Blood-Oxygenation-Level-Dependent-(BOLD-) Based R2′ MRI Study in Monkey Model of Reversible Middle Cerebral Artery Occlusion

Objective. To investigate the value of BOLD-based reversible transverse relaxation rate (R2′) MRI in detecting ischemic penumbra (IP) in a monkey model of reversible middle cerebral artery occlusion (MCAO) and time evolution of relative R2′ (rR2′) in infarcted core, IP, and oligemia. Materials and Methods. 6 monkeys were used to make MCAO by the microcatheter method. MR scans were performed at 0 h (1 h after MCAO), 1 h, 3 h, 6 h, 12 h, 24 h, and 48 h after reperfusion. R2′ was calculated using quantitative T2 and T2* maps. Ischemic area was subdivided into infracted core, IP and oligemia. rR2′ was calculated respectively. Results. Reversible MCAO model for 4/6 monkeys was made successfully. rR2′ values were significantly different at each time point, being highest in oligemia followed by IP and infarcted core (P < .05). With reperfusion time evolution, rR2′ in infarcted core showed a decreased trend: sharply decreased within 6 hours and maintained at 0 during 6–48 hours (P < .05). rR2′ values in IP and oligemia showed similar increased trend: sharply increased within 6 hours, maintained a plateau during 6–24 hours, and slightly increased until 48 hours. Conclusion. BOLD-based R2′ MRI can be used to describe changes of cerebral oxygen extract in acute ischemic stroke, and it can provide additional information in detecting IP. The time evolution rR2′ in infarcted core, IP, and oligemia is in accordance with the underlying pathophysiology

In-Vitro Study on the Antibacterial and Antioxidant Activity of Four Commercial Essential Oils and In-Situ Evaluation of Their Effect on Quality Deterioration of Pacific White Shrimp (Litopenaeus vannamei) during Cold Storage

The antioxidant and antibacterial properties of four essential oils (oregano essential oil (OEO), tea tree essential oil (TTEO), wild orange essential oil (WOEO), and clove leaf essential oil (CLEO)) were determined. The in-vitro experiment indicated that CLEO had the highest total phenolic content and DPPH scavenging activity, and OEO displayed the highest antibacterial effect, so they were applied to maintain the quality of shrimp for further study. In-situ study, the total viable counts of shrimp were inhibited from 9.05 log CFU/g to 8.18 and 8.34 log CFU/g by 2% of OEO and CLEO treated alone on 10 d. The melanosis ratio was also retarded from 38.16% to 28.98% and 26.35% by the two essential oils. The inhibitory effects of OEO and CLEO on the increase of PPO activity, weight loss, and TCA-soluble peptides, and the decreasing tendency of whiteness, the contents of myofibrillar and sarcoplasmic proteins were also founded. The samples treated with 1% OEO + 1% CLEO had better quality than those treated alone. Therefore, the combination of OEO and CLEO had a synergistic effect, which displayed the highest efficiency to prevent the melanosis, bacterial growth, and protein hydrolysis of shrimp.Peer reviewe

Enhanced inhibitory synaptic transmission in the spinal dorsal horn mediates antinociceptive effects of TC-2559

<p>Abstract</p> <p>Background</p> <p>TC-2559 is a selective α4β2 subtype of nicotinic acetylcholine receptor (nAChR) partial agonist and α4β2 nAChR activation has been related to antinociception. The aim of this study is to investigate the analgesic effect of TC-2559 and its underlying spinal mechanisms.</p> <p>Results</p> <p>1) <it>In vivo </it>bioavailability study: TC-2559 (3 mg/kg) had high absorption rate in rats with maximal total brain concentration reached over 4.6 μM within first 15 min after administration and eliminated rapidly with brain half life of about 20 min after injection. 2) <it>In vivo </it>behavioral experiments: TC-2559 exerts dose dependent antinociceptive effects in both formalin test in mice and chronic constriction injury (CCI) model in rats by activation of α4β2 nAChRs; 3) Whole-cell patch-clamp studies in the superficial dorsal horn neurons of the spinal cord slices: perfusion of TC-2559 (2 μM) significantly increased the frequency, but not amplitude of spontaneous inhibitory postsynaptic currents (sIPSCs). The enhancement of sIPSCs was blocked by pre-application of DHβE (2 μM), a selective α4β2 nicotinic receptor antagonist. Neither the frequency nor the amplitude of spontaneous excitatory postsynaptic currents (sEPSCs) of spinal dorsal horn neurons were affected by TC-2559.</p> <p>Conclusions</p> <p>Enhancement of inhibitory synaptic transmission in the spinal dorsal horn via activation of α4β2 nAChRs may be one of the mechanisms of the antinociceptive effects of TC-2559 on pathological pain models. It provides further evidence to support the notion that selective α4β2 subtype nAChR agonist may be developed as new analgesic drug for the treatment of neuropathic pain.</p

Grp94 Protein Delivers γ-Aminobutyric Acid Type A (GABAA) Receptors to Hrd1 Protein-mediated Endoplasmic Reticulum-associated Degradation

This research was originally published in the Journal of Biological Chemistry. Xiao-Jing Di, Ya-Juan Wang, Dong-Yun Han, Yan-Lin Fu, Adam S. Duerfeldt, Brian S. J. Blagg and Ting-Wei Mu.Grp94 Protein Delivers γ-Aminobutyric Acid Type A (GABAA) Receptors to Hrd1 Protein-mediated Endoplasmic Reticulum-associated Degradation. Journal of Biological Chemistry. 2016; 291, 9526-9539.Proteostasis maintenance of γ-aminobutyric acid type A (GABAA) receptors dictates their function in controlling neuronal inhibition in mammalian central nervous systems. However, as a multisubunit, multispan, integral membrane protein, even wild type subunits of GABAA receptors fold and assemble inefficiently in the endoplasmic reticulum (ER). Unassembled and misfolded subunits undergo ER-associated degradation (ERAD), but this degradation process remains poorly understood for GABAA receptors. Here, using the α1 subunits of GABAA receptors as a model substrate, we demonstrated that Grp94, a metazoan-specific Hsp90 in the ER lumen, uses its middle domain to interact with the α1 subunits and positively regulates their ERAD. OS-9, an ER-resident lectin, acts downstream of Grp94 to further recognize misfolded α1 subunits in a glycan-dependent manner. This delivers misfolded α1 subunits to the Hrd1-mediated ubiquitination and the valosin-containing protein-mediated extraction pathway. Repressing the initial ERAD recognition step by inhibiting Grp94 enhances the functional surface expression of misfolding-prone α1(A322D) subunits, which causes autosomal dominant juvenile myoclonic epilepsy. This study clarifies a Grp94-mediated ERAD pathway for GABAA receptors, which provides a novel way to finely tune their function in physiological and pathophysiological conditions

Method of determining cosmological parameter ranges with samples of candles with an intrinsic distribution

In this paper, the effect of the intrinsic distribution of cosmological candles is investigated. We find that, in the case of a narrow distribution, the deviation of the observed modulus of sources from the expected central value could be estimated within a ceratin range. We thus introduce a lower and upper limits of $\chi ^{2}$, $\chi_{\min}^{2}$ and $\chi_{\max}^{2}$, to estimate cosmological parameters by applying the conventional minimizing $\chi ^{2}$ method. We apply this method to a gamma-ray burst (GRB) sample as well as to a combined sample including this GRB sample and an SN Ia sample. Our analysis shows that: a) in the case of assuming an intrinsic distribution of candles of the GRB sample, the effect of the distribution is obvious and should not be neglected; b) taking into account this effect would lead to a poorer constraint of the cosmological parameter ranges. The analysis suggests that in the attempt of constraining the cosmological model with current GRB samples, the results tend to be worse than what previously thought if the mentioned intrinsic distribution does exist.Comment: 6 pages,4 figures,1 tables.Data updated. Main conclusion unchange

Screening therapeutic EMT blocking agents in a three-dimensional microenvironment

Epithelial–mesenchymal transition (EMT) plays a critical role in the early stages of dissemination of carcinoma leading to metastatic tumors, which are responsible for over 90% of all cancer-related deaths. Current therapeutic regimens, however, have been ineffective in the cure of metastatic cancer, thus an urgent need exists to revisit existing protocols and to improve the efficacy of newly developed therapeutics. Strategies based on preventing EMT could potentially contribute to improving the outcome of advanced stage cancers. To achieve this goal new assays are needed to identify targeted drugs capable of interfering with EMT or to revert the mesenchymal-like phenotype of carcinoma to an epithelial-like state. Current assays are limited to examining the dispersion of carcinoma cells in isolation in conventional 2-dimensional (2D) microwell systems, an approach that fails to account for the 3-dimensional (3D) environment of the tumor or the essential interactions that occur with other nearby cell types in the tumor microenvironment. Here we present a microfluidic system that integrates tumor cell spheroids in a 3D hydrogel scaffold, in close co-culture with an endothelial monolayer. Drug candidates inhibiting receptor activation or signal transduction pathways implicated in EMT have been tested using dispersion of A549 lung adenocarcinoma cell spheroids as a metric of effectiveness. We demonstrate significant differences in response to drugs between 2D and 3D, and between monoculture and co-culture.Singapore. National Research Foundation (Singapore MIT Alliance for Research and Technology's BioSystems and Micromechanics Inter-Disciplinary Research programme)National University of Singapore (Cancer Science Institute)Singapore. Agency for Science, Technology and ResearchSingapore. Institute of Molecular and Cell Biology (IMCB core funding A*STAR

Atherosclerosis and Helminths Infection

Atherosclerosis is a chronic disease that causes various cardiovascular complications. Plaque formation in atherosclerosis is considered similar to the pathogenesis of other autoimmune diseases; thus, immunomodulation and immunosuppression may present strategies for the treatment and prevention of these diseases. Interestingly helminth infection was found to inhibit T helper 1-mediated autoimmune diseases and T helper 2-mediated allergy and asthma, indicating significant potential for clinical application. Some study even found that therapeutic efficacy of the viable tapeworm was superior to dexamethasone treatment. Recently, some studies have shown an inverse association between helminth infections and inflammatory diseases, including diabetes mellitus, lipid abnormality, and atherosclerosis. Will the underlying mechanism bring us a new idea on the treatment for these diseases? We tried to find an answer by reviewing recent articles

Screening therapeutic EMT blocking agents in a three-dimensional microenvironment

Epithelial–mesenchymal transition (EMT) plays a critical role in the early stages of dissemination of carcinoma leading to metastatic tumors, which are responsible for over 90% of all cancer-related deaths. Current therapeutic regimens, however, have been ineffective in the cure of metastatic cancer, thus an urgent need exists to revisit existing protocols and to improve the efficacy of newly developed therapeutics. Strategies based on preventing EMT could potentially contribute to improving the outcome of advanced stage cancers. To achieve this goal new assays are needed to identify targeted drugs capable of interfering with EMT or to revert the mesenchymal-like phenotype of carcinoma to an epithelial-like state. Current assays are limited to examining the dispersion of carcinoma cells in isolation in conventional 2-dimensional (2D) microwell systems, an approach that fails to account for the 3-dimensional (3D) environment of the tumor or the essential interactions that occur with other nearby cell types in the tumor microenvironment. Here we present a microfluidic system that integrates tumor cell spheroids in a 3D hydrogel scaffold, in close co-culture with an endothelial monolayer. Drug candidates inhibiting receptor activation or signal transduction pathways implicated in EMT have been tested using dispersion of A549 lung adenocarcinoma cell spheroids as a metric of effectiveness. We demonstrate significant differences in response to drugs between 2D and 3D, and between monoculture and co-culture.Singapore. National Research Foundation (Singapore MIT Alliance for Research and Technology's BioSystems and Micromechanics Inter-Disciplinary Research programme)National University of Singapore (Cancer Science Institute)Singapore. Agency for Science, Technology and ResearchSingapore. Institute of Molecular and Cell Biology (IMCB core funding A*STAR

ESC-Derived Basal Forebrain Cholinergic Neurons Ameliorate the Cognitive Symptoms Associated with Alzheimer’s Disease in Mouse Models

SummaryDegeneration of basal forebrain cholinergic neurons (BFCNs) is associated with cognitive impairments of Alzheimer’s disease (AD), implying that BFCNs hold potentials in exploring stem cell-based replacement therapy for AD. However, studies on derivation of BFCNs from embryonic stem cells (ESCs) are limited, and the application of ESC-derived BFCNs remains to be determined. Here, we report on differentiation approaches for directing both mouse and human ESCs into mature BFCNs. These ESC-derived BFCNs exhibit features similar to those of their in vivo counterparts and acquire appropriate functional properties. After transplantation into the basal forebrain of AD model mice, ESC-derived BFCN progenitors predominantly differentiate into mature cholinergic neurons that functionally integrate into the endogenous basal forebrain cholinergic projection system. The AD mice grafted with mouse or human BFCNs exhibit improvements in learning and memory performances. Our findings suggest a promising perspective of ESC-derived BFCNs in the development of stem cell-based therapies for treatment of AD