Redefining differential roles of MAO-A in dopamine degradation and MAO-B in tonic GABA synthesis

Monoamine oxidase (MAO) is believed to mediate the degradation of monoamine neurotransmitters, including dopamine, in the brain. Between the two types of MAO, MAO-B has been believed to be involved in dopamine degradation, which supports the idea that the therapeutic efficacy of MAO-B inhibitors in Parkinson’s disease can be attributed to an increase in extracellular dopamine concentration. However, this belief has been controversial. Here, by utilizing in vivo phasic and basal electrochemical monitoring of extracellular dopamine with fast-scan cyclic voltammetry and multiple-cyclic square wave voltammetry and ex vivo fluorescence imaging of dopamine with GRAB_DA2m, we demonstrate that MAO-A, but not MAO-B, mainly contributes to striatal dopamine degradation. In contrast, our whole-cell patch-clamp results demonstrated that MAO-B, but not MAO-A, was responsible for astrocytic GABA-mediated tonic inhibitory currents in the rat striatum. We conclude that, in contrast to the traditional belief, MAO-A and MAO-B have profoundly different roles: MAO-A regulates dopamine levels, whereas MAO-B controls tonic GABA levels.Subject terms: Neurotransmitters, Biosensors, Astrocyte, Inhibition, Transporters in the nervous system     

Development of a hollow fiber membrane module for using implantable artificial lung

This study examined the effects of multiple mechanical forces on gas exchange and hemolysis in intravascular lung assist devices (IVLAD). Specific attention was paid to on the effect of membrane vibration. This study adhered to the recommended practice for the assessment of hemolysis described by the American Society of Testing and Materials (ASTM). The results showed a higher oxygen (O₂) transfer rate and carbon dioxide (CO₂) removal rate in each excited frequency bandwidth than those without vibration. The maximum oxygen transfer and carbon dioxide removal rate occurred at frequency band of 7 Hz. The gas exchange improved maximum 52%. The plasma-free hemoglobin was 11.2 ± 0.57 and 14.4 ± 0.74 mg/100 ml by exciting a piezo-vibrator with a sinusoidal wave magnitude of DC10 V and DC50 V, respectively. The NIHO value was determined to be 59 ± 2.76 and 95 ± 4.32 mg/100 ml by exciting a piezo-vibrator with a sinusoidal wave magnitude of DC10 V and DC50 V, respectively. In conclusion, piezoelectric lead zirconate titanate (PZT) materials are exciting systems for improving the oxygen transfer efficiency and blood suitability of hollow fiber membrane in the development of new IVLAD.     

Quantification of nimesulide in human plasma by high-performance liquid chromatography with ultraviolet detector (HPLC-UV): application to pharmacokinetic studies in 28 healthy Korean subjects

Nimesulide is a selective COX-2 inhibitor that is as effective as the classical non-acidic nonsteroidal anti-inflammatory drugs in the relief of various pain and inflammatory conditions, but is better tolerated with lower incidences of adverse effects than other drugs. After oral dose of 100 mg nimesulide to western subjects, a mean maximal concentration (C(max)) of 2.86 ～ 6.5 μg/mL was reached at 1.22 ～ 2.75 h and mean t(1/2β) of 1.8 ～ 4.74 h. This study developed a robust method for quantification of nimesulide for the pharmacokinetics and suitability of its dosage in Korea and compared its suitability with other racial populations. Nimesulide and internal standard were extracted from acidified samples with methyl tert-butyl ether and analyzed by high-performance liquid chromatography with ultraviolet detection (HPLC-UV). The 28 healthy volunteers took 2 tablets of 100 mg nimesulide and blood concentrations were analyzed during the 24 h post dose. Several pharmacokinetic parameters were represented: AUC(0-infinity) = 113.0 mg-h/mL, C(max) = 12.06 mg/mL, time for maximal concentrations (T(max)) = 3.19 h and t(1/2β) = 4.51 h. These were different from those of western populations as follows: AUC was 14.5% and C(max) was 28% that of of Korean subjects and T(max) and t(1/2β) were also different. The validated HPLC-UV method was successfully applied for the pharmacokinetic studies of nimesulide in Korean subjects. Because the pharmacokinetics of nimesulide were different from western populations, its dosage regimen needs to be adjusted for Koreans.     

Regulation of anti-inflammatory cytokines IL-10 and TGF-β in mouse dendritic cells through treatment with Clonorchis sinensis crude antigen

Dendritic cells (DCs), which are regarded as the most potent antigen-presenting cells, are involved in innate and adaptive immunity. Upon uptake of pathogens, DCs express cell surface markers and secrete cytokines. In this study, we analyzed production of cytokines and found that interleukin (IL)-10 and transforming growth factor (TGF)-β production significantly increased in bone marrow-derived DCs and a mouse DC line, DC2.4, after treatment with crude antigen (CA) from liver fluke, Clonorchis sinensis. However, expression patterns of several activation molecules did not change. In addition, following treatment of DC2.4 cells with antigen from the lung fluke, Paragonimus westermani, production of IL-10 and TGF-β significantly increased compared with groups treated with other parasite antigens, Spirometra erinacei plerocercoid CA and Echinococcus granulosus hydatid cystic fluid. We also found that treatment of DC2.4 cells with C. sinensis CA resulted in rapid and significant phosphorylation of extracellular signal-regulated kinase 1/2, a mitogen-activated protein kinase. Following treatment of DC2.4 cells with C. sinensis CA, treatment with an inhibitor specific to an extracellular signal-regulated kinase inhibited production of IL-10 and TGF-β. Our results suggest that CA from C. sinensis has a role in the anti-inflammatory function of DC cells by inducing IL-10 and TGF-β through activation of extracellular signal-regulated kinase 1/2.     

Quantification and Metabolite Identification of Sulfasalazine in Mouse Brain and Plasma Using Quadrupole-Time-of-Flight Mass Spectrometry

Sulfasalazine (SAS), an anti-inflammatory drug with potent cysteine/glutamate antiporter system xc-(SXC) inhibition has recently shown beneficial effects in brain-related diseases. Despite many reports related to central nervous system (CNS) effect of SAS, pharmacokinetics (PK) and metabolite identification studies in the brain for SAS were quite limited. The aim of this study was to investigate the pharmacokinetics and metabolite identification of SAS and their distributions in mouse brain. Using in vivo brain exposure studies (neuro PK), the PK parameters of SAS was calculated for plasma as well as brain following intravenous and oral administration at 10 mg/kg and 50 mg/kg in mouse, respectively. In addition, in vivo metabolite identification (MetID) studies of SAS in plasma and brain were also conducted. The concentration of SAS in brain was much lower than that in plasma and only 1.26% of SAS was detected in mouse brain when compared to the SAS concentration in plasma (brain to plasma ratio (%): 1.26). In the MetID study, sulfapyridine (SP), hydroxy-sulfapyridine (SP-OH), and N-acetyl sulfapyridine (Ac-SP) were identified in plasma, whereas only SP and Ac-SP were identified as significant metabolites in brain. As a conclusion, our results suggest that the metabolites of SAS such as SP and Ac-SP might be responsible for the pharmacological effect in brain, not the SAS itself.     

Liquid chromatography-high resolution mass spectrometric method for the quantification of monomethyl auristatin E (MMAE) and its preclinical pharmacokinetics

MMAE is a potent antimitotic drug used as payload of an antibody-drug conjugate which shows potent activity in preclinical and clinical studies against a range of lymphomas, leukemia and solid tumors. Liquid chromatography-high resolution mass spectrometric method was developed for the quantification of MMAE and its preclinical pharmacokinetics. The method consisted of protein precipitation using acetonitrile (ACN) for sample preparation and liquid chromatography – quadrupole – time-of-flight – tandem mass spectrometry (LC-qTOF-MS/MS) analysis in the positive ion mode. A quadratic regression (weighted 1/concentration²), with an equation y = ax² + bx + c, was used to fit calibration curves over the concentration range of 1.01–2,200 ng/mL for MMAE. The qualification run met the acceptance criteria of ±25% accuracy and precision values for QC samples. Recovery was 42.84%. The dilution integrity was determined for 5-fold dilution and the accuracy and precision ranged within ±25%. The stability results indicated that MMAE was stable for the following conditions: short-term (4 h), long-term (4 weeks), freeze/thaw (3 cycles) and post-preparative stability (12 h). This qualified method was successfully applied to a pharmacokinetic study of MMAE in rat as a preclinical animal model. The PK results suggest that MMAE has moderate CL and low BA.Also, these results would be helpful in having a comprehensive understanding of the PK characteristics of MMAE and developing ADC in future.     

A selective array activation method for the generation of a focused source considering listening position

A focused source can provide an auditory illusion of a virtual source placed between the loudspeaker array and the listener. When a focused source is generated by time-reversed acoustic focusing solution, its use as a virtual source is limited due to artifacts caused by convergent waves traveling towards the focusing point. This paper proposes an array activation method to reduce the artifacts for a selected listening point inside an array of arbitrary shape. Results show that energy of convergent waves can be reduced up to 60 dB for a large region including the selected listening point.     

Copper-glucosamine microcubes: synthesis, characterization, and C-reactive protein detection

Cubelike microstructures of glucosamine-functionalized copper (GlcN-CuMC's) have been fabricated by the integration of injection pump and ultrasonochemistry. Although bulk microstructures and the nanostructure of metallic copper exhibit distinct applications, the amino sugar surface-functionalized copper is almost biocompatible and exhibits advanced features such as more crystallinity, high thermal stability, and electrochemical feasibility toward biomolecule (C-reactive protein, CRP) detection. An electrochemical test of this GlcN-CuMC's was demonstrated by immobilization on a conventional gold-PCB (Au-PCB) electrode. The combination of a biointerface membrane, from glucosamine functionalization, and electroactive sites of metallic copper provides a very efficient electrochemical response against various concentration of CRP. A perfect scaling of steady-state currents with r(2) values of 0.9862 (I(pa)) and 0.9972 (I(pc)) indicate the promise of this kind of biofunctionalized microstructure electrode for many surface and interface applications.