RESUMO
Polyethylene glycol modification (PEGylation) is a widely used strategy to improve the physicochemical properties of various macromolecules, especially protein drugs. However, its application in enhancing the performance of enzymes for molecular biology remains underexplored. This study explored the PEGylation of Bst DNA polymerase, determining optimal modification reaction conditions. In comparison to the unmodified wild-type counterpart, the modified Bst DNA polymerase exhibited significantly improved activity, thermal stability, and inhibitor tolerance during loop-mediated isothermal amplification. When applied for the detection of Salmonella in crude samples, the modified enzyme demonstrated a notably accelerated reaction rate. Therefore, PEGylation emerges as a viable strategy for refining DNA polymerases, helping in the development of novel molecular diagnostic reagents.
Assuntos
DNA Polimerase Dirigida por DNA , Estabilidade Enzimática , Polietilenoglicóis , Polietilenoglicóis/química , Polietilenoglicóis/farmacologia , DNA Polimerase Dirigida por DNA/metabolismo , DNA Polimerase Dirigida por DNA/química , Temperatura , Salmonella/genética , Salmonella/enzimologia , Salmonella/efeitos dos fármacos , Técnicas de Amplificação de Ácido Nucleico/métodos , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/químicaRESUMO
INTRODUCTION: Heimler syndrome (HS) is a rare disorder that includes sensorineural hearing loss (SNHL), nail abnormalities, and enamel hypoplasia. Patients with this syndrome can also exhibit ocular manifestations. At present, only a few cases of HS have been reported, existing knowledge of this syndrome is limited, and many cases have been misdiagnosed or even missed. This is the first report of Heimler syndrome with blurred vision as the first complaint, which was diagnosed by genetic analysis in the ophthalmology department. CASE DESCRIPTION: An 8-year-old girl complained of bilateral visual blur and night blindness from birth. Ophthalmic examinations revealed bilateral retinitis pigmentosa with cystoid macular edema, visual impairment with hyperopia and astigmatism. Hearing test revealed bilateral severe sensorineural hearing loss. Dental examinations revealed enamel hypoplasia. In addition, whole-exome sequencing (WES) identified two pathogenic variants in PEX1: the previously reported missense variant c.2966T > C (p.I989â T), and the novel frameshift variant c.1671_1672del (p.G558Sfs*33). CONCLUSION: Heimler syndrome is caused by compound heterozygous PEX1 pathogenic variants, c.2966T > C (p.I989â T) and c.1671_1672del (p.G558Sfs*33), which contributed to the diversity of clinical and genetic profiles in this patient. The main clinical manifestations include bilateral retinitis pigmentosa with cystoid macular edema, sensorineural hearing loss, and enamel hypoplasia. Systemic examinations are suggested for patients suspected of having pigmentary retinal dystrophy, especially combined with hearing-related impairments. Genetic testing can help us to make a definitive diagnosis.
Assuntos
Amelogênese Imperfeita , Perda Auditiva Neurossensorial , Unhas Malformadas , Humanos , Feminino , Criança , Perda Auditiva Neurossensorial/genética , Perda Auditiva Neurossensorial/diagnóstico , Unhas Malformadas/genética , Unhas Malformadas/diagnóstico , Amelogênese Imperfeita/genética , Amelogênese Imperfeita/diagnóstico , Acuidade Visual , Heterozigoto , Análise Mutacional de DNA , Sequenciamento do Exoma , Tomografia de Coerência Óptica , Transtornos da Visão/genética , Transtornos da Visão/diagnóstico , Proteínas de Membrana/genética , DNA/genética , ATPases Associadas a Diversas Atividades CelularesRESUMO
Oil leakage is a global environmental issue and happens frequently, resulting in a waste of oil resources and even threatening the safety of marine creatures and humans. Because of unidirectional oil transportation performance, "oil-diode" Janus membranes have attracted lots of attention for oil/water separation. However, the hydrophobic side of traditional "oil-diode" Janus membrane is completely hydrophobic, resulting in an easy permeation of oil, which hampers light oil recycling. Herein, we provide a facile approach to develop "oil-diode" Janus membranes with the special wettable structure for fast oil refining. The material characteristics and surface wettability of the membranes that generate superimposed efforts are vital to fabricate "oil-diode" Janus membranes. Interestingly, the manufactured membranes exhibit extra-high oil intrusion pressure up to 12 kPa and present high permeance of about 2993 L m-2 h-1 bar-1 in separating stable water-in-oil emulsion containing surfactant and separation efficiency up to 99.6%, thereby showing promising potential in oil recovery and refining.