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1.
J Comp Pathol ; 187: 27-39, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34503652

RESUMO

Akabane, Aino and Peaton viruses are closely related arthropod-borne viruses in the genus Orthobunyavirus of the family Peribunyaviridae that can cause congenital abnormalities in cattle, sheep and goats. East Asian Akabane virus strains are subdivided into genogroups Ⅰ and Ⅱ, and the former can also cause non-suppurative encephalomyelitis in post-natal animals. Specific detection of the infecting virus in tissues is essential for accurate diagnosis. Immunohistochemistry (IHC) has been used to identify viral antigen but cannot always detect specific viruses due to potential cross-reactivity of the primary antisera. We compared in-situ hybridization (ISH), based on the use of cocktail probe sets targeted at the RNA of each virus, with IHC for the detection of the specific viruses in tissues of suckling rats inoculated intracerebrally with Akabane (KM-1 or OBE-1 strains), Aino or Peaton viruses at 3 or 7 days of age. Most inoculated rats developed severe neurological signs and histopathological brain lesions including necrosis, spongy degeneration and non-suppurative inflammation. A rabbit polyclonal antiserum immunolabelled antigen of all three viruses within the lesions, whereas ISH specifically detected RNA of each individual virus. The distribution of viral RNA was comparable to that of viral antigens, but tended to be more widespread, especially in immature nervous tissue. Viral antigen and RNA were detected in skeletal muscle and heart of the rats infected with the KM-1 strain of Akabane virus but not with any of the other viruses. This study demonstrates the value of ISH detection of these viruses in a rat model and may prove useful for clarification of the pathogenesis of post-natal arbovirus infection.

2.
BMC Microbiol ; 21(1): 215, 2021 07 17.
Artigo em Inglês | MEDLINE | ID: mdl-34273946

RESUMO

BACKGROUND: Legionellosis is caused by the inhalation of aerosolized water contaminated with Legionella bacteria. In this study, we investigated the prevalence of Legionella species in aerosols collected from outdoor sites near asphalt roads, bathrooms in public bath facilities, and other indoor sites, such as buildings and private homes, using amoebic co-culture, quantitative PCR, and 16S rRNA gene amplicon sequencing. RESULTS: Legionella species were not detected by amoebic co-culture. However, Legionella DNA was detected in 114/151 (75.5%) air samples collected near roads (geometric mean ± standard deviation: 1.80 ± 0.52 log10 copies/m3), which was comparable to the numbers collected from bathrooms [15/21 (71.4%), 1.82 ± 0.50] but higher than those collected from other indoor sites [11/30 (36.7%), 0.88 ± 0.56] (P < 0.05). The amount of Legionella DNA was correlated with the monthly total precipitation (r = 0.56, P < 0.01). It was also directly and inversely correlated with the daily total precipitation for seven days (r = 0.21, P = 0.01) and one day (r = - 0.29, P < 0.01) before the sampling day, respectively. 16S rRNA gene amplicon sequencing revealed that Legionella species were detected in 9/30 samples collected near roads (mean proportion of reads, 0.11%). At the species level, L. pneumophila was detected in 2/30 samples collected near roads (the proportion of reads, 0.09 and 0.11% of the total reads number in each positive sample). The three most abundant bacterial genera in the samples collected near roads were Sphingomonas, Streptococcus, and Methylobacterium (mean proportion of reads; 21.1%, 14.6%, and 1.6%, respectively). In addition, the bacterial diversity in outdoor environment was comparable to that in indoor environment which contains aerosol-generating features and higher than that in indoor environment without the features. CONCLUSIONS: DNA from Legionella species was widely present in aerosols collected from outdoor sites near asphalt roads, especially during the rainy season. Our findings suggest that there may be a risk of exposure to Legionella species not only in bathrooms but also in the areas surrounding asphalt roads. Therefore, the possibility of contracting legionellosis in daily life should be considered.

3.
J Vet Med Sci ; 83(9): 1378-1388, 2021 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-34248104

RESUMO

Epizootic hemorrhagic disease (EHD) is an arthropod-borne disease of wild and domestic ruminants caused by the EHD virus (EHDV). To date, seven EHDV serotypes have been identified. In Japan, strain Ibaraki of EHDV serotype 2 has caused outbreaks of Ibaraki disease in cattle. In addition, EHDV serotype 7 (EHDV-7) has caused large-scale EHD epizootics. In mid-September 2016, eight cattle at a breeding farm in Fukuoka Prefecture, Japan developed fever. Since EHDV-7 was detected in sentinel cattle in western Japan in 2016, we suspected that the cause of this fever might be an EHDV-7 infection. In this study, we tested cattle for EHDV-7 and some other viruses. Consequently, EHDV was isolated from washed blood cells collected from three of the eight cattle, and genetic analysis of genome segment 2 revealed that this isolate was EHDV-7. Moreover, all affected cattle tested positive for anti-EHDV-7 neutralizing antibodies. Our results suggest that the fever was caused by EHDV-7 infection. In addition, we modified a conventional reverse transcription polymerase chain reaction assay for the specific detection of EHDV. This modified assay could detect various strains of EHDV isolated in Japan, Australia, and North America. Furthermore, the assay permitted the detection of EHDV-7 in blood cells collected from seven of the eight cattle. We believe that this modified assay will be a useful tool for the diagnosis of EHD.


Assuntos
Doenças dos Bovinos , Vírus da Doença Hemorrágica Epizoótica , Infecções por Reoviridae , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologia , Vírus da Doença Hemorrágica Epizoótica/genética , Japão/epidemiologia , Infecções por Reoviridae/diagnóstico , Infecções por Reoviridae/epidemiologia , Infecções por Reoviridae/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Transcrição Reversa , Sorogrupo
4.
Vet Microbiol ; 258: 109128, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34058522

RESUMO

Although RNA viruses exhibit extensive sequence diversity, the mutation rate must be limited to ensure protein functions that maintain the viral life cycle. Here, we compared the whole genome sequences of 150 isolates of classical swine fever virus (CSFV), obtained from a single epidemic that occurred in Japan during 2018-2020. After the detection of the first case, the disease spread among both farm pigs and wild boars and caused severe impact on the pig industry. To evaluate the diversification of the CSFV genome that eliminated mutations negatively affecting viral transmission, the substitution sets inherited by at least two isolates were separately evaluated as shared single nucleotide variants (SNVs) or shared single amino acid variants (SAVs). Comparisons of 12 protein-coding regions indicated that the percentages of SNVs and SAVs in the multifunctional nonstructural protein NS3 were the lowest, and shared SAVs were not detected in another nonstructural protein, NS4A. This demonstrated purifying negative selection suppressing changes in the protein sequences of NS3 and NS4A during virus transmission in the field. In contrast, a high possibility of nonsynonymous substitution among shared SNVs was detected only in genes encoding the secreted protein Erns and the nonstructural protein NS2, suggesting positive selection during the epidemic. Mapping of shared SAVs to the three-dimensional structure of Erns revealed that shared SAVs were not present in the substrate-binding sites but were instead localized to the peripheral region of the protein. These data will support efforts toward the development of diagnostic methods, recombinant vaccines, and antiviral agents targeting conserved and indispensable viral genes.


Assuntos
Vírus da Febre Suína Clássica/genética , Peste Suína Clássica/virologia , Variação Genética , Sequência de Aminoácidos , Animais , Peste Suína Clássica/epidemiologia , Regulação Viral da Expressão Gênica , Japão/epidemiologia , Modelos Moleculares , Conformação Proteica , Sus scrofa/virologia , Suínos , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/metabolismo
5.
Transbound Emerg Dis ; 2021 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-33890426

RESUMO

After 26 years, another classical swine fever virus (CSFV) outbreak in domestic pigs and wild boars occurred in Japan 2018. Herein, we investigated the entry and the spatial dynamics of the CSFV outbreak in Japan using the nearly complete genomes of strains isolated from both wild boars and domestic pigs during this epidemic. Phylogenetic analysis showed that the most recent common ancestor (MRCA) of the Japanese lineage emerged 146 days (95% highest posterior density (HPD): 85-216 days) before the index case was detected. Based on epidemiological analysis, the period for the 95% HPD was 1 month earlier than the time of virus introduction into the index farm. The disease mainly spreads to the adjoining regions during the epidemic, with no spread to the nonadjacent regions. This result indicates that human activities, such as the movement of vehicles, contributed to the infection spread. As cases occurred in nonadjacent regions, the MRCA for the epidemic in the Saitama prefecture was estimated to have emerged 93 days before the date of detection in the initial farm in this region. Similarly, the MRCA for the epidemic in Okinawa prefecture, more than 1,300 km away from the other infected regions, was estimated to have emerged 34 days before the date of detection in the region's primary farm. Therefore, our results indicate that if exotic diseases emerge after a long period of absence or in a disease-free country, a longer period of time will elapse before detection, resulting in further spread. Additionally, subsequent infections occurring in regions distant from the original infected region will require less time for detection than in the original region. This study provides valuable insights into a CSFV outbreak that occurred in a previously CSFV-free country and thus beneficial in enhancing producers' awareness and allow for better preparation for infections.

6.
Transbound Emerg Dis ; 2021 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-33730417

RESUMO

Classical swine fever (CSF) is a worldwide devastating disease of the pig industry caused by classical swine fever virus (CSFV). In September 2018, an outbreak of CSF occurred in Japan where the disease had been eradicated and was officially designated a CSF-free country since 2015. Following the detection of the first 2018 case on a farm in Gifu Prefecture, the disease spread among both farm pigs and wild boars and still continues. Epigenome analysis using whole-genome information is helpful in identifying the infection route, but the current approaches provide an insufficient resolution. In this study, a novel method of using single-nucleotide variants (SNVs) was employed to identify the associations among 158 isolates (65 from farms and 93 from wild boars). The identified groups of CSFV strains were plotted in different colours on a map, identifying the location where each strain was collected. The lack of an SNV set shared between the index case and the other strains suggested the first infection in Japan during the outbreak occurred in wild boars, not at the index farm. For the Atsumi Peninsula outbreaks, where nine farms were found infected within a 10-km radius area, the farm strains were assembled into three groups, suggesting these outbreaks resulted from at least three different infection events in this area. For the infections in the area around Saitama Prefecture, an area remote from the epicentre, strains from both the farms and wild boars were identified as being in the same group, suggesting they resulted from one viral introduction. Likewise, seven infected farms in Okinawa Prefecture, almost 1,500 km from Gifu Prefecture, were identified as being in a common, but separate group. By demonstrating the variety of transmission routes and possibility of long-distance infection, these results will help improve disease control measures.

7.
Biomaterials ; 270: 120686, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33540171

RESUMO

The accumulation of ß-amyloid (Aß) aggregates in the brain occurs early in the progression of Alzheimer's disease (AD), and non-fibrillar soluble Aß oligomers are particularly neurotoxic. During binding to Aß fibrils, curcumin, which can exist in an equilibrium state between its keto and enol tautomers, exists predominantly in the enol form, and binding activity of the keto form to Aß fibrils is much weaker. Here we described the strong binding activity the keto form of curcumin derivative Shiga-Y51 shows for Aß oligomers and its scant affinity for Aß fibrils. Furthermore, with imaging mass spectrometry we revealed the blood-brain barrier permeability of Shiga-Y51 and its accumulation in the cerebral cortex and the hippocampus, where Aß oligomers were mainly localized, in a mouse model of AD. The keto form of curcumin derivatives like Shiga-Y51 could be promising seed compounds to develop imaging probes and therapeutic agents targeting Aß oligomers in the brain.


Assuntos
Doença de Alzheimer , Curcumina , Doença de Alzheimer/diagnóstico por imagem , Doença de Alzheimer/tratamento farmacológico , Peptídeos beta-Amiloides/metabolismo , Animais , Encéfalo/diagnóstico por imagem , Encéfalo/metabolismo , Modelos Animais de Doenças , Camundongos , Fragmentos de Peptídeos
9.
J Mol Endocrinol ; 66(1): 11-22, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33151898

RESUMO

Cholecystokinin (CCK) is secreted from enteroendocrine I cells in response to fat, carbohydrate, and protein ingestion. Gene expression of nutrient-sensing molecules in I cells remains unclear, primarily due to the difficulty in distinguishing I cells from intestinal epithelial cells in vivo. In this study, we generated CCK reporter male mice in which the red fluorescence protein tdTomato (Tomato) is produced by activation of the native murine Cck promoter. Fluorescence microscopy revealed the presence of Tomato-positive cells in upper small intestine (SI), lower SI, and colon. Flow cytometer analysis revealed that Tomato-positive cells among epithelial cells of upper SI, lower SI, and colon occurred at the rate of 0.95, 0.54, and 0.06%, respectively. In upper SI and lower SI, expression levels of Cck mRNA were higher in Tomato-positive cells than those in Tomato-negative cells. The fatty acid receptors Gpr120, Gpr40, and Gpr43 and the oleoylethanolamide receptor Gpr119 were highly expressed in Tomato-positive cells isolated from SI, but were not found in Tomato-positive cells from colon. The glucose and fructose transporters Sglt1, Glut2, and Glut5 were expressed in both Tomato-positive cells and -negative cells, but these expression levels tended to be decreased in Tomato-positive cells from upper SI to colon. The peptide transporter Pept1 and receptor Gpr93 were expressed in both Tomato-positive cells and -negative cells, whereas Casr was expressed only in Tomato-positive cells isolated from SI. Thus, this transgenic mouse reveals that I cell number and gene expression in I cells vary according to region in the gastrointestinal tract.

10.
Echocardiography ; 37(11): 1749-1756, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32959402

RESUMO

BACKGROUND: Increased body mass index (BMI) is a major risk factor for heart failure with preserved ejection fraction (HFpEF), and HFpEF is more prevalent in elderly females than males. We hypothesized that there may be gender differences in the association between BMI and echocardiographic left ventricular (LV) diastolic parameters. METHODS: We enrolled 456 subjects (243 males) without overt cardiac diseases, all of whom underwent a health checkup. Early (E) and late (A) diastolic transmitral flow velocity, early diastolic mitral annular velocity (e'), and left atrial (LA) volume index were measured by echocardiography to assess LV diastolic function. To examine gender differences in the association between BMI and LV diastolic function, we analyzed the interaction effects of gender on the association between BMI and echocardiographic LV diastolic parameters. RESULTS: Although there were significant gender differences in the association between BMI and E/A and e' in the crude model (interaction effect 0.037 and 0.173, respectively; P = .006 and .022, respectively), these differences were not statistically significant after adjustment for factors related to LV diastolic function. On the other hand, there were significant associations between BMI and LV diastolic parameters in each gender, even after adjustment. CONCLUSIONS: Our findings suggest there is no gender difference in the association between BMI and echocardiographic LV diastolic parameters. However, the association between BMI and LV diastolic parameters was significant in both genders. Controlling body weight might be beneficial for both women and men to prevent progression of LV diastolic dysfunction and development of HFpEF.


Assuntos
Insuficiência Cardíaca , Disfunção Ventricular Esquerda , Idoso , Índice de Massa Corporal , Diástole , Feminino , Insuficiência Cardíaca/diagnóstico por imagem , Humanos , Japão/epidemiologia , Masculino , Caracteres Sexuais , Volume Sistólico , Disfunção Ventricular Esquerda/diagnóstico por imagem , Função Ventricular Esquerda
12.
Genet Mol Biol ; 43(2): e20190017, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32251494

RESUMO

The transcription factor DMRT1 (doublesex and mab-3 related transcription factor) has two distinct functions, somatic-cell masculinization and germ-cell development in some vertebrate species, including mouse and the African clawed frog Xenopus laevis. However, its transcriptional regulation remains unclear. We tried to identify DMRT1-interacting proteins from X. laevis testes by immunoprecipitation with an anti-DMRT1 antibody and MS/MS analysis, and selected three proteins, including PACT/PRKRA (Interferon-inducible double-stranded RNA dependent protein kinase activator A) derived from testes. Next, we examined the effects of PACT/PRKRA and/or p53 on the transcriptional activity of DMRT1. In transfected 293T cells, PACT/PRKRA and p53 significantly enhanced and repressed DMRT1-driven luciferase activity, respectively. We also observed that the enhanced activity by PACT/PRKRA was strongly attenuated by p53. Moreover, in situ hybridization analysis of Pact/Prkra mRNA in tadpole gonads indicated high expression in female and male germline stem cells. Taken together, these findings suggest that PACT/PRKRA and p53 might positively and negatively regulate the activity of DMRT1, respectively, for germline stem cell fate.

14.
J Virol Methods ; 275: 113736, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31669454

RESUMO

A silver amplification immunochromatography (SAI) kit for the detection of all seven serotypes of foot-and-mouth disease virus (FMDV)-FMDV-Ag SAI-was developed using the monoclonal antibody 1H5 recognizing the highly conserved N terminus region of VP2. The FMDV-Ag SAI can be used under conditions of high biosecurity containment as it does not require any apparatus. The FMDV-Ag SAI exhibited 10-100 times higher sensitivity against the five serotypes (O, A, Asia1, C, and SAT1) and similar sensitivity against SAT2 and SAT3, compared with the Svanodip® FMDV-Ag kit immunochromatography kit. The Svanodip kit showed inhibitory results with several saliva samples but not with the FMDV-Ag SAI kit. In a validation study using clinical samples (n = 132; vesicular epithelium = 92, vesicular lesion swabs = 20, saliva = 20) in Mongolia, the sensitivity of FMDV-Ag SAI in comparison with real-time reverse transcription-polymerase chain reaction revealed the following data: vesicular epithelium, 85.4% (76/89); vesicular lesion swab, 46.7% (7/17); and saliva, 36.8% (7/19). No cross-reactivity with the non-FMDV vesicular-forming viruses and taxonomically related viruses of the Picornaviridae family occurred. The FMDV-Ag SAI is a highly sensitive diagnostic tool that enables pen-side diagnosis without requiring the use of any equipment.


Assuntos
Antígenos Virais/isolamento & purificação , Cromatografia de Afinidade/instrumentação , Vírus da Febre Aftosa/isolamento & purificação , Kit de Reagentes para Diagnóstico , Prata/química , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Linhagem Celular , Febre Aftosa/diagnóstico , Vírus da Febre Aftosa/classificação , Sensibilidade e Especificidade , Sorogrupo
15.
Microbiol Resour Announc ; 8(37)2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31515349

RESUMO

Sapelovirus A (SV-A) is currently spreading as an enteric pathogen of pigs worldwide. We isolated SV-A strain XTND/2018 from the small intestine of a dead pig with severe diarrhea in the north of Vietnam and determined the genomic sequence. This is the first report of the genomic sequence of SV-A circulating in Vietnam.

16.
Sci Rep ; 9(1): 12560, 2019 08 29.
Artigo em Inglês | MEDLINE | ID: mdl-31467356

RESUMO

SRY (sex-determining region Y)-box 9 (SOX9) is a transcription factor regulating both chondrogenesis and sex determination. Among vertebrates, SOX9's functions in chondrogenesis are well conserved, while they vary in sex determination. To investigate the conservation of SOX9's regulatory functions in chondrogenesis and gonad development among species, we performed chromatin immunoprecipitation sequencing (ChIP-seq) using developing limb buds and male gonads from embryos of two vertebrates, mouse and chicken. In both mouse and chicken, SOX9 bound to intronic and distal regions of genes more frequently in limb buds than in male gonads, while SOX9 bound to the proximal upstream regions of genes more frequently in male gonads than in limb buds. In both species, SOX palindromic repeats were identified more frequently in SOX9 binding regions in limb bud genes compared with those in male gonad genes. The conservation of SOX9 binding regions was significantly higher in limb bud genes. In addition, we combined RNA expression analysis (RNA sequencing) with the ChIP-seq results at the same stage in developing chondrocytes and Sertoli cells and determined SOX9 target genes in these cells of the two species and disclosed that SOX9 targets showed high similarity of targets in chondrocytes, but not in Sertoli cells.


Assuntos
Condrócitos/metabolismo , Sequência Conservada , Fatores de Transcrição SOX9/metabolismo , Células de Sertoli/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Galinhas , Condrócitos/citologia , Masculino , Camundongos , Ligação Proteica , Células de Sertoli/citologia , Especificidade por Substrato
17.
Curr Cardiol Rep ; 21(6): 53, 2019 05 10.
Artigo em Inglês | MEDLINE | ID: mdl-31076948

RESUMO

PURPOSE OF REVIEW: Noninvasive hemodynamic assessments in patients with heart failure (HF) are essential for appropriate diagnosis and establishment of the best treatment strategies. Recently, the impact of pulmonary circulation and right ventricular function on prognosis in HF patients has drawn increasing attention. In this article, we explore the usefulness of cardiac imaging for hemodynamic assessments, mainly focusing on echocardiographic evaluation. RECENT FINDINGS: The reliability of Doppler echocardiography as a noninvasive alternative to Swan-Ganz catheterization has been well investigated with higher than 80% accuracy for estimating pulmonary artery pressure. Strain measurement and three-dimensional echocardiography are useful for evaluating right ventricular function together with pulmonary circulation. The accuracy of analyzing left and right ventricular functions by cardiac computed tomography and cardiac magnetic resonate imaging has also been established. These modalities can provide myocardial tissue information and allow calculation of the extracellular volume fraction as well. According to the rapid improvement of technologies, cardiac imaging has become an essential tool for hemodynamic evaluation in HF management.


Assuntos
Técnicas de Imagem Cardíaca/métodos , Insuficiência Cardíaca/diagnóstico por imagem , Insuficiência Cardíaca/fisiopatologia , Pneumopatias/fisiopatologia , Pulmão/fisiopatologia , Cateterismo Cardíaco , Hemodinâmica , Humanos , Reprodutibilidade dos Testes
18.
Vet Ital ; 55(1): 81-89, 2019 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-30951185

RESUMO

Viruses of the Simbu serogroup are arboviruses that are known to cause outbreaks of abortion, stillbirth and congenitally deformed neonates. This study presents the results of antibody screening of Simbu serogroup viruses in heifers born in Israel after October 2013, and in adult milking cows born before May 2012. Thirteen dairy cattle farms in five regions, and one sheep flock, entered this study. Serum samples that were found to be positive by ELISA were further tested by specific virus- neutralization test against a panel of Simbu serogroup viruses including Akabane, Aino, Sathuperi, Shamonda, and Peaton viruses. Antibody detection in lactating adult cows revealed that several viruses were circulating in Israel between 2008-2014. Moreover, during autumn 2014 the heifers became serum-positive after being exposed to more than one Simbu serogroup virus concurrently. The results of this study shed new light on Simbu virus infections in Israel, and may contribute to the epidemiology of the Simbu serogroup around the Mediterranean Basin in general.


Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/virologia , Vírus Simbu/fisiologia , Animais , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/virologia , Bovinos , Doenças dos Bovinos/epidemiologia , Feminino , Israel/epidemiologia , Sorogrupo , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/virologia , Vírus Simbu/classificação , Vírus Simbu/genética
19.
J Diabetes Investig ; 10(6): 1430-1437, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31002464

RESUMO

AIMS/INTRODUCTION: Incretin hormone glucose-dependent insulinotropic polypeptide/gastric inhibitory polypeptide (GIP) plays a key role in high-fat diet-induced obesity and insulin resistance. GIP is strongly secreted from enteroendocrine K cells by oil ingestion. G protein-coupled receptor (GPR)120 and GPR40 are two major receptors for long chain fatty acids, and are expressed in enteroendocrine K cells. In the present study, we investigated the effect of the two receptors on oil-induced GIP secretion using GPR120- and GPR40-double knockout (DKO) mice. MATERIALS AND METHODS: Global knockout mice of GPR120 and GPR40 were crossbred to generate DKO mice. Oral glucose tolerance test and oral corn oil tolerance test were carried out. For analysis of the number of K cells and gene expression in K cells, DKO mice were crossbred with GIP-green fluorescent protein knock-in mice in which visualization and isolation of K cells can be achieved. RESULTS: Double knockout mice showed normal glucose-induced GIP secretion, but no GIP secretion by oil. We then investigated the number of K cells and gene characteristics in K cells isolated from GIP-green fluorescent protein knock-in mice. Deficiency of both receptors did not affect the number of K cells in the small intestine or expression of GIP messenger ribonucleic acid in K cells. Furthermore, there was no significant difference in the expression of the genes associated with lipid absorption or GIP secretion in K cells between wild-type and DKO mice. CONCLUSIONS: Oil-induced GIP secretion is triggered by the two major fatty acid receptors, GPR120 and GPR40, without changing K-cell number or K-cell characteristics.


Assuntos
Óleo de Milho/farmacologia , Ácidos Graxos/metabolismo , Polipeptídeo Inibidor Gástrico/metabolismo , Receptores Acoplados a Proteínas G/fisiologia , Animais , Polipeptídeo Inibidor Gástrico/efeitos dos fármacos , Teste de Tolerância a Glucose , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout
20.
J Med Chem ; 62(10): 4915-4935, 2019 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-31009559

RESUMO

Anaplastic lymphoma kinase (ALK), a member of the receptor tyrosine kinase family, is predominantly expressed in the brain and implicated in neuronal development and cognition. However, the detailed function of ALK in the central nervous system (CNS) is still unclear. To elucidate the role of ALK in the CNS, it was necessary to discover a potent, selective, and brain-penetrant ALK inhibitor. Scaffold hopping and lead optimization of N-(2,4-difluorobenzyl)-3-(1 H-pyrazol-5-yl)imidazo[1,2- b]pyridazin-6-amine 1 guided by a cocrystal structure of compound 1 bound to ALK resulted in the identification of (6-(1-(5-fluoropyridin-2-yl)ethoxy)-1-(5-methyl-1 H-pyrazol-3-yl)-1 H-pyrrolo[2,3- b]pyridin-3-yl)((2 S)-2-methylmorpholin-4-yl)methanone 13 as a highly potent, selective, and brain-penetrable compound. Intraperitoneal administration of compound 13 significantly decreased the phosphorylated-ALK (p-ALK) levels in the hippocampus and prefrontal cortex in the mouse brain. These results suggest that compound 13 could serve as a useful chemical probe to elucidate the mechanism of ALK-mediated brain functions and the therapeutic potential of ALK inhibition.


Assuntos
Quinase do Linfoma Anaplásico/antagonistas & inibidores , Encéfalo/efeitos dos fármacos , Descoberta de Drogas/métodos , Inibidores de Proteínas Quinases/síntese química , Animais , Transporte Biológico , Encéfalo/metabolismo , Cristalografia por Raios X , Células HEK293 , Humanos , Concentração Inibidora 50 , Células LLC-PK1 , Camundongos , Camundongos Endogâmicos ICR , Estrutura Molecular , Fosforilação , Inibidores de Proteínas Quinases/sangue , Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacologia , Relação Estrutura-Atividade , Suínos
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