Food preference-based screening method for identification of effectors of substance use disorders using Caenorhabditis elegans.

Substance use disorder (SUD) affects over 48 million Americans aged 12 and over. Thus, identifying novel chemicals contributing to SUD will be critical for developing efficient prevention and mitigation strategies. Considering the complexity of the actions and effects of these substances on human behavior, a high-throughput platform using a living organism is ideal. We developed a quick and easy screening assay using Caenorhabditis elegans. C. elegans prefers high-quality food (Escherichia coli HB101) over low-quality food (Bacillus megaterium), with a food preference index of approximately 0.2, defined as the difference in the number of worms at E. coli HB101 and B. megaterium over the total worm number. The food preference index was significantly increased by loperamide, a µ-opioid receptor (MOPR) agonist, and decreased by naloxone, a MOPR antagonist. These changes depended on npr-17, a C. elegans homolog of opioid receptors. In addition, the food preference index was significantly increased by arachidonyl-2'-chloroethylamide, a cannabinoid 1 receptor (CB1R) agonist, and decreased by rimonabant, a CB1R inverse agonist. These changes depended on npr-19, a homolog of CB1R. These results suggest that the conserved opioid and endocannabinoid systems modulate the food preference behaviors of C. elegans. Finally, the humanoid C. elegans strains where npr-17 was replaced with human MOPR and where npr-19 was replaced with human CB1R phenocopied the changes in food preference by the drug treatment. Together, the current results show that this method can be used to rapidly screen the potential effectors of MOPR and CB1R to yield results highly translatable to humans.

Trifuhalol A, a phlorotannin from the brown algae Agarum cribrosum, reduces adipogenesis of human primary adipocytes through Wnt/ß-catenin and AMPK-dependent pathways.

Trifuhalol A, a fucol-type phlorotannin, was extracted and identified from the brown algae Agarum cribrosum. The total yield and purity of trifuhalol A from A. cribrosum were 0.98% and 86%, respectively. Trifuhalol A at 22 and 44 µM inhibited lipid accumulation in human primary adipocytes. Consistently trifuhalol A suppressed the expression of adipogenesis-related genes, such as proliferator-activated receptor-gamma (PPAR-γ), CCAAT/enhancer-binding protein-alpha (C/EBP-α), fatty acid synthase (FAS), and sterol regulatory element-binding protein-1 (SREBP-1), in a dose-dependent manner. Trifuhalol A increased the level of proteins such as wingless/integrated (Wnt)10b, nuclear-ß-catenin, total-ß-catenin, phospho-AMP-activated protein kinase (pAMPK), and phospho-liver kinase B1 (pLKB1) as well as the expression of genes such as Wnt10b, Frizzled 1, and low-density lipoprotein receptor-related protein 6 (LRP6). Additionally, trifuhalol A decreased the expression of the glycogen synthase kinase-3beta (GSK3ß) gene. These results suggest that trifuhalol A reduces fat accumulation in human adipocytes via the Wnt/ß-catenin- and AMPK-dependent pathways.

Anti-Allergic Inflammatory Effect of Agarum cribrosum and Its Phlorotannin Component, Trifuhalol A, against the Ovalbumin-Induced Allergic Asthma Model.

Asthma is a chronic inflammatory disease involving structural changes to the respiratory system and severe immune responses mediated by allergic cytokines and pro-inflammatory mediators. Agarum cribrosum (AC) is a kind of seaweed which contains a phlorotannin, trifuhalol A. To evaluate its anti-allergic inflammatory effect against asthma, an ovalbumin inhalation-induced mouse asthma model was used. Histologic observations proved that trifuhalol A is minimizing the lung and tracheal structure changes as well as the infiltration of eosinophils and mast cells against ovalbumin inhalation challenge. From the serum and bronchoalveolar lavage fluid, ovalbumin-specific IgE and Th2-specific cytokines, IL-4, -5, and -13, were reduced with trifuhalol A treatment. In addition, IL-1ß, IL-6, and TNF-α concentrations in lung homogenate were also significantly reduced via trifuhalol A treatment. Taken together, trifuhalol A, isolated from AC, was able to protect lung and airways from Th2-specific cytokine release, and IgE mediated allergic inflammation as well as the attenuation of IL-1ß, IL-6, and TNF-α in lung, which results in the suppression of eosinophils and the mast cells involved asthmatic pathology.

The osteogenic effects of sponges synthesized with biomaterials and nano-hydroxyapatite.

Artificial bone substitutes have been developed using various biomaterials for use in medicine. Silk fibroin (SF) displays excellent mechanical properties and cell compatibility. Nonetheless, the mechanical properties of silk fibroin scaffolds used in artificial bone substitutes are weaker than those of natural bone, and silk fibroin is deficient as an osteogenic agent. This limits their effectiveness in bone tissue engineering. We added nano-hydroxyapatite (nHAp) particles to an existing cell-based artificial bone substitute with a silk fibroin scaffold, which will improve its mechanical properties and osteogenic efficacy, leading to significant bone regeneration. The mechanical characters of silk fibroin modifying with nHAp were measured by Atomic Force Microscopy Analysis, dispersive x-ray spectroscopy, Porosity measurement, and Microcomputed Tomography. The proliferation and toxicity of a fibroin/dextran/collagen sponge (FDS) containing nHAp were evaluatedin vitro, and its osteogenic efficacy was evaluated using nude mouse and rabbit radius defect models. The defect area was repaired and showed callus formation of new bone in the rabbit radius defect models of the nHAp-FDS-treated group, whereas the defect area was unchanged in the FDS-treated group. The nHAp-FDS manufactured in this study showed significant bone regeneration owing to the synergistic effects of the components, such as those due to the broad range of pore sizes in the sponge and protein adsorbability of the nHAp, which could be suggested as a better supportive material for bone tissue engineering.

Deep learning-based reconstruction on cardiac CT yields distinct radiomic features compared to iterative and filtered back projection reconstructions.

We aimed to determine the effects of deep learning-based reconstruction (DLR) on radiomic features obtained from cardiac computed tomography (CT) by comparing with iterative reconstruction (IR), and filtered back projection (FBP). A total of 284 consecutive patients with 285 cardiac CT scans that were reconstructed with DLR, IR, and FBP, were retrospectively enrolled. Radiomic features were extracted from the left ventricular (LV) myocardium, and from the periprosthetic mass if patients had cardiac valve replacement. Radiomic features of LV myocardium from each reconstruction were compared using a fitting linear mixed model. Radiomics models were developed to diagnose periprosthetic abnormality, and the performance was evaluated using the area under the receiver characteristics curve (AUC). Most radiomic features of LV myocardium (73 of 88) were significantly different in pairwise comparisons between all three reconstruction methods (P < 0.05). The radiomics model on IR exhibited the best diagnostic performance (AUC 0.948, 95% CI 0.880-1), relative to DLR (AUC 0.873, 95% CI 0.735-1) and FBP (AUC 0.875, 95% CI 0.731-1), but these differences did not reach significance (P > 0.05). In conclusion, applying DLR to cardiac CT scans yields radiomic features distinct from those obtained with IR and FBP, implying that feature robustness is not guaranteed when applying DLR.

Trifuhalol A Suppresses Allergic Inflammation through Dual Inhibition of TAK1 and MK2 Mediated by IgE and IL-33.

The activation and degranulation of immune cells play a pivotal role in allergic inflammation, a pathological condition that includes anaphylaxis, pruritus, and allergic march-related diseases. In this study, trifuhalol A, a phlorotannin isolated from Agarum cribrosum, inhibited the degranulation of immune cells and the biosynthesis of IL-33 and IgE in differentiated B cells and keratinocytes, respectively. Additionally, trifuhalol A suppressed the IL-33 and IgE-mediated activation of RBL-2H3 cells through the regulation of the TAK1 and MK2 pathways. Hence, the effect of trifuhalol A on allergic inflammation was evaluated using a Compound 48/80-induced systemic anaphylaxis mouse model and a house dust mite (HDM)-induced atopic dermatitis (AD) mouse model. Trifuhalol A alleviated anaphylactic death and pruritus, which appeared as an early-phase reaction to allergic inflammation in the Compound 48/80-induced systemic anaphylaxis model. In addition, trifuhalol A improved symptoms such as itching, edema, erythema, and hyperkeratinization in HDM-induced AD mice as a late-phase reaction. Moreover, the expression of IL-33 and thymic stromal lymphopoietin, inflammatory cytokines secreted from activated keratinocytes, was significantly reduced by trifuhalol A administration, resulting in the reduced infiltration of immune cells into the skin and a reduction in the blood levels of IgE and IL-4. In summarizing the above results, these results confirm that trifuhalol A is a potential therapeutic candidate for the regulation of allergic inflammation.

Injectable Click Fibroin Bioadhesive Derived from Spider Silk for Accelerating Wound Closure and Healing Bone Fracture.

Wound closure is a critical step in postoperative wound recovery. Substantial advancements have been made in many different means of facilitating wound closure, including the use of tissue adhesives. Compared to conventional methods, such as suturing, tissue bioadhesives better accelerate wound closure. However, several existing tissue adhesives suffer from cytotoxicity, inadequate tissue adhesive strength, and high costs. In this study, a series of bioadhesives was produced using non-swellable spider silk-derived silk fibroin protein and an outer layer of swellable polyethylene glycol and tannic acid. The gelation time of the spider silk-derived silk fibroin protein bioadhesive is less than three minutes and thus can be used during rapid surgical wound closure. By adding polyethylene glycol (PEG) 2000 and tannic acid as co-crosslinking agents to the N-Hydroxysuccinimide (NHS), and 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) reaction, the adhesive strength of the bioadhesive became 2.5 times greater than that of conventional fibrin glue adhesives. Silk fibroin bioadhesives do not show significant cytotoxicity in vitro compared with other bioadhesives. In conclusion, silk fibroin bioadhesive is promising as a new medical tool for more effective and efficient surgical wound closure, particularly in bone fractures.

Effect of Preirradiation Fluoride Treatment on the Physical Properties of Dentin.

Objective: To determine the effects of preirradiation fluoride treatments on the Knoop hardness of dentin. Materials and Methods: Human posterior teeth mounted into acrylic resin molds were polished with silicon carbide (SiC) abrasives and 3-micron diamond paste. The Knoop hardness of dentin was measured with a Leco hardness instrument. The teeth were divided into groups of ten teeth per group as follows: no treatment (control), treatment with silver diamine fluoride (SDF), MI varnish (MI), and cavity shield (CS). The teeth were exposed to 2 Gy of daily radiation for six weeks using an X-Rad 320ix biological irradiator. Hardness was measured weekly, before, during, and after irradiation. The teeth were stored in artificial saliva at 37oC between radiation treatments. Results: In preirradiation dentin, a Knoop hardness value of 58.8 (14.1) KHN was obtained. Treatment with SDF significantly increased KHN before irradiation. Immediately after radiation treatment, hardness was significantly reduced in all experimental groups. Postirradiation fluoride treatments increased the hardness of dentin to varying degrees. Conclusions: Preirradiation fluoride treatment does not provide protection from decreases in the hardness of dentin. Treatment of teeth with fluoride formulations after radiation progressively restores the hardness of dentin to different degrees.

[Toxic epidermal necrolysis: idiosyncratic reaction to drugs]. / Necrólisis epidérmica tóxica: reacción idiosincrática a neurofármacos.

Toxic epidermal necrolysis (NET) and Steven Johnson syndrome (SJS) are infrequent mucocutaneous hypersensitivity reactions with systemic involvement. They are predominantly caused by drugs. We report the case of a patient over 60 years of age who presented with extensive mucocutaneous and ophthalmic injury with hemodynamic failure, associated with the rapid onset of lamotrigine in a short period of time. Although the incidence is low, the mortality rate is high. It requires early suspicious and diagnosis in addition to an interdisciplinary therapeutic approach.

Antioxidant and antidiabetic activities of vanadium-binding protein and trifuhalol A.

The antioxidant and antidiabetic activities of vanadium-binding protein (VBP) and trifuhalol A, alone or combined, were investigated. Both VBP and trifuhalol A showed potent radical scavenging activity (RSA) on 2,2'-azinobis (3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), hydrogen peroxide, and ferric-reducing antioxidant power. Their combination at a concentration of 100 µg/ml VBP and 40 µg/ml trifuhalol A exhibited more than 99% RSA against ABTS. Additionally, VBP and trifuhalol A, alone or combined, displayed potential antidiabetic activities against Saccharomyces cerevisiae α-glucosidase. The highest inhibition of 70.26% against S. cerevisiae α-glucosidase was observed in the case of the combination of 250 µg/ml VBP and 1.75 µg/ml trifuhalol A. Kinetics study revealed that VBP and trifuhalol A were noncompetitive inhibition type against S. cerevisiae α-glucosidase, while VBP and trifuhalol A combined treatment was a mixed inhibition type against S. cerevisiae α-glucosidase. These results indicated that VBP and trifuhalol A, alone or combined, had high free radical scavenging activity and inhibitory activity against S. cerevisiae a-glucosidase, suggesting that VBP and trifuhalol A could be used as candidates for the development of natural antidiabetic drugs or functional food. PRACTICAL APPLICATIONS: The present study showed that VBP and trifuhalol A, alone or combined, had potential antioxidant and antidiabetic activities, suggesting that VBP and trifuhalol A could be developed to a novel nutraceutical or natural antidiabetic drugs in the management of obesity or diabetes. This finding will be beneficial for all peoples who are directly or indirectly associated with obesity or diabetes.

Enrichment of Polyglucosylated Isoflavones from Soybean Isoflavone Aglycones Using Optimized Amylosucrase Transglycosylation.

Isoflavones in soybeans are well-known phytoestrogens. Soy isoflavones present in conjugated forms are converted to aglycone forms during processing and storage. Isoflavone aglycones (IFAs) of soybeans in human diets have poor solubility in water, resulting in low bioavailability and bioactivity. Enzyme-mediated glycosylation is an efficient and environmentally friendly way to modify the physicochemical properties of soy IFAs. In this study, we determined the optimal reaction conditions for Deinococcus geothermalis amylosucrase-mediated α-1,4 glycosylation of IFA-rich soybean extract to improve the bioaccessibility of IFAs. The conversion yields of soy IFAs were in decreasing order as follows: genistein > daidzein > glycitein. An enzyme quantity of 5 U and donor:acceptor ratios of 1000:1 (glycitein) and 400:1 (daidzein and genistein) resulted in high conversion yield (average 95.7%). These optimal reaction conditions for transglycosylation can be used to obtain transglycosylated IFA-rich functional ingredients from soybeans.

The effects of dopamine D4 receptor ligands on operant alcohol self-administration and cue- and stress-induced reinstatement in rats.

Dopamine, a neurotransmitter with 5 receptor subtypes, is critical to the dependence-forming properties of drugs of abuse. The role of the dopamine D4 receptor subtype in substance use disorders has remained somewhat elusive but the recent development of selective ligands holds promise for future investigations of this receptor subtype in substance use disorders, including alcohol use disorder. The purpose of the present study was to further elucidate the effects of a selective antagonist (L-745,870) and agonist (PD 168,077) on alcohol self-administration and reinstatement induced either by cues or stress. It was found that the D4 antagonist, but not agonist, reduced alcohol intake at the highest doses. Further, the D4 antagonist reduced stress-induced reinstatement, with no effects on cue-induced reinstatement; the agonist was without effect on either form of reinstatement. The dopamine D4 receptor antagonist was without effect on food reinforcement. This work deepens existing lines of evidence that the dopamine D4 receptor is involved in substance use disorders and suggests that dopamine D4 receptor blockade diminishes motivation for alcohol-taking without influencing natural food rewards. Furthermore, there appears to be a plausible effect of dopamine D4 receptor blockade interfering with stress- but not cue-induced alcohol-seeking.

Anti-inflammatory effects of vanadium-binding protein from Halocynthia roretzi in LPS-stimulated RAW264.7 macrophages through NF-κB and MAPK pathways.

Vanadium-binding protein (VBP) was separated from the blood of the fresh sea urchin Halocynthia roretzi through (NH4)2SO4 precipitation, Diethylaminoethyl Sepharose fast-flow ion-exchange chromatography, and Sephacryl S-200 high-resolution size-exclusion chromatography. The protein size and purification yield of VBP were 27 kDa and 5.5%, respectively. VBP exerted anti-inflammatory effects in lipopolysaccharide-stimulated RAW264.7 macrophages by downregulating iNOS expression and inhibiting nitric oxide production. VBP also suppressed the expression of pro-inflammatory mediators such as COX-2, IL-1ß, IL-6, and TNF-α. The anti-inflammatory activity of VBP was further demonstrated in the NF-κB and MAPK inflammation pathways, in which VBP inhibited phosphorylation of signaling proteins such as p65, JNK, ERK1/2, and p38. Therefore, VBP from H. roretzi has anti-inflammatory effects and could potentially be used to treat inflammation.

Inhibitory Effects of Vanadium-Binding Proteins Purified from the Sea Squirt Halocynthia roretzi on Adipogenesis in 3T3-L1 Adipocytes.

The inhibitory effects of vanadium-binding proteins (VBPs) from the blood plasma and the intestine of sea squirt on adipogenesis in 3T3-L1 adipocytes were examined. 3T3L-1 cells treated with VBP blood plasma decreased markedly the lipid content in maturing pre-adipocytes in a dose-dependent manner, whereas VBP intestine did not show significant effects on lipid accumulation. Both VBPs did not have significant effect on cell viability. In order to demonstrate the anti-adipogenic effects of VBP blood plasma, the expressions of several adipogenic transcription factors and enzymes were investigated by Reverse Transcriptase-Polymerase Chain Reaction. VBP blood plasma down-regulated the expressions of transcription factors; PPAR-γ, C/EBP-α, SREBP1, and FAS, but did not have significant effects on the expressions of lipolytic enzymes; HSL and LPL. Both the crude and purified VBPs significantly increased the mRNA levels of Wnt10b, FZ1, LRP6, and ß-catenin, while decreased the expression of GSK-3ß. Hence, VBP blood plasma inhibited adipogenesis by activating WNT/ß-catenin pathway via the activation of Wnt10b. Based on the findings, VBP blood plasma decreased lipid accumulation which was mediated by decreasing adipogenesis, not by lipolysis. Therefore, VBP blood plasma could be used to treat obesity.

The effects of buspirone on occupancy of dopamine receptors and the rat gambling task.

BACKGROUND: The dopamine D3 receptor (DRD3) has been proposed as a target for drug development for the treatment of addictive disorders. Recently, the anxiolytic buspirone has been shown to have affinity for DRD3 and DRD4, and interest in repurposing it for addictive disorders has grown. METHODS: Binding of [3H]-(+)-PHNO in the rat cerebellum and striatum was used to measure occupancy by buspirone of DRD3 or DRD2, respectively. Effects of buspirone in the rat gambling task (rGT) and the five-choice serial reaction time task (5-CSRTT) were examined. RESULTS: Buspirone occupied both the DRD2 and DRD3 at high doses and the DRD3, but not the DRD2, in the narrow dose range of 3 mg/kg. At 10 mg/kg, a disruption of performance on rGT was observed. All measures of performance on the rGT, except for perseverations, were affected at 3 mg/kg. On the 5-CSRTT, omissions were increased. Impairments in the rGT were not mimicked by the effects induced by satiation. Further, buspirone did not impair food-maintained responding under a progressive ratio schedule of reinforcement at any dose, suggesting that the effects of buspirone on the rGT cannot be explained by non-selective actions. CONCLUSIONS: Although buspirone had effects on the rGT at the dose that selectively occupied the DRD3, the effects found do not parallel those found in previous studies of the effects of selective DRD3 antagonists on the rGT. Thus, buspirone may impair performance on the rGT through actions at multiple receptor sites.

Twelve-year survival of 2-surface composite resin and amalgam premolar restorations placed by dental students.

STATEMENT OF PROBLEM: Composite resin and amalgam restorations are indicated for the restoration of posterior teeth. With increased esthetic demands, long-term clinical studies are required to evaluate the restorative success and reasons for failure of these materials. PURPOSE: The purpose of this retrospective study was to determine the survival and reasons for failure of directly placed 2-surface composite resin restorations and directly placed 2-surface amalgam restorations on premolars placed by Canadian dental students. MATERIAL AND METHODS: Using The University of Manitoba's dental management software and paper charts, all 2-surface composite resin and 2-surface amalgam restorations placed on premolars between January 1, 2002, and May 30, 2014, were included. Short-term failure (within 2 years), long-term failure, and reasons for failure were collected. A Kaplan-Meier survival estimate with an associated P value comparing composite resin to amalgam restoration curves was performed using SPSS statistical software. RESULTS: Over 12 years, 1695 composite resin and 1125 amalgam 2-surface premolar restorations were placed. Of these restorations, 134 composite resins (7.9%) and 66 amalgams (5.9%) failed. Short-term failures (2 years or less) consisted of 57 composite resin (4%) and 23 amalgam (2.3%) restorations. Long-term failures (greater than 2 years) consisted of 77 composite resin (4.5%) and 43 amalgam (3.8%) restorations. After 12 years of service, the survival probability of composite resin restorations was 86% and that of amalgam restorations 91.5%. The differences in composite resin and amalgam survival curves were also found to be statistically significant (P=.009 for Log-rank test). The main reasons for failure were recurrent caries and fracture of the tooth being restored. CONCLUSIONS: Within the limitations of this study, both composite resin and amalgam restorations had acceptable success rates and similar failure modes. Recurrent caries was still the most common reason for failure.

Overview of Intraoperative Neurophysiological Monitoring During Spine Surgery.

Intraoperative neurophysiologic monitoring has had major advances in the past few decades. During spine surgery, the use of multimodality monitoring enables us to assess the integrity of the spinal cord, nerve roots, and peripheral nerves. The authors present a practical approach to the current modalities in use during spine surgery, including somatosensory evoked potentials, motor evoked potentials, spinal D-waves, and free-run and triggered electromyography. Understanding the complementary nature of these modalities will help tailor monitoring to a particular procedure to minimize postoperative neurologic deficit during spine surgery.

The Impact of Selective Dopamine D2, D3 and D4 Ligands on the Rat Gambling Task.

Gambling is an addictive disorder with serious societal and personal costs. To-date, there are no approved pharmacological treatments for gambling disorder. Evidence suggests a role for dopamine in gambling disorder and thus may provide a therapeutic target. The present study therefore aimed to investigate the effects of selective antagonists and agonists of D2, D3 and D4 receptors in a rodent analogue of the Iowa gambling task used clinically. In this rat gambling task (rGT), animals are trained to associate different response holes with different magnitudes and probabilities of food pellet rewards and punishing time-out periods. As in the Iowa gambling task, the optimal strategy is to avoid the tempting high-risk high-reward options, and instead favor those linked to smaller per-trial rewards but also lower punishments, thereby maximizing the amount of reward earned over time. Administration of those selective ligands did not affect decision making under the rGT. Only the D4 drug had modest effects on latency measures suggesting that D4 may contribute in some ways to decision making under this task.