High adherence to anti-tuberculosis treatment among patients attending a hospital and slum health centre in Nairobi,Kenya

Abstract

We conducted a study among patients with tuberculosis (TB) attending two health facilities—a hospital and a slum health centre—in Nairobi, in order to: (a) assess adherence to anti-TB treatment; and (b) identify reasons for non-adherence.

Urine Isoniazid (INH), used as a proxy for overall adherence, was detected in 142 (97% {95% CI 92–99}) of the 147 patients involved in the study. Five patients had no INH detected in urine and had run out of pills within the previous three days. The reasons included: not having enough pills to last until the appointment date (1); delays due to work or family reasons (2); needing to seek money for transport (1); and losing some pills (1).

Anti-TB treatment adherence is high, and this is reassuring information as Kenya plans to change to a superior first-line regimen based on rifampicin throughout the course of anti-TB treatment. Providing patients with a three-day “excess stock” of pills would provide a “safety net” for continued treatment.     

HPLC法测定人体血浆中利福平和异烟肼的浓度

目的:建立一种用HPLC同时测定人体血浆中利福平和异烟肼浓度的方法。方法:色谱柱为TSK-gel ODS-80(5μm,250mm×4.6mm),流动相0.02mol·L~(-1)磷酸二氢钾-乙腈-三乙胺(40:60:0.15,磷酸调pH 4.30),流速0.9mL·min~(-1),柱温30℃,检测波长为264 nm,内标为氟哌啶醇。结果:利福平和异烟肼的线性范围分别为0.25～32 mg·L~(-1)(r=0.999 8)和0.125～16 mg·L~(-1)(r=0.999 8);高、中、低3个浓度的萃取回收率分别为86.6％～91.8％和88.0％～90.2％: 日内RSD为1.1％～9.0％和1.6％～4.2％,日间RSD为1.2％～8.2％和2.9％～7.2％,n=5。结论:方法简便、准确,可为利福平复方制剂药代动力学研究提供—理想的检测方法。     

Molecular characteristics of rifampin and isoniazid resistant Mycobacterium tuberculosis strains from Beijing, China

Background China is one of the high burden countries of Mycobacterium tuberculosis （TB） infection globally, with high incidence and mortality. We studied the molecular characteristics of rifampin （RIF） and isoniazid （INH） resistant Mycobacterium tuberculosis strains from Beijing, China, in order to find out the genetic marker for rapid detection of specific drug resistance. Methods Forty pansusceptible and 81 resistant strains of Mycobacterium tuberculosis isolated from Beijing, China during 2002-2005 were analyzed. The modified rifampin oligonucleotide （RIFO） assay based on reverse line blot hybridization was used to detect mutations in the 81 bp hot-spot region of rpoB gene, which is associated with RIF resistance. The INH resistance associated genes, regulatory region mab-inhA （-15C/T） and structural gene katG S315T were detected by reverse line blot hybridization and PCR-restriction fragment length polymorphism （RFLP） method respectively. All the strains were typed by spoligotying and the Beijing genotype was further subdivided by NTF locus analysis. The distribution of drug resistance associated mutations in the above genes was compared in these groups. Results Sixty-five （91.5%） of 71 RIF resistant and 52 （92.9%） of 56 multidrug-resistant （MDR, i.e. resistant to at least RIF and INH） strains were found to harbor mutations in the rpoB hot-spot region. No mutation was detected in RIF sensitive strains. The specificity and sensitivity of the modified RIFO assay were 100% and 91.5%, respectively, katG315 AGC〉ACC and inhA-15C〉T mutations were found in 40 （60.6%） and 10 （15.2%） of 66 INH resistant strains, respectively; 7.6% of INH-resistant strains had mutations in both of these genes. Therefore, a combined use of both katG315 and inhA-15 identified 68.2% of INH-resistant strains. The Beijing genotype accounted for 91.7% of total strains and was further subdivided into ＂modern＂ （76.6%） and ＂ancestral＂ （23.4%） group. There is no significant difference between ＂ancestral＂ and ＂modern＂ group in prevalance of drug resistance-associated gene mutations. Conclusions The hot-spot region of rpoB gene can be used as genetic marker for detection of RIF resistant strains; a combined use of both katG315 and inhA-15 can improve the detection rate of I NH resistant strains; the Beijing genotype is prevalent in Beijing, China; the modified RIFO assay can be a practical tool for rapid detection of RIF resistant and MDR isolates in the routine diagnostic work.     

槲皮素对异烟肼诱导肝细胞毒性的保护作用及其机制

目的： 探讨活性氧（ROS）介导的线粒体氧化损伤在异烟肼（INH）诱导肝细胞毒性中的作用及槲皮素对INH肝细胞毒性的保护效应及机制。方法： 将L-02细胞随机分为5组：异烟肼组（10 mmol/L INH）、槲皮素处理组（10 mmol/L INH和50 μmol/L槲皮素）、谷胱甘肽（GSH）预处理组（20 mg/mL GSH、10 mmol/L INH和50 μmol/L槲皮素）、谷胱甘肽组（20 mg/mL GSH）、对照组（等体积的无血清培养基）,作用24 h后,采用差速离心法制备细胞线粒体,荧光探针DCFH-DA和Rho-123检测细胞线粒体ROS水平及膜电位;TBA比色法测定丙二醛（MDA）含量;DPNH比色法测定蛋白质羰基含量;ELISA法检测8-羟基脱氧鸟嘌呤核苷（8-OHdG）含量。结果： 与对照组相比,INH处理细胞后细胞线粒体ROS水平升高（P < 0.01）,膜电位降低（P < 0.01）。与异烟肼组相比,槲皮素处理组细胞线粒体ROS水平降低（P < 0.01）,膜电位升高（P < 0.05）;谷胱甘肽预处理组细胞线粒体ROS水平低于槲皮素处理组（P < 0.05）,线粒体膜电位高于槲皮素处理组（P < 0.05）。与对照组相比,细胞经INH处理后MDA、蛋白质羰基及8-OhdG的含量增加（P < 0.01）。与异烟肼组比较,应用槲皮素后可使MDA、蛋白羰基、8-OHdG的含量减少（P < 0.01）;谷胱甘肽预处理组MDA、蛋白羰基、8-OHdG含量低于槲皮素处理组（P < 0.05）。结论： 在本实验条件下,INH可诱导L-02细胞线粒体氧化损伤,且ROS参与了此过程;槲皮素对INH诱导的线粒体氧化损伤具有保护效应,其机制可能与其抑制ROS水平有关。     

Nrf2/ARE信号通路在槲皮素抑制异烟肼诱导的肝细胞线粒体氧化损伤中的作用

目的：探讨核因子E2相关因子2/抗氧化反应元件（Nrf2/ARE）信号通路在槲皮素抑制异烟肼（INH）诱导的L-02细胞线粒体氧化损伤中的作用。方法：将L-02细胞随机分为阴性对照组、INH组（10 mmol/L INH）、单独槲皮素组（50 μmol/L槲皮素）、槲皮素保护组（10 mmol/L INH+50 μmol/L槲皮素）,各组细胞处理24 h后,采用四甲基偶氮唑蓝（MTT）法测定细胞存活率;荧光探针DCFH-DA检测细胞线粒体活性氧（ROS）水平;比色法测定细胞内丙二醛（MDA）含量、谷胱甘肽（GSH）含量和超氧化物歧化酶（SOD）活性,评价细胞线粒体氧化损伤状态。Western blot检测Nrf2、血红素氧合酶1（HO-1）蛋白表达水平,评价细胞内Nrf2/ARE信号通路的变化。结果：与阴性对照组比较,INH组细胞存活率显著降低（P < 0.01）,线粒体ROS水平和MDA含量显著升高（P < 0.01）,GSH含量及SOD活性明显降低（P < 0.01）;与INH组相比较,槲皮素保护组细胞存活率明显增加（P < 0.01）,线粒体ROS水平和MDA含量显著减少（P < 0.01）,GSH含量及SOD活性明显升高（P < 0.05或P < 0.01）。INH组细胞质中HO-1蛋白及细胞核中Nrf2蛋白表达较阴性对照组明显增加（P < 0.01）;槲皮素保护组细胞质中HO-1蛋白及细胞核中Nrf2蛋白表达明显高于INH组（P < 0.01）。结论：槲皮素能够抑制INH诱导的肝细胞线粒体氧化损伤,这可能与槲皮素对Nrf2/ARE信号通路的活性调节相关。     

Nanodiagnostics for tuberculosis detection

Introduction: Tuberculosis (TB) is a leading killer worldwide. End TB strategy aims at ending the TB epidemic by 2030. Early, accurate, and affordable diagnosis represents a cornerstone to achieve this goal. Innovative strategies for TB diagnostics have been introduced. However, the ideal assay is yet unavailable and conventional methods remain necessary for diagnosis. Unique properties of nanoparticles (NPs) have allowed their utilization in TB detection via targeting disease biomarkers.

Area covered: Until now, around thirty-five TB NP-based assays have been partially or fully characterized. Accuracy, low-cost, and short time-to-result represent the common properties of proposed platforms. TB nanodiagnostics now encompass almost all clinical aspects of the disease including active TB, non-tuberculous mycobacteria, rifampicin resistant TB, TB/HIV co-infection, latent TB, and extra-pulmonary TB. This review summarizes state-of-the-art knowledge of TB nanodiagnostics for the last 10 years. Special consideration is given for fabrication concepts, detection strategies, and clinical performance using various clinical specimens. The potential of TB nanodiagnostics to fulfill the need for ideal MTB testing is assessed.

Expert commentary: TB nanodiagnostics show promise to be ideal detection tools that can meet the rigorous demands to end the TB epidemic by 2030.     

Risk of hepatitis with various reintroduction regimens of anti-tubercular therapy: a systematic review and network meta-analysis

ABSTRACT

Objective: To compare risk of hepatotoxicity between various regimens for reintroduction of antitubercular therapy (ATT) in patients with previous episode of ATT hepatitis.

Methods: We searched various databases (PubMed, Embase, CENTRAL, Scopus, WoS and LILACS) for studies comparing ATT reintroduction regimens using terms ‘drug-induced liver injury’ and ‘antitubercular drugs’ AND ‘reintroduction’. The reintroduction regimens i.e concomitant (all drugs introduced together), sequential (reintroduction of one drug in full dose followed by another) or incremental (one drug in a low dose and then higher dose followed by next drug) were compared using Bayesian approach for network meta-analysis with random-effect model. Cochrane revised tool was used to assess risk of bias in included studies (RoB 2.0).

Results: Four randomized studies with 577 patients were eligible for analysis. Compared with concomitant regimen (baseline comparator), incremental regimen appeared to have lower risk of ATT hepatitis (odds ratio [OR] 0.24; 95% CrI 0.017, 1.2) as also the sequential regimen (OR 0.33; 95% CrI 0.033, 1.7). Rifampicin first and isoniazid first reintroduction regimens were similar via-a-vis recurrence of hepatotoxicity.

Conclusion: The sequential and incremental regimen may be better than concomitant regimen in reducing risk of ATT hepatitis although the odds did not achieve statistical significance.     

Follow-up of compliance with tuberculosis treatment in children: monitoring by urine tests

This study was designed to follow up patient compliance by detection of antituberculous drugs in urine during the course of treatment. It was conducted in the Outpatient Clinic of Pediatric Infectious Diseases, Sisli Etfal Hospital (Istanbul, Turkey). In total, 45 children with pulmonary tuberculosis participated. Patients were seen twice in the first month and once a month thereafter during the 6-month course of treatment. The second urine of the day was collected at each visit. Urine was tested for isoniazid (INH), rifampicin (RIF), and pyrazinamide (PZA). In the presence of these drugs or their metabolites, the addition of certain chemicals caused a color change in the urine. On day 15 of treatment, urine tested positive for INH in 82% of patients, for RIF in 67%, and for PZA in 73%. At the end of the second month, the ratio of adherence was 96, 89, and 96% for each drug, respectively. All patients were found to be adherent at months 5 and 6. We recommend detection of antituberculous drugs in urine to assess compliance to treatment. Once the defaulting patients were identified, adherence was improved by repeatedly providing patient education throughout the treatment.     

评价大鼠离体肝灌流模型稳定性的方法

以自建的大鼠原位循环肝灌流模型为研究模型,建立一套实用的模型稳定性评价方法。首先进行一般稳定性评价,包括肉眼观察肝脏形态,监测门脉压、灌流液pH,定时检测灌流液K⁺浓度、丙氨酸转氨酶(ALT)、天冬氨酸氨基转移酶(AST)水平,肝组织病理检查等;要求在整个灌流过程中肝脏色泽均匀,无斑块状改变,以门静脉压始终无骤变,稳定在8~14 mmHg范围内,变化不超过0.5 mmHg/30 min,pH始终处于7.4~7.2,变化不超过0.1/30 min,K⁺浓度无突然升高现象,灌流液中ALT、AST水平随时间无显著性变化,肝组织病理检查不出现除空泡性变以外的病变为标准,确定模型可维持稳定的时间。在满足一般稳定性的基础上,根据不同的研究目的对模型进行功能稳定性评估,要求研究所用的指标能在空白灌流组和阳性药灌流组之间体现显著性差异;本研究以药物肝损伤研究为例,以灌流液ALT、AST、LDH(乳酸脱氢酶)水平为指标,并进行肝组织病理检查,结果两组出现极显著性差异,故判定用所建立的灌流模型进行肝损伤研究可真实反映受试药的肝损伤情况,一般不会出现假阴性和假阳性结果。     

Tuberculosis screening and isoniazid preventive therapy implementation: a Brazilian experience

Evaluation of: Durovni B, Saraceni V, Moulton LH, et al. Effect of improved tuberculosis screening and isoniazid preventive therapy on incidence of tuberculosis and death in patients with HIV in clinics in Rio de Janeiro, Brazil: a stepped wedge, cluster-randomised trial. Lancet Infect Dis 2013;13(10):852-8

Tuberculosis Preventive therapy has been used since the 1960s for both mass treatment of populations and for targeted therapy to high-risk groups such as children. The World Health Organization (WHO) recommends the use of Isoniazid preventive therapy (IPT) as one of the strategies to reduce the TB burden among people living with HIV infection. However, its uptake in countries with high TB burden has been poor. One of the main barriers to its widespread implementation is concerns around the ruling out of active TB. WHO, in its revised guidelines on IPT, recommends the use of a symptom-based algorithm for TB screening. The randomized clinical trial evaluated here explores the effect of an intervention, in the form of training the clinic staff to screen for TB, use of tuberculin skin test and IPT to treat latent TB infection, and assess its effect on rates of TB and death in HIV infected.