Deep Learning-Based 6-DoF Object Pose Estimation Considering Synthetic Dataset.

Due to the difficulty in generating a 6-Degree-of-Freedom (6-DoF) object pose estimation dataset, and the existence of domain gaps between synthetic and real data, existing pose estimation methods face challenges in improving accuracy and generalization. This paper proposes a methodology that employs higher quality datasets and deep learning-based methods to reduce the problem of domain gaps between synthetic and real data and enhance the accuracy of pose estimation. The high-quality dataset is obtained from Blenderproc and it is innovatively processed using bilateral filtering to reduce the gap. A novel attention-based mask region-based convolutional neural network (R-CNN) is proposed to reduce the computation cost and improve the model detection accuracy. Meanwhile, an improved feature pyramidal network (iFPN) is achieved by adding a layer of bottom-up paths to extract the internalization of features of the underlying layer. Consequently, a novel convolutional block attention module-convolutional denoising autoencoder (CBAM-CDAE) network is proposed by presenting channel attention and spatial attention mechanisms to improve the ability of AE to extract images' features. Finally, an accurate 6-DoF object pose is obtained through pose refinement. The proposed approach is compared to other models using the T-LESS and LineMOD datasets. Comparison results demonstrate the proposed approach outperforms the other estimation models.

Characterization of the ability of a, second-generation SST-DA chimeric molecule, TBR-065, to suppress GH secretion from human GH-secreting adenoma cells.

CONTEXT: Somatostatin (SST) and dopamine (DA) inhibit growth hormone (GH) secretion and proliferation of GH-secreting pituitary adenomas (GHomas) through binding to SSTR2 and D2R receptors. Chimeric SST-DA compounds (Dopastatins) display increased potency in inhibiting GH secretion, as compared with individual SST or DA analogs (alone or combined). OBJECTIVE: To assess the efficacy of a second-generation dopastatin, TBR-065, in suppressing GH secretion from human GH- and GH/prolactin(PRL)-omas. DESIGN: We compared the ability of TBR-065 to inhibit GH secretion from primary cultures of human GH- or GH/PRLoma cells to that of the first generation dopastatin, TBR-760 (formerly BIM-23A760), octreotide (OCT) and cabergoline (CAB), the later either alone or combined. We investigated whether there was any impact of BIM-133, the metabolite of TBR-065, on the ability of TBR-065 to inhibit GH in these cultures. METHODS: 17 GH- and GH/PRLomas were included in this study. Inhibition of GH secretion by TBR-065, TBR-760, OCT and CAB (0.1 pM to 0.1 µM) was assessed over a period of 8 h. RESULTS: All tumors expressed SSTR2 and D2R mRNAs. GH suppression was higher with TBR-065 as compared with TBR-760 (Emax = 57 ± 5.6% vs. 41.1 ± 12.5%, respectively, p < 0.001) or with OCT + CAB (Emax = 56.8 ± 7.2% vs. 44.4 ± 9.4%, p < 0.001). BIM-133 did not have any impact on the activity of TBR-065. CONCLUSION: TBR-065 has significantly improved efficacy in suppressing GH secretion as compared to current available therapies and may represent a new promising option for the treatment of acromegaly.

Interaction of imidazolium-based room-temperature ionic liquids with DOPC phospholipid monolayers: electrochemical study.

To test the biocompatible character of room-temperature ionic liquids (ILs), the interaction of various ILs with biological membrane (biomembrane) models was studied in this work. Dioleoyl phosphatidylcholine (DOPC) adsorbed on a mercury (Hg) electrode forms an impermeable defect-free monolayer which is a well established biomembrane model, prone to be studied by electrochemical techniques. We have monitored the modifications of the Hg supported monolayer caused by ILs using rapid cyclic voltammetry (RCV), alternating current voltammetry (ACV), and electrochemical impedance spectroscopy (EIS). A series of imidazolium-based ILs were investigated whose interaction highlighted the role of anion and lateral side chain of cation during the interaction with DOPC monolayers. It was shown that the hydrophobic and lipophilic character of the IL cations is a primary factor responsible for this interaction. Hg-supported monolayers provide an accurate analysis of the behavior of ILs at the interface of a biomembrane leading to a comprehensive understanding of the interaction mechanisms involved. At the same time, these experiments show that the Hg-phospholipid model is an effective toxicity sensing technique as shown by the correlation between literature in vivo toxicity data and the data from this study.

The novel somatostatin receptor 2/dopamine type 2 receptor chimeric compound BIM-23A758 decreases the viability of human GOT1 midgut carcinoid cells.

The majority of neuroendocrine tumors (NETs) of the gastroenteropancreatic system coexpress somatostatin receptors (SSTRs) and dopamine type 2 receptors (D2R), thus providing a rationale for the use of novel SSTR2/D2R chimeric compounds in NET disease. Here we investigate the antitumor potential of the SSTR2/D2R chimeric compounds BIM-23A760 and BIM-23A758 in comparison to the selective SSTR2 agonist BIM-23023 and the selective D2R agonist BIM-53097 on human NET cell lines of heterogeneous origin. While having only minor effects on human pancreatic and bronchus carcinoid cells (BON1 and NCI-H727), BIM-23A758 induced significant antitumor effects in human midgut carcinoid cells (GOT1). These effects involved apoptosis induction as well as inhibition of mitogen-activated protein kinase and Akt signaling. Consistent with their antitumor response to BIM-23A758, GOT1 cells showed relatively high expression levels of SSTR2 and D2R mRNA. In particular, GOT1 cells highly express the short transcript variant of D2R. In contrast to BIM-23A758, the SSTR2/D2R chimeric compound BIM-23A760 as well as the individual SSTR2 and D2R agonistic compounds BIM-23023 and BIM-53097 induced no or only minor antitumor responses in the examined NET cell lines. Taken together, our findings suggest that the novel SSTR2/D2R chimeric compound BIM-23A758 might be a promising substance for the treatment of NETs highly expressing SSTR2 and D2R. In particular, a sufficient expression of the short transcript variant of DR2 might play a pivotal role for effective treatment.

Experimental Study on Shear Behavior of Rock Composite Material under Normal Unloading Conditions.

As a composite material, the stability of rock mass is usually controlled by a joint. During the process of excavation, the normal stress of the joint decreases continuously, and then the shear strength of the joint decreases, which may eventually lead to the instability and failure of rock mass. Previous studies have mainly focused on the shear behavior of joints under constant normal stress, but have rarely considered the unloading of normal stress. In this paper, a direct shear test of joints with different roughness was carried out, in which the shear stress remained unchanged while the normal stress decreased. The strength characteristics of joints were explored, and the deformation and acoustic emission-counting characteristics of joints were analyzed by digital image correlation (DIC) techniques and acoustic emission (AE). A new method for predicting the instability of joints under normal unloading was proposed based on the evolution law of normal deformation energy (Un), tangential deformation energy (Us) and total deformation energy (U0). The results show the following: (1) The unloading amount of normal stress was enlarged for greater initial normal stress and roughness, while it decreased with an increase in initial shear stress. (2) AE events reached their maximum when the normal stress was equal to the failure normal stress, and the b-value fluctuated more frequently in stable development periods under normal unloading conditions. (3) U0 would change with the loading and unloading of stress, and this may be used to predict the unloading instability of rock mass using the abrupt change of U0.

Constructing a cellulose based chiral liquid crystal film with high flexibility, water resistance, and optical property.

Cellulose nanocrystal (CNC) derived from cellulose can form a liquid crystal film with bright structural color by evaporative-induced self-assembly (EISA). As a new class of photonic liquid crystals material, it has attracted much attention because of its intrinsic unique structural characteristics and excellent optical properties. However, the brittleness and water sensitivity of CNC film have hindered its practical application. Herein, multiple cross-linked networks CNC/(polyethylene glycol diacrylate:polyethylene oxide) (PEGDA:PEO) composite film was prepared through EISA and UV irradiation strategies. The as-prepared film exhibits high-flexibility with a fracture strain of up to 36.40 % and strong water resistance, with water absorption at an equilibrium of only 17.41 %. Moreover, the film retains its structural color in aqueous solution for a long time due to its water stability. The outstanding flexibility and water resistance of CNC composite film are attributed to multiple crosslinked networks (i.e. PEGDA, PEO, and PEDGA-PEO networks), which endow the film with excellent stress dispersion and transferability when stretched and limit film swelling in water without affecting chiral nematic structures of CNC. Overall, this work provides a promising strategy to prepare CNC-based film with high-flexibility, water resistance, and optical property for applications like decoration, sensor, and anti-counterfeiting.

Genome editing without nucleases confers proliferative advantage to edited hepatocytes and corrects Wilson disease.

Application of classic liver-directed gene replacement strategies is limited in genetic diseases characterized by liver injury due to hepatocyte proliferation, resulting in decline of therapeutic transgene expression and potential genotoxic risk. Wilson disease (WD) is a life-threatening autosomal disorder of copper homeostasis caused by pathogenic variants in copper transporter ATP7B and characterized by toxic copper accumulation, resulting in severe liver and brain diseases. Genome editing holds promise for the treatment of WD; nevertheless, to rescue copper homeostasis, ATP7B function must be restored in at least 25% of the hepatocytes, which surpasses by far genome-editing correction rates. We applied a liver-directed, nuclease-free genome editing approach, based on adeno-associated viral vector-mediated (AAV-mediated) targeted integration of a promoterless mini-ATP7B cDNA into the albumin (Alb) locus. Administration of AAV-Alb-mini-ATP7B in 2 WD mouse models resulted in extensive liver repopulation by genome-edited hepatocytes holding a proliferative advantage over nonedited ones, and ameliorated liver injury and copper metabolism. Furthermore, combination of genome editing with a copper chelator, currently used for WD treatment, achieved greater disease improvement compared with chelation therapy alone. Nuclease-free genome editing provided therapeutic efficacy and may represent a safer and longer-lasting alternative to classic gene replacement strategies for WD.

Freezing or wrapping: the role of particle size in the mechanism of nanoparticle-biomembrane interaction.

Understanding the interactions between nanoparticles (NPs) and biological matter is a high-priority research area because of the importance of elucidating the physical mechanisms underlying the interactions leading to NP potential toxicity as well as NP viability as therapeutic vectors in nanomedicine. Here, we use two model membrane systems, giant unilamellar vesicles (GUVs) and supported monolayers, to demonstrate the competition between adhesion and elastic energy at the nanobio interface, leading to different mechanisms of NP-membrane interaction relating to NP size. Small NPs (18 nm) cause a "freeze effect" of otherwise fluid phospholipids, significantly decreasing the phospholipid lateral mobility. The release of tension through stress-induced fracture mechanics results in a single microsize hole in the GUVs after interaction. Large particles (>78 nm) promote membrane wrapping, which leads to increased lipid lateral mobility and the eventual collapse of the vesicles. Electrochemical impedance spectroscopy on the supported monolayer model confirms that differently sized NPs interact differently with the phospholipids in close proximity to the electrode during the lipid desorption process. The time scale of these processes is in accordance with the proposed NP/GUV interaction mechanism.

Novel AAV-mediated genome editing therapy improves health and survival in a mouse model of methylmalonic acidemia.

Methylmalonic acidemia (MMA) is an inborn error of metabolism mostly caused by mutations in the mitochondrial methylmalonyl-CoA mutase gene (MMUT). MMA patients suffer from frequent episodes of metabolic decompensation, which can be life threatening. To mimic both the dietary restrictions and metabolic decompensation seen in MMA patients, we developed a novel protein-controlled diet regimen in a Mmut deficient mouse model of MMA and demonstrated the therapeutic benefit of mLB-001, a nuclease-free, promoterless recombinant AAV GeneRideTM vector designed to insert the mouse Mmut into the endogenous albumin locus via homologous recombination. A single intravenous administration of mLB-001 to neonatal or adult MMA mice prevented body weight loss and mortality when challenged with a high protein diet. The edited hepatocytes expressed functional MMUT protein and expanded over time in the Mmut deficient mice, suggesting a selective growth advantage over the diseased cells. In mice with a humanized liver, treatment with a human homolog of mLB-001 resulted in site-specific genome editing and transgene expression in the transplanted human hepatocytes. Taken together, these findings support the development of hLB-001 that is currently in clinical trials in pediatric patients with severe forms of MMA.

The effects of substituent grafting on the interaction of pH-responsive polymers with phospholipid monolayers.

pH-responsive amphiphilic polymers with suitable graftings have demonstrated highly efficient cell membrane activity and hence are promising applicants for drug-delivery. Grafting the hydrophobic amino acid l-phenylalanine and the hydrophilic methoxy poly(ethylene glycol) amine onto the pendant carboxylic acid moieties of a linear polyamide, poly(l-lysine isophthalamide), can effectively modify the amphiphilicity and conformation of the amphiphilic polymers. Here, the interactions of these polymers with phospholipid monolayers adsorbed on mercury (Hg) electrodes have been studied. AC voltammetry (ACV), rapid cyclic voltammetry (RCV), and electrochemical impedance spectroscopy (EIS) have been applied to monitor phospholipid monolayer associations with different polymer concentrations under different pH values. The polymers interact reversibly with the monolayer shown by altering the monolayer capacitance and inhibiting the phospholipid reorientation in electric field. Polymer grafting enhances the pH-mediated conformational change of the polymers which in turn increases their phospholipid monolayer activity. The most significant monolayer interactions have been observed with the polymer grafted with hydrophobic l-phenylalanine. A low level of PEGylation of the backbone also increases the monolayer activity. The polymer/DOPC interactions have been represented with an impedance model, which takes account of the interaction giving rise to an increase in monolayer capacitance and inhomogeneity and a Debye type dielectric relaxation. The extent of penetration of the polymers into the monolayer is inversely related to the electrical resistance they give rise to during the Debye relaxation. The cell membrane activities of these amphiphilic polymers have been successfully mirrored in this supported DOPC monolayer system, isolating the key parameters for biomembrane activities and giving insight into the mechanism of the interactions. The conclusions from this study provide strategic directions in material design catering to different requirements in biomedical applications.

Layer-by-layer synthesis of multilayer core-shell latex and the film formation properties.

Polymer latex particles were synthesized with multilayer core-shell structure via surface cross-linking emulsion polymerization. The latex core is coated with a five-layer shell. The polymerization was done in a semicontinuous fashion monitored by a dynamic laser scattering (DLS). The copolymer in each layer is designed with alternating high and low glass transition temperature (T(g)). Divinylbenzene (DVB) was added as the cross-linking agent in the synthesis of the "hard" layers to prevent the molecular diffusion from the adjacent "soft" layers. The layer-by-layer increment on the latex core is proved by the alternating changes on the film-formation capabilities of different latex emulsions at room temperature in correspondence with the variance in the T(g) of the outermost polymer layer. The detailed morphologies of the films formed by the latex with different number of layers were characterized by atom force microscopy (AFM). The deformation of the latex particles is largely depended on the nature of the polymer in the outermost layer of the latex particles. Further characterization carried out by multifrequency temperature-modulated differential scanning calorimetry (TOPEM-DSC) confirmed the layer-by-layer structure of the particles, although the molecular redistribution and the interlayer structures were observed. The work provides a routine toward the synthesis of multilayer polymer latexes.

A Somatostatin Receptor Subtype-3 (SST3) Peptide Agonist Shows Antitumor Effects in Experimental Models of Nonfunctioning Pituitary Tumors.

PURPOSE: Somatostatin analogues (SSA) are efficacious and safe treatments for a variety of neuroendocrine tumors, especially pituitary neuroendocrine tumors (PitNET). Their therapeutic effects are mainly mediated by somatostatin receptors SST2 and SST5. Most SSAs, such as octreotide/lanreotide/pasireotide, are either nonselective or activate mainly SST2. However, nonfunctioning pituitary tumors (NFPTs), the most common PitNET type, mainly express SST3 and finding peptides that activate this particular somatostatin receptor has been very challenging. Therefore, the main objective of this study was to identify SST3-agonists and characterize their effects on experimental NFPT models. EXPERIMENTAL DESIGN: Binding to SSTs and cAMP level determinations were used to screen a peptide library and identify SST3-agonists. Key functional parameters (cell viability/caspase activity/chromogranin-A secretion/mRNA expression/intracellular signaling pathways) were assessed on NFPT primary cell cultures in response to SST3-agonists. Tumor growth was assessed in a preclinical PitNET mouse model treated with a SST3-agonist. RESULTS: We successfully identified the first SST3-agonist peptides. SST3-agonists lowered cell viability and chromogranin-A secretion, increased apoptosis in vitro, and reduced tumor growth in a preclinical PitNET model. As expected, inhibition of cell viability in response to SST3-agonists defined two NFPT populations: responsive and unresponsive, wherein responsive NFPTs expressed more SST3 than unresponsive NFPTs and exhibited a profound reduction of MAPK, PI3K-AKT/mTOR, and JAK/STAT signaling pathways upon SST3-agonist treatments. Concurrently, SSTR3 silencing increased cell viability in a subset of NFPTs. CONCLUSIONS: This study demonstrates that SST3-agonists activate signaling mechanisms that reduce NFPT cell viability and inhibit pituitary tumor growth in experimental models that expresses SST3, suggesting that targeting this receptor could be an efficacious treatment for NFPTs.

Use of CeO2 Nanoparticles to Enhance UV-Shielding of Transparent Regenerated Cellulose Films.

The major challenge in preparing polymer nanocomposites is to prevent the agglomeration of inorganic nanoparticles (NPs). Here, with regenerated cellulose (RC) films as supporting medium, UV-shielding and transparent nanocomposite films with hydrophobicity were fabricated by in situ synthesis of CeO2 NPs. Facilitated through the interaction between organic and inorganic components revealed by X-ray diffraction (XRD) and Fourier transformation infrared spectroscopy (FTIR) characterization, it was found that CeO2 NPs were uniformly dispersed in and immobilized by a cellulose matrix. However some agglomeration of CeO2 NPs occurred at higher precursor concentrations. These results suggest that the morphology and particle size of CeO2 and the corresponding performance of the resulting films are affected by the porous RC films and the concentrations of Ce(NO3)3·6H2O solutions. The optimized nanocomposite film containing 2.95 wt% CeO2 NPs had more than 75% light transmittance (550 nm), high UV shielding properties, and a certain hydrophobicity.

The type III neurofilament peripherin is expressed in the tuberomammillary neurons of the mouse.

BACKGROUND: Peripherin, a type III neuronal intermediate filament, is widely expressed in neurons of the peripheral nervous system and in selected central nervous system hindbrain areas with projections towards peripheral structures, such as cranial nerves and spinal cord neurons. Peripherin appears to play a role in neurite elongation during development and axonal regeneration, but its exact function is not known. We noticed high peripherin expression in the posterior hypothalamus of mice, and decided to investigate further the exact location of expression and function of peripherin in the mouse posterior hypothalamus. RESULTS: In situ hybridization indicated expression of peripherin in neurons with a distribution reminiscent of the histaminergic neurons, with little signal in any other part of the forebrain. Immunocytochemical staining for histidine decarboxylase and peripherin revealed extensive colocalization, showing that peripherin is produced by histaminergic neurons in all parts of the tuberomammillary nucleus. We next used histamine immunostaining in peripherin knockout, overexpressing and wild type mice to study if altered peripherin expression affects these neurons, but could not detect any visible difference in the appearance of these neurons or their axons. Peripherin knockout mice and heterozygotic littermates were used for measurement of locomotor activity, feeding, drinking, and energy expenditure. Both genotypes displayed diurnal rhythms with all the parameters higher during the dark period. The respiratory quotient, an indicator of the type of substrate being utilized, also exhibited a significant diurnal rhythm in both genotypes. The diurnal patterns and the average values of all the recorded parameters for 24 h, daytime and night time were not significantly different between the genotypes, however. CONCLUSION: In conclusion, we have shown that peripherin is expressed in the tuberomammillary neurons of the mouse hypothalamus. Monitoring of locomotor activity, feeding, drinking, and energy expenditure in mice either lacking or overexpressing peripherin did not reveal any difference, so the significance of peripherin in these neurons remains to be determined. The complete overlap between histidine decarboxylase and peripherin, both the protein and its mRNA, renders peripherin a useful new marker for histaminergic neurons in the hypothalamus.

Interpenetrating polymer networks in polyvinyl alcohol/cellulose nanocrystals hydrogels to develop absorbent materials.

Polyvinyl alcohol/cellulose nanocrystals/poly(2-Hydroxyethyl methacrylate) (PVA/CNC/polyHEMA) and PVA/CNC/poly(N'-methylenebisacrylamide) (PVA/CNC/polyMBA) hydrogels were prepared by photo-crosslinking followed by freezing/thawing (F-T) cycle and this novel preparation method was reported. The formation of interpenetrating polymer networks (IPN) resulted from the addition of crosslinking HEMA or MBA monomers displayed improved interfacial adhesion. The produced hydrogels were measured by scanning electron microscopy (SEM), real-time fourier transform infrared (RTIR), thermogravimetric analysis (TGA), mechanical, swelling and adsorption tests. The results showed both PVA/CNC/polyHEMA with semi-IPN and PVA/CNC/polyMBA with dual network (DN) hydrogels had higher thermal stability, lower water loss rate and better swelling and reswelling and mechanical properties, comparing to PVA and PVA/CNC hydrogels. The adsorption behaviors of hydrogels using xylenol orange (XO) and methylene blue (MB) as model dyes were evaluated, indicating that PVA/CNC/polyHEMA and PVA/CNC/polyMBA hydrogels could hold some dyes. Overall, this work provided a good way for increasing mechanical, swelling, reswelling, thermal, and adsorption properties of PVA/CNC, which will be a promising water-manageable material for agriculture application and a candidate for dye carrier.

Preparation and characterization of melamine-formaldehyde/Ag composite microspheres with surface-enhanced Raman scattering and antibacterial activities.

A facile and environmentally friendly approach was proposed to decorate Ag nanoparticles on melamine-formaldehyde (MF) colloidal particles (MF/Ag composite microspheres). In this approach, monodisperse MF colloidal particles were prepared via a two-step organic sol-gel process and served as the active templates for the decoration of Ag nanoparticles. Then, the [Ag(NH3)2]+ ions as the Ag precursors can be adsorbed onto the surfaces of the MF colloidal particles and were in situ reduced into metallic Ag nanoparticles, forming MF/Ag composite microspheres. During this synthesis, neither presurface activation nor extra reductants were necessary. These MF/Ag composite microspheres can be used as the surface-enhanced Raman scattering (SERS) active substrates for the trace detection of organic compounds, e.g., 4-aminobenzenethiol (4-ABT) and penicillin G sodium. Furthermore, these MF/Ag composite microspheres also showed excellent antibacterial activities against both Escherichia coli (E. coli, gram-negative bacteria) and Staphylococcus aureus (S. aureus, gram-positive bacteria).

Effects of Melanin on Optical Behavior of Polymer: From Natural Pigment to Materials Applications.

Melanin is a kind of ubiquitous natural pigment, which serves a variety of protective functions in many organisms. In the present study, natural melanin and synthetic melanin nanoparticles (NPs) were systematically investigated for its potential application in polymeric optical materials. A significant short-wavelength shielding and high visible light transparency polymer nanocomposite was easily obtained via tuning the melanin particle size. In particular, the nanocomposite film with melanin NPs (diameter ≈ 15 nm) loading even as low as 1 wt % blocks most ultraviolet light below 340 nm and still keeps high visible light transparency (83%) in the visible spectrum. More importantly, because of the excellent photoprotection and radical scavenging capabilities of melanin, the resulting polymer nanocomposite exhibits outstanding photostability. In effect, such fantastic melanin NPs is promising for applications in various optical materials.

Artificial Nacre from Supramolecular Assembly of Graphene Oxide.

Inspired by the "brick-and-mortar" structure and remarkable mechanical performance of nacre, many efforts have been devoted to fabricating nacre-mimicking materials. Herein, a class of graphene oxide (GO) based artificial nacre material with quadruple hydrogen-bonding interactions was fabricated by functionalization of polydopamine-capped graphene oxide (PDG) with 2-ureido-4[1 H]-pyrimidinone (UPy) self-complementary quadruple hydrogen-bonding units followed by supramolecular assembly process. The artificial nacre displays a strict "brick-and-mortar" structure, with PDG nanosheets as the brick and UPy units as the mortar. The resultant nanocomposite shows an excellent balance of strength and toughness. Because of the strong strengthening via quadruple hydrogen bonding, the tensile strength and toughness can reach 325.6 ± 17.8 MPa and 11.1 ± 1.3 MJ m-3, respectively, thus exceeding natural nacre, and reaching 3.6 and 10 times that of a pure GO artificial nacre. Furthermore, after further H2O treatment, the resulting H2O-treated PDG-UPy actuator displays significant bending actuations when driven by heat. This work provides a pathway for the development of artificial nacre for their potential applications in energy conversion, temperature sensor, and thermo-driven actuator.

Sex difference in body weight gain and leptin signaling in hypocretin/orexin deficient mouse models.

Recent studies in human and animal models of narcolepsy have suggested that obesity in narcolepsy may be due to deficiency of hypocretin signaling, and is also under the influence of environmental factors and the genetic background. In the current study, using two hypocretin/orexin deficient narcoleptic mouse models (i.e. preproorexin knockout (KO) and orexin/ataxin-3 transgenic (TG) mice) with cross-sectional assessments, we have further analyzed factors affecting obesity. We found that both KO and TG narcoleptic mice with mixed genetic backgrounds (N4-5, 93.75-96.88% genetic composition of C57BL/6) tended to be heavier than wild type (WT) mice of 100-200 days old. The body weight of heterozygous mice was intermediate between those of KO and WT mice. Obesity was more prominent in females in both KO and TG narcoleptic mice and was associated with higher serum leptin levels, suggesting a partial leptin resistance. Obesity is less prominent in the congenic TG narcoleptic mice, but is still evident in females. Our results confirmed that hypocretin/orexin ligand deficiency is one of the critical factors for the obese tendency in narcolepsy. However, multiple factors are also likely to affect this phenotype, and a sex difference specific alteration of leptin-hypocretin signaling may be involved.

Insulin signaling inhibits the 5-HT2C receptor in choroid plexus via MAP kinase.

BACKGROUND: G protein-coupled receptors (GPCRs) interact with heterotrimeric GTP-binding proteins (G proteins) to modulate acute changes in intracellular messenger levels and ion channel activity. In contrast, long-term changes in cellular growth, proliferation and differentiation are often mediated by tyrosine kinase receptors and certain GPCRs by activation of mitogen-activated protein (MAP) kinases. Complex interactions occur between these signaling pathways, but the specific mechanisms of such regulatory events are not well-understood. In particular it is not clear whether GPCRs are modulated by tyrosine kinase receptor-MAP kinase pathways. RESULTS: Here we describe tyrosine kinase receptor regulation of a GPCR via MAP kinase. Insulin reduced the activity of the 5-HT2C receptor in choroid plexus cells which was blocked by the MAP kinase kinase (MEK) inhibitor, PD 098059. We demonstrate that the inhibitory effect of insulin and insulin-like growth factor type 1 (IGF-1) on the 5-HT2C receptor is dependent on tyrosine kinase, RAS and MAP kinase. The effect may be receptor-specific: insulin had no effect on another GPCR that shares the same G protein signaling pathway as the 5-HT2C receptor. This effect is also direct: activated MAP kinase mimicked the effect of insulin, and removing a putative MAP kinase site from the 5-HT2C receptor abolished the effect of insulin. CONCLUSION: These results show that insulin signaling can inhibit 5-HT2C receptor activity and suggest that MAP kinase may play a direct role in regulating the function of a specific GPCR.