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Long-term inhibition of kappa opioid receptor (KOR) signaling in peripheral pain-sensing neurons is a potential obstacle for development of peripherally-restricted KOR agonists that produce analgesia. Such a long-term inhibitory mechanism is invoked from activation of c-Jun N-terminal kinase (JNK) that follows a single injection of the KOR antagonist norbinaltorphimine (norBNI). This effect requires protein synthesis of an unknown mediator in peripheral pain-sensing neurons. Using 2D difference gel electrophoresis with tandem mass spectrometry, we have identified that the scaffolding protein 14-3-3γ is upregulated in peripheral sensory neurons following activation of JNK with norBNI. Knockdown of 14-3-3γ by siRNA eliminates the long-term reduction in KOR-mediated cAMP signaling by norBNI in peripheral sensory neurons in culture. Similarly, knockdown of 14-3-3γ in the rat hind paw abolished the norBNI-mediated long-term reduction in peripheral KOR-mediated antinociception. Further, overexpression of 14-3-3γ in KOR expressing CHO cells prevented KOR-mediated inhibition of cAMP signaling. These long-term effects are selective for KOR as heterologous regulation of other receptor systems was not observed. These data suggest that 14-3-3γ is both necessary and sufficient for the long-term inhibition of KOR by norBNI in peripheral sensory neurons.

The capacity of three inocula (sewer biofilm, mangrove and estuary sediment) to utilise typical fermentation products of municipal solid waste for biological sulfate reduction was investigated. Each inoculum was used in two reactors, one fed a mixture of volatile fatty acids and another fed glucose to provide a suite of fermentation products via naturally occurring fermentation. Following 228 days of reactor operation, reactors inoculated with mangrove and estuary sediments exhibited higher sulfate reducing efficiencies (80-88%) compared to the biofilm-inoculated reactors (32-49%). Minimal use of acetate and its accumulation in the biofilm-inoculated reactors pointed to the high abundance of incomplete-oxidising sulfate reducing bacteria (SRB)

Pain and pain management in the elderly population is a significant social and medical problem. Pain sensation is a complex phenomenon that typically involves activation of peripheral pain-sensing neurons (nociceptors) which send signals to the spinal cord and brain that are interpreted as pain, an unpleasant sensory experience. In this work, young (4-5 months) and aged (26-27 months) Fischer 344 x Brown Norway (F344xBN) rats were examined for nociceptor sensitivity to activation by thermal (cold and heat) and mechanical stimulation following treatment with inflammatory mediators and activators of transient receptor potential (TRP) channels. Unlike other senses that decrease in sensitivity with age, sensitivity of hindpaw nociceptors to thermal and mechanical stimulation was not different between young and aged F344xBN rats. Intraplantar injection of bradykinin (BK) produced greater thermal and mechanical allodynia in aged versus young rats, whereas only mechanical allodynia was greater in aged rats following injection of prostaglandin E

Opioid overdose is a leading cause of death in the United States. The only treatment available currently is the competitive antagonist, naloxone (Narcan®). Although naloxone is very effective and has saved many lives, as a competitive antagonist it has limitations. Due to the short half‐life of naloxone, renarcotization can occur if the ingested opioid agonist remains in the body longer. Moreover, because antagonism by naloxone is surmountable, renarcotization can also occur in the presence of naloxone if a relatively larger dose of opioid agonist is taken. In such circumstances, a long‐lasting, non‐surmountable antagonist would offer an improvement in overdose treatment. Methocinnamox (MCAM) has been reported to have a long duration of antagonist action at mu opioid receptors in vivo. In HEK cells expressing the human mu opioid receptor, MCAM antagonism of mu agonist‐inhibition of cAMP production was time‐dependent, non‐surmountable and non‐reversible, consistent with (pseudo)‐irreversible binding. In vivo, MCAM injected locally into the rat hindpaw antagonized mu agonist‐mediated inhibition of thermal allodynia for up to 96 h. By contrast, antagonism by MCAM of delta or kappa agonists in HEK cells and in vivo was consistent with simple competitive antagonism. Surprisingly, MCAM also shifted the concentration‐response curves of mu agonists in HEK cells in the absence of receptor reserve in a ligand‐dependent manner. The shift in the [D‐Ala2,N‐MePhe4,Gly‐ol5]‐enkephalin (DAMGO) concentration‐response curve by MCAM was insensitive to naloxone, suggesting that in addition to (pseudo)‐irreversible orthosteric antagonism, MCAM acts allosterically to alter the affinity and/or intrinsic efficacy of mu agonists., Methocinnamox (MCAM) is a (pseudo‐)irreversible, orthosteric antagonist at mu opioid receptors. In addition, MCAM produces a naloxone insensitive shift in agonist concentration‐response curves which suggests that MCAM acts allosterically to alter the affinity and/or intrinsic efficacy of mu agonists.

The concept of biorefinery expands the possibilities to extract value from organic matter in form of either bespoke crops or organic waste. The viability of biorefinery schemes depends on the recovery of higher-value chemicals with potential for a wide distribution and an untapped marketability. The feasibility of biorefining organic waste is enhanced by the fact that the biorefinery will typically receive a waste management fee for accepting organic waste. The development and implementation of waste biorefinery concepts can open up a wide array of possibilities to shift waste management towards higher sustainability. However, barriers encompassing environmental, technical, economic, logistic, social and legislative aspects need to be overcome. For instance, waste biorefineries are likely to be complex systems due to the variability, heterogeneity and low purity of waste materials as opposed to dedicated biomasses. This article discusses the drivers that can make the biorefinery concept applicable to waste management and the possibilities for its development to full scale. Technological, strategic and market constraints affect the successful implementations of these systems. Fluctuations in waste characteristics, the level of contamination in the organic waste fraction, the proximity of the organic waste resource, the markets for the biorefinery products, the potential for integration with other industrial processes and disposal of final residues are all critical aspects requiring detailed analysis. Furthermore, interventions from policy makers are necessary to foster sustainable bio-based solutions for waste management. peer-reviewed 2022-07-16

The impact of the starting inoculum on long-term anaerobic digestion performance, process functionality and microbial community composition remains unclear. To understand the impact of starting inoculum, active microbial communities from four different full-scale anaerobic digesters were each used to inoculate four continuous lab-scale anaerobic digesters, which were operated identically for 295 days. Digesters were operated at 15 days solid retention time, an organic loading rate of 1 g COD Lr−1 d−1 (75:25 - cellulose:casein) and 37 °C. Results showed that long-term process performance, metabolic rates (hydrolytic, acetogenic, and methanogenic) and microbial community are independent of the inoculum source. Digesters process performance converged after 80 days, while metabolic rates and microbial communities converged after 120–145 days. The convergence of the different microbial communities towards a core-community proves that the deterministic factors (process operational conditions) were a stronger driver than the initial microbial community composition. Indeed, the core-community represented 72% of the relative abundance among the four digesters. Moreover, a number of positive correlations were observed between higher metabolic rates and the relative abundance of specific microbial groups. These correlations showed that both substrate consumers and suppliers trigger higher metabolic rates, expanding the knowledge of the nexus between microorganisms and functionality. Overall, these results support that deterministic factors control microbial communities in bioreactors independently of the inoculum source. Hence, it seems plausible that a desired microbial composition and functionality can be achieved by tuning process operational conditions.

Constitutive receptor activity/inverse agonism and functional selectivity/biased agonism are 2 concepts in contemporary pharmacology that have major implications for the use of drugs in medicine and research as well as for the processes of new drug development. Traditional receptor theory postulated that receptors in a population are quiescent unless activated by a ligand. Within this framework ligands could act as agonists with various degrees of intrinsic efficacy, or as antagonists with zero intrinsic efficacy. We now know that receptors can be active without an activating ligand and thus display “constitutive” activity. As a result, a new class of ligand was discovered that can reduce the constitutive activity of a receptor. These ligands produce the opposite effect of an agonist and are called inverse agonists. The second topic discussed is functional selectivity, also commonly referred to as biased agonism. Traditional receptor theory also posited that intrinsic efficacy is a single drug property independent of the system in which the drug acts. However, we now know that a drug, acting at a single receptor subtype, can have multiple intrinsic efficacies that differ depending on which of the multiple responses coupled to a receptor is measured. Thus, a drug can be simultaneously an agonist, an antagonist, and an inverse agonist acting at the same receptor. This means that drugs have an additional level of selectivity (signaling selectivity or “functional selectivity”) beyond the traditional receptor selectivity. Both inverse agonism and functional selectivity need to be considered when drugs are used as medicines or as research tools.

An examination of the processes contributing to the production of landfill greenhouse gas (GHG) emissions is required, as the actual level to which waste degrades anaerobically and aerobically beneath covers has not been differentiated. This paper presents a methodology to distinguish between the rate of anaerobic digestion (rAD), composting (rCOM) and CH4 oxidation (rOX) in a landfill environment, by means of a system of mass balances developed for molecular species (CH4, CO2) and stable carbon isotopes (δ13C-CO2 and δ13C-CH4). The technique was applied at two sampling locations on a sloped area of landfill. Four sampling rounds were performed over an 18 month period after a 1.0 m layer of fresh waste and 30–50 cm of silty clay loam had been placed over the area. Static chambers were used to measure the flux of the molecular and isotope species at the surface and soil gas probes were used to collect gas samples at depths of approximately 0.5, 1.0 and 1.5 m. Mass balances were based on the surface flux and the concentration of the molecular and isotopic species at the deepest sampling depth. The sensitivity of calculated rates was considered by randomly varying stoichiometric and isotopic parameters by ±5% to generate at least 500 calculations of rOX, rAD and rCOM for each location in each sampling round. The resulting average value of rAD and rCOM indicated anaerobic digestion and composting were equally dominant at both locations. Average values of rCOM: ranged from 9.8 to 44.5 g CO2 m−2 d−1 over the four sampling rounds, declining monotonically at one site and rising then falling at the other. Average values of rAD: ranged from 10.6 to 45.3 g CO2 m−2 d−1. Although the highest average rAD value occurred in the initial sampling round, all subsequent rAD values fell between 10 and 20 g CO2 m−2 d−1. rOX had the smallest activity contribution at both sites, with averages ranging from 1.6 to 8.6 g CO2 m−2 d−1. This study has demonstrated that for an interim cover, composting and anaerobic digestion of shallow landfill waste can occur simultaneously.

There is abundant evidence for formation of G protein coupled receptor heteromers in heterologous expression systems, however, little is known of the function of heteromers in native systems. Heteromers of delta and kappa opioid receptors (DOR-KOR heteromers) have been identified in native systems. We previously reported that activation of DOR-KOR heteromers expressed by rat pain-sensing neurons (nociceptors) produces robust, peripherally-mediated antinociception. Moreover, DOR agonist potency and efficacy is regulated by KOR antagonists via allosteric interactions within the DOR-KOR heteromer in a ligand-dependent manner. Here we assessed the reciprocal regulation of KOR agonist function by DOR antagonists in adult rat nociceptors in culture and in a behavioral assay of nociception. Naltrindole enhanced the potency of the KOR agonist ICI-199441 (10-20 fold), but did not alter responses to U50488. By contrast, the potency of U50488 was enhanced (20 fold) by 7-benzylidenenaltrexone. The efficacy of 69-guanidinonaltrindole (69-GNTI) to inhibit nociceptors was blocked by siRNA knock-down of DOR or KOR. Replacing 69-GNTI occupancy of DOR with either naltrindole or 7-benzylidenenaltrexone abolished 69-GNTI efficacy. Further, peptides derived from DOR transmembrane segment 1, fused to the cell membrane penetrating HIV transactivator of transcription peptide, also blocked 69-GNTI-mediated responses ex vivo and in vivo, suggesting that 69-GNTI efficacy in nociceptors is due to its positive allosteric regulation of KOR via occupancy of DOR in a DOR-KOR heteromer. Together, these results provide evidence for the existence of functional DOR-KOR heteromers in rat peripheral sensory neurons and that reciprocal, ligand-dependent, allosteric interactions occur between the DOR and KOR protomers.

Although CH oxidation in landfill soil covers is widely studied, the extent of composting and CH oxidation in underlying waste layers has been speculated but not measured. The objective of this study was to develop and validate a mass balance model to estimate the simultaneous rates of anaerobic digestion (r), CH oxidation (r) and composting (r) in environments where O penetration is variable and zones of aerobic and anaerobic activity are intermingled. The modelled domain could include, as an example, a soil cover and the underlying shallow waste to a nominated depth. The proposed model was demonstrated on a blend of biogas from three separate known sources of gas representing the three reaction processes: (i) a bottle of laboratory grade 50:50% CH:CO gas representing anaerobic digestion biogas; (ii) an aerated 250mL bottle containing food waste that represented composting activity; and (iii) an aerated 250mL bottle containing non-degradable graphite granules inoculated with methanotrophs and incubated with CH and O to represent methanotrophic activity. CO, CH, O and the stable isotope C-CO were chosen as the components for the mass balance model. The three reaction rates, r (=r, r, r) were calculated as fitting parameters to the overdetermined set of 4mass balance equations with the net flux of these components from the bottles q (= [Formula: see text] , [Formula: see text] , [Formula: see text] and [Formula: see text] ) as inputs to the model. The coefficient of determination (r) for observed versus modelled values of r were 1.00, 0.97, 0.98 when the stoichiometry of each reaction was based on gas yields measured in the individual bottles and q was calculated by summing yields from the three bottles. r deteriorated to 0.95, 0.96, 0.87 when using an average stoichiometry from 11 incubations of each of the composting and methane oxidation processes. The significant deterioration in the estimation of r showed that this output is highly sensitive to the evaluated stoichiometry coefficients for the reactions. r deteriorated further to 0.86, 0.77, 0.74 when using the average stoichiometry and experimental measurement of the composition and volume of the blended biogas to determine q. This was primarily attributed to average errors of 8%, 7%, 11% and 14% in the measurement of [Formula: see text] , [Formula: see text] , [Formula: see text] and [Formula: see text] relative to the measurement of the same quantities from the individual bottles.

Tuning of operational variables is a common practice to control the anaerobic digestion process and, in advanced applications, to promote the accumulation of fermentation products. However, process variables are interrelated. In this study, the hydraulic retention time (HRT) was decoupled from the organic loading rate (OLR) in order to isolate the effect of HRT as a selective pressure on: process performance, metabolic rates (hydrolytic, acetogenic, and methanogenic) and the microbial community. Four mesophilic anaerobic digesters were subjected to a sequential decrease in HRT from 15 to 8, 4 and 2 days while keeping the OLR constant at chemical oxygen demand of 1 gCOD L r−1 d−1. The results showed that HRT alone was insufficient to washout methanogens from the digesters, which in turn prevented the accumulation of volatile fatty acids (VFA). Methanosaeta was the dominant genus in the four digesters at all HRTs. Metabolic rates showed that process performance was controlled by hydrolysis, with a clear shift in acetogenic rates, from butyrate and propionate degradation to ethanol degradation at 4 and 2d HRT. The change in acetogenic pathways was attributed to a shift in the fermentation pathways co-current with changes in fermentative bacteria. At 2d HRT, biofilm was formed on the walls and paddles of the digesters, probably as a survival strategy. Most of the taxa in the biofilm were also present in the digester media. Overall, it is the combination of HRT with other operational parameters which promotes the washout of methanogens and the accumulation of VFA.

The occurrence of opioid overdose is reaching epidemic proportions; an estimated 115 Americans die daily from an opioid overdose. The current pharmacological treatment relies exclusively on naloxon...

Biological iodine cycling has been widely studied in marine environments, but rarely considered in terrestrial waters. In this work, a microalgae biofilm, with an iodine content of 350 ± 29 mg kg −1 , is shown to potentially be a significant iodine cycler in a groundwater holding pond. Cultivation of the biofilm in extracted groundwater was carried out at bench scale. In parallel, iodine release from an iodine-rich biofilm kept in the dark was monitored. Iodine uptake and release were found to be proportional to algal growth and decay, with the maximum growth rate and specific decay rate being 0.53 ± 0.05 g m −2 d −1 and 0.12 ± 0.02 d −1 respectively. Iodine update and release in an engineered groundwater holding pond (1 m 3 :1 m 2 × 1 m) was simulated. The results indicated that bioaccumulation causes non-steady state conditions in such ponds, and in extreme circumstances decay events can elevate iodine concentrations to problematic levels.

This paper compares the digestion of a packed bed of shredded municipal waste using a flood and drain regime against a control digestion of similarly prepared material using a trickle flow regime. All trials were performed on shallow (2m) beds of the sub-8cm fraction of shredded mixed MSW, encapsulated in a polyethylene bladder. The control cell (Cell 1) was loaded with 1974 tonnes shredded municipal waste and produced 76±9m(3) CH4dryt(-1) (45±2m(3) CH4 'as received't(-1)) over 200days in response to a daily recirculation of the leachate inventory which was maintained at 60m(3). The flood and drain operation was performed on two co-located cells (Cell 2 and Cell 3) that were loaded simultaneously with 1026 and 915 tonnes of the sub-8cm fraction of shredded mixed MSW, with a third empty cell used as a reservoir for 275m(3) of mature landfill leachate. Cell 2 was first digested in isolation by flooding and draining once per week to avoid excessive souring. Gas production from Cell 2 peaked and declined to a steady residual level in 150days. Cell 3 was flooded and drained for the first time 186days after the commencement of Cell 2, using the same inventory of leachate which was now exchanged between the cells, such that each cell was flooded and drained twice per week. Biogas production from Cell 3 commenced immediately with flooding, peaking and reducing to a residual level within 100days. The average CH4 yield from Cells 2 and 3 was 123±15m(3)dryt(-1) (92±2m(3) 'as received't(-1), equal to 95% of the long term (2month) BMP yield.

Paclitaxel (PTX), a microtubule-targeting anticancer agent, produces a debilitating peripheral neuropathy that is accompanied by neuropathic pain. Currently, there are only marginally effective therapeutic interventions available. Consequently, patients are forced to reduce or discontinue life-saving chemotherapy to cope with the pain. Recently, a newly identified agent, P7C3-A20, was found to be protective in several models of neurodegeneration, including Parkinson's disease and traumatic brain injury. Given that PTX triggers progressive degeneration of peripheral afferent neurons, this study was performed to evaluate the potential neuroprotective efficacy of P7C3-A20 in rodent models of PTX-induced peripheral neuropathy. P7C3-A20 (10 mg/kg) or vehicle (Cremophor EL/DMSO/5% Dextrose; 1:1:3) was administered intraperitoneally (i.p.) to Sprague-Dawley rats (250-300 g) everyday over a 28-day experimental paradigm. Following two days of treatment with P7C3-A20 or vehicle, rats also received 3 injections of PTX (11.7 mg/kg, i.p.) or vehicle (Cremophor EL/DMSO/5% Dextrose; i.p.) administered every other day. Treatment with P7C3-A20 did not alter body weights or leukocyte counts in control or PTX-treated rats. PTX treatment increased sensitivity to mechanical and cold stimulation (allodynia) of the hindpaw, evidence of peripheral neuropathy. Notably, P7C3-A20 treatment prevented the development of allodynia in response to PTX. Immunohistochemical analysis of paw biopsies indicated that P7C3-A20 prevented PTX-mediated degeneration of terminal nerve endings. Collectively, these data suggest that P7C3-A30 prevented the neurotoxic effects of PTX on peripheral sensory neurons. A xenograft tumor model of triple negative breast cancer was then used to determine whether P7C3-A20 diminished the antitumoral efficacy of PTX. MDA-MB-231 tumors were bilaterally implanted into flanks of female athymic nude mice and allowed to grow for 3 weeks prior to treatment. Using the same treatment protocol as the rats, tumor-bearing mice received daily treatment with P7C3-A20 (20 mg/kg, i.p.) or vehicle for 16 days. P7C3-A20 did not alter PTX-mediated inhibition of tumor growth. Furthermore, P7C3-A20 prevented PTX-induced mechanical allodynia in the mice. Taken together, this work indicates that P7C3-A20 prevents PTX-induced neuropathic damage without diminishing its antitumoral efficacy. P7C3-A20 may be an exciting new candidate to prevent peripheral neuropathy in patients undergoing cancer treatment with PTX. Citation Format: LoCoco PM, Risinger AL, Mooberry SL, Berg KA, Clarke WP. The neuroprotective aminopropyl carbazole, P7C3-A20, prevents paclitaxel-induced pain. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-15-07.

There is a growing trend to consider organic wastes as potential sources of renewable energy and value-add products. Fermentation products have emerged as attractive value-add option due to relative easy production and broad application range. However, pre-fermentation and extraction of soluble products may impact down-stream treatment processes, particularly energy recovery by anaerobic digestion. This paper investigates primary sludge pre-fermentation at different temperatures (20, 37, 55, and 70°C), treatment times (12, 24, 48, and 72h), and oxygen availability (semi-aerobic, anaerobic); and its impact on anaerobic digestion. Pre-fermentation at 20 and 37°C succeeded for VFA production with acetate and propionate being major products. Pre-fermentation at 37, 55, and 70°C resulted in higher solubilisation yield but it reduced sludge methane potential by 20%. Under semi-aerobic conditions, pre-fermentation allowed both VFA recovery (43gCODVFAkg(-1)VS) and improved methane potential. The latter phenomenon was linked to fungi that colonised the sludge top layer during pre-fermentation.

Receptor heteromers often display distinct pharmacological and functional properties compared to the individual receptor constituents. In this study, we compared the properties of the DOP-KOP heteromer agonist, 6ʹ-guanidinonaltrindole (6′-GNTI), with agonists for DOP ([D-Pen2,5]-enkephalin [DPDPE]) and KOP (U50488) in peripheral sensory neurons in culture and in vivo. In primary cultures, all three agonists inhibited PGE2-stimulated cAMP accumulation as well as activated extracellular signal-regulated kinase 1/2 (ERK) with similar efficacy. ERK activation by U50488 was Gi-protein mediated but that by DPDPE or 6′-GNTI was Gi-protein independent (i.e., pertussis toxin insensitive). Brief pretreatment with DPDPE or U50488 resulted in loss of cAMP signaling, however, no desensitization occurred with 6′-GNTI pretreatment. In vivo, following intraplantar injection, all three agonists reduced thermal nociception. The dose-response curves for DPDPE and 6′-GNTI were monotonic whereas the curve for U50488 was an inverted U-shape. Inhibition of ERK blocked the downward phase and shifted the curve for U50488 to the right. Following intraplantar injection of carrageenan, antinociceptive responses to either DPDPE or U50488 were transient but could be prolonged with inhibitors of 12/15-lipoxgenases (LOX). By contrast, responsiveness to 6′-GNTI remained for a prolonged time in the absence of LOX inhibitors. Further, pretreatment with the 12/15-LOX metabolites, 12- and 15- hydroxyeicosatetraenoic acid, abolished responses to U50488 and DPDPE but had no effect on 6′-GNTI-mediated responses either in cultures or in vivo. Overall, these results suggest that DOP-KOP heteromers exhibit unique signaling and functional regulation in peripheral sensory neurons and may be a promising therapeutic target for the treatment of pain. This article is part of the Special Issue entitled ‘New Vistas in Opioid Pharmacology’.

Heavy metals present in landfill leachate have infrequently been related to complete anaerobic degradation municipal solid waste (MSW) due to discrete ages of deposited MSW layers and leachate channelling in landfills. In this study, anaerobic digestion of MSW was performed in two enclosed 1000 tonne bioreactors using a unique flood and drain process. Leachates were characterised in terms of pH, soluble chemical oxygen demand, volatile fatty acids (VFAs), ammonium nitrogen and heavy metals over the entire course of digestion. All parameters, including pH, fluctuated during acidogenesis, acetogenesis and methanogenesis, which strongly impacted on the dynamics of dissolved heavy metal concentrations. The simulation of dissolution and precipitation processes indicated that metal sulphide precipitation was not a factor as metal concentrations exceeded solubility limits. The correlation of pH and dissolved heavy metal concentrations indicated that other, mechanisms were involved in the homogenised conditions within the bioreactors. Beside dissolution and precipitation, the main processes most likely involved in metal distributions were adsorption (Zn, Cu, Ni, Pb and Cd), complexation (Cr) or combinations of both process (As and Co).

It is now well accepted that receptors can regulate cellular signaling pathways in the absence of a stimulating ligand, and inverse agonists can reduce this ligand-independent or "constitutive" receptor activity. Both the serotonin 5-HT2A and 5-HT2C receptors have demonstrated constitutive receptor activity in vitro and in vivo. Each has been identified as a target for treatment of schizophrenia. Further, most, if not all, atypical antipsychotic drugs have inverse agonist properties at both 5-HT2A and 5-HT2C receptors. This paper describes our current knowledge of inverse agonism of atypical antipsychotics at 5-HT2A/2C receptor subtypes in vitro and in vivo. Exploiting inverse agonist properties of APDs may provide new avenues for drug development.

Paclitaxel (PTX), a microtubule-targeting anticancer agent, produces a debilitating peripheral neuropathy that is accompanied by neuropathic pain. Patients develop localized pain in their distal extremities, and more frequently complain of increased sensitivity to cold rather than hot temperatures. These clinical observations led us to hypothesize that PTX differentially damages subpopulations of peripheral sensory neurons. The purpose of this study was to evaluate the effects of PTX on bradykinin (BK) and eicosanoid (EP) receptor-expressing sensory neurons. PTX (11.7 mg/kg) or vehicle (Cremophor EL/EtOH/PBS; 1:1:6) was administered intraperitoneally to Sprague-Dawley rats (250-300 g) every other day for 5 days. Following treatment, PTX-treated rats exhibited increased sensitivity to cold and mechanical stimuli, but decreased sensitivity to heat stimulation as compared to vehicle-treated rats. Intraplantar (i.pl.) injection with BK (5 μg) produced a prolonged allodynia to cold, but a diminished allodynia to heat in PTX-treated rats. Interestingly, following treatment with PGE2 (0.3 μg, i.pl.), PTX-treated rats displayed no statistically significant differences in the allodynia to cold or heat compared to vehicle-treated rats. Efficacy of BK (1 pM - 1 μM) to stimulate inositol phosphate (IP) production in cultured primary sensory neurons from PTX-treated rats was significantly reduced. By contrast, stimulation of cAMP production by PGE2 (0.01 pM - 100 nM) was unaffected by PTX treatment. Preliminary mechanistic studies treating naïve cultures of rat primary sensory neurons directly with PTX (5 nM, 0 - 48 hr) yielded a time-dependent, tri-phasic effect on BK-stimulated IP accumulation and reduced PGE2-stimulated cAMP production, suggesting PTX differentially disrupts BK- and PGE2-mediated signaling. Collectively, these data provide evidence that subpopulations of peripheral sensory neurons are differentially sensitive to PTX. Understanding why particular subclasses are less sensitive, or resistant, to damage could reveal novel approaches to protect sensory neurons from chemotoxicity and ultimately prevent chemotherapy-induced peripheral neuropathy. Supported by USPS grant #8ULITR000149, Greehey Fellowship, and Translational Science Training Across Disciplines Program at UTHSCSA The project described was supported by Award Number 8UL1TR000149 from the National Center for Advancing Translational Sciences. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Advancing Translational Sciences. Citation Format: Peter M LoCoco, Teresa C Chavera, Raehannah J Jamshidi, Susan L Mooberry, Kelly A Berg, William P Clarke. Subpopulations of peripheral sensory neurons are differentially sensitive to the microtubule-targeting agent, paclitaxel [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-12-06.

It is well established that metabolic pathways in the fermentation of organic waste are primarily controlled by dissolved H concentrations, but there is no reported study that compares observed and predicted shifts in fermentation pathways induced by manipulating the dissolved H concentration. A perfusion system is presented that was developed to control dissolved H concentrations in the continuous fermentation of glucose by a culture highly enriched towards Thermoanaerobacterium thermosaccharolyticum (86±9% relative abundance) from an originally diverse consortia in the leachate of a laboratory digester fed with municipal solid waste. Media from a 2.5L CSTR was drawn through sintered steel membrane filters to retain biomass, allowing vigorous sparging in a separate chamber without cellular disruption. Through a combination of sparging and variations in glucose feeding rate from 0.8 to 0.2g/L/d, a range of steady state fermentations were performed with dissolved H concentrations as low as an equivalent equilibrated H partial pressure of 3kPa. Trends in product formation rates were simulated using a H regulation partitioning model. The model correctly predicted the direction of products redistribution in response to H concentration changes and the acetate and butyrate formation rates when H concentrations were less than 6kPa. However, the model over-estimated acetate, ethanol and butanol productions at the expense of butyrate production at higher H concentrations. The H yield at the lowest dissolved H concentration was 2.67±0.08mol H/mol glucose, over 300% higher than the yield achieved in a CSTR operated without sparging.

Chemotherapy-induced peripheral neuropathy (CIPN) arises from collateral damage to peripheral afferent sensory neurons by anticancer pharmacotherapy, leading to debilitating neuropathic pain. No effective treatment for CIPN exists, short of dose-reduction which worsens cancer prognosis. Here, we report that stimulation of nicotinamide phosphoribosyltransferase (NAMPT) produced robust neuroprotection in an aggressive CIPN model utilizing the frontline anticancer drug, paclitaxel (PTX). Daily treatment of rats with the first-in-class NAMPT stimulator, P7C3-A20, prevented behavioral and histologic indicators of peripheral neuropathy, stimulated tissue NAD recovery, improved general health, and abolished attrition produced by a near maximum-tolerated dose of PTX. Inhibition of NAMPT blocked P7C3-A20-mediated neuroprotection, whereas supplementation with the NAMPT substrate, nicotinamide, potentiated a subthreshold dose of P7C3-A20 to full efficacy. Importantly, P7C3-A20 blocked PTX-induced allodynia in tumored mice without reducing antitumoral efficacy. These findings identify enhancement of NAMPT activity as a promising new therapeutic strategy to protect against anticancer drug-induced peripheral neurotoxicity.

An examination of the processes contributing to the production of landfill greenhouse gas (GHG) emissions is required, as the actual level to which waste degrades anaerobically and aerobically beneath covers has not been differentiated. This paper presents a methodology to distinguish between the rate of anaerobic digestion (r

There is abundant evidence for formation of G protein-coupled receptor heteromers in heterologous expression systems, but little is known of the function of heteromers in native systems. Heteromers of δ and κ opioid receptors (DOR-KOR heteromers) have been identified in native systems. We previously reported that activation of DOR-KOR heteromers expressed by rat pain-sensing neurons (nociceptors) produces robust, peripherally mediated antinociception. Moreover, DOR agonist potency and efficacy is regulated by KOR antagonists via allosteric interactions within the DOR-KOR heteromer in a ligand-dependent manner. Here we assessed the reciprocal regulation of KOR agonist function by DOR antagonists in adult rat nociceptors in culture and in a behavioral assay of nociception. Naltrindole enhanced the potency of the KOR agonist 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-phenyl-2-pyrrolidin-1-ylethyl]acetamide (ICI-199441) 10- to 20-fold, but did not alter responses to 2-(3,4-dichlorophenyl)-N-methyl-N-[(1R,2R)-2-pyrrolidin-1-ylcyclohexyl]acetamide (U50488). By contrast, the potency of U50488 was enhanced 20-fold by 7-benzylidenenaltrexone. The efficacy of 6ʹ-guanidinonaltrindole (6ʹ-GNTI) to inhibit nociceptors was blocked by small interfering RNA knockdown of DOR or KOR. Replacing 6ʹ-GNTI occupancy of DOR with either naltrindole or 7-benzylidenenaltrexone abolished 6ʹ-GNTI efficacy. Further, peptides derived from DOR transmembrane segment 1 fused to the cell membrane–penetrating HIV transactivator of transcription peptide also blocked 6ʹ-GNTI–mediated responses ex vivo and in vivo, suggesting that 6ʹ-GNTI efficacy in nociceptors is due to its positive allosteric regulation of KOR via occupancy of DOR in a DOR-KOR heteromer. Together, these results provide evidence for the existence of functional DOR-KOR heteromers in rat peripheral sensory neurons and that reciprocal, ligand-dependent allosteric interactions occur between the DOR and KOR protomers.

The aim of this paper was to apply and validate a model for measuring the rate and extent of anaerobic digestion, composting and CH4 oxidation in laboratory scale beds. Degradation studies were performed in four reactors each packed with shredded unsorted municipal solid waste, with one bed covered with a 100 mm layer of soil. The rates of production of CH4, CO2, 13C-CO2 and the rate of consumption of O2 were measured and used as inputs to a mass balance expressions for these components to calculate the rates of anaerobic digestion, composting and CH4 oxidation. The results showed that anaerobic digestion, composting and CH4 oxidation occurred simultaneously in both the covered and uncovered beds. The analysis showed that 50 ± 4% of the solids (COD basis) in the uncovered beds degraded anaerobically, with the generated CH4 subsequently oxidized, and that 32 ± 4% of the solids degraded aerobically in the covered bed.

Methanotrophs are bacteria capable of using methane as a carbon source. They can lower atmospheric methane emissions, remove N in environmental and wastewater treatment systems and even transform organic pollutants in soils. Methanotrophic methane mitigation technologies have been demonstrated beyond the laboratories as adaptable field-scale systems that may be engineered to meet site-specific climatic variations and ensure minimal atmospheric methane emission. In agricultural sediments and soils, methanotrophs sequester methane but are affected by fertiliser applications, while in wastewater treatment systems they can lower the costs associated with N removal. Finally, the methanotrophs are particularly appealing as bioremediation agents in methane-containing environments, as their primary enzymes have a broad substrate range that can transform various hydrocarbons, including aromatic compounds and halogenated aliphatics. These diverse bacteria are an important global methane sink and this importance is set to increase as anthropogenic emissions increase over the coming decades.