elevated expression of the EP3 receptor in females-which we leverage to illustrate the behavioral and pharmacologic relevance of these findings-and of Slc6a15 and Lin28b, both major depressive disorder (MDD)-associated genes. Broadly, we present a means of transcriptionally profiling locus coeruleus under baseline and experimental conditions. Our findings underscore the need for preclinical work to include both sexes and suggest that sex differences in noradrenergic neurons may underlie behavioral differences relevant to disease.

Novel Small-Molecule Inhibitors of Protein Kinase C Epsilon Reduce Ethanol Consumption in Mice.

BACKGROUND: Despite the high cost and widespread prevalence of alcohol use disorders, treatment options are limited, underscoring the need for new, effective medications. Previous results using protein kinase C epsilon (PKCε) knockout mice, RNA interference against PKCε, and peptide inhibitors of PKCε predict that small-molecule inhibitors of PKCε should reduce alcohol consumption in humans. METHODS: We designed a new class of PKCε inhibitors based on the Rho-associated protein kinase (ROCK) inhibitor Y-27632. In vitro kinase and binding assays were used to identify the most potent compounds. Their effects on ethanol-stimulated synaptic transmission; ethanol, sucrose, and quinine consumption; ethanol-induced loss of righting; and ethanol clearance were studied in mice. RESULTS: We identified two compounds that inhibited PKCε with Ki <20 nM, showed selectivity for PKCε over other kinases, crossed the blood-brain barrier, achieved effective concentrations in mouse brain, prevented ethanol-stimulated gamma-aminobutyric acid release in the central amygdala, and reduced ethanol consumption when administered intraperitoneally at 40 mg/kg in wild-type but not in Prkce-/- mice. One compound also reduced sucrose and saccharin consumption, while the other was selective for ethanol. Both transiently impaired locomotion through an off-target effect that did not interfere with their ability to reduce ethanol intake. One compound prolonged recovery from ethanol-induced loss of righting but this was also due to an off-target effect since it was present in Prkce-/- mice. Neither altered ethanol clearance. CONCLUSIONS: These results identify lead compounds for development of PKCε inhibitors that reduce alcohol consumption.

Consistent sex-dependent effects of PKMζ gene ablation and pharmacological inhibition on the maintenance of referred pain.

BACKGROUND: Persistently active PKMζ has been implicated in maintaining spinal nociceptive sensitization that underlies pain hypersensitivity. However, evidence for PKMζ in the maintenance of pain hypersensitivity comes exclusively from short-term studies in males using pharmacological agents of questionable selectivity. The present study examines the contribution of PKMζ to long-lasting allodynia associated with neuropathic, inflammatory, or referred visceral and muscle pain in males and females using pharmacological inhibition or genetic ablation. RESULTS: Pharmacological inhibition or genetic ablation of PKMζ reduced mild formalin pain and slowly developing contralateral allodynia in nerve-injured rats, but not moderate formalin pain or ipsilateral allodynia in models of neuropathic and inflammatory pain. Pharmacological inhibition or genetic ablation of PKMζ also effectively reduced referred visceral and muscle pain in male, but not in female mice and rats. CONCLUSION: We show pharmacological inhibition and genetic ablation of PKMζ consistently attenuate long-lasting pain hypersensitivity. However, differential effects in models of referred versus inflammatory and neuropathic pain, and in males versus females, highlight the roles of afferent input-dependent masking and sex differences in the maintenance of pain hypersensitivity.

Association of a polyadenylation polymorphism in the serotonin transporter and panic disorder.

BACKGROUND: Genetic markers in the serotonin transporter are associated with panic disorder (PD). The associated polymorphisms do not include the serotonin transporter-linked polymorphic region and display no obvious functional attributes. A common polymorphism (rs3813034) occurs in one of the two reported polyadenylation signals for the serotonin transporter and is in linkage disequilibrium with the PD-associated markers. If functional, rs3813034 might be the risk factor that explains the association of the serotonin transporter and PD. METHODS: Quantitative polymerase chain reaction on human brain samples (n = 65) and lymphoblast cultures (n = 71) was used to test rs3813034 for effects on expression of the polyadenylation forms of the serotonin transporter. rs3813034 was also tested for association in a sample of PD cases (n = 307) and a control sample (n = 542) that has similar population structure. RESULTS: The balance of the two polyadenylation forms of the serotonin transporter is associated with rs3813034 in brain (p < .001) and lymphoblasts (p < .001). The balance of the polyadenylation forms is also associated with gender in brain only (p < .05). Association testing of rs3813034 in PD identified a significant association (p = .0068) with a relative risk of 1.56 and 1.81 for the heterozygous and homozygous variant genotypes, respectively. CONCLUSIONS: rs3813034 is a functional polymorphism in the serotonin transporter that alters the balance of the two polyadenylation forms of the serotonin transporter. rs3813034 is a putative risk factor for PD and other behavioral disorders that involve dysregulation of serotonergic neurotransmission.

Prenatal cocaine administration increases glutathione and alpha-tocopherol oxidation in fetal rat brain.

Recent findings suggest that prenatal cocaine exposure results in significant attenuation of uterine and placental blood flow. The extent of blood flow reduction to fetuses positively correlates with reductions in glial-derived neurotrophic factor (GDNF) and dopamine (DA). However, whether such changes in uterine blood flow are sufficient to induce oxidative stress have yet to be determined. In the following experiments, the impact of prenatal cocaine exposure on fetal brain levels of the endogenous antioxidant glutathione (GSH and its oxidized form GSSG) or the exogenous antioxidant alpha-tocopherol (alpha-T and its oxidized quinone form) was investigated. It was hypothesized that cocaine exposure would result in greater oxidation of both GSH and alpha-T. Results indicated that a single injection of cocaine to a drug-naive pregnant dam results in significant (-16.38%) reductions in the levels of GSH. GSSG can be either raised or reduced as a result of fetal uterine position: fetuses at the ovarian extremes show significant increases in GSSG in response to cocaine (+64.73%), whereas cervically situated fetuses show decreased GSSG (-47.91%). Additionally, cocaine significantly decreased the levels of alpha-T (-15.9%) and increased the levels of its oxidative product alpha-Tquinone (alpha-Tq, +34.05%). Levels of alpha-T were not affected by fetal uterine position. These data collectively suggest that cocaine exposure increases the utilization of both endogenous and exogenous anti-oxidants in the fetal rat brain. Along with previous studies, these data support the hypothesis that cocaine-induced vasoconstriction results in oxidative stress in the gestating fetus.

Prenatal cocaine exposure induces an attenuation of uterine blood flow in the rat.

We have previously demonstrated that maternal cocaine injections result in a gradient of fetal brain cocaine levels that decrease as a function of the fetuses' proximity to the ovaries at embryonic (E) day 15. Our prior data suggest that cocaine-induced vasoconstriction may (1) limit cocaine's entry into the brain and (2) cause damage to DA neurons through injury associated with hypoxia or ischemia of the utero-placental junction. Therefore, using the microsphere technique (labeled with Ru(103)), the following study sought to determine whether the previously observed pattern of cocaine distribution among fetuses in the uterus were due to position-specific reductions in uterine or placental blood flow. On day 15, a single subcutaneous injection of 30 mg/kg cocaine HCl was administered to each rat. Thirty minutes after the cocaine injection, reference blood samples were drawn from the ventral tail artery. Uterine segments and placentae were removed and subjected to gamma counting. While results regarding placental blood flow were equivocal, cocaine significantly reduced average uterine blood flow by 54.6%. In addition, as one moves more proximal to the ovaries, cocaine progressively attenuates blood flow in uterine tissue segments. These data support the hypothesis that the pattern of drug distribution and subsequent brain alterations from prenatal cocaine exposure in our previous reports are likely due to differences in uterine blood flow.