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A functional nerve growth factor NGF–Tropomyosin Receptor kinase A (TrkA) system is an essential requisite for the generation and maintenance of long-lasting thermal and mechanical hyperalgesia in adult mammals. Indeed, mutations in the gene encoding for TrkA are responsible for a rare condition, named Hereditary Sensory and Autonomic Neuropathy type IV (HSAN IV), characterized by the loss of response to noxious stimuli, anhidrosis and cognitive impairment. However, to date, there is no available mouse model to properly understand how the NGF–TrkA system can lead to pathological phenotypes that are distinctive of HSAN IV. Here, we report the generation of a knock-in mouse line carrying the HSAN IV TrkAR649W mutation. First, by in vitro biochemical and biophysical analyses, we show that the pathological R649W mutation leads to kinase-inactive TrkA also affecting its membrane dynamics and trafficking. In agreement with the HSAN IV human phenotype, TrkAR649W/m mice display a lower response to thermal and chemical noxious stimuli, correlating with reduced skin innervation, in addition to decreased sweating in comparison to TrkAh/m controls. Moreover, the R649W mutation decreases anxiety-like behavior and compromises cognitive abilities, by impairing spatial-working and social memory. Our results further uncover unexplored roles of TrkA in thermoregulation and sociability. In addition to accurately recapitulating the clinical manifestations of HSAN IV patients, our findings contribute to clarifying the involvement of the NGF–TrkA system in pain sensation.

SummaryCellular exposure to free fatty acids (FFA) is implicated in the pathogenesis of obesity-associated diseases. However, studies to date have assumed that a few select FFAs are representative of broad structural categories, and there are no scalable approaches to comprehensively assess the biological processes induced by exposure to diverse FFAs circulating in human plasma. Furthermore, assessing how these FFA- mediated processes interact with genetic risk for disease remains elusive. Here we report the design and implementation of FALCON (Fatty Acid Library for Comprehensive ONtologies) as an unbiased, scalable and multimodal interrogation of 61 structurally diverse FFAs. We identified a subset of lipotoxic monounsaturated fatty acids (MUFAs) with a distinct lipidomic profile associated with decreased membrane fluidity. Furthermore, we developed a new approach to prioritize genes that reflect the combined effects of exposure to harmful FFAs and genetic risk for type 2 diabetes (T2D). Importantly, we found that c-MAF inducing protein (CMIP) protects cells from exposure to FFAs by modulating Akt signaling and we validated the role of CMIP in human pancreatic beta cells. In sum, FALCON empowers the study of fundamental FFA biology and offers an integrative approach to identify much needed targets for diverse diseases associated with disordered FFA metabolism.HighlightsFALCON (Fatty Acid Library for Comprehensive ONtologies) enables multimodal profiling of 61 free fatty acids (FFAs) to reveal 5 FFA clusters with distinct biological effectsFALCON is applicable to many and diverse cell typesA subset of monounsaturated FAs (MUFAs) equally or more toxic than canonical lipotoxic saturated FAs (SFAs) leads to decreased membrane fluidityNew approach prioritizes genes that represent the combined effects of environmental (FFA) exposure and genetic risk for diseaseC-Maf inducing protein (CMIP) is identified as a suppressor of FFA-induced lipotoxicity via Akt-mediated signaling

Aim: Analyze changes on body composition in obese and T2DM patients treated with semaglutide. Methods: Observational, prospective study. Body composition was studied by bioimpedance (BIO) . Changes were evaluated over a median of 26 wk. Paired Wilcoxon test was used to analyze the differences. Results: 54 patients (53.7% women, mean age 53.5+/-21yr) . Table 1 shows data at baseline and follow up. We found a reduction in body weigh (-3.5 kg (2.7-4.3) p=0.00) mainly related to fat mass (-6 kg (4.1-8.5) p=0.00) with a little loss of lean (-2.6 (1.3-4.1) p=0.0004) and muscle mass (-0.8 (0.0004-1.6) p=0.04) (Fig 1) Discussion: Sarcopenic Obesity is highly prevalent in T2DM associated with comorbidities like HBP, CVD or HF. Semaglutide induces significant weight loss due to a decrease in fat mass with preservation of muscle mass.

Background Huntington’s disease (HD) is a fatal neurodegenerative disorder characterized by progressive motor dysfunction and loss of medium spiny neurons (MSNs) in dorsal striatum. Brain-derived neurotrophic factor (BDNF) sustains functionality and integrity of MSNs, and thus reduced BDNF signaling is integral to the disease. Mutations in BDNF receptor SorCS2 were recently identified in HD patients. Our study investigates the role of SorCS2 in MSNs biology and in HD progression. Methods We derived a double transgenic line by crossbreeding SorCS2 deficient (KO) mice with the HD mouse model R6/1. Subsequently, we characterized the SorCS2 KO; R6/1 line by a set of behavioral and biochemical studies to evaluate phenotypes related to HD. Moreover, in combination with electrophysiology and super resolution microscopy techniques, we addressed the molecular mechanism by which SorCS2 controls synaptic activity in MSNs neurons. Results We show that SorCS2 is expressed in MSNs with reduced levels in R6/1 HD model, and that SorCS2 deficiency exacerbates the disease progression in R6/1 mice. Furthermore, we find that SorCS2 binds TrkB and the NMDA receptor subunit GluN2B, which is required to control neurotransmission in corticostriatal synapses. While BDNF stimulates SorCS2-TrkB complex formation to enable TrkB signaling, it disengages SorCS2 from GluN2B, leading to enrichment of the subunit at postsynaptic densities. Consequently, long-term potentiation (LTP) is abolished in SorCS2 deficient mice, despite increased striatal TrkB and unaltered BDNF expression. However, the addition of exogenous BDNF rescues the phenotype. Finally, GluN2B, but not GluN2A, currents are also severely impaired in the SorCS2 KO mice. Conclusions We formulate a novel molecular mechanism by which SorCS2 acts as a molecular switch. SorCS2 targets TrkB and GluN2B into postsynaptic densities to enable BDNF signaling and NMDAR dependent neurotransmission in the dorsal striatum. Remarkably, the binding between SorCS2 and TrkB or GluN2B, respectively, is mutually exclusive and controlled by BDNF. This mechanism provides an explanation why deficient SorCS2 signaling severely aggravates HD progression in mice. Moreover, we provide evidence that this finding might represent a general mechanism of SorCS2 signaling found in other brain areas, thus increasing its relevance for other neurological and psychiatric impairments.

Water reuse is a safe and often the least energy-intensive method of providing water from non-conventional sources in water stressed regions. Although public perception can be a challenge, water reuse is gaining acceptance. Recent advances in membrane technology allow for reclamation of wastewater through the production of high-quality treated water, including potable reuse. This study takes an in-depth evaluation of a combination of membrane-based tertiary processes for its application in reuse of brewery wastewater, and is one of the few studies that evaluates long-term membrane performance at the pilot-scale. Two different advanced tertiary treatment trains were tested with secondary wastewater from a brewery wastewater treatment plant (A) ultrafiltration (UF) and reverse osmosis (RO), and (B) ozonation, coagulation, microfiltration with ceramic membranes (MF) and RO. Three specific criteria were used for membrane comparison: 1) pilot plant optimisation to identify ideal operating conditions, 2) Clean-In-Place (CIP) procedures to restore permeability, and 3) final water quality obtained. Both UF and Micro-Filtration membranes were operated at increasing fluxes, filtration intervals and alternating phases of backwash (BW) and chemically enhanced backwash (CEB) to control fouling. Operation of polymeric UF membranes was optimized at a flux of 25–30 LMH with 15–20 min of filtration time to obtain longer production periods and avoid frequent CIP membrane cleaning procedures. Combination of ozone and coagulation with ceramic MF membranes resulted in high flux values up to 120 LMH with CEB:BW ratios of 1:4 to 1:10. Coagulation doses of 3–6 ppm were required to deal with the high concentrations of polyphenols (coagulation inhibitors) in the feed, but higher concentrations led to increasing fouling resistance of the MF membrane. Varying the ozone concentration stepwise from 0 to 25 mg/L had no noticeable effect on coagulation. The most effective cleaning strategy was found to be a combination of 2000 mg/L NaOCl followed by 5% HCl which enabled to recover permeability up to 400 LMH·bar−1. Both polymeric UF and ceramic MF membranes produced effluents that fulfil the limits of the national regulatory framework for reuse in industrial services (RD 1620/2007). Coupling to the RO units in both tertiary trains led to further water polishing and an improved treated water quality.

This chapter presents the construction, operation, and validation of all the MIDES systems, including water pre-treatment, wastewater pre-treatment, the microbial desalination cell (MDC), low-pressure reverse osmosis (RO), and post-treatment (remineralization and disinfection). MIDES technology has been validated with different water sources: brackish water from Demo Site 1, (Racons Brackish Water Desalination Plant (BWDP), located in Denia, Spain) and seawater from Demo Site 2 (Fonsalía Seawater Desalination Plant (SWDP), located in Guía de Isora, Spain). In this chapter, the preparation of both demo sites for the reception and installation of the pilot plants is also presented.

This chapter presents the full design of two microbial desalination cells (MDCs) at pilot-plant scale from the MIDES project. The final MDC pilot unit design was based on the knowledge gained through up scaling of the MDC from lab- to prepilot scale. The MDC pilot plant consists of one stack of 15 MDC pilot units with 0.4 m2 electrode area. This chapter also presents the piping and instrumentation diagram (P&ID) and layout of the MDC pilot plant. The MIDES pilot plants are comprised of an MDC pilot plant housed in a 40-ft container with the rest of the peripheral elements. Finally, this chapter presents the improvement made from the first to the second MDC stack in terms of stability and the chemical compatibility of the end plates.

This chapter presents desalination as one of the technologies to alleviate water scarcity and its contribution to the sustainable development goals. An overview of the world and regional desalination capacity is presented and areas where desalination has potential for development are identified. The overall concept of the microbial desalination cells is presented and the areas where key innovations were developed is presented. A discussion on the energy costs and production costs in seawater reverse osmosis is briefly discussed.

Ubiquitylation of receptor tyrosine kinases (RTKs) regulates both the levels and functions of these receptors. The neurotrophin receptor TrkB (also known as NTRK2), a RTK, is ubiquitylated upon activation by brain-derived neurotrophic factor (BDNF) binding. Although TrkB ubiquitylation has been demonstrated, there is a lack of knowledge regarding the precise repertoire of proteins that regulates TrkB ubiquitylation. Here, we provide mechanistic evidence indicating that ubiquitin carboxyl-terminal hydrolase 8 (USP8) modulates BDNF- and TrkB-dependent neuronal differentiation. USP8 binds to the C-terminus of TrkB using its microtubule-interacting domain (MIT). Immunopurified USP8 deubiquitylates TrkB in vitro, whereas knockdown of USP8 results in enhanced ubiquitylation of TrkB upon BDNF treatment in neurons. As a consequence of USP8 depletion, TrkB levels and its activation are reduced. Moreover, USP8 protein regulates the differentiation and correct BDNF-dependent dendritic formation of hippocampal neurons in vitro and in vivo. We conclude that USP8 positively regulates the levels and activation of TrkB, modulating BDNF-dependent neuronal differentiation. This article has an associated First Person interview with the first author of the paper.

The main source of potable water in high water-stress areas is commonly produced in brackish and seawater desalination plants. Owing to the presence of high concentration of suspended solids, organic matter and colloidal particles in raw water, pretreatment processes are needed for a stable operation of desalination plants. A submerged membrane ultrafiltration pilot plant has been operated as pretreatment of complex brackish surface water to study the filtration performance. The results show the membrane performance, chemical reagent requirements, water quality and cleaning procedures efficiency of an ultrafiltration pilot plant used as pretreatment for a reverse osmosis system. Alternative chemical cleaning procedures have been satisfactorily implemented, which maximize permeability recovery and allow a stable operation.

The development of the nervous system is a temporally and spatially coordinated process that relies on the proper regulation of the genes involved. Neurotrophins and their receptors are directly responsible for the survival and differentiation of sensory and sympathetic neurons; however, it is not fully understood how genes encoding Trk neurotrophin receptors are regulated. Here, we show that rat Bex3 protein specifically regulates TrkA expression by acting at the trkA gene promoter level. Bex3 dimerization and shuttling to the nucleus regulate the transcription of the trkA promoter under basal conditions and also enhance nerve growth factor (NGF)-mediated trkA promoter activation. Moreover, qChIP assays indicate that Bex3 associates with the trkA promoter within a 150 bp sequence, immediately upstream from the transcription start site, which is sufficient to mediate the effects of Bex3. Consequently, the downregulation of Bex3 using shRNA increases neuronal apoptosis in NGF-dependent sensory neurons deprived of NGF and compromises PC12 cell differentiation in response to NGF. Our results support an important role for Bex3 in the regulation of TrkA expression and in NGF-mediated functions through modulation of the trkA promoter., This work was supported by Ministerio de Ciencia e Innovación Grant BFU2008-00162, by Ministerio de Economía y Competitividad Grant (BFU2011-22898 to J.C.A. and BFU2013-39394-P to P.P.), by a Marie Curie International Reintegration Grant within the European Community 7th Framework Programme, by the Consejería de Educación Grant SA074A08, and by a Consejería de Sanidad de Junta Castilla y León grant to J.C.A. This work was in part supported by the FP7-PAINCAGE integrative project. D.M.-Z. is funded by a grant from the Spanish “Instituto de Saludo Carlos III” PI1202847. L.C. has been the recipient of a Predoctoral Fellowship from the University of Salamanca. J.C.A. was a “Ramón y Cajal” Investigator from the University of Salamanca and a NARSAD 2009 Young Investigator Awardee.

Commercial cellulase production has increased in recent years and consistent research has been carried out to improve levels of β-glucosidase. Bioprocesses have been successfully adapted to produce this enzyme, with solid-state fermentations as the best-suited technique involving fungi. The aim of this study was to use leaves of tarbush (Flourensia cernua), an abundant shrub of the Chihuahuan Desert, as a carbon source for β-glucosidase production by Aspergillus niger. During the solid bioprocess, this enzyme reached its peak production at 36 h of culture with 3876.6 U/L. There is a particular interest in the substrate composition because of the possibility of phenolic glycosides having an important role in β-glucosidase production. HPLC-MS analyses showed that glycosides were present with the highest accumulation at 36 h of fungal culture. Luteolin and apigenin glycosides [1.8 and 2.4 absorbance units, respectively] were also detected and showed their highest point of detection alongside the highest β-glucosidase activity. No apparent changes in cellulose were observed, while hemicellulose content decreased, which could be related to production and activity of β-glucosidase. This study shows that leaves of F. cernua are an important raw material for β-glucosidase production and give a source of compounds of added value which also may have an important role for β-glucosidase production.