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Intrarenal dopamine plays a protective role against the development of diabetic nephropathy during the early stages of the disease. In streptozotocin-induced diabetic mice with a renal-specific catechol-O-methyl transferase knockout, intrarenal dopamine was found to suppress glomerular hyperfiltration, reduce oxidative stress and inflammation, and inhibit fibrosis. However, while dopamine activation in streptozotocin-induced diabetic models has been shown to provide renal protection, the role of dopamine in models of naturally induced diabetes mellitus is still unclear. In the present study, we administered 10 mg/kg p.o. benserazide, a peripheral decarboxylase inhibitor, to Spontaneously Diabetic Torii rats daily, in order to investigate the activation of the renal dopaminergic system during diabetic nephropathy progression. Our findings show that peripheral dopamine decreased urinary 8-iso-prostaglandin F2a and suppressed increases in plasma cystatin C levels. This study demonstrates that a reduction in peripheral dopamine can exacerbate renal dysfunction, even in the early stages of diabetic nephropathy characterized by glomerular hyperfiltration, thereby clarifying the pivotal role of endogenous peripheral dopamine in modulating oxidative stress and kidney performance.
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PURPOSE: It was reported that maternal-infant bonding failure predicts abusive parenting. Maternal-infant bonding is important to prevent child abuse. This study aimed to investigate the association between prenatal depressive symptoms, anxiety, cortisol, and oxytocin levels, and postnatal maternal-infant bonding. METHODS: The participants completed a self-report prenatal questionnaire that included the Edinburgh Postnatal Depression Scale (EPDS) and State-Trait Anxiety Inventory (STAI) in the second trimester. Blood and saliva were collected in the second trimester. Cortisol levels were measured in plasma, while oxytocin levels were measured in saliva. Postnatal questionnaires, including the Mother-to-Infant Bonding Scale (MIBS), were administered at 2-5 days, 1 month, and 3 months postpartum. Multiple linear regression and generalized estimating equation (GEE) were conducted for analysis. RESULTS: Sixty-six primiparas participated in the study. Prenatal depressive symptoms (EPDS ≥ 9) and anxiety (STAI-S ≥ 42) were observed in 21.2% and 28.8% of the participants, respectively. The median cortisol and oxytocin levels were 21.0 µg/dL and 30.4 pg/mL, respectively. Multivariate linear regression showed that postnatal social support, prenatal depressive symptoms, anxiety, and salivary oxytocin levels predicted MIBS scores at 2-5 days postpartum. At 1 month postpartum, household income, history of miscarriage, postnatal social support, and prenatal anxiety predicted MIBS scores. At 3 months postpartum, only postnatal social support predicted MIBS scores. The results of GEE showed that prenatal anxiety, oxytocin levels, postpartum period, household income, and postpartum social support were associated with MIBS scores. CONCLUSION: Prenatal depressive symptoms, anxiety, and lower salivary oxytocin levels were predicted to worsen maternal-infant bonding at 2-5 days postpartum. Prenatal anxiety was predicted to cause the same 1 month postpartum. Measuring prenatal depressive symptoms, anxiety, and salivary oxytocin levels may render the assessment of the risk of maternal-infant bonding failure during the early postpartum period and intervene during pregnancy possible.
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Ansiedad , Depresión , Hidrocortisona , Relaciones Madre-Hijo , Apego a Objetos , Oxitocina , Saliva , Humanos , Femenino , Oxitocina/análisis , Oxitocina/sangre , Oxitocina/metabolismo , Embarazo , Saliva/química , Adulto , Ansiedad/psicología , Estudios Longitudinales , Relaciones Madre-Hijo/psicología , Hidrocortisona/análisis , Depresión/psicología , Depresión/metabolismo , Japón , Periodo Posparto/psicología , Encuestas y Cuestionarios , Madres/psicología , Depresión Posparto/psicología , Lactante , Segundo Trimestre del Embarazo , Escalas de Valoración Psiquiátrica , Paridad , Pueblos del Este de AsiaRESUMEN
Effective early-stage markers for predicting which patients are at risk of developing SARS-CoV-2 infection have not been fully investigated. Here, we performed comprehensive serum metabolome analysis of a total of 83 patients from two cohorts to determine that the acceleration of amino acid catabolism within 5 days from disease onset correlated with future disease severity. Increased levels of de-aminated amino acid catabolites involved in the de novo nucleotide synthesis pathway were identified as early prognostic markers that correlated with the initial viral load. We further employed mice models of SARS-CoV2-MA10 and influenza infection to demonstrate that such de-amination of amino acids and de novo synthesis of nucleotides were associated with the abnormal proliferation of airway and vascular tissue cells in the lungs during the early stages of infection. Consequently, it can be concluded that lung parenchymal tissue remodeling in the early stages of respiratory viral infections induces systemic metabolic remodeling and that the associated key amino acid catabolites are valid predictors for excessive inflammatory response in later disease stages.
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COVID-19 , Neumonía , Humanos , Animales , Ratones , SARS-CoV-2 , ARN Viral , AminoácidosRESUMEN
Dyskinesia is involuntary movement caused by long-term medication with dopamine-related agents: the dopamine agonist 3,4-dihydroxy-L-phenylalanine (L-DOPA) to treat Parkinson's disease (L-DOPA-induced dyskinesia [LID]) or dopamine antagonists to treat schizophrenia (tardive dyskinesia [TD]). However, it remains unknown why distinct types of medications for distinct neuropsychiatric disorders induce similar involuntary movements. Here, we search for a shared structural footprint using magnetic resonance imaging-based macroscopic screening and super-resolution microscopy-based microscopic identification. We identify the enlarged axon terminals of striatal medium spiny neurons in LID and TD model mice. Striatal overexpression of the vesicular gamma-aminobutyric acid transporter (VGAT) is necessary and sufficient for modeling these structural changes; VGAT levels gate the functional and behavioral alterations in dyskinesia models. Our findings indicate that lowered type 2 dopamine receptor signaling with repetitive dopamine fluctuations is a common cause of VGAT overexpression and late-onset dyskinesia formation and that reducing dopamine fluctuation rescues dyskinesia pathology via VGAT downregulation.
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Discinesia Inducida por Medicamentos , Trastornos Parkinsonianos , Ratones , Animales , Agonistas de Dopamina/efectos adversos , Levodopa/efectos adversos , Dopamina , Antiparkinsonianos/efectos adversos , Trastornos Parkinsonianos/inducido químicamente , Trastornos Parkinsonianos/patología , Discinesia Inducida por Medicamentos/etiología , Discinesia Inducida por Medicamentos/tratamiento farmacológico , Discinesia Inducida por Medicamentos/patología , Oxidopamina/efectos adversos , Ácido gamma-Aminobutírico/efectos adversosRESUMEN
BACKGROUND: Epidermoid cysts in intrapancreatic accessory spleen (ECIPAS) are a rare lesion. Its pathogenesis, including the origin of cystic epithelium, is not well established. We aimed to elucidate new aspects of the pathological features of ECIPAS to clarify its pathogenesis. METHODS: Six cases of ECIPAS were included in this study. As well as histopathological analysis, to elucidate the features and nature of cystic epithelial cells, immunohistochemical analysis including Pbx1 and Tlx1 and imaging mass spectrometry was performed. RESULTS: Histologically, the cysts were covered by either monolayered or multilayered epithelium. Immunohistochemistry revealed that the epithelial cells in multilayered epithelium exhibited different attributes between the basal and superficial layers. Few epithelial cells had abundant clear cytoplasm and were immunohistochemically positive for adipophilin, suggesting lipid-excreting function. The intracystic fluid contained cholesterol clefts and foamy macrophages, and imaging mass spectrometry revealed the accumulation of lipids. Immunohistochemical analysis indicated that the epithelial cells were positive for Pbx1 in some cases. CONCLUSION: Novel histological features of epithelial cells of ECIPAS were indicated. Although more cases need to be evaluated, we propose that the cause of ECIPAS may be different from that of pancreatic ductal origin. J. Med. Invest. 70 : 251-259, February, 2023.
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Quiste Epidérmico , Enfermedades Pancreáticas , Humanos , Quiste Epidérmico/patología , Bazo/patología , Enfermedades Pancreáticas/patología , Células Epiteliales , InmunohistoquímicaRESUMEN
Plants produce the volatile hormone ethylene to regulate many developmental processes and to deal with (a)biotic stressors. In seed plants, ethylene is synthesized from 1-aminocyclopropane-1-carboxylic acid (ACC) by the dedicated enzyme ACC oxidase (ACO). Ethylene biosynthesis is tightly regulated at the level of ACC through ACC synthesis, conjugation and transport. ACC is a non-proteinogenic amino acid, which also has signaling roles independent from ethylene. In this work, we investigated the biological function of an uncharacterized ACC dipeptide. The custom-synthesized di-ACC molecule can be taken up by Arabidopsis in a similar way as ACC, in part via Lysine Histidine Transporters (e.g., LHT1). Using Nano-Particle Assisted Laser Desoprtion/Ionization (Nano-PALDI) mass-spectrometry imaging, we revealed that externally fed di-ACC predominantly localizes to the vasculature tissue, despite it not being detectable in control hypocotyl segments. Once taken up, the ACC dimer can evoke a triple response phenotype in dark-grown seedlings, reminiscent of ethylene responses induced by ACC itself, albeit less efficiently compared to ACC. Di-ACC does not act via ACC-signaling, but operates via the known ethylene signaling pathway. In vitro ACO activity and molecular docking showed that di-ACC can be used as an alternative substrate by ACO to form ethylene. The promiscuous nature of ACO for the ACC dimer also explains the higher ethylene production rates observed in planta, although this reaction occurred less efficiently compared to ACC. Overall, the ACC dipeptide seems to be transported and converted into ethylene in a similar way as ACC, and is able to augment ethylene production levels and induce subsequent ethylene responses in Arabidopsis.
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Simultaneous imaging of l-dihydroxyphenylalanine (l-DOPA), dopamine (DA) and norepinephrine (NE) in the catecholamine metabolic pathway is particularly useful because l-DOPA is a neurophysiologically important metabolic intermediate. In this study, we found that 2,4,6-trimethylpyrillium tetrafluoroborate (TMPy) can selectively and efficiently react with target catecholamine molecules. Specifically, simultaneous visualization of DA and NE as metabolites of l-DOPA with high steric hinderance was achieved by derivatized-imaging mass spectrometry (IMS). Interestingly, l-DOPA showed strong localization in the brainstem, in contrast to the pattern of DA and NE, which co-localized with tyrosine hydroxylase (TH). In addition, to identify whether the detected molecules were endogenous or exogenous l-DOPA, mice were injected with l-DOPA deuterated in three positions (D3-l-DOPA), which was identifiable by a mass shift of 3Da. TMPy-labeled l-DOPA, DA and NE were detected at m/z 302.1, 258.1 and 274.1, while their D3 versions were detected at 305.0, 261.1 and 277.1 in mouse brain, respectively. l-DOPA and D3-l-DOPA were localized in the BS. DA and NE, and D3-DA and D3-NE, all of which are metabolites of L-DOPA and D3-l-DOPA, were localized in the striatum (STR) and locus coeruleus (LC). These findings suggest a mechanism in the brainstem that allows l-DOPA to accumulate without being metabolized to monoamines downstream of the metabolic pathway.
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Dopamina , Levodopa , Animales , Catecolaminas , Dopamina/metabolismo , Espectrometría de Masas , Ratones , Norepinefrina/metabolismoRESUMEN
To demonstrate the accurate analysis of catecholamines and amino acid using derivatization reagents, we investigated the reaction conditions for 2,4,6-triethyl-3,5-dimethyl pyrylium trifluoromethanesulfonate (Py-Tag), derivatization of the targets dopamine (DA) and γ-aminobutyric acid (GABA) on tissue sections, and constructed an optimized reaction compartment. Ten different Py-Tag reaction conditions with the targets were considered. The optimal condition for the Py-Tag reaction with the targets was identified as a 70% methanol with 5% trimethylamine (v/v) solution at 60 °C under homogenous conditions. To reproduce this reaction on tissue sections, we constructed a reaction compartment to maintain humidity levels and facilitate the derivatization reaction. Moreover, visualization of DA and GABA was archived by derivatized-imaging mass spectrometry. Brain sections of unilateral 6-OHDA lesioned Parkinson's disease model rats showed Py-Tag DA (m/z 328.3) in the unilateral striatum and Py-Tag GABA (m/z 278.3) in the cerebral cortex, striatum, hippocampus and hypothalamus. Using the Parkinson's disease model rat brain, images with left-right differences were obtained for the localization of DA and GABA. These findings indicate that it is important to consider the reaction conditions that allow high reaction efficiency between DA or GABA and Py-Tag as well as high quality imaging of sections.
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Enfermedad de Parkinson , Animales , Dopamina/análisis , Dopamina/metabolismo , Indicadores y Reactivos , Espectrometría de Masas , Mesilatos , Enfermedad de Parkinson/metabolismo , Ratas , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Plant hormones can act in synergistic and antagonistic ways in response to biotic and abiotic stresses and during plant growth and development. Thus, a technique is needed to simultaneously determine the distribution and concentration of several plant hormones. A relatively new technology, mass spectrometry imaging (MSI), enables the direct mapping and imaging of biomolecules on tissue sections. MSI permits simultaneous detection of multiple analytes on a single section of plant tissue, even in the absence of target-specific markers such as antibodies. Recently, MSI has been used to localize multiple, small molecule (m/z < 500) plant hormones by the nanoparticle-assisted laser desorption/ionization (Nano-PALDI) mass spectrometry (MS) method. Here, we illustrate a technology for multiple-hormone imaging using Nano-PALDI MSI and discuss its potential in investigating the role of hormone signaling in plant development and stress responses.
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Nanopartículas , Reguladores del Crecimiento de las Plantas , Rayos Láser , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Zebrafish models have been developed for several studies involving lipid metabolism and lipid-related diseases. In the present study, the migration of dietary docosahexaenoic acid (DHA) in whole-body zebrafish was estimated by stable-isotope tracer and matrix-assisted laser desorption/ionization mass spectrometry imaging. Administration of 1-13C-2,2-D2-labeled DHA ((+3)DHA) ethyl ester to male zebrafish was conducted to evaluate its accumulation, migration, and distribution in the body. The (+3)DHA content in the body of zebrafish after administering (+3)DHA for 10 and 15 d was significantly higher than that in the control group. (+3)DHA was observed as a constituent of phosphatidylcholine (PC) in the intestine of zebrafish that were administered (+3)DHA for 5 and 10 d. (+3)DHA-containing PC tended to accumulate in the intestines of zebrafish administered (+3)DHA for 1 d, indicating that recombination of (+3)DHA from ethyl ester to PC occurs quickly at intestine. After administration for 15 d, (+3)DHA-containing PC accumulated in the intestine, liver, and muscle of whole-body zebrafish. In contrast, (+3)DHA-containing PC was not detected in the brain. These results showed that dietary DHA is initially constructed into PC as a structural component of intestinal cell membranes and gradually migrates into peripheral tissues such as muscle.
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Ácidos Docosahexaenoicos/metabolismo , Pez Cebra/metabolismo , Animales , Encéfalo/metabolismo , Dieta , Ácidos Docosahexaenoicos/administración & dosificación , Intestinos/metabolismo , Metabolismo de los Lípidos , Hígado/metabolismo , Masculino , Modelos Animales , Músculos/metabolismo , Fosfatidilcolinas/metabolismo , Fosfolípidos/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Ammonium is a prominent source of inorganic nitrogen for plant nutrition, but excessive amounts can be toxic for many species. However, most conifers are tolerant to ammonium, a relevant physiological feature of this ancient evolutionary lineage. For a better understanding of the molecular basis of this trait, ammonium-induced changes in the transcriptome of maritime pine (Pinus pinaster Ait.) root apex have been determined by laser capture microdissection and RNA sequencing. Ammonium promoted changes in the transcriptional profiles of multiple transcription factors, such as SHORT-ROOT, and phytohormone-related transcripts, such as ACO, involved in the development of the root meristem. Nano-PALDI-MSI and transcriptomic analyses showed that the distributions of IAA and CKs were altered in the root apex in response to ammonium nutrition. Taken together, the data suggest that this early response is involved in the increased lateral root branching and principal root growth, which characterize the long-term response to ammonium supply in pine. All these results suggest that ammonium induces changes in the root system architecture through the IAA-CK-ET phytohormone crosstalk and transcriptional regulation.
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Compuestos de Amonio , Pinus , Compuestos de Amonio/metabolismo , Pinus/genética , Pinus/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Raíces de Plantas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
While the interaction of cells such as macrophages and hepatic stellate cells is known to be involved in the generation of fibrosis in nonalcoholic steatohepatitis (NASH), the mechanism remains unclear. This study employed a high-fat/cholesterol/cholate (HFCC) diet to generate a model of NASH-related fibrosis to investigate the pathogenesis of fibrosis. Two mouse strains: C57BL/6J, the one susceptible to obesity, and A/J, the one relatively resistant to obesity, developed hepatic histologic features of NASH, including fat deposition, intralobular inflammation, hepatocyte ballooning, and fibrosis, after 9 weeks of HFCC diet. The severity of hepatic inflammation and fibrosis was greater in A/J mice than in the C57BL/6J mice. A/J mice fed HFCC diet exhibited characteristic CD204-positive lipid-laden macrophage aggregation in hepatic parenchyma. Polarized light was used to visualize the Maltese cross, cholesterol crystals within the aggregated macrophages. Fibrosis developed in a ring shape from the periphery of the aggregated macrophages such that the starting point of fibrosis could be visualized histologically. Matrix-assisted laser desorption/ionization mass spectrometry imaging analysis detected a molecule at m/z 772.462, which corresponds to the protonated ion of phosphatidylcholine [P-18:1 (11Z)/18:0] and phosphatidylethanolamine [18:0/20:2 (11Z, 14Z)], in aggregated macrophages adjacent to the fibrotic lesions. In conclusion, the HFCC diet-fed A/J model provides an ideal tool to study fibrogenesis and enables novel insights into the pathophysiology of NASH-related fibrosis.
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Lípidos/química , Macrófagos/metabolismo , Enfermedad del Hígado Graso no Alcohólico/patología , Animales , Peso Corporal , Modelos Animales de Enfermedad , Ingestión de Energía , Regulación de la Expresión Génica , Lípidos/sangre , Hígado/patología , Masculino , Ratones Endogámicos C57BL , Modelos Biológicos , Enfermedad del Hígado Graso no Alcohólico/sangre , Enfermedad del Hígado Graso no Alcohólico/genética , Tamaño de los Órganos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Recent epidemiological and intervention studies have suggested that polyphenol-rich plant food consumption reduced the risk of cognitive decline. However, the findings were tentative and by no means definitive. In the present study, we examined the impact of short-term oral administration of cinnamtannin A2 (A2), an (-)-epicatechin tetramer, on adult hippocampal neurogenesis and cognitive function in mice. Mice received supplementation with vehicle (20% glycerol) or 100 µg/kg A2 for 10 days. Then, we conducted the open field test, the object location test, and the novel object test. In the open field test, the A2-treated group tended to spend more time in the center of the arena, compared to the vehicle-treated group. The A2-treated group spent significantly more time exploring objects placed in different locations, compared to the vehicle-treated group. There were no significant differences between groups in the object preference index or in the novel object test. In addition, A2 administration significantly increased the number of hippocampal bromodeoxyuridine-labeled cells in the dentate gyrus, but not in the CA1 or CA3 regions. These results suggested that short-term administration of A2 may impact spatial memory by enhancing neurogenesis in the dentate gyrus of adult mice.
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Antocianinas/farmacología , Catequina/farmacología , Hipocampo/efectos de los fármacos , Neurogénesis/efectos de los fármacos , Memoria Espacial/efectos de los fármacos , Administración Oral , Animales , Antocianinas/administración & dosificación , Antocianinas/química , Bromodesoxiuridina/metabolismo , Catequina/administración & dosificación , Catequina/química , Giro Dentado/citología , Giro Dentado/metabolismo , Conducta Exploratoria/efectos de los fármacos , Conducta Exploratoria/fisiología , Hipocampo/citología , Hipocampo/fisiología , Ratones Endogámicos C57BL , Estructura Molecular , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Memoria Espacial/fisiología , Factores de TiempoRESUMEN
The localization of essential oils, including flavor components, in perilla herb (Perilla frutescens var. crispa) were visually determined using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) imaging. The surface of a perilla leaf was peeled using a cyanoacrylate adhesion compound and contained oil glands that retained their morphology and chemical properties. We imaged the three essential oils perillaldehyde, ß-caryophyllene, and rosmarinic acid (RA). Perillaldehyde was derivatized using glycine to prevent evaporation and allow its detection and imaging while localized in oil glands. ß-caryophyllene also localized in the oil glands and not in the epidermis region. RA was detected throughout the leaf, including the oil glands. Quantitative data for the three essential oils were obtained by gas chromatography- or liquid chromatography-MS. The concentrations of perillaldehyde, ß-caryophyllene, and RA were 12.6 ± 0.62, 0.27 ± 0.02, and 0.16 ± 0.02 [mg/g] in the paste sample of perilla herb. Peeling using a cyanoacrylate adhesion compound, and derivatization of a target such as an aroma component have great potential for mass spectrometry imaging for multiple essential oils.
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Transcriptome and metabolome analysis in tomato (Solanum lycopersicum) fruits cultivated under drought conditions showed that drought stress promoted fatty acid synthesis and increased the content of fatty acids in fruits. The accumulation of some phospholipids composed of palmitic acid and oleic acid also was significantly increased, especially in seeds. Moreover, inositol, which is a component of cell membranes and cell walls, was increased through the activity of the myoinositol monophosphatase 1-mediated pathway. In mature fruits, the levels of metabolic regulators such as ß-alanine and 4-aminobutyric acid were elevated. These results showed that these compounds are drought-responsive and enhance drought tolerance and subsequently they could enhance the nutritional value and health benefits of tomato fruit.
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Solanum lycopersicum , Sequías , Ácidos Grasos , Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Fosfolípidos , Proteínas de Plantas/metabolismo , Transcriptoma , Regulación hacia ArribaRESUMEN
The drying process used for persimmon fruit (Diospyros kaki) can alter the composition of nutrients, and especially vitamins. We visually determined whether the amounts of vitamin A1, vitamin B6 and vitamin C vary after drying persimmon fruit, using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) imaging. Drying altered the amount of moisture between the fruit interior and surface. Vitamin A1 is lipophilic and localized at the desiccated outer regions (pericarp) and not in the inner region (mesocarp and endocarp), and its concentration was increased 3.4 times in dried fruit compared with raw persimmon. Vitamin B1 and B6 are water-soluble and concentrated in the moist mesocarp. The vitamin C content of dried persimmon is decreased by drying in the sun. The drying process affected the localizations and amounts of all the vitamins. The observed opposite localization of vitamin A1 compared to B1 and B6 was due to vitamin A1 being lipophilic and B1 and B6 being water soluble. Multiplevitamin imaging using MALDI-MSI has great potential for enhancing commodity value and for visually investigating the effects of manufacturing processes.
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Desecación/métodos , Diospyros/química , Análisis de los Alimentos/métodos , Manipulación de Alimentos , Vitaminas/análisis , Interacciones Hidrofóbicas e Hidrofílicas , Valor Nutritivo , Solubilidad , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Tiamina/análisis , Vitamina A/análisis , Vitamina B 6/análisis , AguaRESUMEN
We developed a new method for monitoring the distribution of administrated fatty acids in the body by combination of a stable isotope-labeling technique and imaging mass spectrometry (IMS). The developed stable isotope-labeling technique is very simple and able to adapt to all the fatty acid species. In this study, we synthesized stable isotope-labeled arachidonic acid (AA) and docosahexaenoic acid (DHA), and they were simultaneously administrated to mice to examine their migrations and distributions in the brain. The administrated AA and DHA have two more molecular weights compared to the originals and apparently were distinguished from the originally accumulated AA and DHA in the brain using IMS. As a result, we reveal that the administered AA and DHA first accumulated in the hippocampus and cerebellar cortex in the brain. This technique does not use radio isotopes and would appear to elucidate the role of all kinds of fatty acid species in the body.
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Ácido Araquidónico/análisis , Encéfalo/metabolismo , Ácidos Docosahexaenoicos/análisis , Espectrometría de Masas/métodos , Animales , Corteza Cerebelosa/química , Corteza Cerebelosa/metabolismo , Deuterio/química , Ácidos Grasos/análisis , Femenino , Cromatografía de Gases y Espectrometría de Masas , Hipocampo/química , Hipocampo/metabolismo , Marcaje Isotópico , Ratones , Ratones Endogámicos ICR , Peso Molecular , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Plant hormones can act in synergistic and antagonistic ways in response to biotic and abiotic stresses and in plant growth and development. Thus, a technique is needed to simultaneously determine the distributions and concentrations of several plant hormones. Previously, we reported that localizations of two plant hormones [cytokinin (CK) and abscisic acid (ABA)] can be simultaneously visualized in a plant tissue using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). In MALDI-MS, however, self-ionization of an organic matrix occasionally interferes with ionizations of small molecules (<500 m/z) including most plant hormones. Another technique, nanoparticle-assisted laser desorption/ionization (Nano-PALDI), can avoid matrix self-ionization using nanoparticles to assist the ionization of analytes. Here, we compared the ionization efficiencies of common plant hormones by MALDI-MS and Nano-PALDI-MS. For the comparison, we prepared a standard mix of seven plant hormones [ABA, auxin (IAA), brassinosteroid (Br), two CKs (trans-zeatin, tZ, and 6-(γ,γ-dimethylallylamino) purine, iP), jasmonic acid, and salicylic acid (SA)], an ethylene precursor (1-aminocyclopropane-1-carboxylic acid, ACC), and a heavy hydrogen-labeled ABA (D6-ABA). Basic MALDI-MS detected all compounds except IAA, Br, and D6-ABA, while Nano-PALDI-MS detected all nine compounds. By Nano-PALDI-MS imaging (MSI), each of the abovementioned hormones and ACC were also detected in root cross sections of rice which were incubated in the hormone mix for 2 h. In the elongation zone of untreated roots, Nano-PALDI-MSI revealed high levels of ABA and CKs in the outer part of roots and much lower levels in the stele, but Br, SA, and ACC were broadly distributed in the cross section. IAA seemed to be distributed in the epidermis, cortex, and stele. Multiple-hormone imaging using Nano-PALDI-MS has great potential for investigating the roles of hormone signaling in crop development and stress responses.
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Espectrometría de Masas/métodos , Oryza/química , Reguladores del Crecimiento de las Plantas/química , Rayos Láser , Espectrometría de Masas/instrumentación , Nanopartículas/química , Oryza/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Raíces de Plantas/química , Raíces de Plantas/metabolismoRESUMEN
Cinnamtannin A2, an (-)-epicatechin tetramer, was reported to have potent physiological activity. Cinnamtannin A2 is rarely absorbed from the gastrointestinal tract into the blood and the mechanisms of its beneficial activities are unknown. Cinnamtannin A2 reported to increase sympathetic nervous activity, which was induced by various stressors. In present study, we examined the stress response in the mouse paraventricular nucleus following a single oral dose of cinnamtannin A2 by monitoring mRNA expression of corticotropin-releasing hormone (CRH) and c-fos using in situ hybridization. Corticotropin-releasing hormone mRNA showed a tendency to increase at 15 min and significantly increased at 60 min following a single oral administration of 100 µg/kg cinnamtannin A2. After a single dose of 10 µg/kg cinnamtannin A2, there was significant upregulation of CRH mRNA at 60 min. These results suggested that cinnamtannin A2 was recognized as a stressor in central nervous system and this may lead to its beneficial effects on circulation and metabolism.