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1.
Arq Bras Oftalmol ; 83(2): 149-152, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32159596

RESUMO

Gyrate atrophy is a rare metabolic autosomal recessive disorder caused by ornithine aminotransferase enzyme deficiency that leads to characteristic progressive, degenerative chorioretinal findings. Patients complain mostly of low vision, night blindness, and peripheral vision loss. Posterior subcapsular cataract, myopia, choroid neovascularization, and intraretinal cysts may be accompanying factors related to vision loss. We encountered a patient with vision loss secondary to posterior subcapsular cataract and intraretinal cysts. After treatment with topical brinzolamide and nepafenac (and without any diet mo dification and/or supplementation), we observed 143- and 117-mm macular thickness resolutions with 2 and 1 Snellen lines of visual gain in his right and left eyes, respectively. Also, we detected a novel homozygous mutation in the ornithine aminotransferase gene: c.1253T>C (p.Leu418Pro). Carbonic anhydrase inhibitors and/or non-steroid anti-inflammatory drugs can control macular edema in patients with gyrate atrophy-associated intraretinal cysts. The genetic variants may also be a determinant in the responsiveness to the therapy type.


Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Benzenoacetamidas/administração & dosagem , Inibidores da Anidrase Carbônica/administração & dosagem , Atrofia Girata/genética , Edema Macular/tratamento farmacológico , Fenilacetatos/administração & dosagem , Sulfonamidas/administração & dosagem , Tiazinas/administração & dosagem , Administração Oftálmica , Adulto , Angiofluoresceinografia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Edema Macular/diagnóstico por imagem , Masculino , Mutação , Ornitina-Oxo-Ácido Transaminase/genética , Tomografia de Coerência Óptica
2.
Nat Commun ; 11(1): 504, 2020 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-31980629

RESUMO

Endogenous homeostatic mechanisms can restore normal neuronal function following cocaine-induced neuroadaptations. Such mechanisms may be exploited to develop novel therapies for cocaine addiction, but a molecular target has not yet been identified. Here we profiled mouse gene expression during early and late cocaine abstinence to identify putative regulators of neural homeostasis. Cocaine activated the transcription factor, Nr4a1, and its target gene, Cartpt, a key molecule involved in dopamine metabolism. Sustained activation of Cartpt at late abstinence was coupled with depletion of the repressive histone modification, H3K27me3, and enrichment of activating marks, H3K27ac and H3K4me3. Using both CRISPR-mediated and small molecule Nr4a1 activation, we demonstrated the direct causal role of Nr4a1 in sustained activation of Cartpt and in attenuation of cocaine-evoked behavior. Our findings provide evidence that targeting abstinence-induced homeostatic gene expression is a potential therapeutic target in cocaine addiction.


Assuntos
Comportamento Animal/efeitos dos fármacos , Cocaína/farmacologia , Epigênese Genética , Homeostase/efeitos dos fármacos , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Animais , Sistemas CRISPR-Cas/genética , Cocaína/administração & dosagem , Epigênese Genética/efeitos dos fármacos , Feminino , Histonas/metabolismo , Homeostase/genética , Masculino , Camundongos Endogâmicos C57BL , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fenilacetatos/farmacologia , Regiões Promotoras Genéticas/genética , Processamento de Proteína Pós-Traducional , Sinapsinas/metabolismo
3.
Medicine (Baltimore) ; 99(4): e18683, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31977858

RESUMO

Vicagrel is a new antiplatelet pro-drug based on clopidogrel sulfur lactone metabolites. The purpose of the study was to evaluate the safety, tolerability, and pharmacodynamics (PD) of vicagrel in healthy Chinese subjects.This study was designed as a single-center, randomized, double-blind, placebo-controlled, single oral ascending dose study. Fifty nine subjects were assigned to 6 vicagrel dose cohorts (5, 10, 20, 40, 60, and 75 mg), and 8 subjects were assigned to 75 mg clopidogrel. Within each vicagrel dose cohort, the 10 subjects (9 in the 75 mg cohort) were randomized 4:1 to receive vicagrel or placebo. Platelet function was assessed using VerifyNow P2Y12. ΔP2Y12 reaction units (ΔPRU) and percent inhibition platelet aggregation (%IPA) were used to evaluate the PD of vicagrel.Although the number of adverse events (AEs) increased with vicagrel dose, none were considered serious, suggesting that vicagrel is safe and well-tolerated. The ΔPRU and %IPA patterns suggest that inhibition of ADP-induced platelet aggregation increased in a dose-dependent manner across the 10 to 40 mg dose range. The inhibitory effect was nearly complete at 4 hours (mean %IPA 87.9%-93.0%, mean ΔPRU 206.6-240.0) for doses of 40 to 75 mg of vicagrel. In contrast, for 5 mg vicagrel and 75 mg clopidogrel, there were no measurable effects on platelet aggregation throughout the study.The results suggest that vicagrel at 40 to 75 mg inhibits ADP-induced platelet aggregation, with a fast onset of action and significantly greater potency than clopidogrel. These findings indicate that vicagrel may be a highly effective and well-tolerated antiplatelet agent.


Assuntos
Fenilacetatos/farmacologia , Inibidores da Agregação de Plaquetas/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Tiofenos/farmacologia , Adulto , Clopidogrel/farmacologia , Relação Dose-Resposta a Droga , Método Duplo-Cego , Feminino , Humanos , Masculino , Fenilacetatos/administração & dosagem , Fenilacetatos/efeitos adversos , Ativação Plaquetária/efeitos dos fármacos , Inibidores da Agregação de Plaquetas/administração & dosagem , Inibidores da Agregação de Plaquetas/efeitos adversos , Testes de Função Plaquetária , Tiofenos/administração & dosagem , Tiofenos/efeitos adversos , Adulto Jovem
5.
J Pharm Biomed Anal ; 177: 112883, 2020 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-31546136

RESUMO

The article is devoted to the application of modern sample preparation technique - microextraction by packed sorbent (MEPS) - in conjunction with non-conventional type of sorbent - hypercrosslinked polystyrene, that was investigated for the first time in this work. Their combination was used to extract phenylcarboxylic acid-type aromatic microbial metabolites from serum samples of a healthy volunteer with following derivatization and GC-MS detection. As barrel insert and needle for MEPS with hypercrosslinked polystyrene is not produced, we designed a device to imitate the commercial MEPS system with packed granular biporous hypercrosslinked polystyrene. Nine aromatic microbial metabolites, including sepsis associated phenyllactic, 4-hydroxyphenyllactic and 4-hydroxyphenylacetic acids, were extracted from serum samples (recoveries were 20-70%) and a linear dependence was revealed in the most clinically significant range of concentrations (0.5-18 µM). The results obtained demonstrate the perspective of the applying of hypercrosslinked polystyrene in commercial devices for MEPS for the future analyses of biological samples, in particular for the early diagnosis of sepsis and treatment effectiveness control.


Assuntos
Bactérias/metabolismo , Fenilacetatos/sangue , Poliestirenos/química , Sepse/diagnóstico , Microextração em Fase Sólida/métodos , Reagentes para Ligações Cruzadas/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Voluntários Saudáveis , Humanos , Limite de Detecção , Fenilacetatos/metabolismo , Sepse/sangue , Sepse/microbiologia
7.
Genes (Basel) ; 11(1)2019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31878002

RESUMO

Vulpinic acid, a naturally occurring methyl ester of pulvinic acid, has been reported to exert anti-fungal, anti-cancer, and anti-oxidative effects. However, its metabolic action has not been implicated yet. Here, we show that vulpinic acid derived from a mushroom, Pulveroboletus ravenelii controls the cell fate of mesenchymal stem cells and preadipocytes by inducing the acetylation of histone H3 and α-tubulin, respectively. The treatment of 10T1/2 mesenchymal stem cells with vulpinic acid increased the expression of Wnt6, Wnt10a, and Wnt10b, which led to osteogenesis inhibiting the adipogenic lineage commitment, through the upregulation of H3 acetylation. By contrast, treatment with vulpinic acid promoted the terminal differentiation of 3T3-L1 preadipocytes into mature adipocytes. In this process, the increase in acetylated tubulin was accompanied, while acetylated H3 was not altered. As excessive generation of adipocytes occurs, the accumulation of lipid drops was not concentrated, but dispersed into a number of adipocytes. Consistently, the expressions of lipolytic genes were upregulated and inflammatory factors were downregulated in adipocytes exposed to vulpinic acid during adipogenesis. These findings reveal the multiple actions of vulpinic acid in two stages of differentiation, promoting the osteogenesis of mesenchymal stem cells and decreasing hypertrophic adipocytes, which can provide experimental evidence for the novel metabolic advantages of vulpinic acid.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Furanos/farmacologia , Fenilacetatos/farmacologia , Células-Tronco/efeitos dos fármacos , Células 3T3-L1 , Adipócitos/metabolismo , Adipogenia/fisiologia , Animais , Furanos/metabolismo , Lipólise/fisiologia , Células-Tronco Mesenquimais , Camundongos , Osteogênese/fisiologia , Fenilacetatos/metabolismo , Transdução de Sinais , Células-Tronco/metabolismo
8.
Respir Res ; 20(1): 288, 2019 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-31856838

RESUMO

BACKGROUND: Although allergic asthma is a complex area with many interacting factors involved, the 'hygiene hypothesis' proposes that a lack of exposure to infection during childhood may polarise the immune system towards allergen-reactive Th2-type responses in genetically susceptible individuals. Toll-like receptors (TLRs) play a key role within the innate immune system and TLR7 agonists have previously been shown to up-regulate Th1 responses and down-regulate Th2 responses to allergens in murine models of allergic or chronic asthma. This study aimed to examine the efficacy and safety of the novel TRL7 agonist AZD8848, which has been developed as an antedrug. METHODS: In this double-blind, randomised, parallel-group study, AZD8848 60 µg or placebo was administered intranasally once-weekly for 8 weeks in patients with mild-to-moderate allergic asthma (NCT00999466). Efficacy assessments were performed at 1 and 4 weeks after the last dose. The primary outcome was the late asthmatic response (LAR) fall in forced expiratory volume in 1 s (FEV1) after allergen challenge at 1-week post-treatment. RESULTS: AZD8848 significantly reduced average LAR fall in FEV1 by 27% vs. placebo at 1 week after treatment (p = 0.035). This effect was sustained at 4 weeks post-treatment; however, it did not reach clinical significance. AZD8848 reduced post-allergen challenge methacholine-induced airway hyper-responsiveness (AHR) vs. placebo at 1 week post-dosing (treatment ratio: 2.20, p = 0.024), with no effect at 4 weeks. There was no significant difference between the two groups in plasma cytokine, sputum Th2 cytokine or eosinophil responses post-allergen challenge at 1 week after treatment. The incidence of adverse events was similar in the two groups. AZD8848 was generally well tolerated. CONCLUSIONS AND CLINICAL RELEVANCE: In patients with allergic asthma, TLR7 agonists could potentially reduce allergen responsiveness by stimulating Type 1 interferon responses to down-regulate the dominant Th2 responses. TRIAL REGISTRATION: clinicaltrials.gov identifier NCT00999466.


Assuntos
Adenosina/análogos & derivados , Alérgenos/efeitos adversos , Antiasmáticos/administração & dosagem , Asma/tratamento farmacológico , Testes de Provocação Brônquica/métodos , Fenilacetatos/administração & dosagem , Receptor 7 Toll-Like/agonistas , Adenosina/administração & dosagem , Administração Intranasal , Adolescente , Adulto , Asma/induzido quimicamente , Asma/metabolismo , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Adulto Jovem
9.
ACS Chem Biol ; 14(12): 2876-2886, 2019 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-31689071

RESUMO

Numerous aromatic compounds are aerobically degraded in bacteria via the central intermediate phenylacetic acid (paa). In one of the key steps of this widespread catabolic pathway, 1,2-epoxyphenylacetyl-CoA is converted by PaaG into the heterocyclic oxepin-CoA. PaaG thereby elegantly generates an α,ß-unsaturated CoA ester that is predisposed to undergo ß-oxidation subsequent to hydrolytic ring-cleavage. Moreover, oxepin-CoA serves as a precursor for secondary metabolites (e.g., tropodithietic acid) that act as antibiotics and quorum-sensing signals. Here we verify that PaaG adopts a second role in aromatic catabolism by converting cis-3,4-didehydroadipoyl-CoA into trans-2,3-didehydroadipoyl-CoA and corroborate a Δ3,Δ2-enoyl-CoA isomerase-like proton shuttling mechanism for both distinct substrates. Biochemical and structural investigations of PaaG reveal active site adaptations to the structurally different substrates and provide detailed insight into catalysis and control of stereospecificity. This work elucidates the mechanism of action of unusual isomerase PaaG and sheds new light on the ubiquitous enoyl-CoA isomerases of the crotonase superfamily.


Assuntos
Bactérias/metabolismo , Coenzima A/metabolismo , Isomerases/metabolismo , Oxepinas/metabolismo , Catálise , Isomerases/química , Ligantes , Fenilacetatos/metabolismo , Conformação Proteica , Metabolismo Secundário
10.
Biomed Res Int ; 2019: 2612849, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31781604

RESUMO

Background: Sepsis-associated encephalopathy (SAE) is a transient and reversible brain dysfunction, that occurs when the source of sepsis is located outside of the central nervous system; SAE affects nearly 30% of septic patients at admission and is a risk factor for mortality. In our study, we sought to determine whether metabolite changes in plasma could be a potential biomarker for the early diagnosis and/or the prediction of the prognosis of sepsis. Method: A total of 31 SAE patients and 28 healthy controls matched by age, gender, and body mass index (BMI) participated in our study. SAE patients were divided into four groups according to the Glasgow Coma Score (GCS). Plasma samples were collected and used to detect metabolism changes by gas chromatography-mass spectrometry (GC-MS). Analysis of variance was used to determine which metabolites significantly differed between the control and SAE groups. Results: We identified a total of 63 metabolites that showed significant differences among the SAE and control groups. In particular, the 4 common metabolites in the four groups were 4-hydroxyphenylacetic acid; carbostyril, 3-ethyl-4,7-dimethoxy (35.8%); malic acid peak 1; and oxalic acid. The concentration of 4-hydroxyphenylacetic acid in sepsis patients decreased with a decrease of the GCS. Conclusions: According to recent research on SAE, metabolic disturbances in tissue and cells may be the main pathophysiology of this condition. In our study, we found a correlation between the concentration of 4-hydroxyphenylacetic acid and the severity of consciousness disorders. We suggest that 4-hydroxyphenylacetic acid may be a potential biomarker for SAE and useful in predicting patient prognosis.


Assuntos
Biomarcadores/sangue , Metabolômica , Encefalopatia Associada a Sepse/sangue , Sepse/sangue , Idoso , Índice de Massa Corporal , Diagnóstico Precoce , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Escala de Coma de Glasgow , Humanos , Hidroxiquinolinas/sangue , Unidades de Terapia Intensiva , Malatos/sangue , Masculino , Pessoa de Meia-Idade , Ácido Oxálico/sangue , Fenilacetatos/sangue , Prognóstico , Quinolonas/sangue , Sepse/complicações , Sepse/patologia , Encefalopatia Associada a Sepse/complicações , Encefalopatia Associada a Sepse/patologia
11.
Int J Mol Sci ; 20(20)2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31658607

RESUMO

The molecular recognition process and the ability to form multicomponent supramolecular systems have been investigated for the amide of triphenylacetic acid and l-tyrosine (N-triphenylacetyl-l-tyrosine, TrCOTyr). The presence of several supramolecular synthons within the same amide molecule allows the formation of various multicomponent crystals, where TrCOTyr serves as a chiral host. Isostructural crystals of solvates with methanol and ethanol and a series of binary crystalline molecular complexes with selected organic diamines (1,5-naphthyridine, quinoxaline, 4,4'-bipyridyl, and DABCO) were obtained. The structures of the crystals were planned based on non-covalent interactions (O-H···N or N-H+···O- hydrogen bonds) present in a basic structural motif, which is a heterotrimeric building block consisting of two molecules of the host and one molecule of the guest. The complex of TrCOTyr with DABCO is an exception. The anionic dimers built off the TrCOTyr molecules form a supramolecular gutter, with trityl groups located on the edge and filled by DABCO cationic dimers. Whereas most of the racemic mixtures crystallize as racemic crystals or as conglomerates, the additional tests carried out for racemic N-triphenylacetyl-tyrosine (rac-TrCOTyr) showed that the compound crystallizes as a solid solution of enantiomers.


Assuntos
2,2'-Dipiridil/química , Diaminas/química , Fenilacetatos/química , Sais/química , Tirosina/química , Amidas , Varredura Diferencial de Calorimetria , Cátions , Cristalização , Cristalografia por Raios X , Ligação de Hidrogênio , Modelos Moleculares , Estrutura Molecular , Soluções/química , Estereoisomerismo
12.
Eur J Med Chem ; 183: 111673, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31536894

RESUMO

Cryptosporidiosis is a human gastrointestinal disease caused by protozoans of the genus Cryptosporidium, which can be fatal in immunocompromised individuals. The essential enzyme, thymidylate synthase (TS), is responsible for de novo synthesis of deoxythymidine monophosphate. The TS active site is relatively conserved between Cryptosporidium and human enzymes. In previous work, we identified compound 1, (2-amino-4-oxo-4,7-dihydro-pyrrolo[2,3-d]pyrimidin-methyl-phenyl-l-glutamic acid), as a promising selective Cryptosporidium hominis TS (ChTS) inhibitor. In the present study, we explore the structure-activity relationship around 1 glutamate moiety by synthesizing and biochemically evaluating the inhibitory activity of analogues against ChTS and human TS (hTS). X-Ray crystal structures were obtained for compounds bound to both ChTS and hTS. We establish the importance of the 2-phenylacetic acid moiety methylene linker in optimally positioning compounds 23, 24, and 25 within the active site. Moreover, through the comparison of structural data for 5, 14, 15, and 23 bound in both ChTS and hTS identified that active site rigidity is a driving force in determining inhibitor selectivity.


Assuntos
Cryptosporidium/enzimologia , Glutamatos/química , Fenilacetatos/química , Pirimidinas/química , Pirróis/química , Timidilato Sintase/antagonistas & inibidores , Domínio Catalítico , Desenho de Fármacos , Humanos , Modelos Moleculares , Estrutura Molecular , Ligação Proteica , Timidilato Sintase/química
13.
Nat Commun ; 10(1): 4127, 2019 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-31511507

RESUMO

Substrate channeling is a mechanism for the internal transfer of hydrophobic, unstable or toxic intermediates from the active site of one enzyme to another. Such transfer has previously been described to be mediated by a hydrophobic tunnel, the use of electrostatic highways or pivoting and by conformational changes. The enzyme PaaZ is used by many bacteria to degrade environmental pollutants. PaaZ is a bifunctional enzyme that catalyzes the ring opening of oxepin-CoA and converts it to 3-oxo-5,6-dehydrosuberyl-CoA. Here we report the structures of PaaZ determined by electron cryomicroscopy with and without bound ligands. The structures reveal that three domain-swapped dimers of the enzyme form a trilobed structure. A combination of small-angle X-ray scattering (SAXS), computational studies, mutagenesis and microbial growth experiments suggests that the key intermediate is transferred from one active site to the other by a mechanism of electrostatic pivoting of the CoA moiety, mediated by a set of conserved positively charged residues.


Assuntos
Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimologia , Metaboloma , Fenilacetatos/metabolismo , Sítios de Ligação , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/ultraestrutura , Modelos Moleculares , Fenilacetatos/química , Domínios Proteicos , Especificidade por Substrato
14.
J Vet Med Sci ; 81(9): 1379-1384, 2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31366852

RESUMO

In healthy dogs, amino acid infusion significantly attenuates the decrease in body temperature during anesthesia by facilitating insulin secretion, suggesting that such an increase in insulin secretion is related to increased heat production. In dogs, selective cyclooxygenase-2 (COX-2) inhibitors, which are used for pain relief in veterinary medicine, possess anti-pyretic action. And, in mice and humans, selective COX-2 inhibitors increase insulin secretion and sensitivity. Therefore, treatment with COX-2 inhibitors may negate or accelerate the attenuating effect on decreased body temperature during anesthesia by amino acid infusion. In the present study, influences on insulin secretion and body temperature by treatment with meloxicam or robenacoxib at therapeutic dose were evaluated in healthy dogs. Treatment with meloxicam or robenacoxib did not affect insulin secretion in the unanesthetized and anesthetized dogs, and did not affect body temperature and heart rate under the anesthetized condition with amino acid infusion. In conclusion, COX-2 inhibitors at therapeutic doses did not affect body temperature during anesthesia in dogs administered amino acids.


Assuntos
Anestesia/veterinária , Inibidores de Ciclo-Oxigenase 2/farmacologia , Difenilamina/análogos & derivados , Meloxicam/farmacologia , Fenilacetatos/farmacologia , Aminoácidos/administração & dosagem , Anestesia/efeitos adversos , Animais , Temperatura Corporal/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase 2/administração & dosagem , Difenilamina/administração & dosagem , Difenilamina/farmacologia , Cães , Feminino , Teste de Tolerância a Glucose/veterinária , Frequência Cardíaca/efeitos dos fármacos , Infusões Intravenosas/veterinária , Secreção de Insulina/efeitos dos fármacos , Masculino , Meloxicam/administração & dosagem , Fenilacetatos/administração & dosagem
15.
Immunohorizons ; 3(8): 402-411, 2019 08 22.
Artigo em Inglês | MEDLINE | ID: mdl-31439624

RESUMO

Chronically activated CD4+ T cells drive uncontrolled inflammation, leading to tissue damage in various autoimmune disorders, such as rheumatoid arthritis (RA). Investigation of the molecular mechanisms involved in RA and recent analysis of transcriptomic profiles has implicated members of the nuclear receptor (NR) superfamily in RA. NRs are required for the development, differentiation, and effector function of CD4+ T cells; therefore, it is thought that NRs are important in shaping the CD4+ T cell repertoire and associated inflammation in RA. Despite their relevance, the full potential of the NR superfamily in RA, either as biomarkers or disease targets, has not been harnessed. To gain insight on the NR members that are closely associated with RA disease activity, we generated an expression atlas for the NR superfamily in CD4+ T cells isolated either in a steady state or over the course of collagen-induced arthritis mouse model of RA. We observed discrete expression patterns among the NR superfamily during the disease stages. NRs that instigate anti-inflammatory programs underwent major downregulation during disease onset; however, during the fully developed disease stage we noticed that NRs that induce proinflammatory programs had reduced transcript levels. These animal findings corroborated well with the expression patterns of NRs in clinical samples obtained from RA patients. Furthermore, we observed that targeting NRs using synthetic ligands alleviates the progression of collagen-induced arthritis. Overall, our data demonstrates the potential of the NR superfamily as novel therapeutic targets for the treatment of autoimmune disorders.


Assuntos
Artrite Experimental/metabolismo , Artrite Reumatoide/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Receptores Citoplasmáticos e Nucleares/genética , Receptores Citoplasmáticos e Nucleares/metabolismo , Animais , Anti-Inflamatórios/uso terapêutico , Anticorpos/imunologia , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/patologia , Colágeno Tipo II/imunologia , Colágeno Tipo II/farmacologia , Citocinas/metabolismo , Feminino , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos DBA , Fenilacetatos/uso terapêutico , Retinoides/uso terapêutico , Líquido Sinovial/metabolismo , Tiazóis/uso terapêutico , Tiossemicarbazonas/uso terapêutico , Transcrição Genética
16.
Metab Eng ; 55: 258-267, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31390538

RESUMO

Microbial conversions known as "biological funneling" have attracted attention for their ability to upgrade heterogeneous mixtures of low-molecular-weight aromatic compounds obtained by chemical lignin depolymerization. ß-hydroxypropiovanillone (HPV) and its analogs can be obtained by chemoselective catalytic oxidation of lignin using 2,3-dichloro-5,6-dicyano-1,4-benzoquinone/tert-butyl nitrite/O2, followed by cleavage of arylglycerol-ß-aryl ether with zinc. Sphingobium sp. strain SYK-6 can degrade HPV generated by the catabolism of arylglycerol-ß-aryl ether through 2-pyrone-4,6-dicarboxylate (PDC), a promising platform chemical. Therefore, production of PDC from HPV can be achieved using the HPV catabolic pathway. However, the pathway and genes involved in the catabolism of vanilloyl acetic acid (VAA) generated during HPV catabolism have not been investigated. In the present study, we isolated SLG_24960 (vceA), which encodes an enzyme that converts VAA into a coenzyme A (CoA) derivative of vanillate (vanilloyl-CoA) from SYK-6, by shotgun cloning. The analysis of a vceA mutant indicated that this gene is not required for VAA conversion in vivo, but it encodes a major enzyme catalyzing CoA-dependent VAA conversion in vitro. We also identified SLG_12450 (vceB), whose product can convert vanilloyl-CoA to vanillate. Enzyme genes besides vceA and vceB, which are necessary for the conversions of HPV to VAA and of vanillate to PDC, were introduced and expressed in Pseudomonas putida. The resulting engineered strain completely converted 1  mM HPV into PDC after 24  h. Our results suggest that the enzyme genes that are not required for the catabolic pathway in microorganisms but can be used for the conversion of target substrates are buried in microbial genomes. These genes are, thus, useful for designing metabolic pathways to produce value-added metabolites.


Assuntos
Proteínas de Bactérias , Genes Bacterianos , Lignina , Redes e Vias Metabólicas , Fenilacetatos/metabolismo , Sphingomonadaceae , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Éteres , Lignina/genética , Lignina/metabolismo , Oxirredução , Sphingomonadaceae/enzimologia , Sphingomonadaceae/genética
17.
Endocrine ; 66(1): 105-114, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31359245

RESUMO

Several metabolic products that derive from L-thyroxine (T4) and 3,3'5-L-triiodothyronine (T3), the main thyroid hormones secreted by the thyroid gland, possess biologic activities. Among these metabolites or derivatives showing physiological actions some have received greater attention: diiodothyronines, iodothyronamines, acetic acid analogues. It is known that increased thyroid hormone (T3 and T4) levels can improve serum lipid profiles and reduce body fat. These positive effects are, however, counterbalanced by adverse effects on the heart, muscle and bone, limiting their use. In addition to the naturally occurring metabolites, thyroid hormone analogues have been developed that either have selective effects on specific tissues or bind selectively to thyroid hormone receptor (TR) isoform. Among these GC-1, KB141, KB2115, and DITPA were deeply investigated and displayed promising therapeutic results in the potential treatment of conditions such as dyslipidemias and obesity. In this review, we summarize the current knowledge of metabolites and analogues of T4 and T3 with reference to their possible clinical application in the treatment of human diseases.


Assuntos
Hormônios Tireóideos/metabolismo , Acetatos/uso terapêutico , Anilidas/uso terapêutico , Animais , Di-Iodotironinas/uso terapêutico , Humanos , Fenóis/uso terapêutico , Éteres Fenílicos/uso terapêutico , Fenilacetatos/uso terapêutico , Propionatos/uso terapêutico , Hormônios Tireóideos/química
18.
Optom Vis Sci ; 96(7): 500-506, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31274738

RESUMO

SIGNIFICANCE: Central serous chorioretinopathy (CSCR) is still a therapeutic challenge with no criterion standard treatment. However, anatomic changes at the level of the retinal pigment epithelium could prove of predictive value in the course of the disease for selective treatment in cases of increased risk of chronicity. PURPOSE: This pilot study analyzes the efficacy for treating acute CSCR with combined systemic acetazolamide 250 mg twice a day and nepafenac 0.1% eye drops three times a day in comparison with an untreated control group. It also evaluates the presence a pigment epithelial detachment (PED) as a risk factor for chronic CSCR. METHODS: Nineteen consecutive patients (group 1) with new or new onset of recurrent CSCR were treated with oral acetazolamide and nepafenac eye drops for at least 2 months. A control group of 14 patients (group 2) with new or new onset of recurrent CSCR were untreated while under regular observation for 4 months. Primary end points were central macular thickness and best-corrected visual acuity after 4 months. Secondary end points were complete regression of subretinal fluid at 3 months and association of PED at baseline with recurrent or chronic CSCR imaged by optical coherence tomography. RESULTS: Group 1 showed significantly faster resolution of subretinal fluid with a mean central macular thickness at 4 months of 271 ± 85 µm compared with 322 ± 79 µm for group 2 (P < .05), but with no functional benefit with a best-corrected visual acuity at 4 months of 0.8 ± 0.2 for group 1 compared with 0.9 ± 0.1 for the control group (P < .05). Patients with a small flat PED were at a higher risk of developing chronic CSCR compared with patients with a dome-shaped or no PED (P < .05). CONCLUSIONS: Central serous chorioretinopathy remains a therapeutic challenge. This pilot study shows faster resolution of subretinal fluid with treatment but without functional benefit compared with observation. The presence of small, flat PED was associated with development of chronic CSCR.


Assuntos
Acetazolamida/uso terapêutico , Anti-Inflamatórios não Esteroides/uso terapêutico , Benzenoacetamidas/uso terapêutico , Inibidores da Anidrase Carbônica/uso terapêutico , Coriorretinopatia Serosa Central/tratamento farmacológico , Fenilacetatos/uso terapêutico , Administração Oftálmica , Administração Oral , Adulto , Coriorretinopatia Serosa Central/diagnóstico , Coriorretinopatia Serosa Central/fisiopatologia , Quimioterapia Combinada , Feminino , Angiofluoresceinografia , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Epitélio Pigmentado da Retina , Líquido Sub-Retiniano , Tomografia de Coerência Óptica/métodos , Acuidade Visual/fisiologia
19.
J Biosci Bioeng ; 128(6): 677-682, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31235413

RESUMO

Recently, we reported a fusion-protein-based immunodetection system comprising the two domains of an antibody variable region as the detectors, each tethered to an interface mutant ß-glucuronidase (GUSm) as the reporter, for detecting small molecules via dimerization of dimer activation. However, the poor stability of GUSm and background signal propagation possibly due to spontaneous proteolysis undermined its performance. To solve these problems, we attempted thermostabilization of GUSm by using a previously isolated thermostable mutant GUSIV5 as a backbone. After screening several interface mutants, we selected one with M516K/Y517W mutation because it exhibited higher activity after dimerization than the wild-type GUS, while maintaining very low background activity. By using this improved immunosensor, we achieved a two-fold improvement in terms of sensitivity in the detection of 4-hydroxy-3-nitrophenyl acetyl. Moreover, by constructing a new biosensor tethered to a nanobody for caffeine as the detector, we could achieve noncompetitive signal-on detection of caffeine in a practically useful concentration range.


Assuntos
Antígenos/imunologia , Glucuronidase/metabolismo , Região Variável de Imunoglobulina/imunologia , Antígenos/química , Técnicas Biossensoriais , Glucuronidase/química , Imunoensaio , Região Variável de Imunoglobulina/química , Modelos Moleculares , Nitrofenóis/metabolismo , Fenilacetatos/metabolismo , Estrutura Quaternária de Proteína , Estrutura Terciária de Proteína
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