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1.
J Clin Immunol ; 38(6): 712-716, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30039354

RESUMO

PURPOSE: Mendelian suceptibility to mycobacterial disease (MSMD) is a rare primary immunodeficiency predisposing to severe disease caused by mycobacteria and other intracellular pathogens. Delay in diagnosis can have an impact on the patient's prognosis. METHODS: We evaluated the IFN-γ circuit by studying IFN-γ production after mycobacterial challenge as well as IL-12Rß1 expression and STAT4 phosphorylation in response to IL-12p70 stimulation in whole blood of a 6-year-old Peruvian girl with disseminated recurrent mycobacterial infection diagnosed as multidrug-resistant tuberculosis. Genetic studies with Sanger sequencing were used to identify the causative mutation. Microbiological studies based on PCR reactions were used to diagnose the specific mycobacterial species. RESULTS: We identified a homozygous mutation in the IL12RB1 gene (p. Arg211*) causing abolished expression of IL-12Rß1 and IL-12 response. MSMD diagnosis led to a microbiological reevaluation of the patient, revealing a BCG vaccine-related infection instead of tuberculosis. Treatment was then adjusted, with good response. CONCLUSIONS: We report the first Peruvian patient with IL-12Rß1 deficiency. Specific mycobacterial species diagnosis within Mycobacterium tuberculosis complex is still challenging in countries with limited access to PCR-based microbiological diagnostic techniques. Awareness of MSMD warning signs and accurate microbiological diagnosis of mycobacterial infections are of the utmost importance for optimal diagnosis and management of affected patients.


Assuntos
Vacina BCG/imunologia , Suscetibilidade a Doenças , Mycobacterium tuberculosis/imunologia , Receptores de Interleucina-12/deficiência , Tuberculose Resistente a Múltiplos Medicamentos/genética , Tuberculose Resistente a Múltiplos Medicamentos/imunologia , Antituberculosos/farmacologia , Antituberculosos/uso terapêutico , Criança , Análise Mutacional de DNA , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Mutação , Mycobacterium tuberculosis/efeitos dos fármacos , Peru , Prognóstico , Índice de Gravidade de Doença , Tuberculose Resistente a Múltiplos Medicamentos/diagnóstico , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia
2.
Toxicon ; 110: 27-34, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26615828

RESUMO

Parotoid gland secretions of toad species are a vast reservoir of bioactive molecules with a wide range of biological properties. Herein, for the first time, it is described the isolation by preparative reversed-phase HPLC and the structure elucidation by NMR spectroscopy and/or mass spectrometry of nine major bufadienolides from parotoid gland secretions of the Cuban endemic toad Peltophryne fustiger: ψ-bufarenogin, gamabufotalin, bufarenogin, arenobufagin, 3-(N-suberoylargininyl) marinobufagin, bufotalinin, telocinobufagin, marinobufagin and bufalin. In addition, the secretion was analyzed by UPLC-MS/MS which also allowed the identification of azelayl arginine. The effect of arenobufagin, bufalin and ψ-bufarenogin on Na(+)/K(+)-ATPase activity in a human kidney preparation was evaluated. These bufadienolides fully inhibited the Na(+)/K(+)-ATPase in a concentration-dependent manner, although arenobufagin (IC50 = 28.3 nM) and bufalin (IC50 = 28.7 nM) were 100 times more potent than ψ-bufarenogin (IC50 = 3020 nM). These results provided evidence about the importance of the hydroxylation at position C-14 in the bufadienolide skeleton for the inhibitory activity on the Na(+)/K(+)-ATPase.


Assuntos
Venenos de Anfíbios/toxicidade , Bufanolídeos/toxicidade , Bufonidae/metabolismo , Rim/efeitos dos fármacos , Moduladores de Transporte de Membrana/toxicidade , Glândula Parótida/metabolismo , ATPase Trocadora de Sódio-Potássio/antagonistas & inibidores , Venenos de Anfíbios/química , Venenos de Anfíbios/isolamento & purificação , Venenos de Anfíbios/metabolismo , Animais , Bufanolídeos/química , Bufanolídeos/isolamento & purificação , Bufanolídeos/metabolismo , Bufonidae/crescimento & desenvolvimento , Cromatografia Líquida de Alta Pressão , Cuba , Humanos , Hidroxilação , Rim/enzimologia , Cinética , Espectroscopia de Ressonância Magnética , Masculino , Moduladores de Transporte de Membrana/química , Moduladores de Transporte de Membrana/isolamento & purificação , Moduladores de Transporte de Membrana/metabolismo , Estrutura Molecular , Rios , ATPase Trocadora de Sódio-Potássio/metabolismo , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massa de Íon Secundário , Espectrometria de Massas em Tandem
3.
Planta Med ; 81(17): 1609-13, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26132850

RESUMO

Verbascoside is a phenylethanoid glycoside widely distributed in nature, especially among the order Lamiales, occurring in numerous plants that are constituents of folk medicine preparations. This natural compound, previously isolated by our group from the ethyl acetate extract of Lantana trifolia using the gradient approach in countercurrent chromatography, was now isolated from the butanol extract of the same plant and from Lippia alba f. intermedia (Verbenaceae) using countercurrent chromatography in either gradient or isocratic elution modes. The ethyl acetate extract of L. alba, rich in phenylethanoids and flavonoids, was fractionated using countercurrent chromatography in the step-gradient elution approach. The four-step solvent system was composed of n-hexane-ethyl acetate-n-butanol-water (4 : 10 : X : 10), where X = 1 (solvent system A), 3 (solvent system B), 5 (solvent system C), and 7 (solvent system D), and allowed for the isolation of verbascoside along with other phenylethanoids and flavonoids from both plants. Verbascoside and 2'-O-ß-apiosylverbascoside were further isolated from the n-butanol extract of L. trifolia using the solvent system ethyl acetate-n-butanol-water 10 : 2 : 10 on an isocratic run. The difference in the complexity of the two plant extracts demanded different purification steps, which included a second high-speed countercurrent chromatography purification using the isocratic elution mode.


Assuntos
Distribuição Contracorrente , Glucosídeos/isolamento & purificação , Lantana/química , Lippia/química , Fenóis/isolamento & purificação , Extratos Vegetais/isolamento & purificação , 1-Butanol , Acetatos , Brasil , Flavonoides/isolamento & purificação , Estrutura Molecular , Solventes
4.
Rev. cuba. farm ; 40(2)mayo-ago. 2006. tab
Artigo em Espanhol | CUMED | ID: cum-32823

RESUMO

Se le realizó un tamizaje fitoquímico a las hojas de la especie Pluchea carolinensis Jacq. G. Don in Sweet y se sometieron a un fraccionamiento con el empleo de solventes de polaridad creciente. En cada uno de los extractos obtenidos se verificó la presencia de flavonoides y se evaluó la actividad antimicrobiana frente a 7 microorganismos. Se detectó la presencia de flavonoides en los extractos CHCl3, AcOEt y n-BuOH, los que presentaron actividad antibacteriana. El extracto CHCl3 presentó actividad biológica frente a Bacillus subtilis a una concentración de 1 000 µg/mL, mientras que los extractos AcOEt y n-BuOH mostraron resultados positivos frente a las bacterias Staphylococcus aureus y Bacillus subtilis a la misma concentración(AU)


Assuntos
Salvia/química , Programas de Rastreamento , Extratos Vegetais/análise , Testes de Sensibilidade Microbiana/métodos
5.
Rev. cuba. farm ; 40(2)mayo-ago. 2006. tab
Artigo em Espanhol | LILACS | ID: lil-465269

RESUMO

Se le realizó un tamizaje fitoquímico a las hojas de la especie Pluchea carolinensis Jacq. G. Don in Sweet y se sometieron a un fraccionamiento con el empleo de solventes de polaridad creciente. En cada uno de los extractos obtenidos se verificó la presencia de flavonoides y se evaluó la actividad antimicrobiana frente a 7 microorganismos. Se detectó la presencia de flavonoides en los extractos CHCl3, AcOEt y n-BuOH, los que presentaron actividad antibacteriana. El extracto CHCl3 presentó actividad biológica frente a Bacillus subtilis a una concentración de 1 000 µg/mL, mientras que los extractos AcOEt y n-BuOH mostraron resultados positivos frente a las bacterias Staphylococcus aureus y Bacillus subtilis a la misma concentración


Assuntos
Extratos Vegetais , Testes de Sensibilidade Microbiana/métodos , Salvia , Peneiramento de Líquidos
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