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Performance of empirical and model-based classifiers for detecting sucrase-isomaltase inhibition using the13C-sucrose breath test. 利用 13C 蔗糖呼气试验检测蔗糖酶-异麦芽糖酶抑制的经验分类器和基于模型的分类器的性能。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-10 DOI: 10.1088/1752-7163/ad748d
Hannah Van Wyk, Gwenyth O Lee, Robert J Schillinger, Christine A Edwards, Douglas J Morrison, Andrew F Brouwer

The13C-sucrose breath test (13C-SBT) has been proposed to estimate sucrase-isomaltase (SIM) activity and is a promising test for SIM deficiency, which can cause gastrointestinal symptoms, and for intestinal mucosal damage caused by gut dysfunction or chemotherapy. We previously showed how various summary measures of the13C-SBT breath curve reflect SIM inhibition. However, it is uncertain how the performance of these classifiers is affected by test duration. We leveraged13C-SBT data from a cross-over study in 16 adults who received 0, 100, and 750 mg of Reducose, an SIM inhibitor. We evaluated the performance of a pharmacokinetic-model-based classifier,ρ, and three empirical classifiers (cumulative percent dose recovered at 90 min (cPDR90), time to 50% dose recovered, and time to peak dose recovery rate), as a function of test duration using receiver operating characteristic (ROC) curves. We also assessed the sensitivity, specificity, and accuracy of consensus classifiers. Test durations of less than 2 h generally failed to accurately predict later breath curve dynamics. The cPDR90 classifier had the highest ROC area-under-the-curve and, by design, was robust to shorter test durations. For detecting mild SIM inhibition,ρhad a higher sensitivity. We recommend13C-SBT tests run for at least a 2 h duration. Although cPDR90 was the classifier with highest accuracy and robustness to test duration in this application, concerns remain about its sensitivity to misspecification of the CO2production rate. More research is needed to assess these classifiers in target populations.

背景:13C-蔗糖呼气试验(13C-SBT)已被提出来估测蔗糖酶-异麦芽糖酶(SIM)的活性,它是检测SIM缺乏症(SIM缺乏症可引起胃肠道症状)以及肠道功能障碍或化疗引起的肠粘膜损伤的一种很有前途的方法。我们以前曾展示过 13C-SBT 呼气曲线的各种汇总指标如何反映 SIM 的抑制情况。但是,这些分类器的性能如何受到测试持续时间的影响尚不确定:我们利用了一项交叉研究中的 13C-SBT 数据,研究对象是 16 名成人,他们分别服用了 0、100 和 750 毫克的 SIM 抑制剂红糖。我们利用接收器操作特征曲线(ROC)评估了基于药代动力学模型的分类器 ρ 和三种经验分类器(90 分钟内累积剂量恢复百分比(cPDR90)、剂量恢复到 50% 的时间和剂量恢复率达到峰值的时间)的性能与试验持续时间的关系。我们还评估了共识分类器的灵敏度、特异性和准确性:测试持续时间少于 2 小时通常无法准确预测后期的呼吸曲线动态。cPDR90 分类器具有最高的 ROC 曲线下面积,并且在设计上对较短的测试时间具有稳健性。对于检测轻度 SIM 抑制,ρ 的灵敏度更高:我们建议 13C-SBT 检测至少持续 2 小时。虽然 cPDR90 是本应用中准确率最高、对测试持续时间最稳健的分类器,但它对二氧化碳产生率的错误指定的敏感性仍然令人担忧。需要进行更多的研究来评估这些分类器在目标人群中的应用。
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引用次数: 0
Systematic study of polymer gas sampling bags for offline analysis of exhaled breath. 用于离线分析呼出气体的聚合物气体采样袋的系统研究
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-13 DOI: 10.1088/1752-7163/ad6a31
Mateusz Fido, Simone Hersberger, Andreas T Güntner, Renato Zenobi, Stamatios Giannoukos

Polymeric bags are a widely applied, simple, and cost-effective method for the storage and offline analysis of gaseous samples. Various materials have been used as sampling bags, all known to contain impurities and differing in their cost, durability, and storage capabilities. Herein, we present a comparative study of several well-known bag materials, Tedlar (PVF), Kynar (PVDF), Teflon (PTFE), and Nalophan (PET), as well as a new material, ethylene vinyl copolymer (EVOH), commonly used for storing food. We investigated the influences of storage conditions, humidity, bag cleaning, and light exposure on volatile organic compound concentration (acetone, acetic acid, isoprene, benzene, limonene, among others) in samples of exhaled human breath stored in bags for up to 48 h. Specifically, we show high losses of short-chain fatty acids (SCFAs) in bags of all materials (for most SCFAs, less than 50% after 8 h of storage). We found that samples in Tedlar, Nalophan, and EVOH bags undergo changes in composition when exposed to UV radiation over a period of 48 h. We report high initial impurity levels in all the bags and their doubling after a period of 48 h. We compare secondary electrospray ionization and proton transfer reaction mass spectrometry in the context of offline analysis after storage in sampling bags. We provide an analytical perspective on the temporal evolution of bag contents by presenting the intensity changes of all significantm/zfeatures. We also present a simple, automated, and cost-effective offline sample introduction system, which enables controlled delivery of collected gaseous samples from polymeric bags into the mass spectrometer. Overall, our findings suggest that sampling bags exhibit high levels of impurities, are sensitive to several environmental factors (e.g. light exposure), and provide low recoveries for some classes of compounds, e.g. SCFAs.

聚合袋是一种应用广泛、操作简单、成本效益高的气体样本存储和离线分析方法。各种材料都被用作采样袋,它们都含有杂质,在成本、耐用性和存储能力方面也各不相同。在此,我们对几种著名的袋子材料进行了比较研究,包括 Tedlar(PVF)、Kynar(PVDF)、Teflon(PTFE)和 Nalophan(PET),以及一种常用于储存食品的新型材料乙烯-乙烯共聚物(EVOH)。我们研究了储存条件、湿度、袋子清洁和光照对储存在袋子中长达 48 小时的人体呼出气体样本中挥发性有机化合物(丙酮、乙酸、异戊二烯、苯、柠檬烯等)浓度的影响。具体来说,我们发现短链脂肪酸(SCFAs)在所有材料的袋子中都有较高的损失(对于大多数 SCFAs 来说,储存 8 小时后的损失率低于 50%)。我们发现,Tedlar、Nalophan 和 EVOH 袋中的样品在暴露于紫外线辐射 48 小时后,其成分会发生变化。我们发现所有袋子中的初始杂质含量都很高,48 小时后杂质含量会翻倍。我们比较了二级电喷雾离子化 (SESI) 和质子转移反应 (PTR) 质谱法在采样袋储存后的离线分析中的应用。我们通过展示所有重要 m/z 特征的强度变化,从分析角度展示了采样袋内容物的时间演变。我们还介绍了一种简单、自动化且经济高效的离线样品引入系统,该系统可将收集的气态样品从聚合物袋中可控地送入质谱仪。总之,我们的研究结果表明,采样袋中的杂质含量很高,对一些环境因素(如光照)很敏感,对某些类别的化合物(如短链脂肪酸)的回收率很低。
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引用次数: 0
Evaluation of tetrachloroethylene (PCE) and its degradation products in human exhaled breath and indoor air in a community setting. 评估社区环境中人体呼出气体和室内空气中的四氯乙烯 (PCE) 及其降解产物。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-08 DOI: 10.1088/1752-7163/ad67fd
Jung Hyun Lee, Alaina K Bryant, Marwan Alajlouni, Brandon E Boor, Antonios Tasoglou, Sa Liu

Tetrachloroethylene (PCE) is a widely utilized volatile chemical in industrial applications, including dry cleaning and metal degreasing. Exposure to PCE potentially presents a significant health risk to workers as well as communities near contamination sites. Adverse health effects arise not only from PCE, but also from PCE degradation products, such as trichloroethylene (TCE) and vinyl chloride (VC). PCE, TCE, and VC can contaminate water, soil, and air, leading to exposure through multiple pathways, including inhalation, ingestion, and dermal contact. This study focused on a community setting in Martinsville, Indiana, a working-class Midwestern community in the United States, where extensive PCE contamination has occurred due to multiple contamination sites (referring to 'plumes'), including a Superfund site. Utilizing proton transfer reaction time-of-flight mass spectrometry (PTR-TOF-MS), PCE, TCE, and VC concentrations were measured in the exhaled breath of 73 residents from both within and outside the plume areas. PCE was detected in 66 samples, TCE in 26 samples, and VC in 68 samples. Our results revealed a significant positive correlation between the concentrations of these compounds in exhaled breath and indoor air (Pearson correlation coefficients: PCE = 0.75, TCE = 0.71, and VC = 0.89). This study confirms the presence of PCE and its degradation products in exhaled breath in a community exposure investigation, demonstrating the potential of using exhaled breath analysis in monitoring exposure to environmental contaminants. This study showed the feasibility of utilizing PTR-TOF-MS in community investigations to assess exposure to PCE and its degradation products by measuring these compounds in exhaled breath and indoor air.

四氯乙烯(PCE)是一种广泛应用于工业领域的挥发性化学品,包括干洗和金属脱脂。接触 PCE 可能会对工人以及污染场地附近的社区造成严重的健康风险。对健康的不利影响不仅来自于 PCE,还来自于 PCE 的降解产物,如三氯乙烯 (TCE) 和氯乙烯 (VC)。PCE、TCE 和 VC 可污染水、土壤和空气,导致通过多种途径(包括吸入、摄入和皮肤接触)接触这些物质。这项研究的重点是印第安纳州马丁斯维尔的一个社区环境,这是美国中西部的一个工人阶级社区,由于存在多个污染点(指 "羽流"),包括一个超级基金场地,该社区受到了广泛的 PCE 污染。利用质子转移反应飞行时间质谱法(PTR-TOF-MS),测量了羽流区内外 73 位居民呼出气体中的 PCE、TCE 和 VC 浓度。在 66 个样本中检测到了 PCE,在 26 个样本中检测到了 TCE,在 68 个样本中检测到了 VC。我们的研究结果表明,这些化合物在呼出气体和室内空气中的浓度之间存在明显的正相关关系(皮尔逊相关系数:PCE = 0.75,TCE = 0.71,VC = 0.89)。这项研究证实,在一项社区接触调查中,呼出的气体中含有五氯乙烯及其降解产物,这证明了利用呼出气体分析监测环境污染物接触情况的潜力。这项研究表明,在社区调查中利用 PTR-TOF-MS 测量呼出气体和室内空气中的五氯乙烯及其降解产物是可行的。
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引用次数: 0
Volatile organic compounds in exhaled breath: a promising approach for accurate differentiation of lung adenocarcinoma and squamous cell carcinoma. 呼出气体中的挥发性有机化合物:准确区分肺腺癌和鳞癌的有效方法
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-05 DOI: 10.1088/1752-7163/ad6474
Xian Li, Lin Shi, Yijing Long, Chunyan Wang, Cheng Qian, Wenwen Li, Yonghui Tian, Yixiang Duan

Lung cancer subtyping, particularly differentiating adenocarcinoma (ADC) from squamous cell carcinoma (SCC), is paramount for clinicians to develop effective treatment strategies. In this study, we aimed: (i) to discover volatile organic compound (VOC) biomarkers for precise diagnosis of ADC and SCC, (ii) to investigated the impact of risk factors on ADC and SCC prediction, and (iii) to explore the metabolic pathways of VOC biomarkers. Exhaled breath samples from patients with ADC (n= 149) and SCC (n= 94) were analyzed by gas chromatography-mass spectrometry. Both multivariate and univariate statistical analysis method were employed to identify VOC biomarkers. Support vector machine (SVM) prediction models were developed and validated based on these VOC biomarkers. The impact of risk factors on ADC and SCC prediction was investigated. A panel of 13 VOCs was found to differ significantly between ADC and SCC. Utilizing the SVM algorithm, the VOC biomarkers achieved a specificity of 90.48%, a sensitivity of 83.50%, and an area under the curve (AUC) value of 0.958 on the training set. On the validation set, these VOC biomarkers attained a predictive power of 85.71% for sensitivity and 73.08% for specificity, along with an AUC value of 0.875. Clinical risk factors exhibit certain predictive power on ADC and SCC prediction. Integrating these risk factors into the prediction model based on VOC biomarkers can enhance its predictive accuracy. This work indicates that exhaled breath holds the potential to precisely detect ADCs and SCCs. Considering clinical risk factors is essential when differentiating between these two subtypes.

肺癌亚型鉴定,尤其是腺癌(ADC)与鳞癌(SCC)的鉴别,对于临床医生制定有效的治疗策略至关重要。本研究旨在:(i) 发现用于精确诊断 ADC 和 SCC 的挥发性有机化合物生物标志物;(ii) 研究风险因素对 ADC 和 SCC 预测的影响;(iii) 探索挥发性有机化合物生物标志物的代谢途径。气相色谱-质谱法(GC-MS)分析了 ADC 患者(149 人)和 SCC 患者(94 人)的呼气样本。采用多变量和单变量统计分析方法确定挥发性有机化合物生物标志物。根据这些挥发性有机化合物生物标志物建立并验证了支持向量机(SVM)预测模型。研究了风险因素对 ADC 和 SCC 预测的影响。研究发现,13 种挥发性有机化合物在 ADC 和 SCC 之间存在显著差异。利用 SVM 算法,VOC 生物标记物在训练集上的特异性达到 90.48%,灵敏度达到 83.50%,AUC 值达到 0.958。在验证集上,这些 VOC 生物标记物的灵敏度和特异度分别达到了 85.71% 和 73.08%,AUC 值为 0.875。临床风险因素对 ADC 和 SCC 预测具有一定的预测能力。将这些风险因素整合到基于挥发性有机化合物生物标志物的预测模型中可提高其预测准确性。这项研究表明,呼出气体具有精确检测 ADC 和 SCC 的潜力。在区分这两种亚型时,考虑临床风险因素至关重要。
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引用次数: 0
Management of functional constipation-associated halitosis: a retrospective study. 功能性便秘相关口臭的治疗:一项回顾性研究。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-25 DOI: 10.1088/1752-7163/ad63c4
Xiao Xian Qian 钱孝先

The features of functional constipation (FC)-associated halitosis were identified in the author's previous report. In this report, the author aimed to further investigate its treatment and efficacy. This retrospective study reviewed 100 FC patients, including 82 (82%) halitosis patients and 18 (18%) non-halitosis patients. They underwent the organoleptic test (OLT) to diagnose halitosis, and the organoleptic score (OLS) (0-5) was used to evaluated halitosis severity. The Cleveland Clinical Constipation Score (CCCS) (0-30) was used to evaluate FC severity. Patients were treated with the laxative polyethylene glycol electrolyte powder (PGEP) for four weeks. These tests were performed before and after treatment. The author found that, before treatment, the CCCS was 20.00 (18.00-23.00) for all patients, 21.00 (19.00-24.00) for halitosis patients, and 18.00 (17.00-18.25) for non-halitosis patients. A significant difference was observed between halitosis patients and non-halitosis patients (P< 0.001). The OLS for halitosis patients was 3.00 (3.00-4.00). A positive correlation (r= 0.814, 95% CI: 0.732-0.872,P< 0.001) was found between OLS and CCCS. A CCCS ⩾18 predicted over 50% probability of halitosis. After treatment, the CCCS significantly decreased to 11.50 (6.00-14.75) (P< 0.001), and OLS significantly decreased to 1.00 (0.00-2.00) (P< 0.001). A positive correlation (r= 0.770, 95% CI: 0.673-0.841,P< 0.001) persisted between OLS and CCCS. A pre-treatment CCCS ⩾21 predicted over 50% probability of post-treatment halitosis, while a post-treatment CCCS ⩾12 predicted over 50% probability of post-treatment halitosis. The author concludes that the severity of FC parallels the severity of FC-associated halitosis, and can predict the probability of halitosis. Laxative treatment with PGEP is effective in improving FC-associated halitosis.

作者在之前的报告中指出了功能性便秘(FC)相关口臭的特征。在本报告中,作者旨在进一步研究其治疗和疗效。这项回顾性研究回顾了 100 例功能性便秘患者,包括 82 例(82%)口臭患者和 18 例(18%)非口臭患者。他们接受了感官测试(OLT)来诊断口臭,并用感官评分(OLS)(0-5)来评估口臭的严重程度。克利夫兰临床便秘评分(CCCS)(0-30)用于评估 FC 的严重程度。患者接受为期四周的泻药聚乙二醇电解质粉(PGEP)治疗。这些测试分别在治疗前后进行。作者发现,在治疗前,所有患者的 CCCS 为 20.00(18.00-23.00),口臭患者为 21.00(19.00-24.00),非口臭患者为 18.00(17.00-18.25)。口臭患者与非口臭患者之间存在明显差异(P < 0.001)。口臭患者的 OLS 为 3.00(3.00-4.00)。OLS 与 CCCS 之间呈正相关(r = 0.814,95% CI:0.732-0.872,P <0.001)。CCCS≥18 预测口臭的可能性超过 50%。治疗后,CCCS 显著降至 11.50(6.00-14.75)(P < 0.001),OLS 显著降至 1.00(0.00-2.00)(P < 0.001)。OLS 和 CCCS 之间持续存在正相关(r = 0.770,95% CI:0.673-0.841,P <0.001)。治疗前 CCCS≥21 预测治疗后口臭的概率超过 50%,而治疗后 CCCS≥12 预测治疗后口臭的概率超过 50%。作者的结论是,FC 的严重程度与 FC 相关口臭的严重程度相似,可以预测口臭的概率。使用 PGEP 进行通便治疗可有效改善 FC 相关性口臭。
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引用次数: 0
Cannabinoids detected in exhaled breath condensate after cannabis use. 吸食大麻后呼出的冷凝物中检测到大麻素。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-23 DOI: 10.1088/1752-7163/ad6347
Jennifer L Berry, Ashley Brooks-Russell, Cheryle N Beuning, Sarah A Limbacher, Tara M Lovestead, Kavita M Jeerage

Cannabinoids can be detected in breath after cannabis use, but different breath matrices need to be explored as studies to date with filter-based devices that collect breath aerosols have not demonstrated that breath-based measurements can reliably identify recent cannabis use. Exhaled breath condensate (EBC) is an unexplored aqueous breath matrix that contains condensed volatile compounds and water vapor in addition to aerosols. EBC was collected from participants both before and at two time points (0.7 ± 0.2 h and 1.7 ± 0.3 h) after observed cannabis use. Eleven different cannabinoids were monitored with liquid chromatography tandem mass spectrometry. Five different cannabinoids, including Δ9-tetrahydrocannabinol (THC), were detected in EBC collected from cannabis users. THC was detected in some EBC samples before cannabis use, despite the requested abstinence period. THC was detected in all EBC samples collected at 0.7 h post use and decreased for all participants at 1.7 h. Non-THC cannabinoids were only detected after cannabis use. THC concentrations in EBC samples collected at 0.7 h showed no trend with sample metrics like mass or number of breaths. EBC sampling devices deserve further investigation with respect to modes of cannabis use (e.g, edibles), post use time points, and optimization of cannabinoid recovery.

使用大麻后可在呼气中检测到大麻素,但需要探索不同的呼气基质,因为迄今为止使用基于过滤的设备收集呼气气溶胶的研究并未证明基于呼气的测量能够可靠地识别近期使用大麻的情况。呼出气体冷凝物(EBC)是一种尚未探索的含水呼气基质,除气溶胶外还含有冷凝挥发性化合物和水蒸气。在观察到吸食大麻之前和之后的两个时间点(0.7 h ± 0.2 h 和 1.7 h ± 0.3 h)收集参与者的 EBC。使用液相色谱串联质谱法(LC-MS/MS)监测了 11 种不同的大麻素。从大麻使用者体内收集的 EBC 中检测到五种不同的大麻素,包括 Δ9-四氢大麻酚(THC)。尽管规定了禁食期,但在使用大麻前的一些 EBC 样本中还是检测到了四氢大麻酚。在使用大麻后 0.7 小时采集的所有 EBC 样本中都检测到了四氢大麻酚,而在使用大麻后 1.7 小时,所有参与者的 EBC 样本中检测到的四氢大麻酚浓度都有所下降。在 0.7 小时采集的 EBC 样本中,四氢大麻酚浓度与样本质量或呼吸次数等指标没有趋势关系。EBC 采样设备在大麻使用方式(如食用)、使用后时间点和大麻素回收优化方面值得进一步研究。
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引用次数: 0
Methane gas in breath test is associated with non-alcoholic fatty liver disease. 呼气测试中的甲烷气体与非酒精性脂肪肝有关。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-17 DOI: 10.1088/1752-7163/ad5faf
Sanggwon An, Eui-Young Cho, Junho Hwang, Hyunseong Yang, Jungho Hwang, Kyusik Shin, Susie Jung, Bom-Taeck Kim, Kyu-Nam Kim, Wooyoung Lee

Although the associations between a patient's body mass index (BMI) and metabolic diseases, as well as their breath test results, have been studied, the relationship between breath hydrogen/methane levels and metabolic diseases needs to be further clarified. We aimed to investigate how the composition of exhaled breath gases relates to metabolic disorders, such as diabetes mellitus, dyslipidemia, hypertension, and nonalcoholic fatty liver disease (NAFLD), and their key risk factors. An analysis was performed using the medical records, including the lactulose breath test (LBT) data of patients who visited the Ajou University Medical Center, Suwon, Republic of Korea, between January 2016 and December 2021. The patients were grouped according to four different criteria for LBT hydrogen and methane levels. Of 441 patients, 325 (72.1%) had positive results for methane only (hydrogen < 20 parts per million [ppm] and methane ⩾ 3 ppm). BMIs and NAFLD prevalence were higher in patients with only methane positivity than in patients with hydrogen and methane positivity (hydrogen ⩾ 20 ppm and methane ⩾ 3 ppm). According to a multivariate analysis, the odds ratio of only methane positivity was 2.002 (95% confidence interval [CI]: 1.244-3.221,P= 0.004) for NAFLD. Our results demonstrate that breath methane positivity is related to NAFLD and suggest that increased methane gas on the breath tests has the potential to be an easily measurable biomarker for NAFLD diagnosis.

虽然已经研究了身体质量指数(BMI)水平与代谢性疾病之间的关系,以及呼气测试结果与 BMI 水平之间的关系,但呼出气体中氢气/甲烷水平与代谢性疾病之间的关系仍有待进一步明确。本研究旨在探讨呼出气体的成分与代谢性疾病及其主要风险因素之间的关系。体重指数(BMI)水平升高会大大增加患代谢性疾病的风险;本研究将其纳入研究范围,以发现两者之间的关联。本研究还包括糖尿病、血脂异常、高血压和非酒精性脂肪肝(NAFLD)等代谢性疾病。 研究人员对2016年1月至2021年12月期间在大韩民国水原市安州大学医疗中心就诊的患者的医疗记录(包括乳果糖呼气试验(LBT)数据)进行了分析。受试者按照乳果糖呼气试验氢气和甲烷水平的四种不同标准分组:1)正常(N)(氢气< 20 ppm,甲烷< 3 ppm);2)仅氢气(H+)(氢气≥ 20 ppm,甲烷< 3 ppm);3)甲烷阳性(M+)(氢气< 20 ppm,甲烷≥ 3 ppm);4)甲烷和氢气阳性(M+/H+)(氢气≥ 20 ppm,甲烷≥ 3 ppm)。在 441 名受试者中,325 人(72.1%)的甲烷检测结果呈阳性(M+)。M+受试者的体重指数和非酒精性脂肪肝发病率高于氢气和甲烷阳性(H+/M+)受试者。根据多变量分析,M+ 与非酒精性脂肪肝的几率比(OR)为 2.002(95% CI:1.244-3.221,P = 0.004)。我们的研究结果表明,呼气甲烷阳性与非酒精性脂肪肝有关,并表明呼气检测中甲烷气体的增加有可能成为诊断非酒精性脂肪肝的一种易于测量的生物标志物。
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引用次数: 0
Detection of pathogenic bacteria and biomarkers in lung specimens from cystic fibrosis patients. 检测囊性纤维化患者肺部标本中的致病菌和生物标记物。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-16 DOI: 10.1088/1752-7163/ad56bc
James J Tolle, Samadhan Jadhao, Brijesh Patel, Heying Sun, Susan Eastman, Tina Hartert, David N Ku, Larry J Anderson

Diagnosing lung infections is often challenging because of the lack of a high-quality specimen from the diseased lung. Since persons with cystic fibrosis are subject to chronic lung infection, there is frequently a need for a lung specimen. In this small, proof of principle study, we determined that PneumoniaCheckTM, a non-invasive device that captures coughed droplets from the lung on a filter, might help meet this need. We obtained 10 PneumoniaCheckTMcoughed specimens and 2 sputum specimens from adult CF patients hospitalized with an exacerbation of their illness. We detected amylase (upper respiratory tract) with an enzymatic assay, surfactant A (lower respiratory tract) with an immunoassay, pathogenic bacteria by PCR, and markers of inflammation by a Luminex multiplex immunoassay. The amylase and surfactant A levels suggested that 9/10 coughed specimens were from lower respiratory tract with minimal upper respiratory contamination. The PCR assays detected pathogenic bacteria in 7 of 9 specimens and multiplex Luminex assay detected a variety of cytokines or chemokines. These data indicate that the PneumoniaCheckTMcoughed specimens can capture good quality lower respiratory tract specimens that have the potential to help in diagnosis, management and understanding of CF exacerbations and other lung disease.

由于缺乏高质量的病肺标本,诊断肺部感染往往具有挑战性。由于囊性纤维化患者会受到慢性肺部感染,因此经常需要肺部标本。在这项小型原理验证研究中,我们确定 PneumoniaCheckTM 是一种非侵入性设备,可通过过滤器捕捉肺部咳出的液滴,从而帮助满足这一需求。我们从因病情加重而住院的成年 CF 患者身上获取了 10 份 PneumoniaCheckTM 咳嗽标本和 2 份痰标本。我们用酶法检测了淀粉酶(上呼吸道),用免疫测定法检测了表面活性物质 A(下呼吸道),用 PCR 检测了致病菌,用 Luminex 多重免疫测定法检测了炎症标记物。淀粉酶和表面活性物质 A 水平表明,9/10 的咳嗽标本来自下呼吸道,上呼吸道污染极少。PCR 检测在 9 份标本中的 7 份中检测到了病原菌,而多重 Luminex 检测则检测到了多种细胞因子或趋化因子。这些数据表明,PneumoniaCheckTM 咳嗽标本能采集到高质量的下呼吸道标本,有可能帮助诊断、管理和了解 CF 恶化和其他肺部疾病。
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引用次数: 0
Stability of selected exhaled breath volatiles stored in Tenax®TA adsorbent tubes at -80 °C. 部分呼出气体挥发物在 -80°C 温度下储存于 Tenax® TA 吸附管中的稳定性。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-12 DOI: 10.1088/1752-7163/ad5dee
Pawel Mochalski, Chris A Mayhew

Preservation of the breath sample integrity during storage and transport is one of the biggest challenges in off-line exhaled breath gas analysis. In this context, adsorbent tubes are frequently used as storage containers for use with analytical methods employing gas chromatography with mass spectrometric detection. The key objective of this short communication is to provide data on the recovery of selected breath volatiles from Tenax®TA adsorbent tubes that were stored at -80 °C for up to 90 d. For this purpose, an Owlstone Medical's ReCIVA®Breath Sampler was used for exhaled breath collection. The following fifteen compounds, selected to cover a range of chemical properties, were monitored for their stability: isoprene, n-heptane, n-nonane, toluene, p-cymene, allyl methyl sulfide, 1-(methylthio)-propane, 1-(methylthio)-1-propene,α-pinene, DL-limonene,β-pinene,γ-terpinene, 2-pentanone, acetoin and 2,3 butanedione. All compounds, but one (acetoin), were found to be stable during the first 4 weeks of storage (recovery within ± 2 × RSD). Furthermore, n-nonane was stable during the whole of the investigated period.

在离线呼出气体分析中,如何在储存和运输过程中保持呼气样本的完整性是最大的挑战之一。在这种情况下,吸附管经常被用作储存容器,用于采用气相色谱和质谱检测的分析方法。本短文的主要目的是提供在 -80 C 温度下储存长达 90 天的 Tenax® TA 吸附管中所选呼气挥发物的回收数据。为此,我们使用 Owlstone Medical 的 ReCIVA® 呼吸采样器收集呼出气体。监测了以下 15 种化合物的稳定性,所选化合物涵盖了一系列化学特性:异戊二烯、正庚烷、正壬烷、甲苯、对甲苯、烯丙基甲基硫醚、1-(甲硫基)-丙烷、1-(甲硫基)-1-丙烯、-蒎烯、DL-柠檬烯、β-蒎烯、-萜品烯、2-戊酮、乙炔和 2,3-丁二酮。除一种化合物(乙炔)外,所有化合物在最初 4 周的储藏期间都很稳定(回收率在  2×RSD 范围内)。此外,正壬烷在整个调查期间都很稳定。
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引用次数: 0
Extraction and characterization of exosomes from the exhaled breath condensate and sputum of lung cancer patients and vulnerable tobacco consumers-potential noninvasive diagnostic biomarker source. 从肺癌患者和易感烟草消费者呼出的冷凝液和痰中提取外泌体并确定其特征--潜在的无创诊断生物标记源。
IF 3.7 4区 医学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-11 DOI: 10.1088/1752-7163/ad5eae
Afsareen Bano, Pooja Yadav, Megha Sharma, Deepika Verma, Ravina Vats, Dhruva Chaudhry, Pawan Kumar, Rashmi Bhardwaj

Noninvasive sample sources of exosomes, such as exhaled breath and sputum, which are in close proximity to the tumor microenvironment and may contain biomarkers indicative of lung cancer, are far more permissive than invasive sample sources for biomarker screening. Standardized exosome extraction and characterization approaches for low-volume noninvasive samples are critically needed. We isolated and characterized exhaled breath condensate (EBC) and sputum exosomes from healthy nonsmokers (n= 30), tobacco smokers (n= 30), and lung cancer patients (n= 40) and correlated the findings with invasive sample sources. EBC samples were collected by using commercially available R-Tubes. To collect sputum samples the participants were directed to take deep breaths, hold their breath, and cough in a collection container. Dynamic light scattering, nanoparticle tracking analysis, and transmission electron microscopy were used to evaluate the exosome morphology. Protein isolation, western blotting, exosome quantification via EXOCET, and Fourier transform infrared spectroscopy were performed for molecular characterization. Exosomes were successfully isolated from EBC and sputum samples, and their yields were adequate and sufficiently pure for subsequent downstream processing and characterization. The exosomes were confirmed based on their size, shape, and surface marker expression. Remarkably, cancer exosomes were the largest in size not only in the plasma subgroups, but also in the EBC (p < 0.05) and sputum (p= 0.0036) subgroups, according to our findings. A significant difference in exosome concentrations were observed between the control sub-groups (p < 0.05). Our research confirmed that exosomes can be extracted from noninvasive sources, such as EBC and sputum, to investigate lung cancer diagnostic biomarkers for research, clinical, and early detection in smokers.

外泌体的非侵入性样本来源,如呼气和痰,与肿瘤微环境非常接近,可能含有肺癌的生物标记物,比侵入性样本来源更容易进行生物标记物筛选。我们亟需针对低容量非侵入性样本的标准化外泌体提取和表征方法。我们从健康的非吸烟者(30 人)、烟草吸烟者(30 人)和肺癌患者(40 人)身上分离出呼气冷凝物(EBC)和痰外泌体并对其进行表征,并将结果与侵入性样本来源进行关联。EBC样本使用市售的R-试管收集。收集痰液样本时,参与者要深呼吸、屏住呼吸并在收集容器中咳嗽。采用动态光散射、纳米粒子跟踪分析和透射电子显微镜评估外泌体形态。蛋白质分离、Western 印迹、通过 EXOCET 进行外泌体定量,以及傅立叶变换红外光谱进行分子表征。从 EBC 和痰样本中成功分离出了外泌体,其产量和纯度足以进行后续的下游处理和表征。根据外泌体的大小、形状和表面标记物的表达,对外泌体进行了确认。值得注意的是,根据我们的研究结果,癌症外泌体不仅在血浆亚组中体积最大,在EBC(p p= 0.0036)亚组中也是如此。对照亚组与对照亚组之间的外泌体浓度存在明显差异(p
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Journal of breath research
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