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1.
Biosci Biotechnol Biochem ; 88(2): 203-205, 2024 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-37947260

RESUMO

Licking behavior with various salts in transmembrane channel-like 4 (Tmc4) knockout (KO) mice was observed. In Tmc4 KO mice, a significant decrease in sensitivity to chloride salts, such as NaCl, KCl, and NH4Cl, was observed, while no significant decrease in sensitivity to Na-gluconate was observed. This finding suggests that TMC4 may be involved in the detection of chloride taste.


Assuntos
Cloretos , Sais , Animais , Camundongos , Amilorida , Camundongos Knockout , Cloreto de Potássio/farmacologia , Paladar
2.
Front Neurosci ; 17: 1260655, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37781249

RESUMO

Mastication stimuli have been demonstrated to affect memory function and autonomic nerve activity; however, this process has not been well studied during weaning compared to old age. Previously, we conducted molecular analyses of the thalamus and hippocampus to elucidate the mechanisms underlying this memory-enhancing effect in weaning-stage rats. In this study, we aimed to evaluate the effect of masticatory stimuli on the regulation of heartbeat rate (HR) through the hypothalamic-autonomic system. Three-week-old male rats were administered a powdered diet (P group) or chow-diet (C group) for 10 days. Thereafter, transcriptome analysis was performed. Vasopressin, cocaine-amphetamine-regulated transcript prepropeptide, corticotropin-releasing hormone, and thyrotropin-releasing hormone, which are involved in sympathetic activation of heart rate, were downregulated in the C group. Electrocardiograms were recorded continuously for 12 days under the same condition. Interestingly, rats in the C group had a significantly lower HR than those in the P group on day 11. We checked several parameters representing the autonomic regulation of HR. The C group had higher values for the high-frequency band integration of the HR power spectrum (parasympathetic marker) and root mean square successive difference of R-wave intervals (parasympathetic marker) relative to the P group. Such findings provide a molecular and physiological basis for understanding the regulation of cardiovascular function in response to masticatory stimuli in the autonomic nervous system.

3.
Biosci Biotechnol Biochem ; 87(3): 290-302, 2023 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-36572396

RESUMO

The metabolism of tomato fruits changes when plants experience drought stress. In this study, we investigated changes in microRNA (miRNA) abundance and detected 32 miRNAs whose expression changes in fruit. The candidate target genes for each miRNA were predicted from the differentially expressed genes identified by transcriptome analysis at the same fruit maturation stage. The predicted targeted genes were related to cell wall metabolisms, response to pathogens, and plant hormones. Among these, we focused on cell wall metabolism-related genes and performed a dual luciferase assay to assess the targeting of their mRNAs by their predicted miRNA. As a result, sly-miR10532 and sly-miR7981e suppress the expression of mRNAs of galacturonosyltransferase-10 like encoding the main enzyme of pectin biosynthesis, while sly-miR171b-5p targets ß-1,3-glucosidase mRNAs involved in glucan degradation. These results will allow the systematic characterization of miRNA and their target genes in the tomato fruit under drought stress conditions.


Assuntos
MicroRNAs , Solanum lycopersicum , MicroRNAs/genética , Frutas/metabolismo , Secas , Parede Celular/metabolismo , Regulação da Expressão Gênica de Plantas
4.
Front Behav Neurosci ; 16: 1006359, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36263297

RESUMO

Mastication stimuli are known to relieve senile dementia in human and animal studies. However, few studies have focused on its effect on weaning-stage animals and the underlying molecular processes. In this study, 3-week-old male rats were raised on a powdered (P-group) or chow (C-group) diet for 8 days, and their behavior was examined using the Y-maze and novel object recognition tests. In the Y-maze test, the C-group rats showed a larger alternation ratio than the P-group rats. In the novel object recognition test, the C-group rats exhibited a significantly larger discrimination index for novel objects than for familiar objects, but the P-group rats did not. We then compared the hippocampal neuron morphology and transcriptome between the groups. C-group rats exhibited larger dendrite branch numbers in the apical dendrites of pyramidal cells in the cornu ammonis 1 (CA1) region and a larger spine density in the basal dendrites of CA1 neurons than the P-group rats. Using DNA microarray analysis, we identified 621 (P < C) and 96 (P > C) genes that were differentially expressed between the groups. These genes were enriched in functional terms related to dendrite growth and included the Igf2, RhoA, and Rho GEF genes, most of which were upregulated in the C-group. These results suggest that the mastication stimuli during the weaning period can enhance the learning ability of rats by increasing the dendrite branches of hippocampal CA1 neurons and by regulating genes related to dendrite growth.

5.
BMC Genomics ; 22(1): 347, 2021 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-33985426

RESUMO

BACKGROUND: Curculigo latifolia is a perennial plant endogenous to Southeast Asia whose fruits contain the taste-modifying protein neoculin, which binds to sweet receptors and makes sour fruits taste sweet. Although similar to snowdrop (Galanthus nivalis) agglutinin (GNA), which contains mannose-binding sites in its sequence and 3D structure, neoculin lacks such sites and has no lectin activity. Whether the fruits of C. latifolia and other Curculigo plants contain neoculin and/or GNA family members was unclear. RESULTS: Through de novo RNA-seq assembly of the fruits of C. latifolia and the related C. capitulata and detailed analysis of the expression patterns of neoculin and neoculin-like genes in both species, we assembled 85,697 transcripts from C. latifolia and 76,775 from C. capitulata using Trinity and annotated them using public databases. We identified 70,371 unigenes in C. latifolia and 63,704 in C. capitulata. In total, 38.6% of unigenes from C. latifolia and 42.6% from C. capitulata shared high similarity between the two species. We identified ten neoculin-related transcripts in C. latifolia and 15 in C. capitulata, encoding both the basic and acidic subunits of neoculin in both plants. We aligned these 25 transcripts and generated a phylogenetic tree. Many orthologs in the two species shared high similarity, despite the low number of common genes, suggesting that these genes likely existed before the two species diverged. The relative expression levels of these genes differed considerably between the two species: the transcripts per million (TPM) values of neoculin genes were 60 times higher in C. latifolia than in C. capitulata, whereas those of GNA family members were 15,000 times lower in C. latifolia than in C. capitulata. CONCLUSIONS: The genetic diversity of neoculin-related genes strongly suggests that neoculin genes underwent duplication during evolution. The marked differences in their expression profiles between C. latifolia and C. capitulata may be due to mutations in regions involved in transcriptional regulation. Comprehensive analysis of the genes expressed in the fruits of these two Curculigo species helped elucidate the origin of neoculin at the molecular level.


Assuntos
Curculigo , Paladar , Curculigo/genética , Curculigo/metabolismo , Frutas/genética , Frutas/metabolismo , Perfilação da Expressão Gênica , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Edulcorantes , Transcriptoma
6.
J Agric Food Chem ; 69(9): 2894-2905, 2021 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-33645220

RESUMO

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.


Assuntos
Solanum lycopersicum , Secas , Ácidos Graxos , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Fosfolipídeos , Proteínas de Plantas/metabolismo , Transcriptoma , Regulação para Cima
7.
Photosynth Res ; 150(1-3): 343-356, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33146872

RESUMO

Bacteria exposed to environmental stresses often exhibit superior acclimation abilities to environmental change. Acid treatment causes an increase in the cell length of the cyanobacterium Synechocystis sp. PCC6803 under light conditions. We aimed to elucidate the relationship between acidic stress and cell enlargement. After being synchronized under dark conditions, the cells were cultivated at different pH (pH 8.0 or pH 6.0) levels under light conditions. Synechocystis 6803 cells exhibited only cell growth occurred (cell volume expansion) and slow proliferation under the acidic condition. In the recovery experiment of the enlarged cells, they proliferated normally at pH 8.0, and the cell lengths decreased to the normal cell size under light conditions. Inhibition of cell division might be caused by acidic stress. To understand the effect of acidic stress on cell division, we evaluated the expression of FtsZ via Western blotting. The FtsZ concentration in cells was lower at pH 6.0 than at pH 8.0 and was not sufficient for cell division in the photoautotrophic conditions. ClpXP is well known as a regulator of the Z-ring dynamics in E. coli. The transcriptional level of four clpXP genes was upregulated approximately threefold at pH 6.0 after 24 h compared with that in cells grown at pH 8.0. The lack of FtsZ may be caused by the upregulation of clpXP expression under acidic condition. Therefore, ClpXP may participate in the degradation of FtsZ and be involved in the regulation of cell division via FtsZ under acidic stress in Synechocystis 6803.


Assuntos
Synechocystis , Proteínas de Bactérias/genética , Divisão Celular , Proteínas do Citoesqueleto/genética , Escherichia coli , Células Gigantes , Synechocystis/genética
8.
Int J Mol Sci ; 21(9)2020 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-32354168

RESUMO

Circulating miRNA species are promising symptom markers for various diseases, including cardiovascular disease. However, studies regarding their role in the treatment process are limited, especially concerning cerebral infarction. This study aimed to extract miRNA markers to investigate whether they reflect both onset and treatment process of cerebral infarction. A total of 22 patients (P-group) and 22 control subjects (C-group) were examined for their whole-blood miRNA profiles using DNA GeneChip™ miRNA 4.0 Array, with six patients examined after treatment (T-group). A total of 64 miRNAs were found to be differentially expressed between the C- and P-groups. Out of 64 miRNAs, the expression levels of two miRNAs correlated with hypertension. A total of 155 miRNAs were differentially expressed between the P- and T-groups. Five common miRNAs were found among the 64 and 155 miRNAs identified. Importantly, these common miRNAs were inversely regulated in each comparison (e.g., C < P > T), including miR-505-5p, which was previously reported to be upregulated in aortic stenosis patients. Our previous study using rat cerebral infarction models detected the downregulation of an apoptosis repressor, WDR26, which was repressed by one of the five miRNAs. Our results provide novel information regarding the miRNA-based diagnosis of cerebral infarction in humans. In particular, the five common miRNAs could be useful makers for the onset and the treatment process. Trial registration: This study was registered in the UMIN Clinical Trials Registry (UMIN000038321).


Assuntos
Infarto Cerebral/genética , Hipertensão/genética , MicroRNAs/sangue , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Estudos de Casos e Controles , Infarto Cerebral/sangue , Infarto Cerebral/etiologia , Feminino , Regulação da Expressão Gênica , Predisposição Genética para Doença , Humanos , Hipertensão/complicações , Masculino , Pessoa de Meia-Idade
9.
Biochem Biophys Res Commun ; 498(3): 621-626, 2018 04 06.
Artigo em Inglês | MEDLINE | ID: mdl-29524403

RESUMO

Mastication enhances brain function and mental health, but little is known about the molecular mechanisms underlying the effects of mastication on neural development in early childhood. Therefore, we analysed the gene expression in juvenile neural circuits in rats fed with a soft or chow diet immediately after weaning. We observed that the gene expression patterns in the thalamus varied depending on the diet. Furthermore, gene ontology analysis revealed that two terms were significantly enhanced: chemical synaptic transmission and positive regulation of dendritic spine morphogenesis. With respect to chemical synaptic transmission, glutamate decarboxylase and GABA receptors were upregulated in the chow diet group. The related genes, including vesicular GABA transporter, were also upregulated, suggesting that mastication activates GABAergic signalling. With respect to dendritic spine morphogenesis, Ingenuity Pathway Analysis predicted fewer extension of neurites and neurons and fewer number of branches in the chow diet group. The numbers of spines in the ventral posterolateral and posteromedial regions were significantly decreased. These results suggest that mastication in the early developing period upregulates GABAergic signalling genes, with a decrease of spines in the thalamus.


Assuntos
Espinhas Dendríticas/fisiologia , Mastigação , Transdução de Sinais , Tálamo/fisiologia , Ácido gama-Aminobutírico/metabolismo , Ração Animal/análise , Animais , Espinhas Dendríticas/genética , Espinhas Dendríticas/ultraestrutura , Proteínas da Membrana Plasmática de Transporte de GABA/genética , Proteínas da Membrana Plasmática de Transporte de GABA/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Glutamato Descarboxilase/genética , Glutamato Descarboxilase/metabolismo , Masculino , Neurogênese , Ratos , Ratos Wistar , Receptores de GABA/genética , Receptores de GABA/metabolismo , Transmissão Sináptica , Tálamo/crescimento & desenvolvimento , Transcriptoma , Regulação para Cima , Desmame , Ácido gama-Aminobutírico/genética
10.
Biosci Biotechnol Biochem ; 82(3): 497-506, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29370734

RESUMO

We fed rats noodle (N) -diet containing 30 wt.% instant noodle with a 26% fat-to-energy ratio for 30 days (N-group). Compared with rats that were fed the same amount of nutrients (C-group), the N-group showed lower liver triacylglycerol levels and higher fecal cholesterol levels. We then analyzed transcriptome of the hypothalamic-pituitary (HP), the liver and the white adipose tissue (WAT). Thyroid stimulating hormone (Tshb), and its partner, glycoprotein hormone genes were up-regulated in the HP of N-group. Sterol regulatory element binding transcription factors were activated in the liver of N-group, while an up-regulation of the angiogenic signal occurred in the WAT of N-group. N-group showed higher urine noradrenaline (NA) level suggesting that these tissue signals are regulated by NA and Tshb. The N-diet contains 0.326 wt.% glutamate, 0.00236 wt.% 6-shogaol and Maillard reaction products. Our results suggest that these ingredients may affect lipid homeostasis via the HP axis.


Assuntos
Gorduras na Dieta/análise , Crescimento e Desenvolvimento/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Hipófise/efeitos dos fármacos , Tecido Adiposo Branco/efeitos dos fármacos , Tecido Adiposo Branco/metabolismo , Aminoácidos/sangue , Animais , Catecolaminas/urina , Hipotálamo/fisiologia , Masculino , Hipófise/fisiologia , Ratos , Ratos Wistar , Transcriptoma/efeitos dos fármacos
11.
Int J Mol Sci ; 18(11)2017 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-29113076

RESUMO

Given the abundance of stroke patients and deaths from stroke worldwide, many studies concerning the aftermath of stroke are being carried out. To reveal the precise effect of ischemic infarction, we conducted a comprehensive gene expression analysis. Alongside a middle cerebral artery occlusion (MCAO) Sprague-Dawley rat model, we used a group undergoing sham surgery for comparison, which was the same as MCAO surgery but without blood vessel occlusion. Subsequently, infarction of the brains of MCAO-treated rats occurred, but did not occur in the sham-treated rats. Using whole blood, we carried out DNA microarray analysis, revealing the gene expression alterations caused by stroke. Downregulation of immune pathways and cluster of differentiation (CD) molecules indicated immunodepression. By conducting miRNA microarray analysis, we extracted seven miRNAs as significantly regulated: miR-107-5p, miR-383-5p, miR-24-1-5p, mir-191b, miR-196b-5p, and miR-3552 were upregulated, and mir-194-1 was downregulated. Among these seven miRNAs, three had one target mRNA each that was extracted as differentially expressed, and the expression levels of all pairs were inversely correlated. This indicates the occurrence of miRNA-mRNA regulatory systems in blood: between miR-107-5p and H2A histone family member Z (H2afz), miR-196b-5p and protein tyrosine phosphatase receptor type C (Ptprc), and miR-3552 and serine/arginine-rich splicing factor 2 (Srsf2). Moreover, six miRNAs had matching human miRNAs with similar sequences, which are potential human stroke biomarkers.


Assuntos
Infarto da Artéria Cerebral Média/sangue , MicroRNAs/genética , RNA Mensageiro/genética , Animais , Biomarcadores/sangue , Regulação para Baixo , Histonas/genética , Histonas/metabolismo , Infarto da Artéria Cerebral Média/genética , Infarto da Artéria Cerebral Média/metabolismo , Antígenos Comuns de Leucócito/genética , Antígenos Comuns de Leucócito/metabolismo , Masculino , MicroRNAs/sangue , MicroRNAs/metabolismo , RNA Mensageiro/sangue , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de Processamento de Serina-Arginina/genética , Fatores de Processamento de Serina-Arginina/metabolismo
12.
Genes Nutr ; 12: 10, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28405243

RESUMO

BACKGROUND: To elucidate the effects of altered dietary carbohydrate and fat balance on liver and adipose tissue transcriptomes, 3-week-old rats were fed three kinds of diets: low-, moderate-, and high-fat diets (L, M, and H) containing a different ratio of carbohydrate-fat (C-F) (65:15, 60:20, and 35:45 in energy percent, respectively). METHODS: The rats consumed the diets for 9 weeks and were subjected to biochemical and DNA microarray analyses. RESULTS: The rats in the H-group exhibited lower serum triacylglycerol (TG) levels but higher liver TG and cholesterol content than rats in the L-group. The analysis of differentially expressed genes (DEGs) between each group (L vs M, M vs H, and L vs H) in the liver revealed about 35% of L vs H DEGs that were regulated in the same way as M vs H DEGs, and most of the others were L- vs H-specific. Gene ontology analysis of these L vs H DEGs indicated that those related to fatty acid synthesis and circadian rhythm were enriched. Interestingly, about 30% of L vs M DEGs were regulated in a reverse way compared with L vs H and M vs H DEGs. These reversed liver DEGs included M-up/H-down genes (Sds for gluconeogenesis from amino acids) and M-down/H-up genes (Gpd2 for gluconeogenesis from glycerol, Agpat9 for TG synthesis, and Acot1 for beta-oxidation). We also analyzed L vs H DEGs in white (WAT) and brown (BAT) adipose tissues and found that both oxidation and synthesis of fatty acids were inhibited in these tissues. CONCLUSIONS: These results indicate that the alteration of dietary C-F balance differentially affects the transcriptomes of metabolizing and energy-storing tissues.

13.
Anal Bioanal Chem ; 408(22): 5973-84, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26973237

RESUMO

Enzyme-linked immunosorbent assay (ELISA) is commonly used to determine food allergens in food products. However, a significant number of ELISAs give an erroneous result, especially when applied to highly processed food. Accordingly, an improved ELISA, which utilizes an extraction solution comprising the surfactant sodium lauryl sulfate (SDS) and reductant 2-mercaptoethanol (2-ME), has been specially developed to analyze food allergens in highly processed food by enhancing analyte protein extraction. Recently, however, the use of 2-ME has become undesirable. In the present study, a new extraction solution containing a human- and eco-friendly reductant, which is convenient to use at the food manufacturing site, has been established. Among three chemicals with different reducing properties, sodium sulfite, tris(3-hydroxypropyl)phosphine, and mercaptoethylamine sodium sulfite was selected as a 2-ME substitute. The protein extraction ability of SDS/0.1 M sodium sulfite solution was comparable to that of SDS/2-ME solution. Next, the ELISA performance for egg, milk, wheat, peanut, and buckwheat was evaluated by using model-processed foods and commercially available food products. The data showed that the SDS/0.1 M sulfite ELISA significantly correlated with the SDS/2-ME ELISA for all food allergens examined (p < 0.01), thereby establishing the validity of the SDS/0.1 M sulfite ELISA performance. Furthermore, the new SDS/0.1 M sulfite solution was investigated for its applicability to the lateral-flow (LF) test. The result demonstrated the successful analysis of food allergens in processed food, showing consistency with the SDS/0.1 M sulfite ELISA results. Accordingly, a harmonized analysis system for processed food comprising a screening LF test and a quantitative ELISA with identical extraction solution has been established. The ELISA based on the SDS/0.1 M sulfite extraction solution has now been authorized as the revised official method for food allergen analysis in Japan.


Assuntos
Alérgenos/isolamento & purificação , Fracionamento Químico/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Análise de Alimentos/métodos , Substâncias Redutoras/química , Alérgenos/análise , Animais , Arachis/química , Ovos/análise , Hipersensibilidade Alimentar/diagnóstico , Humanos , Mercaptoetilaminas/química , Leite/química , Fosfinas/química , Sulfitos/química , Triticum/química
14.
Neuron ; 82(2): 365-79, 2014 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-24685175

RESUMO

Thalamocortical (TC) connectivity is reorganized by thalamic inputs during postnatal development; however, the dynamic characteristics of TC reorganization and the underlying mechanisms remain unexplored. We addressed this question using dendritic refinement of layer 4 (L4) stellate neurons in mouse barrel cortex (barrel cells) as a model; dendritic refinement of L4 neurons is a critical component of TC reorganization through which postsynaptic L4 neurons acquire their dendritic orientation toward presynaptic TC axon termini. Simultaneous labeling of TC axons and individual barrel cell dendrites allowed in vivo time-lapse imaging of dendritic refinement in the neonatal cortex. The barrel cells reinforced the dendritic orientation toward TC axons by dynamically moving their branches. In N-methyl-D-aspartate receptor (NMDAR)-deficient barrel cells, this dendritic motility was enhanced, and the orientation bias was not reinforced. Our data suggest that L4 neurons have "fluctuating" dendrites during TC reorganization and that NMDARs cell autonomously regulate these dynamics to establish fine-tuned circuits.


Assuntos
Córtex Cerebral/citologia , Dendritos/fisiologia , Proteínas do Tecido Nervoso/metabolismo , Neurônios/citologia , Receptores de N-Metil-D-Aspartato/metabolismo , Tálamo/fisiologia , Animais , Animais Recém-Nascidos , Córtex Cerebral/fisiologia , Dendritos/efeitos dos fármacos , Agonistas de Aminoácidos Excitatórios/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Técnicas In Vitro , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Modelos Biológicos , N-Metilaspartato/farmacologia , Proteínas do Tecido Nervoso/genética , Vias Neurais/crescimento & desenvolvimento , Vias Neurais/metabolismo , Neurônios/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/genética , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiônico/farmacologia
15.
Retrovirology ; 10: 83, 2013 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-23915234

RESUMO

BACKGROUND: Viral protein R (Vpr), a protein of human immunodeficiency virus type-1 (HIV-1) with various biological functions, was shown to be present in the blood of HIV-1-positive patients. However, it remained unclear whether circulating Vpr in patients' blood is biologically active. Here, we examined the activity of blood Vpr using an assay system by which retrotransposition of long interspersed element-1 (L1-RTP) was detected. We also investigated the in vivo effects of recombinant Vpr (rVpr) by administrating it to transgenic mice harboring human L1 as a transgene (hL1-Tg mice). Based on our data, we discuss the involvement of blood Vpr in the clinical symptoms of acquired immunodeficiency syndrome (AIDS). RESULTS: We first discovered that rVpr was active in induction of L1-RTP. Biochemical analyses revealed that rVpr-induced L1-RTP depended on the aryl hydrocarbon receptor, mitogen-activated protein kinases, and CCAAT/enhancer-binding protein ß. By using a sensitive L1-RTP assay system, we showed that 6 of the 15 blood samples from HIV-1 patients examined were positive for induction of L1-RTP. Of note, the L1-RTP-inducing activity was blocked by a monoclonal antibody specific for Vpr. Moreover, L1-RTP was reproducibly induced in various organs, including the kidney, when rVpr was administered to hL1-Tg mice. CONCLUSIONS: Blood Vpr is biologically active, suggesting that its monitoring is worthwhile for clarification of the roles of Vpr in the pathogenesis of AIDS. This is the first report to demonstrate a soluble factor in patients' blood active for L1-RTP activity, and implies the involvement of L1-RTP in the development of human diseases.


Assuntos
Produtos do Gene vpr/sangue , Produtos do Gene vpr/metabolismo , HIV-1/enzimologia , Elementos Nucleotídeos Longos e Dispersos , Recombinação Genética , Adulto , Animais , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Adulto Jovem
16.
Drug Metab Dispos ; 38(2): 215-22, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19875501

RESUMO

Elucidation of the rate-determining process in the overall hepatic elimination of drugs is critical for predicting their intrinsic hepatic clearance and the impact of variation of sequestration clearance on their systemic concentration. The present study investigated the rate-determining process in the overall hepatic elimination of the HMG-CoA reductase inhibitors pravastatin, pitavastatin, atorvastatin, and fluvastatin both in rats and humans. The uptake of these statins was saturable in both rat and human hepatocytes. Intrinsic hepatic clearance obtained by in vivo pharmacokinetic analysis in rats was close to the uptake clearance determined by the multiple indicator dilution method but much greater than the intrinsic metabolic clearance extrapolated from an in vitro model using liver microsomes. In vivo uptake clearance of the statins in humans (pravastatin, 1.44; pitavastatin, 30.6; atorvastatin, 12.7; and fluvastatin, 62.9 ml/min/g liver), which was obtained by multiplying in vitro uptake clearance determined in cryopreserved human hepatocytes by rat scaling factors, was within the range of overall in vivo intrinsic hepatic clearance (pravastatin, 0.84-1.2; pitavastatin, 14-35; atorvastatin, 11-19; and fluvastatin, 123-185 ml/min/g liver), whereas the intrinsic metabolic clearance of atorvastatin and fluvastatin was considerably low compared with their intrinsic hepatic clearance. Their uptake is the rate-determining process in the overall hepatic elimination of the statins in rats, and this activity likely holds true for humans. In vitro-in vivo extrapolation of the uptake clearance using a cryopreserved human hepatocytes model and rat scaling factors will be effective for predicting in vivo intrinsic hepatic clearance involving active uptake.


Assuntos
Inibidores de Hidroximetilglutaril-CoA Redutases/farmacocinética , Fígado/metabolismo , Algoritmos , Animais , Bile/química , Transporte Biológico Ativo , Cromatografia Líquida de Alta Pressão , Hepatócitos/metabolismo , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/sangue , Inibidores de Hidroximetilglutaril-CoA Redutases/metabolismo , Inibidores de Hidroximetilglutaril-CoA Redutases/urina , Técnicas de Diluição do Indicador , Masculino , Taxa de Depuração Metabólica , Microssomos Hepáticos/metabolismo , Perfusão , Ligação Proteica , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem
17.
J Vet Med Sci ; 71(5): 631-4, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19498290

RESUMO

This study investigated associations of a single-mating occurrence (SMO) with farrowing rate and pigs born alive (PBA) in first-serviced and reserviced female pigs (females), and identified the factors associated with SMO. The data included 111,334 service and 91,233 farrowing records on 117 farms. A mating was defined as any one insemination (mating) of a female during estrus. Mixed-effects models were used to investigate reproductive performance and factors associated with SMO. In the first-service group, single-mated females had a lower farrowing rate and fewer PBA than multiple-mated females (P<0.05). In the reservice group, single-mated females also had a lower farrowing rate than multiple-mated females (P<0.05), but had PBA similar to multiple-mated females. SMO in first-service and reservice groups were 4.1 and 6.0%, respectively. Gilts were 1.030 times more likely to be mated a single time than sows (P<0.05). Gilts with age at first mating 150-224 and > or = 262 days were 1.010-1.016 times more likely to be mated a single time than those with age at first mating 225-260 days (P<0.05). Sows with weaning-to-first-mating interval > or = 7 days were 1.024-1.030 times more likely to be mated a single time than those with weaning-to-first-mating interval < or = 6 days (P<0.05). Factors associated with a higher SMO were a reservice occurrence, being gilts, low or high ages of gilts at first mating, and prolonged weaning-to-first-mating interval.


Assuntos
Cruzamento/métodos , Reprodução/fisiologia , Suínos/fisiologia , Animais , Animais Recém-Nascidos , Feminino , Modelos Logísticos , Masculino
18.
Cell ; 130(4): 742-53, 2007 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-17719550

RESUMO

The ephrin/Eph system plays a central role in neuronal circuit formation; however, its downstream effectors are poorly understood. Here we show that alpha-chimerin Rac GTPase-activating protein mediates ephrinB3/EphA4 forward signaling. We discovered a spontaneous mouse mutation, miffy (mfy), which results in a rabbit-like hopping gait, impaired corticospinal axon guidance, and abnormal spinal central pattern generators. Using positional cloning, transgene rescue, and gene targeting, we demonstrated that loss of alpha-chimerin leads to mfy phenotypes similar to those of EphA4(-/-) and ephrinB3(-/-) mice. alpha-chimerin interacts with EphA4 and, in response to ephrinB3/EphA4 signaling, inactivates Rac, which is a positive regulator of process outgrowth. Moreover, downregulation of alpha-chimerin suppresses ephrinB3-induced growth cone collapse in cultured neurons. Our findings indicate that ephrinB3/EphA4 signaling prevents growth cone extension in motor circuit formation via alpha-chimerin-induced inactivation of Rac. They also highlight the role of a Rho family GTPase-activating protein as a key mediator of ephrin/Eph signaling.


Assuntos
Quimerina 1/metabolismo , Efrina-B3/metabolismo , Transdução de Sinais , Proteínas rac de Ligação ao GTP/fisiologia , Animais , Animais Recém-Nascidos , Células Cultivadas , Córtex Cerebral/citologia , Cromossomos de Mamíferos , Cruzamentos Genéticos , Efrina-B3/genética , Genes Recessivos , Ligação Genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Camundongos Knockout , Camundongos Transgênicos , Dados de Sequência Molecular , Atividade Motora/genética , Mutação , N-Metilaspartato/farmacologia , Neurônios/citologia , Neurônios/metabolismo , Polimorfismo de Nucleotídeo Único , Serotonina/farmacologia , Medula Espinal/efeitos dos fármacos
19.
J Biosci Bioeng ; 101(4): 361-8, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16716946

RESUMO

The production of retroviral vectors using a transient expression system has been improved to obtain a high-titer virus preparation that is difficult to produce using packaging cell lines due to the cytotoxic or cytostatic effect of transgenes. Here, we used one such production method, the so-called Q-vector system, and examined its potential for virus production. The Q-vector system could produce a similar level of viral vectors compared with the packaging cell system but the production seemed to depend on the size and nature of transgenes. In the process of investigation of the quantitative difference in viral components between the transient expression system and the packaging cell system, we found that the Q-vector system could express higher amounts of viral RNA and proteins compared with the packaging cell system. However, this did not lead to a higher virus titer compared with that produced by the packaging cell system. This suggests that retroviral RNA transcribed from the plasmid in the transient system seemed to be used mainly for translation and only some of the RNA molecules were packaged in viral particles.


Assuntos
Técnicas Genéticas , RNA Viral/química , Retroviridae/genética , Animais , Linhagem Celular , Terapia Genética/métodos , Vetores Genéticos , Humanos , Camundongos , Células NIH 3T3 , Plasmídeos/metabolismo , RNA Mensageiro/metabolismo , RNA Viral/genética , Transfecção , Transgenes
20.
J Am Chem Soc ; 124(19): 5274-5, 2002 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-11996561

RESUMO

A second-generation 1,3,5-phenylene-based dendritic porphyrin decorated with flexible alkyl chains exhibited a liquid crystallinity, and the inclusion of fullerene within the nanospace of the dendritic porphyrin strongly affected the mesophase structure in the thermotropic liquid-crystalline phase.

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