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1.
PLoS Pathog ; 18(9): e1010792, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-36173975

RESUMO

When infecting plants, fungal pathogens secrete cell wall-degrading enzymes (CWDEs) that break down cellulose and hemicellulose, the primary components of plant cell walls. Some fungal CWDEs contain a unique domain, named the carbohydrate binding module (CBM), that facilitates their access to polysaccharides. However, little is known about how plants counteract pathogen degradation of their cell walls. Here, we show that the rice cysteine-rich repeat secretion protein OsRMC binds to and inhibits xylanase MoCel10A of the blast fungus pathogen Magnaporthe oryzae, interfering with its access to the rice cell wall and degradation of rice xylan. We found binding of OsRMC to various CBM1-containing enzymes, suggesting that it has a general role in inhibiting the action of CBM1. OsRMC is localized to the apoplast, and its expression is strongly induced in leaves infected with M. oryzae. Remarkably, knockdown and overexpression of OsRMC reduced and enhanced rice defense against M. oryzae, respectively, demonstrating that inhibition of CBM1-containing fungal enzymes by OsRMC is crucial for rice defense. We also identified additional CBM-interacting proteins (CBMIPs) from Arabidopsis thaliana and Setaria italica, indicating that a wide range of plants counteract pathogens through this mechanism.


Assuntos
Arabidopsis , Oryza , Celulose , Cisteína , Proteínas Fúngicas/genética , Oryza/genética , Xilanos
2.
Gan To Kagaku Ryoho ; 50(13): 1405-1407, 2023 Dec.
Artigo em Japonês | MEDLINE | ID: mdl-38303289

RESUMO

Previous reports have shown that a low attenuated area(LAA)in chest CT reflects the prognosis or therapeutic outcomes for chronic obstructive pulmonary disease(COPD). COPD has been reported as one of the important predictive factors for postoperative pulmonary complications(PPCs). In this retrospective study, we analyzed 100 patients who underwent surgery for gastric or colorectal cancer between January 2020 and August 2022, and investigated the impact of the LAA volume ratio(LAAv%)on short term surgical outcomes. Twenty-two cases developed PPCs, and their Clavien-Dindo grades were Grade Ⅰ-Ⅱ(for 21 patients)and Grade Ⅳa(in 1). We found that high-LAAv% and high-BMI were independent risk factors of PPCs in uni- and multi-variate analyses. In a sub analysis of the limited high-LAAv% group, surgical time qA found to be an independent risk factor of PPCs, and it's cut-off value was calculated as 189 minutes(sensitivity 88.2%, specificity 42.4%). The incidence of PPCs might be avoidable in patients with high-LAAv% through selection of the surgical approach or applying respiratory function training when appropriate.


Assuntos
Neoplasias Gastrointestinais , Doença Pulmonar Obstrutiva Crônica , Humanos , Estudos Retrospectivos , Pulmão , Doença Pulmonar Obstrutiva Crônica/complicações , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Fatores de Risco , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Neoplasias Gastrointestinais/complicações , Resultado do Tratamento
3.
Clin Oral Investig ; 26(10): 6187-6193, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35922680

RESUMO

OBJECTIVES: The purpose of this multicenter retrospective study was to investigate the demographic characteristics and treatment outcomes of patients with mucosal malignant melanoma (MM) of the oral cavity. MATERIALS AND METHODS: This was a multicenter study involving 8 Japanese universities. The medical records of 69 patients who were diagnosed with primary oral MM between January 2000 and December 2020 were retrospectively analyzed. Overall survival (OS) and prognostic factors for OS were analyzed statistically. RESULTS: There were 40 (58.0%) males and 29 (42.0%) females, and their mean (range) age was 69.8 ± 14.6 (22-96) years old. The most common primary site was the palate (30 patients, 43.5%). Stage IVA was the most common disease stage (36 patients, 52.2%). Radical therapy was performed in 55 patients (79.7%). The 2-year and 5-year OS rates of the 69 patients were 64.6% and 42.5%, respectively. The 2-year and 5-year OS rates of the stage III patients were 85.9% and 72.5%, respectively, and those of the stage IVA patients were 56.3% and 26.0%, respectively. The 1-year OS rate of the stage IVB/IVC patients was 26.7%. The 2-year and 5-year OS rates of the radical therapy group were 74.1% and 50.5%, respectively, whereas the 2-year OS rate of the non-radical therapy group was 26.0%. An advanced T classification was the only identified prognostic factor for OS (hazard ratio: 6.312, 95% confidence interval: 1.133-38.522, p < 0.05). CONCLUSIONS: Early detection and radical treatment are essential for improving the prognosis of oral MM patients. CLINICAL RELEVANCE: Early detection and adequate radical therapy leads to the better prognosis of oral MM patients.


Assuntos
Melanoma , Neoplasias Bucais , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Japão , Masculino , Melanoma/patologia , Melanoma/terapia , Pessoa de Meia-Idade , Neoplasias Bucais/patologia , Neoplasias Bucais/terapia , Estadiamento de Neoplasias , Prognóstico , Estudos Retrospectivos , Neoplasias Cutâneas , Melanoma Maligno Cutâneo
4.
Molecules ; 27(3)2022 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-35164384

RESUMO

Anemia is a major complication of chronic renal failure. To treat this anemia, prolylhydroxylase domain enzyme (PHD) inhibitors as well as erythropoiesis-stimulating agents (ESAs) have been used. Although PHD inhibitors rapidly stimulate erythropoietin (Epo) production, the precise sites of Epo production following the administration of these drugs have not been identified. We developed a novel method for the detection of the Epo protein that employs deglycosylation-coupled Western blotting. With protein deglycosylation, tissue Epo contents can be quantified over an extremely wide range. Using this method, we examined the effects of the PHD inhibitor, Roxadustat (ROX), and severe hypoxia on Epo production in various tissues in rats. We observed that ROX increased Epo mRNA expression in both the kidneys and liver. However, Epo protein was detected in the kidneys but not in the liver. Epo protein was also detected in the salivary glands, spleen, epididymis and ovaries. However, both PHD inhibitors (ROX) and severe hypoxia increased the Epo protein abundance only in the kidneys. These data show that, while Epo is produced in many tissues, PHD inhibitors as well as severe hypoxia regulate Epo production only in the kidneys.


Assuntos
Eritropoetina/metabolismo , Glicina/análogos & derivados , Isoquinolinas/farmacologia , Inibidores de Prolil-Hidrolase/farmacologia , Biossíntese de Proteínas/efeitos dos fármacos , Animais , Eritropoetina/análise , Eritropoetina/genética , Feminino , Glicina/farmacologia , Hipóxia/genética , Hipóxia/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Regulação para Cima/efeitos dos fármacos
5.
J Biol Chem ; 294(19): 7942-7965, 2019 05 10.
Artigo em Inglês | MEDLINE | ID: mdl-30926603

RESUMO

endo-ß-1,2-Glucanase (SGL) is an enzyme that hydrolyzes ß-1,2-glucans, which play important physiological roles in some bacteria as a cyclic form. To date, no eukaryotic SGL has been identified. We purified an SGL from Talaromyces funiculosus (TfSGL), a soil fungus, to homogeneity and then cloned the complementary DNA encoding the enzyme. TfSGL shows no significant sequence similarity to any known glycoside hydrolase (GH) families, but shows significant similarity to certain eukaryotic proteins with unknown functions. The recombinant TfSGL (TfSGLr) specifically hydrolyzed linear and cyclic ß-1,2-glucans to sophorose (Glc-ß-1,2-Glc) as a main product. TfSGLr hydrolyzed reducing-end-modified ß-1,2-gluco-oligosaccharides to release a sophoroside with the modified moiety. These results indicate that TfSGL is an endo-type enzyme that preferably releases sophorose from the reducing end of substrates. Stereochemical analysis demonstrated that TfSGL is an inverting enzyme. The overall structure of TfSGLr includes an (α/α)6 toroid fold. The substrate-binding mode was revealed by the structure of a Michaelis complex of an inactive TfSGLr mutant with a ß-1,2-glucoheptasaccharide. Mutational analysis and action pattern analysis of ß-1,2-gluco-oligosaccharide derivatives revealed an unprecedented catalytic mechanism for substrate hydrolysis. Glu-262 (general acid) indirectly protonates the anomeric oxygen at subsite -1 via the 3-hydroxy group of the Glc moiety at subsite +2, and Asp-446 (general base) activates the nucleophilic water via another water. TfSGLr is apparently different from a GH144 SGL in the reaction and substrate recognition mechanism based on structural comparison. Overall, we propose that TfSGL and closely-related enzymes can be classified into a new family, GH162.


Assuntos
Proteínas Fúngicas/química , Glicosídeo Hidrolases/química , Microbiologia do Solo , Talaromyces/enzimologia , Relação Estrutura-Atividade , Especificidade por Substrato
6.
Biochem Biophys Res Commun ; 524(3): 542-548, 2020 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-32014251

RESUMO

ES1 homologs are conserved among prokaryotes and eukaryotes, and the gene expression of ES1 homologs has been confirmed in diverse mammalian tissues. However, the localization and function of mammalian ES1 proteins remain poorly understood. ES1 protein was found specifically expressed in the cone cells of zebrafish and was proposed to contribute to the formation of mega mitochondria. We also observed mega mitochondria in the cone cells of porcine retinas, which raised the question regarding the localization of the porcine ES1. Therefore, in the present study, we aimed to determine the localization of ES1 in porcine retinas. We prepared a rabbit polyclonal antibody against the ES1 C-terminal and performed western blotting analysis and immunoelectron microscopy. The ES1 was found to be localized mainly in the mitochondrial intermembrane space of the porcine retinal cells. Immunopositive signals for ES1 were observed in the mitochondria of almost all retinal cells, and not specifically in cone cells. Our results and the ES1 sequences indicated that the glyoxalase III activity of ES1 might contribute to the stable functionality of the active mitochondria in a protective manner.


Assuntos
Proteínas do Olho/metabolismo , Membranas Mitocondriais/metabolismo , Retina/citologia , Homologia de Sequência de Aminoácidos , Suínos/metabolismo , Sequência de Aminoácidos , Animais , Proteínas do Olho/química , Proteínas Mitocondriais/química , Proteínas Mitocondriais/metabolismo , Células Fotorreceptoras de Vertebrados/citologia , Células Fotorreceptoras de Vertebrados/metabolismo , Células Fotorreceptoras de Vertebrados/ultraestrutura , Retina/ultraestrutura , Solubilidade
7.
Biosci Biotechnol Biochem ; 84(12): 2499-2507, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32799730

RESUMO

Di-N-acetylchitobiase (Ctbs) degrades ß-1,4 glycoside bonds of the chitobiose core of free asparagine-linked glycan. This study examined whether Ctbs degrades chitin-oligosaccharides to GlcNAc in mammals. We analyzed Ctbs mRNA and protein expression in mouse tissues and characterized enzymatic activity using recombinant mouse Ctbs expressed in Escherichia coli. Ctbs mRNA and protein were expressed in various tissues of mouse, including the stomach. Optimal conditions for recombinant Ctbs were pH 3.0 and 45°C, and the recombinant enzyme was retained more than 94% activity after incubation at pH 3.0-7.0 and below 37°C. The recombinant Ctbs hydrolyzed (GlcNAc)3 and (GlcNAc)6 at pH 3.0 and produced GlcNAc. The K m of Ctbs was lowest with (GlcNAc)3 as a substrate. k cat/K m was fourfold as high with (GlcNAc)3 and (GlcNAc)4 as substrates than with (GlcNAc)2. These results suggest that Ctbs digests chitin-oligosaccharides or (GlcNAc)2 of reducing-end residues of oligosaccharides and produces GlcNAc in mouse tissues.


Assuntos
Acetilglucosaminidase/metabolismo , Quitina/química , Quitina/metabolismo , Oligossacarídeos/química , Animais , Cinética , Camundongos , Especificidade por Substrato
8.
J Lipid Res ; 60(9): 1491-1502, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31345992

RESUMO

Fatty acids have various physiological effects on melanoma. For example, palmitic acid (PA) increases melanin levels; linoleic acid and DHA decrease melanin levels; and DHA suppresses tumor growth. In this study, we focused on the relationship between the structure of fatty acids and their physiological effects in melanoma to examine the likely mechanisms of action. We showed that saturated fatty acids and PUFAs display opposing effects on melanin content in melanoma cells. Likewise, PA and EPA have opposing effects in terms of actin polymerization. Our findings suggest that PA and EPA change melanin content in melanoma to alter melanosome trafficking by modulating actin polymerization. Here, we also examined the mechanism of the anti-tumor effect of DHA. We found that DHA interacts with receptor for activated C kinase 1 and represses melanoma cell proliferation by suppressing protein kinase C signaling. Our results suggest a new mechanism to explain the physiological effects of fatty acids.


Assuntos
Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Ácidos Graxos/farmacologia , Melanoma/metabolismo , Actinas/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Di-Hidroxifenilalanina/metabolismo , Humanos , Melaninas/metabolismo , Camundongos , Transdução de Sinais/efeitos dos fármacos
9.
Ann Surg Oncol ; 26(2): 555-563, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30515671

RESUMO

BACKGROUND: This multicenter retrospective study aimed to determine whether elective neck dissection (END) can be performed for T1-2N0M0 tongue cancer. METHODS: Patients with T1-2N0M0 tongue squamous cell carcinoma who received treatment between January 2000 and December 2012 were enrolled at 14 multicenter study sites. The 5-year overall survival (OS) and 5-year disease-specific survival (DSS) were compared between the propensity score-matched END and observation (OBS) groups. RESULTS: The results showed that the OS rates among the 1234 enrolled patients were 85.5% in the END group and 90.2% in the OBS group (P = 0.182). The DSS rates were 87.0% in the END group and 94.3% in the OBS group (P = 0.003). Among the matched patients, the OS rates were 87.1% in the END group and 76.2% in the OBS group (P = 0.0051), and the respective DSS rates were 89.2% and 82.2% (P = 0.0335). CONCLUSION: This study showed that END is beneficial for T1-2N0M0 tongue cancer. However, END should be performed for patients with a tumor depth of 4-5 mm or more, which is the depth associated with a high rate of lymph node metastasis. The use of END should be carefully considered for both elderly and young patients.


Assuntos
Carcinoma de Células Escamosas/cirurgia , Procedimentos Cirúrgicos Eletivos/mortalidade , Esvaziamento Cervical/mortalidade , Neoplasias da Língua/cirurgia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/secundário , Feminino , Seguimentos , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Pontuação de Propensão , Estudos Retrospectivos , Taxa de Sobrevida , Neoplasias da Língua/patologia , Adulto Jovem
10.
Bioorg Med Chem ; 27(24): 115161, 2019 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-31732281

RESUMO

Neomacrophorins I-III (1-3) and X have previously been isolated from Trichoderma sp. 1212-03. Their mode of action against cancer cells and the mechanism of biosynthesis of the characteristic [4.4.3] propellane framework in neomacrophorin X have not been reported. The isolation and characterization of neomacrophorins IV (4), V (5), and VI (6) is reported. Epoxyquinones 1, 4, and 6 potently induced apoptotic cell death in human acute promyelocytic leukemia HL60 cells, while epoxysemiquinols 2, 3, and 5 showed weak activity. This indicates that the epoxyquinone moiety is crucial for apoptosis-inducing activities of neomacrophorins. We also found that neomacrophorins inhibit proteasome in vitro, and 1, 4, and 6 induced significant accumulation of ubiquitinated proteins in HL60 cells. These activities were completely suppressed by a nucleophile, N-acetyl-l-cysteine (NAC). The analysis of reaction mechanisms using LC-MS suggested that C2' and C7' of neomacrophorins could be Michael acceptors in the reaction with NAC methyl ester (NACM). These findings indicated that the electrophilic properties of neomacrophorins are responsible for both their potent biological effects and the biosynthesis of unique [4.4.3] propellane framework in neomacrophorin X.


Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Terpenos/química , Terpenos/farmacologia , Trichoderma/metabolismo , Antineoplásicos/administração & dosagem , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células HL-60 , Humanos , Estrutura Molecular , Terpenos/metabolismo , Trichoderma/química
11.
J Chem Ecol ; 45(7): 579-587, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31256322

RESUMO

Domestic cats (Felis silvestris catus) are solitary and territorial, and mark their territories by spraying urine, which emits a strong odor produced by volatile organic compounds (VOCs). Previous studies have focused on identifications of specific VOCs, such as 3-mercpto-3-methyl-1-butanol, a cat-specific VOCs. However, little is known about how whole volatile profiles of their sprayed urine change over time or how the profiles differ among individuals. This study investigated temporal changes and individual differences of volatile profiles produced by whole VOCs in cat urine, and the ability of cats to discriminate between these scent differences. Volatile profiles of fresh and aged cat urine were analyzed by using two-dimensional gas chromatography-mass spectrometry with a VOC preconcentrator comprehensively. Volatile profiles produced by hundreds of VOCs emitted from cat urine were influenced primarily by the age of the urine, and secondarily by individuality. During habituation-dishabituation tests, subjects discriminated between fresh and 24 h-old samples of same individuals, and between odor of different individuals from 0 h-, 3 h-old, and 24 h-old samples. These results strongly suggest that cats can recognize conspecific individuals via olfaction. Since most VOCs varied among individuals but were not stable over time, their urine may contain unknown VOCs that vary among individuals, are stable over time, and act as individual recognition signals.


Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Compostos Orgânicos Voláteis/urina , Animais , Comportamento Animal , Gatos , Análise Discriminante , Masculino , Análise de Componente Principal , Fatores de Tempo , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/isolamento & purificação
12.
Biochem J ; 475(7): 1309-1322, 2018 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-29581147

RESUMO

Gentiobiose, a ß-1,6-linked glycosyl-disaccharide, accumulates abundantly in Gentianaceae and is involved in aspects of plant development, such as fruits ripening and release of bud dormancy. However, the mechanisms regulating the amount of gentio-oligosaccharide accumulation in plants remain obscure. The present study aimed to identify an enzyme that modulates gentio-oligosaccharide amount in gentian (Gentiana triflora). A protein responsible for gentiobiose hydrolysis, GtGen3A, was identified by partial purification and its peptide sequence analysis. The enzyme had a molecular mass of ∼67 kDa without a secretory signal peptide sequence. Sequence analysis revealed that GtGen3A could be a ß-glucosidase member belonging to glycoside hydrolase family 3 (GH3). GtGen3A showed a homology to GH3 ß-glucan exohydrolases, ExoI of Hordeum vulgare, and ExgI from Zea mays, which preferentially hydrolyzed ß-1,3- and ß-1,4-linked oligosaccharides. The purified recombinant GtGen3A (rGtGen3A) produced in Escherichia coli showed optimal reaction at pH 6.5 and 20°C. The rGtGen3A liberated glucose from ß-1,2-, ß-1,3-, ß-1,4-, and ß-1,6-linked oligosaccharides, and showed the highest activity toward gentiotriose among the substrates tested. Kinetic analysis also revealed that rGtGen3A preferentially hydrolyzed gentiotriose. Virus-induced gene silencing of Gtgen3A in gentian plantlets resulted in predominant accumulation of gentiotriose rather than gentiobiose. Furthermore, the expression level of Gtgen3A was almost similar to the amount of gentiobiose in field-grown gentians. These findings suggest that the main function of GtGen3A is the hydrolysis of gentiotriose to gentiobiose, and that GtGen3A plays a role in modulating gentiobiose amounts in gentian.


Assuntos
Dissacarídeos/metabolismo , Gentiana/enzimologia , Glicosídeos/metabolismo , Oligossacarídeos/metabolismo , beta-Glucosidase/metabolismo , Clonagem Molecular , Gentiana/genética , Gentiana/crescimento & desenvolvimento , Hidrólise , Filogenia , Especificidade por Substrato , beta-Glucosidase/genética
13.
J Biol Chem ; 292(18): 7487-7506, 2017 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-28270506

RESUMO

ß-1,2-Glucan is an extracellular cyclic or linear polysaccharide from Gram-negative bacteria, with important roles in infection and symbiosis. Despite ß-1,2-glucan's importance in bacterial persistence and pathogenesis, only a few reports exist on enzymes acting on both cyclic and linear ß-1,2-glucan. To this end, we purified an endo-ß-1,2-glucanase to homogeneity from cell extracts of the environmental species Chitinophaga arvensicola, and an endo-ß-1,2-glucanase candidate gene (Cpin_6279) was cloned from the related species Chitinophaga pinensis The Cpin_6279 protein specifically hydrolyzed linear ß-1,2-glucan with polymerization degrees of ≥5 and a cyclic counterpart, indicating that Cpin_6279 is an endo-ß-1,2-glucananase. Stereochemical analysis demonstrated that the Cpin_6279-catalyzed reaction proceeds via an inverting mechanism. Cpin_6279 exhibited no significant sequence similarity with known glycoside hydrolases (GHs), and thus the enzyme defines a novel GH family, GH144. The crystal structures of the ligand-free and complex forms of Cpin_6279 with glucose (Glc) and sophorotriose (Glc-ß-1,2-Glc-ß-1,2-Glc) determined up to 1.7 Å revealed that it has a large cavity appropriate for polysaccharide degradation and adopts an (α/α)6-fold slightly similar to that of GH family 15 and 8 enzymes. Mutational analysis indicated that some of the highly conserved acidic residues in the active site are important for catalysis, and the Cpin_6279 active-site architecture provided insights into the substrate recognition by the enzyme. The biochemical characterization and crystal structure of this novel GH may enable discovery of other ß-1,2-glucanases and represent a critical advance toward elucidating structure-function relationships of GH enzymes.


Assuntos
Proteínas de Bactérias/química , Bacteroidetes/enzimologia , Celulase/química , Proteínas de Bactérias/isolamento & purificação , Catálise , Domínio Catalítico , Celulase/isolamento & purificação , Cristalografia por Raios X
14.
Biochem Biophys Res Commun ; 504(2): 454-459, 2018 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-30195492

RESUMO

Calpains are Ca2+-dependent cysteine proteases that are widely distributed in animal tissues and modulate a variety of cellular processes. There are 15 members of the calpain family in mammals. In animal cells, there are three types of calpains, viz., calpain-1, calpain-2, and calpain-10 in the mitochondria. The three types of calpains have been shown to play significant roles in pathophysiological conditions, including in apoptosis- and necrosis-like cell death. One of the severe retinal diseases, autosomal dominant neovascular inflammatory vitreoretinopathy, is known to be induced by mutations of the calpain-5 gene. However, the distribution of calpain-5 in the retina has not been elucidated. Therefore, in the present study, we determined the localization of calpain-5 in the porcine retina. We detected calpain-5 in the inner segment of photoreceptor cells using immunohistochemistry. With immunoelectron microscopy, calpain-5 was localized in the mitochondria of photoreceptor cells. Western blot analyses showed that calpain-5 was present in each mitochondrial subfraction. Furthermore, we showed that the molecular weight of mitochondrial calpain-5 was slightly smaller than cytosolic one. Our results demonstrated that a novel mitochondrial calpian, calpain-5, was localized in the mitochondria of retinal photoreceptor cells.


Assuntos
Calpaína/metabolismo , Mitocôndrias/metabolismo , Células Fotorreceptoras/metabolismo , Retina/metabolismo , Animais , Citosol/metabolismo , Elétrons , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Inflamação , Microscopia Imunoeletrônica , Células Fotorreceptoras de Vertebrados/metabolismo , Frações Subcelulares , Suínos
15.
Biochem Biophys Res Commun ; 503(4): 3121-3127, 2018 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-30146260

RESUMO

Erythropoietin has been thought to be secreted to plasma soon after the production because of the difficulty of Western blot analysis and immunohistochemistry. We established the new methods of Western blot analysis and immunohistochemistry. Using the new methods, we investigated the effects of aldosterone and fludrocortisone, an analogue of aldosterone on erythropoietin mRNA and protein production by the kidneys. Aldosterone stimulated Epo and HIF2α mRNA expressions in tubule suspensions and microdissected medullary thick ascending limbs and outer medullary collecting ducts. Western blot analysis showed a recombinant erythropoietin at 34-45 kDa and kidney erythropoietin at 36-40 and 42 kDa, both of which shifted to 22 kDa by deglycosylation. Erythropoietin protein expression was observed in the nephrons but not in the interstitial cells in control condition. Fludrocortisone stimulated erythropoietin mRNA and protein expressions in the distal nephrons, particularly in the intercalated cells of the collecting ducts. These data show that erythropoietin is produced by the nephrons by the regulation of renin-angiotensin-aldosterone system and not by the renal interstitial cells in control condition.


Assuntos
Aldosterona/metabolismo , Eritropoetina/metabolismo , Fludrocortisona/metabolismo , Túbulos Renais Coletores/metabolismo , Néfrons/metabolismo , Animais , Hipóxia Celular , Eritropoetina/genética , Glicosilação , Túbulos Renais Coletores/citologia , Masculino , Néfrons/citologia , RNA Mensageiro/genética , Ratos Sprague-Dawley , Sistema Renina-Angiotensina , Regulação para Cima
16.
J Chem Ecol ; 44(4): 364-373, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29637491

RESUMO

Scents emitted from excretions provide important information about the owner. Volatile compounds with higher levels in a species and/or sex, or that vary among individuals could be odor cues for species and/or sex, or individual recognition. However, such compounds have been identified in only a few vertebrate species. In domestic cats (Felis silvestris catus), it is known that unburied cat feces are territorial markers asserting the border of their home range, but little was known which fecal compounds are scent cues for species, sex, and individual recognition in cats. In the present study, we demonstrated the chemical basis for species, sex, and individual recognition using feces of cats. For males, major contents were fatty acids and 3-mercapto-3-methyl-1-butanol (MMB), a derivative of the unusual amino acid, felinine. MMB emission levels from feces had sex-based differences (male > female) and dynamic temporal changes during aging. Cats distinguished fecal odors with and without MMB, and different fatty acid compositions among individuals. No cat-specific compound, such as MMB, was detectable from their anal odor emitting fatty acids. We concluded that fecal MMB is a male sex recognition pheromone in cats and also provides a temporal trace of the owner. After sensing MMB, they may distinguish individual differences of conspecific feces with variable subsets of fatty acids. In contrast to scent marks, since cats can obtain species information from visual cues before sniffing conspecific anal odors, they may use their efforts to distinguish individual differences of anal odors during sniffing.


Assuntos
Fezes/química , Atrativos Sexuais/análise , Envelhecimento , Animais , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Gatos , Ácidos Graxos/química , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Masculino , Análise de Componente Principal , Atrativos Sexuais/química , Atrativos Sexuais/farmacologia , Compostos Orgânicos Voláteis/química
17.
Biosci Biotechnol Biochem ; 82(2): 232-237, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29327660

RESUMO

The small Indian mongoose (Herpestes auropunctatus) is an invasive species in Okinawa and Amami-Oshima, Japan. Major strategies for their eradication have been the use of baited traps, which suffer from decreasing efficiency with declining populations and the bycatch of native animals. To address these concerns, mongoose-specific lures are required. In this study, we aimed to identify species- and/or sex-specific compounds from anal sac secretions of small Indian mongooses. Volatile compounds emitted from male and female mongoose anal sac secretions were analyzed by thermal desorption-gas chromatography-mass spectrometry. In addition to several fatty acids, 2-phenylethanol was identified as a minor compound, which is uncommon in mammalian secretions but a dominant odorant in roses. Female samples emitted higher levels of 2-phenylethanol than male samples did. These findings indicate that 2-phenylethanol is a female-specific volatile compound of anal sac secretions in small Indian mongooses, and it may be useful as an ingredient of mongoose-specific scent lures.


Assuntos
Herpestidae/metabolismo , Odorantes/análise , Álcool Feniletílico/metabolismo , Rosa/química , Animais , Feminino , Masculino , Álcool Feniletílico/química , Caracteres Sexuais
18.
Clin Oral Investig ; 22(3): 1311-1318, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28986696

RESUMO

OBJECTIVES: The progression level of extracapsular spread (ECS) for cervical lymph node metastasis of oral squamous cell carcinoma (OSCC) was previously divided into three types, and their relationships with the prognosis of patients were re-examined. PATIENTS AND METHODS: The Kaplan-Meier method was used to examine overall survival (OS) and relapse-free survival (RFS) curves. Prognosis factor for recurrence was analyzed with univariate and multivariate analysis. RESULTS: ECS was detected in 216 cases of OSCC and analyzed. The 5-year overall survival and RFS rates of patients with type C, which was microscopically defined as tumor invasion to perinodal fat or muscle tissue, were significantly poor at 40.6 and 37.8%, respectively. The results of a univariate analysis suggested that the prognosis of ECS in OSCC patients is associated with its progression level, particularly type C. The 5-year RFS rate of type C with tumor budding was significantly poor at 31.5%. Type C with tumor budding correlated with local and regional recurrence as well as distant metastasis. In a multivariate analysis, tumor budding was identified as an independent prognostic factor. CONCLUSIONS: These results suggest that the progression level of ECS and tumor budding are useful prognostic factors in OSCC patients. CLINICAL RELEVANCE: This study indicated that the progression level and tumor budding of ECS for cervical lymph node metastasis were useful prognostic factors in OSCC patients.


Assuntos
Carcinoma de Células Escamosas/patologia , Metástase Linfática/patologia , Neoplasias Bucais/patologia , Invasividade Neoplásica/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Taxa de Sobrevida
19.
Odontology ; 106(1): 96-102, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28936637

RESUMO

The purpose of this study was to investigate the prognostic factor in salivary gland carcinoma patients. Clinical and pathological data of 211 consecutive patients who treated with curative intent were analyzed. The overall survival (OS) rate, local control rate, and distant metastasis rate were calculated. To examine a prognostic factor in salivary gland carcinoma patients, a multivariate analysis was performed. The 5-year-OS rate was 84.0%, and 10-year was 69.2%. The 5-year-local control rate was 84.6%, and 10-year was 70.1%. The 5-year-distant metastasis rate was 16.9%, and 10-year was 21.1%. In a multivariate analysis, the OS rate was affected by pN(+), high-grade malignancy, and primary tumor size. The local control was affected by the primary tumor size, high-grade malignancy, and the status of the surgical margin. The primary tumor size and pN(+) were associated with the distant metastasis. The results of this study suggested that pN(+), malignancy grade, primary tumor size, and the margin status might affect the prognosis of salivary gland carcinoma patients. Postoperative radiotherapy and adjuvant chemotherapy were suggested the possibility of contribution to the good prognosis of salivary gland carcinoma patients.


Assuntos
Neoplasias das Glândulas Salivares/terapia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Estudos Retrospectivos , Neoplasias das Glândulas Salivares/patologia , Taxa de Sobrevida , Resultado do Tratamento
20.
Australas J Dermatol ; 58(4): e223-e227, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28809036

RESUMO

BACKGROUND/OBJECTIVES: The aim of the present study was to investigate treatment modalities and outcomes in oral mucosal melanoma. METHODS: The clinical and pathological data of 38 consecutive patients with oral mucosal melanoma were retrospectively analyzed. Patients' characteristics were analyzed and overall survival (OS) rates were calculated. RESULTS: Sixteen patients had stage III (42%), 19 IVA (50%), and three had stage IVC (8%) disease. Among the therapeutic approaches used, 31 patients (82%) received radical therapy (surgery +/- chemotherapy). The 5-year OS rate was 40%. Five-year OS rates according to the clinical stage were 71% for stage III, 24% for stage IVA, and 0% for stage IVC. Five-year OS rates according to therapeutic approaches were 52% in the radical therapy group and 0% in the palliative therapy and best supportive care groups. CONCLUSIONS: The results of this multicentre retrospective analysis of patients with oral mucosal melanoma suggest that radical therapy based on surgical treatments with complete surgical excision with clear margins leads to a better prognosis.


Assuntos
Melanoma/cirurgia , Neoplasias Bucais/cirurgia , Cuidados Paliativos , Adulto , Idoso , Idoso de 80 Anos ou mais , Quimioterapia Adjuvante , Feminino , Humanos , Japão , Masculino , Melanoma/tratamento farmacológico , Melanoma/secundário , Pessoa de Meia-Idade , Mucosa Bucal , Neoplasias Bucais/tratamento farmacológico , Neoplasias Bucais/patologia , Terapia Neoadjuvante , Estadiamento de Neoplasias , Estudos Retrospectivos , Taxa de Sobrevida , Adulto Jovem
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