RESUMO
BACKGROUND: Typically, genotypic resistance testing is recommended at the start of antiretroviral therapy and is even mandatory in cases of virologic failure. The material of choice is plasma viral RNA. However, in patients with low viremia (viral load < 500 copies/ml), resistance testing by population-based sequencing is very difficult. OBJECTIVE: Therefore, we aimed to investigate whether next generation sequencing (NGS) from proviral DNA and RNA could be an alternative. MATERIAL AND METHODS: EDTA blood samples (n = 36) from routine clinical viral load testing were used for the study. Viral loads ranged from 96 to 390,000 copies/mL, with 100% of samples having low viremia. Distribution of subtypes; A (n = 2), B (n = 16), C (n = 4), D (n = 2), G (1), CRF02 AG (n = 5), CRF01 AE (n = 5), undefined/mixed (n = 4). The extracted consensus sequences were uploaded to the Stanford HIV Drug Resistance Data Base and Geno2pheno for online analysis of drug resistance mutations and resistance factors. RESULTS: A total of 2476 variants or drug resistance mutations (DRMs) were detected with Sanger sequencing, compared with 2892 variants with NGS. An average of 822/1008 variants were identified in plasma viral RNA by Sanger or NGS sequencing, 834/956 in cellular viral RNA, and 820/928 in cellular viral DNA. CONCLUSION: Both methods are well suited for the detection of HIV substitutions or drug resistance mutations. Our results suggest that cellular RNA or cellular viral DNA is an informative alternative to plasma viral RNA for variant detection in patients with low viremia, as shown by the high correlation of variants in the different viral pools. We show that by using UDS, a plus of two DRMs per patient becomes visible, which can make a big difference in the assessment of the expected resistance behavior of the virus.
Assuntos
Fármacos Anti-HIV , Infecções por HIV , Soropositividade para HIV , HIV-1 , Animais , Fármacos Anti-HIV/uso terapêutico , DNA Viral/genética , Farmacorresistência Viral/genética , Ácido Edético/uso terapêutico , Genômica , Genótipo , Infecções por HIV/tratamento farmacológico , Soropositividade para HIV/tratamento farmacológico , HIV-1/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Leucócitos Mononucleares , Estágios do Ciclo de Vida , Mutação , Provírus/genética , RNA Viral/genética , Carga Viral , Viremia/tratamento farmacológicoAssuntos
Algoritmos , Transtornos da Coagulação Sanguínea/diagnóstico , Testes de Coagulação Sanguínea/métodos , Coagulação Sanguínea/efeitos dos fármacos , Fator V/antagonistas & inibidores , Fator V/análise , Pneumonia/complicações , Idoso , Antibacterianos/farmacologia , Coagulação Sanguínea/fisiologia , Transtornos da Coagulação Sanguínea/etiologia , Técnicas de Laboratório Clínico/normas , Humanos , Masculino , Pneumonia/tratamento farmacológico , Valor Preditivo dos TestesRESUMO
The thiocarbamates 4-RC(6)H(4)NHC(S)NR(2)' (R=H, Cl; R'=Me, Et), 4-ClC(6)H(4)NHC(S)NR (NR=2-pyridylpiperazine) react with cis-[PtCl(2)(PTA)(2)] (PTA=1,3,5-triaza-7-phosphaadamantane) in the presence of base to afford the monocationic platinum(II) complexes cis-[Pt{SC(NR(2)')=NC(6)H(4)R}(PTA)(2)](+) (R=H, Cl; R'=Me, Et), cis-[Pt{SC(NR)=NC(6)H(4)Cl}(PTA)(2)](+) (NR=2-pyridylpiperazine), which were isolated as their PF(6) salts in high yields. The complexes were fully characterised spectroscopically and also by X-ray crystallography. Cytotoxicity of these complexes was studied in vitro in three human cancer cell lines (CH1, A549 and SW480) using the MTT assay.
Assuntos
Antineoplásicos/farmacologia , Quelantes/química , Compostos Organoplatínicos/farmacologia , Tiocarbamatos/química , Adamantano/análogos & derivados , Adamantano/química , Antineoplásicos/síntese química , Antineoplásicos/química , Cátions Bivalentes , Cristalografia por Raios X , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Concentração Inibidora 50 , Compostos Organofosforados/química , Compostos Organoplatínicos/síntese química , Compostos Organoplatínicos/química , Piperazinas/química , Piridinas/química , Estereoisomerismo , Células Tumorais CultivadasRESUMO
Organometallic ruthenium-arene compounds bearing a maltol ligand have been shown to be nearly inactive in in vitro anticancer assays, presumably due to the formation of dimeric Ru(II) species in aqueous solutions. In an attempt to stabilize such complexes, [Ru(eta(6)-p-cymene)(XY)Cl] (XY=pyrones or thiopyrones) complexes with different substitution pattern of the (thio)pyrone ligands have been synthesized, their structures characterized spectroscopically, and their aquation behavior investigated as well as their tumor-inhibiting potency. The aquation behavior of pyrone systems with electron-donating substituents and of thiopyrone complexes was found to be significantly different from that of the maltol-type complex reported previously. However, the formation of the dimer can be excluded as the primary reason for the inactivity of the complex because some of the stable compounds are not active in cancer cell lines either. In contrast, studies of their reactivity towards amino acids demonstrate different reactivities of the pyrone and thiopyrone complexes, and the higher stability of the latter probably renders them active against human tumor cells.
