The Phytochemical, Quercetin, Attenuates Nociceptive and Pathological Pain: Neurophysiological Mechanisms and Therapeutic Potential.

Although phytochemicals are plant-derived toxins that are primarily produced as a form of defense against insects or microbes, several lines of study have demonstrated that the phytochemical, quercetin, has several beneficial biological actions for human health, including antioxidant and inflammatory effects without side effects. Quercetin is a flavonoid that is widely found in fruits and vegetables. Since recent studies have demonstrated that quercetin can modulate neuronal excitability in the nervous system, including nociceptive sensory transmission via mechanoreceptors and voltage-gated ion channels, and inhibit the cyclooxygenase-2-cascade, it is possible that quercetin could be a complementary alternative medicine candidate; specifically, a therapeutic agent against nociceptive and pathological pain. The focus of this review is to elucidate the neurophysiological mechanisms underlying the modulatory effects of quercetin on nociceptive neuronal activity under nociceptive and pathological conditions, without inducing side effects. Based on the results of our previous research on trigeminal pain, we have confirmed in vivo that the phytochemical, quercetin, demonstrates (i) a local anesthetic effect on nociceptive pain, (ii) a local anesthetic effect on pain related to acute inflammation, and (iii) an anti-inflammatory effect on chronic pain. In addition, we discuss the contribution of quercetin to the relief of nociceptive and inflammatory pain and its potential clinical application.

[A case of recurrent Campylobacter fetus meningitis, occurring two months after the |initial infection was successfully treated].

A 43-year-old man was admitted to our department due to fever and headache. The cerebrospinal fluid analysis confirmed bacterial meningitis. Campylobacter species were isolated from blood cultures on the third day of admission. The patient was treated with meropenem (MEPM) and discharged on the 17th day. However, he experienced a recurrence of meningitis and was readmitted on the 68th day, initiating MEPM therapy. Campylobacter fetus was isolated from cerebrospinal fluid cultures on the 74th day. MEPM was continued until the 81st day, followed by one month of minocycline (MINO) therapy. The patient had an uneventful recovery without further recurrence. This case highlights the potential for recurrence of Campylobacter fetus meningitis approximately two months after the resolution of the initial infection. In addition to carbapenem therapy for at least two weeks, the adjunctive administration of MINO may be beneficial.

Host-to-graft propagation of inoculated α-synuclein into transplanted human induced pluripotent stem cell-derived midbrain dopaminergic neurons.

Introduction: Cell therapeutic clinical trials using fetal mesencephalic tissue provided a proof-of-concept for regenerative therapy in patients with Parkinson's disease. Postmortem studies of patients with fetal grafts revealed that α-synuclein+ Lewy body (LB)-like inclusions emerged in long-term transplantation and might worsen clinical outcomes even if the grafts survived and innervated in the recipients. Various studies aimed at addressing whether host-derived α-synuclein could be transferred to the grafted neurons to assess α-synuclein+ inclusion appearance in the grafts. However, determining whether α-synuclein in the grafted neurons has been propagated from the host is difficult due to the intrinsic α-synuclein expression. Methods: We induced midbrain dopaminergic (mDA) neurons from human induced pluripotent stem cells (hiPSCs) and transplanted them into the striatum of immunodeficient rats. The recombinant human α-synuclein preformed fibrils (PFFs) were inoculated into the cerebral cortex after transplantation of SNCA-/- hiPSC-derived mDA neural progenitors into the striatum of immunodeficient rats to evaluate the host-to-graft propagation of human α-synuclein PFFs. Additionally, we examined the incorporation of human α-synuclein PFFs into SNCA-/- hiPSC-derived mDA neurons using in vitro culture system. Results: We detected human α-synuclein-immunoreactivity in SNCA-/- hiPSC-derived mDA neurons that lacked endogenous α-synuclein expression in vitro. Additionally, we observed host-to-graft α-synuclein propagation into the grafted SNCA-/- hiPSC-derived mDA neurons. Conclusion: We have successfully proven that intracerebral inoculated α-synuclein PFFs are propagated and incorporated from the host into grafted SNCA-/- hiPSC-derived mDA neurons. Our results contribute toward the basic understanding of the molecular mechanisms related to LB-like α-synuclein deposit formation in grafted mDA neurons.

Effect of pre-rolling aging treatment on evolutions of the microstructure and the texture of aluminium alloy 7005 subjected to heavy cold rolling.

Aluminium alloy 7005 is widely used for structural purposes because of its attractive properties such as good weldability and age-hardening capability. However, since the workability of this alloy falls after a short period of natural aging, the application of cold rolling for the production of strain-hardened sheets of this alloy is a challenge. Two solutions proposed to overcome this challenge are as follows: (a) immediate rolling of the alloy after solution treatment and (b) rolling of the alloy after artificial aging. However, there is no comprehensive study comparing the effect of pre-rolling aging treatments on the evolutions of microstructure and texture of the alloy through heavy cold rolling. This subject is the aim of the present study. For this purpose, different pieces of the alloy are subjected to three different heat treatments before rolling, and afterward, they are rolled to obtain a thickness reduction of 80%. Scanning electron microscopy with electron backscattered diffraction observations are applied to study the evolutions of the microstructure and the texture of the alloy. Results show that the progression of pre-rolling aging decreases the incidence of micro-scaled shear bands by rolling. In addition, the rolling texture intensity decreases with the advancement of pre-rolling aging. Mechanisms responsible for this effect are discussed.

Gene deletion of long-chain acyl-CoA synthetase 4 attenuates xenobiotic chemical-induced lung injury via the suppression of lipid peroxidation.

Long-chain acyl-CoA synthetase (ACSL) 4 converts polyunsaturated fatty acids (PUFAs) into their acyl-CoAs and plays an important role in maintaining PUFA-containing membrane phospholipids. Here we demonstrated decreases in various kinds of PUFA-containing phospholipid species in ACSL4-deficient murine lung. We then examined the effects of ACSL4 gene deletion on lung injury by treating mice with two pulmonary toxic chemicals: paraquat (PQ) and methotrexate (MTX). The results showed that ACSL4 deficiency attenuated PQ-induced acute lung lesion and decreased mortality. PQ-induced lung inflammation and neutrophil migration were also suppressed in ACSL4-deficient mice. PQ administration increased the levels of phospholipid hydroperoxides in the lung, but ACSL4 gene deletion suppressed their increment. We further found that ACSL4 deficiency attenuated MTX-induced pulmonary fibrosis. These results suggested that ACSL4 gene deletion might confer protection against pulmonary toxic chemical-induced lung injury by reducing PUFA-containing membrane phospholipids, leading to the suppression of lipid peroxidation. Inhibition of ACSL4 may be promising for the prevention and treatment of chemical-induced lung injury.

Effects of polycaprolactone degradation products on the water flea, Daphnia magna: Carbodiimide additives have acute and chronic toxicity.

Plastics have benefited our lives in many ways, but their long persistence in the environment causes serious problems. Rapid decomposition and detoxification of plastics after use are significant challenges. As a possible solution, biodegradable plastics have attracted attention, and for environmental risk assessment research on polymer toxicity, use of indicator organisms, like water fleas and fish, has increased globally. However, such research often focuses on standardized substances without considering changes in toxicity due to plastic degradation products. Additionally, tests generally focus on acute toxicity, while long-term effects on organismal reproduction and lifespan are largely unknown. Understanding the impact of degraded polymers on biological activities is crucial for accurate risk assessment. In this study, we investigated the biological toxicity of substances generated during degradation of polycaprolactone (PCL), a common biodegradable plastic, using the indicator organism, Daphnia magna. We examined PCL, oligocaprolactones (OCLs), and monomers resulting from polymer cleavage, as well as carbodiimides, added during polyester synthesis. As a result, PCL, which is insoluble in water, reduced individual survival and total number of offspring at an exposure concentration of 100 mg/L, while no toxicity was observed for water-soluble degradation products, OCLs, and monomers. Furthermore, carbodiimides, which are expected to be released during PCL degradation, showed strong toxicity, significantly reducing individual survival and total number of offspring at 0.1-10 mg/L. These findings suggest that changes in physical properties due to polymer degradation and release of additives can significantly alter their toxicity.

Novel role of DONSON in CMG helicase assembly during vertebrate DNA replication initiation.

CMG (Cdc45-MCM-GINS) helicase assembly at the replication origin is the culmination of eukaryotic DNA replication initiation. This process can be reconstructed in vitro using defined factors in Saccharomyces cerevisiae; however, in vertebrates, origin-dependent CMG formation has not yet been achieved partly due to the lack of a complete set of known initiator proteins. Since a microcephaly gene product, DONSON, was reported to remodel the CMG helicase under replication stress, we analyzed its role in DNA replication using a Xenopus cell-free system. We found that DONSON was essential for the replisome assembly. In vertebrates, DONSON physically interacted with GINS and Polε via its conserved N-terminal PGY and NPF motifs, and the DONSON-GINS interaction contributed to the replisome assembly. DONSON's chromatin association during replication initiation required the pre-replicative complex, TopBP1, and kinase activities of S-CDK and DDK. Both S-CDK and DDK required DONSON to trigger replication initiation. Moreover, human DONSON could substitute for the Xenopus protein in a cell-free system. These findings indicate that vertebrate DONSON is a novel initiator protein essential for CMG helicase assembly.

Suppression of the Excitability of Rat Nociceptive Primary Sensory Neurons Following Local Administration of the Phytochemical, Quercetin.

Although the modulatory effect of quercetin on voltage-gated Na, K, and Ca channels has been studied in vitro, the in vivo effect of quercetin on the excitability of nociceptive primary neurons remains to be determined. The aim of the present study was to examine whether acute local quercetin administration to rats attenuates the excitability of nociceptive trigeminal ganglion (TG) neurons in response to mechanical stimulation in vivo. Extracellular single unit recordings were made from TG neurons of anesthetized rats in response to orofacial non-noxious and noxious mechanical stimulation. The mean firing frequency of TG neurons in response to both non-noxious and noxious mechanical stimuli was dose-dependently inhibited by quercetin, and maximum inhibition of the discharge frequency of both non-noxious and noxious mechanical stimuli was seen within 10 min. The inhibitory effect of quercetin lasted for 15 minutes and was reversible. The mean magnitude of inhibition on TG neuronal discharge frequency with 10 mM quercetin was almost equal to that of the local anesthetic, 2% lidocaine. These results suggest that local injection of quercetin into the peripheral receptive field suppresses the excitability of nociceptive primary sensory neurons in the TG, possibly via inhibition of voltage-gated Na channels and opening voltage-gated K channels. PERSPECTIVE: Local administration of the phytochemical, quercetin, as a local anesthetic may provide relief from trigeminal nociceptive pain with smallest side effects, thus contributing to the area of complementary and alternative medicines.

Targeted single-cell genomics reveals novel host adaptation strategies of the symbiotic bacteria Endozoicomonas in Acropora tenuis coral.

BACKGROUND: Endozoicomonas bacteria symbiosis with various marine organisms is hypothesized as a potential indicator of health in corals. Although many amplicon analyses using 16S rRNA gene have suggested the diversity of Endozoicomonas species, genome analysis has been limited due to contamination of host-derived sequences and difficulties in culture and metagenomic analysis. Therefore, the evolutionary and functional potential of individual Endozoicomonas species symbiotic with the same coral species remains unresolved. RESULTS: In this study, we applied a novel single-cell genomics technique using droplet microfluidics to obtain single-cell amplified genomes (SAGs) for uncultured coral-associated Endozoicomonas spp. We obtained seven novel Endozoicomonas genomes and quantitative bacterial composition from Acropora tenuis corals at four sites in Japan. Our quantitative 16S rRNA gene and comparative genomic analysis revealed that these Endozoicomonas spp. belong to different lineages (Clade A and Clade B), with widely varying abundance among individual corals. Furthermore, each Endozoicomonas species possessed various eukaryotic-like genes in clade-specific genes. It was suggested that these eukaryotic-like genes might have a potential ability of different functions in each clade, such as infection of the host coral or suppression of host immune pathways. These Endozoicomonas species may have adopted different host adaptation strategies despite living symbiotically on the same coral. CONCLUSIONS: This study suggests that coral-associated Endozoicomonas spp. on the same species of coral have different evolutional strategies and functional potentials in each species and emphasizes the need to analyze the genome of each uncultured strain in future coral-Endozoicomonas relationships studies. Video Abstract.

Cooperative effects of oocytes and estrogen on the forkhead box L2 expression in mural granulosa cells in mice.

Forkhead box L2 (FOXL2) plays a critical role in the development and function of mammalian ovaries. In fact, the causative effects of FOXL2 misregulations have been identified in many ovarian diseases, such as primary ovarian insufficiency and granulosa cell tumor; however, the mechanism by which FOXL2 expression is regulated is not well studied. Here, we showed that FOXL2 expression in ovarian mural granulosa cells (MGCs) requires stimulation by both oocyte-derived signals and estrogen in mice. In the absence of oocytes or estrogen, expression of FOXL2 and its transcriptional targets, Cyp19a1 and Fst mRNA, in MGCs were significantly decreased. Moreover, expression levels of Sox9 mRNA, but not SOX9 protein, were significantly increased in the FOXL2-reduced MGCs. FOXL2 expression in MGCs was maintained with either oocytes or recombinant proteins of oocyte-derived paracrine factors, BMP15 and GDF9, together with estrogen, and this oocyte effect was abrogated with an ALK5 inhibitor, SB431542. In addition, the FOXL2 level was significantly decreased in MGCs isolated from Bmp15-/- /Gdf9+/- mice. Therefore, oocyte, probably with estrogen, plays a critical role in the regulation of FOXL2 expression in mural granulosa cells in mice.

Early neurogenic properties of iPSC-derived neurosphere formation in Japanese macaque monkeys.

Non-human primates are important models for investigations of neural development and evolution, and the use of Japanese macaque monkeys has especially contributed to the advancement of neuroscience studies. However, these studies are restricted by the number of animals able to be evaluated and the invasiveness of the methodologies. Induced pluripotent stem cells (iPSCs) can provide an alternative strategy for investigating neural development in vitro. We have established direct neurosphere (dNS) formation cultures of primate iPSCs as an in vitro model of early neurodevelopment in primate species. Here, we used dNS formation and neuronal differentiation cultures established from Japanese macaque iPSCs (jm-iPSCs) to investigate the usefulness of these cells as an in vitro model of early neural development. Time-course analyses of developmental potency and gene expression kinetics were performed during dNS formation culture of jm-iPSCs. During a 1-week culture, jm-iPSC-derived dNSs became neurogenic by day 3 and underwent stepwise expression changes of key developmental regulators along early neural development in a similar manner to chimpanzee dNS formation previously reported. Meanwhile, a subset of genes, including CYP26A1 and NPTX1, showed differential expression propensity in Japanese macaque, chimpanzee, and human iPSC-derived dNSs. Spontaneous upregulation of NOTCH signaling-associated genes HES5 and DLL1 was also observed in neuronal differentiation cultures of Japanese macaque but not chimpanzee dNSs, possibly reflecting the earlier neurogenic competence in Japanese macaques. The use of jm-iPSCs provides an alternative approach to neurological studies of primate development. Furthermore, jm-iPSCs can be used to investigate species differences in early neural development that are key to primate evolution.

Effects of oxygen tension and humidity on the preimplantation development of mouse embryos produced by in vitro fertilization: analysis using a non-humidifying incubator with time-lapse cinematography.

To examine the effects of oxygen tension and humidity on early embryonic development, the preimplantation development of mouse embryos produced by in vitro fertilization was assessed by time-lapse cinematography to evaluate morphokinetic development with higher precision. Zygotes were produced from spermatozoa and oocytes from ICR mice and cultured in KSOM under low or high oxygen tension in a non-humidified incubator with time-lapse cinematography (CCM-iBIS). The developmental rates of embryos to the 4-cell and blastocyst stages under lower oxygen tension in CCM-iBIS were significantly higher than those under higher oxygen tension in CCM-iBIS. Ninety-six hours after insemination, a large number of embryos cultured under low oxygen tension developed to the hatching blastocyst stage. Embryonic development was more synchronized under lower oxygen tension. Non-humidified cultures did not affect embryonic development. On average, mouse embryos cultured at lower oxygen tension reached 2-cell at 18 h, 3-cell at 39 h, 4-cell at 40 h, initiation of compaction at 58 h, morula at 69 h, and blastocyst at 82 h after insemination. In conclusion, lower oxygen tension better supports preimplantation development of mouse embryos fertilized in vitro, and non-humidified culture conditions do not influence the embryonic development in vitro.

Expression and regulation of estrogen receptor 2 and its coregulators in mouse granulosa cells.

The cooperative effects of estrogen and oocyte-derived paracrine factors (ODPFs) play critical roles in the normal development of ovarian follicles; however, the mechanism underlying this cooperation has not been well studied. The present study aimed to determine whether ODPFs affect estrogen signaling by regulating the expression of estrogen receptor (ESR) and its coregulators in mouse granulosa cells. Some transcripts encoding ESR coregulators were differentially expressed between cumulus and mural granulosa cells (MGCs). The transcript levels of ESR coregulators, including nuclear receptor corepressor 1 and activator 2, in cumulus cells were significantly suppressed by ODPFs; however, they increased when cumulus cell-oocyte complexes were treated with the transforming growth factor beta receptor I inhibitor, SB431542. Moreover, MGCs exhibited significantly higher ESR2 protein and transcript levels than those in cumulus cells. ODPFs promoted Esr2 expression in cumulus cells but had no effect on that in MGCs. Overall, regulation of the expression of ESR2 and its coregulators in cumulus cells by oocytes seems to be one of the mechanisms underlying estrogen-oocyte cooperation in well-developed antral follicles in mice.

Inelastic Electron Transport and Ortho-Para Fluctuation of Water Molecule in H2O@C60 Single Molecule Transistors.

Light molecules such as H2O are the systems in which we can have access to quantum mechanical information on their constituent atoms. Here, we have investigated electron transport through H2O@C60 single molecule transistors (SMTs). The H2O@C60 SMTs exhibit Coulomb stability diagrams that show multiple tunneling-induced excited states below 30 meV. Furthermore, we have performed terahertz (THz) photocurrent spectroscopy on H2O@C60 SMTs and confirmed the same excitations. From comparison between experiment and theory, the excitations observed below 10 meV are identified to be the quantum rotational excitations of the water molecule. Surprisingly, the quantum rotational excitations of both para- and ortho-water molecule are observed simultaneously even for a single water molecule, indicating that the fluctuation between the ortho- and para-water states takes place in a time scale shorter than our measurement time (∼1 min), probably by the interaction between the encapsulated water molecule and conducting electrons.

The effects of amino acid/protein supplementation in patients undergoing hemodialysis: A systematic review and meta-analysis of randomized controlled trials.

BACKGROUND & AIMS: We evaluated the efficacy of the intervention consisting of amino acid/protein supplementation on muscle mass, muscle strength and physical function in patients on hemodialysis. METHODS: This systematic review and meta-analysis identified potential studies through a systematic search of 4 electronic databases and references from eligible studies from database inception to August 2020. We included only randomized controlled trials reporting the efficacy of amino acid/protein supplementation on muscle mass, muscle strength and physical function in patients on hemodialysis. RESULTS: Of 6529 unique citation records, 4 studies including 243 participants were selected for inclusion in the meta-analysis. Although there were no significant differences in muscle mass and muscle strength between the intervention and control groups, amino acid/protein supplementation was shown to significantly improve physical function (shuttle walk, MD 32.7, 95% CI 21.7 to 43.7, P < 0.001; gait speed, MD 0.07, 95% CI 0.01 to 0.13, P = 0.02; timed up and go, MD -0.42, 95% CI -0.68 to -0.15, P = 0.002) in patients on hemodialysis. CONCLUSIONS: We confirmed the positive effect of amino acid/protein supplementation on physical function in people undergoing hemodialysis. However, there is still insufficient evidence, and more rigorously designed randomized controlled trials with high quality are needed.

Effects of Hydrastis Canadensis, Commiphora Habessinica, Phytolacca Americana, and Echinacea Purpurea on Bacterial Growth.

CONTEXT: With the rise of antibiotic resistance, new strategies are needed to treat minor bacterial infections so that conventional antibiotics may be reserved for more serious conditions. One herbal formula, known as the HMPE formula, is often prescribed for minor infections. It includes Hydrastis canadensis (H. canadensis), Commiphora habessinica (C. habessinica), Phytolacca americana (P. americana), and Echinacea purpurea (E. purpurea). These herbs offer promise as treatments that may inhibit bacterial growth and stimulate the immune system. OBJECTIVE: To investigate the antibacterial effects of the HMPE formula and its constituent herbs against two organisms, Staphylococcus epidermidis and Escherichia coli. DESIGN: The research team performed an in-vitro study. SETTING: The study occurred at the Helfgott Research Institute at the National University of Natural Medicine in Portland, OR, USA. INTERVENTION: The study tested HMPE and each of its ingredients alone for antibacterial properties. OUTCOME MEASURES: The outcome measure was a disc diffusion assay. Sterile paper discs were impregnated with 15 µl of E. purpurea, H. canadensis, C. habessinica , or P. americana as herbal tinctures; with the complete HMPE formula; or with 65% ethanol as the negative control, and dried at room temperature for 40 minutes. Commercially prepared 10 µg ampicillin discs were used as a positive control. RESULTS: H. Canadensis and, to a lesser extent, the complete HMPE formula significantly inhibited the growth of the gram-positive bacteria Staphylococcus epidermidis, but not the gram-negative bacteria Escherichia coli. C. habessinica, P. americana, and E. purpurea alone did not inhibit growth of either bacterial strain. CONCLUSIONS: The results demonstrated that H. canadensis had antibacterial activity against S. epidermidis, but the HMPE formula was not active against S. epidermidis, when a zone of inhibition threshold of 12 millimeters (mm) was used to determine antibiotic activity. Because the HMPE formula was shown to be less effective than H. canadensis alone, the formula might benefit from an increased percentage of H. canadensis.

Neural Decoding of Multi-Modal Imagery Behavior Focusing on Temporal Complexity.

Mental imagery behaviors of various modalities include visual, auditory, and motor behaviors. Their alterations are pathologically involved in various psychiatric disorders. Results of earlier studies suggest that imagery behaviors are correlated with the modulated activities of the respective modality-specific regions and the additional activities of supramodal imagery-related regions. Additionally, despite the availability of complexity analysis in the neuroimaging field, it has not been used for neural decoding approaches. Therefore, we sought to characterize neural oscillation related to multimodal imagery through complexity-based neural decoding. For this study, we modified existing complexity measures to characterize the time evolution of temporal complexity. We took magnetoencephalography (MEG) data of eight healthy subjects as they performed multimodal imagery and non-imagery tasks. The MEG data were decomposed into amplitude and phase of sub-band frequencies by Hilbert-Huang transform. Subsequently, we calculated the complexity values of each reconstructed time series, along with raw data and band power for comparison, and applied these results as inputs to decode visual perception (VP), visual imagery (VI), motor execution (ME), and motor imagery (MI) functions. Consequently, intra-subject decoding with the complexity yielded a characteristic sensitivity map for each task with high decoding accuracy. The map is inverted in the occipital regions between VP and VI and in the central regions between ME and MI. Additionally, replacement of the labels into two classes as imagery and non-imagery also yielded better classification performance and characteristic sensitivity with the complexity. It is particularly interesting that some subjects showed characteristic sensitivities not only in modality-specific regions, but also in supramodal regions. These analyses indicate that two-class and four-class classifications each provided better performance when using complexity than when using raw data or band power as input. When inter-subject decoding was used with the same model, characteristic sensitivity maps were also obtained, although their decoding performance was lower. Results of this study underscore the availability of complexity measures in neural decoding approaches and suggest the possibility of a modality-independent imagery-related mechanism. The use of time evolution of temporal complexity in neural decoding might extend our knowledge of the neural bases of hierarchical functions in the human brain.

Multisystem Langerhans cell histiocytosis in an adult non-smoker treated with steroid therapy.

We describe the case of a 29-year-old female non-smoker who was treated with steroid therapy for a subacute exacerbation of multisystem Langerhans cell histiocytosis (MS-LCH) with worsening lung, skin, and oral mucosal lesions. The patient developed pneumonia, and computed tomography (CT) showed multiple thin-walled cavities. Transbronchial lung cryobiopsy (TBLC) specimens revealed Langerhans cells, which were positive for CD1a and S-100 expression. Similar histological findings were detected in the submandibular gland, skin, and tooth. On the basis of these findings, the patient was diagnosed with MS-LCH and subsequently treated with steroid therapy. From the literature review, case reports of non-smokers with pulmonary lesions that worsened and required treatment are rare. Almost all cases recurred and needed additional treatment. This case study contributes to our understanding of the potential role of steroid therapy in MS-LCH treatment. Additionally, TBLC is a novel, potentially safer, diagnostic tool that has not been previously described for LCH.