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A photoirradiated potassium-doped C60 film has been studied by Fourier transform mass spectrometry (FT-MS) and by in situ high-resolution Fourier transform infrared spectroscopy (FT-IR) in combination with tight-binding IR calculations. The results of FT-MS and FT-IR strongly suggest that C120 bucky peanuts, which have been theoretically predicted to be stable, were formed in the photoirradiated film. [ABSTRACT FROM AUTHOR]

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS). Its early phase is characterized by a relapse-remitting disease course, followed by disability progression in the later stage. While chronic inflammation accompanied with degeneration is well-established as the key pathological feature, the pathogenesis of MS, particularly progressive MS, remains elusive. Sulfatide is a major glycolipid component of myelin, and previous studies in experimental autoimmune encephalomyelitis mouse models have demonstrated it to have immune-protective functions. Notably, sulfatide concentration is increased in the serum and cerebrospinal fluid of patients with MS, particularly those in a progressive disease course. Here, we show that the myelin-glycolipid sulfatide displays an ability to suppress the proliferation of polyclonally activated human T cells. Importantly, this suppressive effect was impaired in T cells obtained from MS patients having higher disability status. Therefore, it is plausible that progression of MS is associated with an escape from the immune-regulatory effect of sulfatide. Our study suggests that, although the precise mechanisms remain unrevealed, an escape of T cells from immunosuppression by sulfatide is associated with disease progression in the advanced stage. Further studies will provide novel insights into the pathogenesis of MS, particularly regarding disease progression, and help develop novel treatment strategies for this challenging disease., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Objective: To clarify functional alterations of follicular helper T cells (Tfh) in myasthenia gravis (MG) because Tfh play important roles in helping B cells generate antibody-producing cells., Methods: A total of 24 immunotherapy-naive patients with anti-acetylcholine receptor (AchR) antibody-positive MG and 18 age-matched healthy subjects (HS) were enrolled. Samples from 6 patients were available for posttreatment analysis. Subsets of circulating Tfh (cTfh) and B cells were identified by flow cytometry analysis of surface molecules. Cytokine production by isolated cTfh subsets from 5 patients with MG and 5 HS was measured in vitro. Analysis was performed to examine the correlation between the frequency of cTfh subsets and that of plasmablasts and between cTfh subsets and the quantitative MG score., Results: cTfh increased with elevated expression of inducible T-cell costimulator (ICOS) in patients with MG. cTfh shifted to Th2 and Th17 over Th1 in MG. ICOS high cTfh produced significantly higher levels of interleukin (IL)-21, IL-4, and IL-17A than ICOS low cTfh only in patients with MG. The frequency of cTfh within CD4 T cells was more closely associated with disease severity than the serum anti-AchR antibody titer and frequency of plasmablasts within B cells. Abnormalities of cTfh were improved after immunotherapy in parallel with clinical improvement., Conclusions: Alternation of cTfh is a key feature in the development of MG and may become a biomarker for disease severity and therapeutic efficacy., Classification of Evidence: This study provides Class II evidence that the level of cTfh is associated with disease severity in patients with MG., (Copyright © 2021 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology.)

A two-dimensional system composed of supercritical fluid chromatography (SFC) and reverse phase liquid chromatography (RPLC) coupled a tandem mass spectrometry (MS) was developed for the quantitative analysis of vitamin D in daily oily supplements. Two six-port switching valves are configured, allowing four different valve positions. When the valve positions were fixed at Position A, this system worked at SFC-MS mode. When the valve positions switched between Position B and C, this system worked at a SFC-LC-MS switching mode. Vitamin D3 in two kinds of oily drops, Baby Ddrops and Vitamin AD drops, was determined at both SFC-MS and SFC-LC-MS switching modes by using the same system. The linearity, repeatability and recovery were investigated using the internal and external standard methods for the two modes. The results obtained from the internal standard method are better than those of the external standard method at either mode. The coefficient of determination (r 2 ) for the internal standard method is more than 0.999, with a linear range of 20-1000 μg/L. Both Baby Ddrops and Vitamin AD drops were analyzed with good repeatability (<3.21%) and recovery (94.1%-116.1%) using internal standard method. When calculated by the external standard method, as compared to SFC-MS mode, SFC-LC-MS switching mode has better linearity (r 2 >0.999), repeatability (Baby Ddrops: 4.45%, Vitamin AD drops: 1.12%) and recovery (86.3%-107.1%). The results indicate that the two-dimensional SFC-MS/SFC-LC-MS system is useful for determination of vitamin D in oily drops. If it works at SFC-MS mode, the internal standard is required. When SFC-LC-MS switching mode is used, external standard method can also obtain the accurate results with good precision., Competing Interests: Declaration of Competing Interest The authors declare that this article content has no conflicts of interest., (Copyright © 2020. Published by Elsevier B.V.)

Triterpenic acids possess rich biological activity. Due to slight differences in structure and polarity, the simultaneous determination of isomeric triterpenic acids is challenging. In the present work, a simple and effective approach to chromatographic separation of such compounds based on conventional C18 stationary phase with gradient elution was developed, which allowed the simultaneous separation of eleven analytes including euscaphic, arjunic, tormentic, arjunolic, asiatic, pomolic, maslinic, corosolic, oleanolic, ursolic and 2-Epi tormentic acid (internal standard). This approach with mass spectrometric detection and ultrasonic extraction was fast, sensitive and accurate for analyzing isomeric triterpenic acids in O. fragrans fruits with a toal duration of the analytical cycle (including pretreatment) within one hour. The LODs lie in ranges of 0.8-12 ng/mL (30 ng/mL for asiatic and corosolic acid). The developed method was validated and successfully applied in ten batches of O. fragrans fruits, which could reflect the detail content difference of triterpenic acid components., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is an autosomal-dominant type of leukoencephalopathy caused by gene mutation of colony stimulating factor 1 receptor, which is expressed mainly on monocyte lineage cells such as monocytes in the peripheral blood and microglia in the brain. Hence, microglial dysfunction is regarded as critical in the pathogenesis of ALSP. However, functional changes in these cells have not been elucidated. In this study, we report the phenotypic and functional alterations of monocytes in four patients with ALSP. Flow cytometric analysis revealed altered expression of antigen presentation- and migration-related molecules, an inflammatory shift in cytokine production and phagocytic impairment in ALSP monocytes. We speculate that the observed altered features of monocytes are mostly shared by microglial cells, leading to the clinical history and pathological characteristics of ALSP. Our analysis of PB monocytes provides novel insights into the pathogenesis of ALSP., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020. Published by Elsevier Inc.)

Astrapterocarpan (AP) is a bioactive constituent of Astragali Radix and was selected as a model compound for investigating the in vitro metabolism of pterocarpans in this study. Its in vitro metabolism was conducted by incubation with rat hepatic 9000g supernatant (S9) in the presence of an NADPH-generating system. At first, four compounds were isolated and their structures were elucidated as 6a-hydroxy-AP (M1), astrametabolin I [M2, 1a-hydroxy-9, 10-dimethoxy-pterocarp-1(2), 4-diene-3-one], 9-demethyl-AP (M3, nissolin) and 4-methoxy-astraisoflavan (M4, 7, 2'dihydroxy-4, 3', 4'-trimethoxy-isoflavan) on the basis of NMR data, respectively. Among them, M1, M2 and M4 were new compounds. Next, the metabolite profile of AP in rat hepatic S9 was obtained via HPLC-DAD-ESI-IT-TOF-MS n , and 40 new metabolites were tentatively identified. These newly identified metabolites included 9 monohydroxylated metabolites, 1 demethylated metabolite, 7 demethylated and monohydroxylated metabolites, 4 dihydroxylated metabolites, 1 hydration metabolite, 1 didemethylated metabolite, 2 glucosylated metabolites, 1 monohydroxylated and dehydrogenated metabolite, 2 monohydroxylated and demethylated and dehydrogenated metabolites, 2 dimerized metabolites, 3 dimerized and monohydroxylated metabolites, 2 dimerized and didemethylated metabolites, and 5 dimerized and demethylated metabolites. Finally, the major metabolic reactions of AP in rat hepatic S9 were summarized and found to be hydroxylation, demethylation, dimerization, hydration, and dehydrogenation. More importantly, the biotransformation from AP to M2 and the dimerization of AP by incubation with hepatic S9 were reported for the first time. In conclusion, this is the first report on the metabolism of a pure pterocarpan in animal tissues, and these findings will provide a solid basis for further studies on the metabolism of other pterocarpans., (Copyright © 2019 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

The light-driven splitting of water to oxygen (O 2 ) is catalyzed by a protein-bound tetra-manganese penta-oxygen calcium (Mn 4 O 5 Ca) cluster in Photosystem II. In the current study, we used a large-scale integration (LSI)-based amperometric sensor array system, designated Bio-LSI, to perform two-dimensional imaging of light-induced O 2 evolution from spinach leaves. The employed Bio-LSI chip consists of 400 sensor electrodes with a pitch of 250 μm for fast electrochemical imaging. Spinach leaves were illuminated to varying intensities of white light (400-700 nm) which induced oxygen evolution and subsequent electrochemical images were collected using the Bio-LSI chip. Bio-LSI images clearly showed the dose-dependent effects of the light-induced oxygen release from spinach leaves which was then significantly suppressed in the presence of urea-type herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Our results clearly suggest that light-induced oxygen evolution can be monitored using the chip and suggesting that the Bio-LSI is a promising tool for real-time imaging. To the best of our knowledge, this report is the first to describe electrochemical imaging of light-induced O 2 evolution using LSI-based amperometric sensors in plants.

Protein aggregation is the principal component of numerous protein misfolding pathologies termed proteinopathies, such as Alzheimer's disease, Parkinson's disease, prion disease, and AA amyloidosis with unmet treatment needs. Protein aggregation inhibitors have great potential for the prevention and treatment of proteinopathies. Here we report the development of an automated real-time microliter-scale high throughput screening (MSHTS) system for amyloid aggregation inhibitors using quantum-dot nanoprobes. Screening 504 crude extracts and 134 low molecular weight aromatic compounds revealed the relationship of amyloid-β (Aβ) aggregation inhibitory activities of plant extracts using a plant-based classification. Within the eudicots, rosids, Geraniales and Myrtales showed higher activity. Screening low molecular weight aromatic compounds demonstrated that the structure of tropolone endows it with potential Aβ aggregation inhibitory activity. The activity of the most active tropolone derivative was higher than that of rosmarinic acid. MSHTS also identified three chaperone molecules as tau aggregation inhibitors. These results demonstrate that our automated MSHTS system is a novel and robust tool that can be adapted to a wide range of compounds and aggregation-prone polypeptides.

Short-chain chlorinated paraffins (SCCPs), recently listed as Persistent Organic Pollutants (POPs) under the Stockholm Convention, are contained in commercial chlorinated paraffin (CP) products, which are used in industries such as metalworking fluids, sealants, and textiles. A novel method to determine SCCPs by comprehensive two-dimensional gas chromatography coupled with low resolution mass spectrometry was developed. Calibration curves of response factor versus chlorine content in two chlorine content ranges (R 2 were 0.9544 and 0.9736, respectively) were used for quantification of SCCP total concentration. Moreover, relative concentrations of 24 congener groups were also theoretically calculated. Results showed that this method was able to detect SCCP concentration in commercial CP products and urban air samples. SCCP contribution varied largely among different CP products, which is highly determined by carbon chain distribution in paraffins. SCCP concentration in urban air ranged between 12.8-49.1 ng m -3 during nine-month sampling period. The highest SCCP concentration appeared in summer, and the lowest concentration occurred in winter. Gas phase was dominantly occupied by lighter congeners such as C 10 group and Cl 6 group, while heavier congeners such as C 13 group and Cl 7 and Cl 8 groups contributed more in particle phase., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A column-switching system, composed of supercritical fluid chromatography (SFC) and reverse phase liquid chromatography/mass spectrometry (RPLC/MS) was developed for the analysis of vitamin D in oily and fatty matrices. The SFC with the similar retention behavior as normal-phase liquid chromatography (NPLC), was applied for an on-line clean-up of oily and fatty samples, then followed by separation and detection using a reverse-phase LC-MS/MS. Three SFC columns packed with materials of different functional groups (Silica, NH 2 , Diol) were compared and the column with diol groups, on which the retention time of vitamin D was the longest, was finally selected for purification of the samples. 100% methanol was chosen to carry vitamin D from the clean-up column to the pre-treatment column. It was also used as the mobile phase for the separation of vitamin D on a reverse phase C 18 column. Vitamin D 2 and D 3 were baseline separated by using this system. The linearity was calculated with a value of coefficient of determination (r 2 ) ≥ 0.998. The linear range is from 20 ng/mL to 200 ng/mL. Two kinds of liquid vitamin D 3 supplements (Baby Ddrops and Vitamin AD drops) were directly analyzed using this system without any fussy preparation procedure. The limit of detection (LOD) for vitamin D 3 in the two oily samples was estimated to be 10 ng/mL. The relative standard deviations (RSD) of intra- and inter-day precision, repeatability were 1.47%, 2.43% and 1.59% for Baby Ddrops and 5.76%, 8.24% and 5.86% for Vitamin AD drops. The recoveries vary between 84.3% and 102.8% with 7.1% RSD for Baby Ddrops and 90.8-109.6% with 5.83% RSD for Vitamin AD drops, respectively. These results suggest that the method based on the SFC-RPLC/MS column-switching system is simple and suitable for analysis of vitamin D in oily and fatty samples., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The epilepsy syndrome of autoimmune etiology, namely, autoimmune epilepsy has attracted attention in recent years, as was reflected in the new etiology of "immunity" in the Epilepsy Classification of the International League Against Epilepsy (2017). However, no specific tests other than neuronal antibodies have been established. We proposed a diagnostic algorithm for autoimmune epilepsy and preliminarily investigated its clinical utility. We applied this algorithm to 70 patients who were suspected as having autoimmune epilepsy from clinical symptoms and laboratory findings in our institute. At least one of the three neuronal antibodies (antibodies to N-methyl-D-aspartic acid receptor (NMDAR), glutamic acid decarboxylase (GAD), and voltage-gated potassium channels (VGKC) complex) was evaluated. In this two-step algorithm, the patients were initially screened by clinical features and then evaluated by laboratory findings (neuronal antibodies, cerebrospinal fluid (CSF), MRI, FDG-PET). The results of preliminary application of the algorithm are described. One of the three neuronal antibodies was positive in 13 patients. In this preliminary investigation, it was suggested that two or more abnormal findings in the diagnostic tests (CSF, MRI, FDG-PET) favors the diagnosis of autoimmune epilepsy. On the other hand, two patients with a positive neuronal antibody test failed the first step (clinical features), partly because epilepsy was not the major manifestation of autoimmune encephalitis (VGKC complex antibody) or due to a relatively low titer of the antibody (GAD antibody). Recruitment of the patient cohort with comprehensive neuronal antibody testing and multivariate analysis of laboratory findings is warranted for validation and modification of the proposed algorithm.

We herein report a 15-year-old girl who developed rapid progressive muscle weakness soon after the third injection of a bivalent human papilloma virus (HPV) vaccine. Although immunotherapies were performed for possible vaccine-related disorders, she died of respiratory failure 14 months after the onset of the disease. A genetic analysis identified a heterozygous p.P525L mutation of the fused in sarcoma (FUS) gene, and a histopathological analysis was also consistent with FUS-associated amyotrophic lateral sclerosis (ALS) without any evidence of neuroinflammation. We concluded the diagnosis to be FUS-ALS, although we cannot completely rule out the possibility that the vaccination worked as a trigger.

Background: B cells play an important role in the development of multiple sclerosis (MS), but can also exhibit regulatory functions through IL-10 production. Toll-like receptors (TLR) and CD40 signaling are likely to be involved in this process., Objective: To investigate the ability of MS B cells to produce IL-10 in response to TLR stimulation in the presence or absence of CD40 co-stimulation., Methods: Peripheral blood mononuclear cells obtained from 34 MS patients and 24 matched healthy participants (HS) were stimulated through either TLR4 or TLR9 alone, or together with CD40. Intracellular cytokine production was analyzed by flow cytometry., Results: The frequency of IL-10-producing cells in total B cells after either TLR9 or CD40 stimulation was significantly lower in MS than HS, regardless of disease phase. The frequency of IL-10 producing B cells after TLR4 stimulation did not differ significantly between HS and MS, regardless of disease phase. TLR4 and CD40 co-stimulation synergistically increased the frequency of IL-10-producing but not pro-inflammatory cytokine-producing B cells at MS relapse. This effect was observed in both CD27 - naïve and CD27 + memory B cells. The frequency of IL-10-producing B cells following CD40 stimulation was significantly higher in interferon-β responders than non-treated MS patients. Finally, we confirmed that the frequency of IL-10-producing B cells positively correlated with IL-10 production quantity by B cells using magnetic-isolated B cells., Conclusions: Cross-talk between TLR4 and CD40 signaling plays a crucial role in regulating IL-10 production by B cells during MS relapses, which may promote recovery from relapse. CD40 signaling in B cells is involved in the response to interferon-β in MS. Collectively, TLR4 and CD40 signaling in B cells may provide a promising target for MS therapy., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Multiple sclerosis (MS) is a T cell-mediated autoimmune disease. Fingolimod, a highly effective disease-modifying drug for MS, retains CCR7 + central memory T cells in which autoaggressive T cells putatively exist, in secondary lymphoid organs, although relapse may still occur in some patients. Here, we analyzed the T cell phenotypes of fingolimod-treated, fingolimod-untreated patients, and healthy subjects. The frequency of CD56 + T cells and granzyme B-, perforin-, and Fas ligand-positive T cells significantly increased during fingolimod treatment. Each T cell subpopulation further increased during relapse. Interestingly, T cells from fingolimod-treated patients exhibited interferon-γ biased production, and more myelin basic protein-reactive cells was noted in CD56 + than in CD56 - T cells. It is likely that the altered T cell phenotypes play a role in MS relapse in fingolimod-treated patients. Further clinical studies are necessary to investigate whether altered T cell phenotypes are a biomarker for relapse under fingolimod therapy.

A rapid and sensitive ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry method was developed and validated for the simultaneous determination of eight major active components (magnoflorine, menisperine, 20-hydroxyecdysone, cepharanthine, columbamine, jatrorrhizine, columbin, and palmatine) in Radix Tinosporae. The separation was performed on an InterSustainSwift C18 column (1.9 μm, 2.1 id × 100 mm) at 40 °C with a gradient elution. A mixture of acetonitrile and methanol (v/v = 1:1) and ammonium acetate buffer (25 mmol/L ammonium acetate with 0.2% formic acid) were used as mobile phases, and the flow rate was set at 0.4 mL/min. The recovery was tested in real samples and calculated to be 86.97-111.28%, and all the compounds showed good linearity (r > 0.998) in relatively wide concentration ranges. The developed method was applied to the determination of eight active compounds in real herb samples, which were collected from four different places. It has been demonstrated that the proposed method has great potential for the quality control of the traditional Chinese medicine Radix Tinosporae., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)