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Managing hemolysis in serum neuron-specific enolase measurements - an automated algorithm for routine practice. 管理血清神经元特异性烯醇化酶测量中的溶血现象--一种用于常规实践的自动算法。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-06-10 DOI: 10.1080/00365513.2024.2359091
Ragnhild V Nome, Elisabeth Paus, Johanna E Gehin, Nils Bolstad, Trine Bjøro

Neuron-specific enolase (NSE) derived from neurons and peripheral neuroendocrine cells is a biomarker for neuroendocrine tumors and for prognostication in comatose cardiac arrest survivors. However, as platelets and erythrocytes contain NSE, hemolysis causes falsely elevated NSE. We used native serum and hemolysate derived from the same patients to make serial dilutions, and subsequently measured NSE (mNSE) and hemolytic index (HI) in each dilution. An algorithm suitable for the laboratory information system was developed based on the mNSE, HI and the estimated gradient of hemolytic interference from 30 patients. We estimated the associated uncertainty of the corrected NSE (cNSE) results based on the observed range of the gradient and derived an equation for cNSE for samples with limited hemolysis (i.e. 5 < HI ≤ 30): cNSE = mNSE - HI × (0.34 ± 0.23) µg/L. By semi-quantitatively grading the contribution from limited hemolysis, a texted result noting the hemolysis-associated degree of uncertainty can accompany the cNSE result. The major challenge of hemolysis when using serum NSE as a biomarker can be managed using an automated algorithm for correction of NSE results based on degree of hemolysis. However, laboratorians and clinicians should be aware of the limitations associated with in vivo hemolysis.

神经元特异性烯醇化酶(NSE)来源于神经元和外周神经内分泌细胞,是神经内分泌肿瘤的生物标记物,也可用于昏迷的心脏骤停幸存者的预后判断。然而,由于血小板和红细胞中含有 NSE,溶血会导致 NSE 假性升高。我们使用来自同一患者的原生血清和溶血液进行连续稀释,然后测量每个稀释液中的 NSE(mNSE)和溶血指数(HI)。根据 30 名患者的 mNSE、HI 和估计的溶血干扰梯度,我们开发了一种适用于实验室信息系统的算法。我们根据观察到的梯度范围估算了校正 NSE(cNSE)结果的相关不确定性,并推导出了溶血有限样本(即 5 个体内溶血样本)的 cNSE 等式。
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引用次数: 0
Diagnostic performance of the CellaVision preclassification neutrophil count - time to bypass the reclassification? CellaVision 预分类中性粒细胞计数的诊断性能--是时候绕过重新分类了吗?
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-07-11 DOI: 10.1080/00365513.2024.2377967
Mikael Christiansen, Anders Abildgaard, Julie Brogaard Larsen, Gitte Tindbæk, Else Marie Vestergaard

Objectives: The objective of this study was to perform a method comparison between the CellaVision preclassification neutrophil count and the reclassification neutrophil count performed by trained laboratory technicians, and to evaluate the diagnostic performance of the preclassification neutrophil count at clinical decision levels.

Methods: We retrospectively identified patient samples through 2019-2022 in which the differential count was performed on Cellavision (n = 4,354). Data on sample characteristics and leukocyte- and differential counts was extracted from the electronic medical journal. For each sample, data containing the pre- and reclassification leukocyte classification, respectively, was extracted from the Cellavision software. Method comparison between the pre-and reclassification neutrophil count was performed using Bland Altman analysis. Diagnostic performance of the preclassification neutrophil count was evaluated according to four pre-specified categories of results with the reclassification as reference method.

Results: The median difference between the pre- and reclassification neutrophil count was 0.044 x 109/L. The preclassification neutrophil count categorised 95.6% of all samples correctly according to the four categories. The sensitivity, specificity, positive predictive value and negative predictive value for detecting neutrophilia > 7.00 x 109/L was 98.8%, 97.2%, 95.8%, and 99.2%, respectively. In samples with leukopenia (n = 543), the sensitivity, specificity, positive predictive value and negative predictive value for detecting severe neutropenia (< 0.50 x 109/L) was 97.7%, 99.1%, 98.6%, and 98.5%, respectively.

Conclusion: The diagnostic performance of the CellaVision preclassification neutrophil count was satisfactory. The preclassification neutrophil count may be released to the electronic medical journal to improve turnaround time and benefit laboratory management.

研究目的本研究旨在对 CellaVision 预分类中性粒细胞计数与训练有素的实验室技术人员进行的再分类中性粒细胞计数进行方法比较,并评估预分类中性粒细胞计数在临床决策层面的诊断性能:我们回顾性地确定了截至 2019-2022 年在 Cellavision 上进行差分计数的患者样本(n = 4354)。样本特征、白细胞计数和差值计数数据均从电子医学期刊中提取。每个样本的白细胞分类前和分类后的数据分别从 Cellavision 软件中提取。使用布兰德-阿尔特曼分析法对分类前和分类后的中性粒细胞计数进行比较。根据预先指定的四个结果类别,以重新分类作为参考方法,评估了重新分类前中性粒细胞计数的诊断性能:结果:预分类和再分类中性粒细胞计数的中位数差异为 0.044 x 109/L。在所有样本中,95.6%的中性粒细胞计数分类结果正确。检测中性粒细胞> 7.00 x 109/L的灵敏度、特异性、阳性预测值和阴性预测值分别为98.8%、97.2%、95.8%和99.2%。在白细胞减少症样本(n = 543)中,检测严重中性粒细胞减少症(< 0.50 x 109/L)的灵敏度、特异性、阳性预测值和阴性预测值分别为 97.7%、99.1%、98.6% 和 98.5%:结论:CellaVision预分类中性粒细胞计数的诊断性能令人满意。预分类中性粒细胞计数可在电子医学期刊上发布,以缩短周转时间,有利于实验室管理。
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引用次数: 0
Correction. 更正。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-06-21 DOI: 10.1080/00365513.2024.2367391
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引用次数: 0
Lipid levels in a cohort of healthy Danish schoolchildren ages 5 to 17 years. 丹麦 5 至 17 岁健康学童的血脂水平。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-07-16 DOI: 10.1080/00365513.2024.2370011
Jens Heller Greve, Freja Mørk, Andreas Kryger Jensen, Simranjeet Kaur, Jens Otto Broby Madsen, Anna Bugge, Malene Heidemann, Niels Wedderkopp, Jesper Johannesen

It is internationally recognized to use clinical decision limits (CDL) when interpreting the lipid levels in both adults and children, even though the evidence for children is scarce. The purpose of this study is to describe how lipid levels progress in healthy Danish children ages 5 to 17 years. This study is based on the Childhood Health, Activity, and Motor Performance School Study Denmark (CHAMPS-study DK) consisting of 1456 observations of schoolchildren aged 5 to 17 years. Participants have been tested for blood levels of total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides, and remnant cholesterol levels are calculated. Finally, sex-specific percentile reference curves are presented. Percentile reference curves stratified by sex were generated for all cholesterols and showed that the total cholesterol level peaks at 4.32 mmol/l in 10-year-old boys and 4.46 mmol/l in nine-year-old girls. HDL levels in boys peak at 1.72 mmol/l in nine-year-old boys. HDL levels in girls and LDL levels in both sexes are nearly constant. Triglycerides kept rising to the age of 17 years in both sexes and remnant cholesterol decreased from age 5 to 17 years in both sexes. BMI z-score adjustment revealed no significant association with total cholesterol in both sexes but a significant association between HDL, LDL, triglycerides, and remnant cholesterol. This study is the first to generate percentile reference curves for blood levels of total cholesterol, LDL, HDL, triglycerides, and remnant cholesterol in a cohort of healthy Danish children aged 5 to 17 years.

国际公认,在解释成人和儿童的血脂水平时应使用临床决策限值(CDL),尽管针对儿童的证据很少。本研究旨在描述 5 至 17 岁丹麦健康儿童血脂水平的变化情况。这项研究以丹麦儿童健康、活动和运动表现学校研究(CHAMPS-study DK)为基础,对 1456 名 5 至 17 岁的学龄儿童进行了观察。参与者的血液中总胆固醇、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)和甘油三酯的水平均已检测,残余胆固醇水平也已计算。最后,列出了按性别划分的百分位数参考曲线。按性别分层生成的百分位数参考曲线显示,10 岁男孩的总胆固醇水平峰值为 4.32 毫摩尔/升,9 岁女孩的总胆固醇水平峰值为 4.46 毫摩尔/升。男孩高密度脂蛋白水平的峰值为 1.72 毫摩尔/升,9 岁男孩为 1.72 毫摩尔/升。女孩的高密度脂蛋白水平和男女的低密度脂蛋白水平几乎保持不变。男女儿童的甘油三酯在 17 岁前持续上升,而残余胆固醇在 5 至 17 岁期间均有所下降。体重指数 z 值调整显示,男女两性的总胆固醇均无显著关联,但高密度脂蛋白、低密度脂蛋白、甘油三酯和残余胆固醇之间存在显著关联。该研究首次在丹麦 5-17 岁健康儿童队列中生成了血液中总胆固醇、低密度脂蛋白、高密度脂蛋白、甘油三酯和残余胆固醇水平的百分位数参考曲线。
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引用次数: 0
Evaluating routine blood and cerebrospinal fluid samples in narcolepsy patients. 评估嗜睡症患者的常规血液和脑脊液样本。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-06-27 DOI: 10.1080/00365513.2024.2369992
Niels Christian Haubjerg Østerby, Niklas Rye Jørgensen, Poul Jørgen Jennum

Cerebrospinal fluid hypocretin-1 is proven to be a precise diagnostic marker of narcolepsy Type 1 (NT1). However other characteristics of cerebrospinal fluid and blood parameters have not yet been described. The objective of this study was to evaluate the differences in routine blood and cerebrospinal fluid analyses between NT1 patients and patients suspected of hypersomnia. We collected retrospectively all measures of cerebrospinal fluid hypocretin-1 between 2019 and 2022. This yielded 612 patients out of which 146 were diagnosed with NT1 and the rest (466 patients) were used as a control group. We selected the most relevant routine samples from both blood, plasma and cerebrospinal fluid and compared the two groups. The only significantly different analytes were plasma lactate dehydrogenase and cerebrospinal fluid hypocretin-1. No other differences were found between the groups including thyroid markers, markers of neuroendocrine function, inflammatory markers in blood or cerebrospinal fluid, markers of permeability of the blood brain barrier or metabolic markers in blood samples. We found no significant differences in routine blood or cerebrospinal fluid components, neuroendocrine function, neuroinflammation and metabolic markers. The results reflect that the hypocretin system does not seem to play a chronic major role in regulation of these markers. None of the parameters routinely measured in blood in these patients could differentiate between NT1 and non-NT1 disorders besides CSF-hcrt-1.

脑脊液降视素-1 被证明是 1 型嗜睡症(NT1)的精确诊断标志物。然而,脑脊液和血液参数的其他特征尚未得到描述。本研究的目的是评估 NT1 患者与嗜睡症疑似患者在常规血液和脑脊液分析方面的差异。我们回顾性地收集了2019年至2022年期间脑脊液降视素-1的所有指标。结果发现有 612 名患者,其中 146 人被确诊为 NT1 患者,其余(466 人)作为对照组。我们从血液、血浆和脑脊液中选择了最相关的常规样本,并对两组样本进行了比较。唯一有明显差异的分析物是血浆中的乳酸脱氢酶和脑脊液中的降视素-1。两组之间没有发现其他差异,包括甲状腺标志物、神经内分泌功能标志物、血液或脑脊液中的炎症标志物、血脑屏障通透性标志物或血液样本中的代谢标志物。我们发现,在常规血液或脑脊液成分、神经内分泌功能、神经炎症和代谢标志物方面没有明显差异。结果表明,视网膜下视素系统在这些指标的调节中似乎并不长期起主要作用。除了 CSF-hcrt-1 之外,这些患者血液中常规测量的参数都无法区分 NT1 和非 NT1 疾病。
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引用次数: 0
Effects of dabigatran, rivaroxaban, and apixaban on fibrin network permeability, thrombin generation, and fibrinolysis. 达比加群、利伐沙班和阿哌沙班对纤维蛋白网络通透性、凝血酶生成和纤维蛋白溶解的影响。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-07-02 DOI: 10.1080/00365513.2024.2369993
Viktor Schutz Taune, Michal Zabczyk, Shu He, Anna Ågren, Margareta Blombäck, Håkan Wallén, Mika Skeppholm

Introduction: There are important pharmacological differences between direct oral anticoagulants (DOAC) and a deeper knowledge of how they influence different aspects of hemostasis in patients on treatment is desirable.

Materials and methods: Blood samples from patients on dabigatran (n = 23), rivaroxaban (n = 26), or apixaban (n = 20) were analyzed with a fibrin network permeability assay, a turbidimetric clotting and lysis assay, the calibrated automated thrombogram (CAT), plasma levels of thrombin-antithrombin complex (TAT) and D-dimer, as well as DOAC concentrations, PT-INR and aPTT. As a comparison, we also analyzed samples from 27 patients on treatment with warfarin.

Results: Patients on dabigatran had a more permeable fibrin network, longer lag time (CAT and turbidimetric assay), and lower levels of D-dimer in plasma, compared with patients on rivaroxaban- and apixaban treatment, and a more permeable fibrin network than patients on warfarin. Clot lysis time was slightly longer in patients on dabigatran than in patients on rivaroxaban. Warfarin patients formed a more permeable fibrin network than patients on apixaban, had longer lag time than patients on rivaroxaban (CAT assay), and lower peak thrombin and ETP compared to patients on treatment with both FXa-inhibitors.

Conclusions: Results from this study indicate dabigatran treatment is a more potent anticoagulant than apixaban and rivaroxaban. However, as these results are not supported by clinical data, they are probably more related to the assays used and highlight the difficulty of measuring and comparing the effect of anticoagulants.

简介:直接口服抗凝血剂(DOAC)之间存在着重要的药理差异,因此需要更深入地了解这些药物如何影响患者治疗过程中止血的不同方面:直接口服抗凝药(DOAC)之间存在着重要的药理差异,因此需要更深入地了解这些药物如何影响正在接受治疗的患者止血的不同方面:对服用达比加群(23 例)、利伐沙班(26 例)或阿哌沙班(20 例)患者的血样进行了纤维蛋白网络渗透性测定、浊度凝血和裂解测定、校准自动血栓图(CAT)、血浆凝血酶-抗凝血酶复合物(TAT)和 D-二聚体水平以及 DOAC 浓度、PT-INR 和 aPTT 分析。作为对比,我们还分析了27名接受华法林治疗的患者的样本:与接受利伐沙班和阿哌沙班治疗的患者相比,服用达比加群的患者血浆中纤维蛋白网络的渗透性更强,滞后时间(CAT和浊度测定法)更长,D-二聚体水平更低,而服用华法林的患者血浆中纤维蛋白网络的渗透性更强。服用达比加群的患者血栓溶解时间略长于服用利伐沙班的患者。与服用两种FXa抑制剂的患者相比,服用华法林的患者形成的纤维蛋白网络比服用阿哌沙班的患者更具渗透性,滞后时间比服用利伐沙班(CAT检测)的患者更长,凝血酶峰值和ETP更低:本研究结果表明,达比加群比阿哌沙班和利伐沙班的抗凝效果更好。然而,由于这些结果没有得到临床数据的支持,它们可能更多地与所使用的检测方法有关,并凸显了测量和比较抗凝剂效果的难度。
{"title":"Effects of dabigatran, rivaroxaban, and apixaban on fibrin network permeability, thrombin generation, and fibrinolysis.","authors":"Viktor Schutz Taune, Michal Zabczyk, Shu He, Anna Ågren, Margareta Blombäck, Håkan Wallén, Mika Skeppholm","doi":"10.1080/00365513.2024.2369993","DOIUrl":"10.1080/00365513.2024.2369993","url":null,"abstract":"<p><strong>Introduction: </strong>There are important pharmacological differences between direct oral anticoagulants (DOAC) and a deeper knowledge of how they influence different aspects of hemostasis in patients on treatment is desirable.</p><p><strong>Materials and methods: </strong>Blood samples from patients on dabigatran (<i>n</i> = 23), rivaroxaban (<i>n</i> = 26), or apixaban (<i>n</i> = 20) were analyzed with a fibrin network permeability assay, a turbidimetric clotting and lysis assay, the calibrated automated thrombogram (CAT), plasma levels of thrombin-antithrombin complex (TAT) and D-dimer, as well as DOAC concentrations, PT-INR and aPTT. As a comparison, we also analyzed samples from 27 patients on treatment with warfarin.</p><p><strong>Results: </strong>Patients on dabigatran had a more permeable fibrin network, longer lag time (CAT and turbidimetric assay), and lower levels of D-dimer in plasma, compared with patients on rivaroxaban- and apixaban treatment, and a more permeable fibrin network than patients on warfarin. Clot lysis time was slightly longer in patients on dabigatran than in patients on rivaroxaban. Warfarin patients formed a more permeable fibrin network than patients on apixaban, had longer lag time than patients on rivaroxaban (CAT assay), and lower peak thrombin and ETP compared to patients on treatment with both FXa-inhibitors.</p><p><strong>Conclusions: </strong>Results from this study indicate dabigatran treatment is a more potent anticoagulant than apixaban and rivaroxaban. However, as these results are not supported by clinical data, they are probably more related to the assays used and highlight the difficulty of measuring and comparing the effect of anticoagulants.</p>","PeriodicalId":21474,"journal":{"name":"Scandinavian Journal of Clinical & Laboratory Investigation","volume":" ","pages":"257-267"},"PeriodicalIF":1.3,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141493237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The diagnostic accuracy of quality control rules. 质量控制规则的诊断准确性。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-05-28 DOI: 10.1080/00365513.2024.2359085
Arne Åsberg, Bjørn Johan Bolann

Internal quality control in clinical chemistry laboratories are based on analyzing samples of stable control materials among the patient samples. The control results are interpreted by using quality control rules that usually are designed to detect systematic errors. The best rules have a high probability of error detection (Ped), i.e. to detect the maximal allowable (critical) systematic error and a low probability of false rejection (Pfr, false alarm). In this work we show that quality control rules can be represented by points on a ROC curve which appears when Ped is plotted against Pfr and only the control limit is varied. Further, we introduce a new method for choosing the optimal control limit, analogous to choosing the optimal operating point on the ROC curve of a diagnostic test. This decision needs knowledge of the pretest probability of a critical systematic error, the benefit of detecting it when it occurs and the cost of false alarm. The ROC curve analysis showed that if rules based on N = 2 are used, mean rules outperform Westgard rules because the ROC curve of the mean rules was lying above the ROC curves of the Westgard rules. A mean rule also had a lower maximum expected increase in the number of unacceptable patient results reported during the presence of an out-of-control error condition (Max E(NUF)) than comparable Westgard rules.

临床化学实验室的内部质量控制以分析病人样本中的稳定对照材料样本为基础。对照结果通过质量控制规则来解释,这些规则通常是为了检测系统误差而设计的。最佳规则具有较高的错误检测概率(Ped),即检测出最大允许(临界)系统误差,以及较低的错误拒绝概率(Pfr,误报)。在这项工作中,我们证明了质量控制规则可以用 ROC 曲线上的点来表示,当 Ped 与 Pfr 相对应时,ROC 曲线上的点就会出现,并且只改变控制限。此外,我们还介绍了一种选择最佳控制限的新方法,类似于选择诊断测试 ROC 曲线上的最佳操作点。这一决策需要了解临界系统误差的测试前概率、发生误差时检测到误差的收益以及误报的成本。ROC 曲线分析表明,如果使用基于 N = 2 的规则,平均值规则优于 Westgard 规则,因为平均值规则的 ROC 曲线位于 Westgard 规则的 ROC 曲线之上。此外,平均值规则在出现失控错误条件时报告的不可接受的病人结果数量的最大预期增加值(Max E(NUF))也低于同类 Westgard 规则。
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引用次数: 0
Diagnosing sucrase-isomaltase deficiency: a comparison of a 13C-sucrose breath test and a duodenal enzyme assay. 诊断蔗糖酶-异麦芽糖酶缺乏症:13C-蔗糖呼气试验与十二指肠酶测定法的比较。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-07-10 DOI: 10.1080/00365513.2024.2377960
Hanna Fjeldheim Dale, Milada Hagen, Chirajyoti Deb, Viggo Skar, Jørgen Valeur

Background: Reduced activity of the sucrase-isomaltase (SI) enzyme can cause gastrointestinal symptoms. Biochemical measurement of SI activity in small intestinal biopsies is presently considered the gold standard for the diagnosis of SI deficiency, but this invasive test is not suitable as a routine diagnostic tool.

Aim: To evaluate a 13C-sucrose-breath test (13CSBT) as a diagnostic tool for SI deficiency in an adult population.

Methods: 13CSBT results were compared to sucrase activity measured in duodenal biopsies.

Results: Forty patients with gastrointestinal symptoms were included in the study, 4 of whom had celiac disease and the rest (n = 36) had normal histological findings. Nine patients (22.5%) had low sucrase activity measured using duodenal biopsies. No correlation was observed between enzymatic sucrase activity and the 13CSBT results. The 13CSBT-curves for the celiac patients versus patients with normal duodenal histology demonstrated that the patients with celiac disease were within the lower range of the distribution.

Conclusion: We observed a mismatch between the 13CSBT results and the biochemically measured sucrase activity, suggesting that SI activity is not uniformly distributed throughout the small intestines. This methodological discrepancy should be acknowledged when diagnosing SI deficiency.

背景:蔗糖异麦芽糖酶(SI)活性降低可导致胃肠道症状。目的:评估 13C-蔗糖呼气试验(13CSBT)作为诊断成人 SI 缺乏症的工具的效果。方法:将 13CSBT 结果与十二指肠活检中测得的蔗糖酶活性进行比较:研究共纳入了 40 名有胃肠道症状的患者,其中 4 人患有乳糜泻,其余患者(n = 36)的组织学检查结果正常。九名患者(22.5%)的十二指肠活检结果显示蔗糖酶活性较低。酶促蔗糖酶活性与 13CSBT 结果之间没有相关性。乳糜泻患者与十二指肠组织学正常患者的 13CSBT 曲线显示,乳糜泻患者的分布范围较低:我们观察到 13CSBT 结果与生化测定的蔗糖酶活性不匹配,这表明蔗糖酶活性在整个小肠的分布并不均匀。在诊断 SI 缺乏症时,应认识到这一方法上的差异。
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引用次数: 0
Evaluation of a flow cytometry-based method for determination of T-lymphocyte subtypes for quality assessment of cell therapy products. 评估基于流式细胞仪的 T 淋巴细胞亚型测定方法,用于细胞治疗产品的质量评估。
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-07-14 DOI: 10.1080/00365513.2024.2377961
Vladimira Rimac, Ines Bojanić, Nikolina Blažević, Koraljka Gojčeta

Chimeric antigen receptor-T (CAR-T) cell therapy is currently the best-known type of immune effector cells therapy. For CAR T-cell therapy, the determination of CD3+ T cells is necessary for the quality control of fresh leukapheresis product as starting material. The aim was to validate analytical method for quantification of percentage and absolute count of T lymphocyte subtypes (CD3+, CD4+ and CD8+ cells) in fresh apheresis products using single-platform method on flow cytometer BD FACS Canto II. Validation study included determination of precision, trueness (bias), assessment of linearity, carryover, comparison of results obtained with two different protocols on flow cytometer for CD3+ cells determination and stability study. For between-run precision coefficients of variation (CVs) were <20%, as well as bias for all T-lymphocyte subtypes. For within-run precision, CVs were <10%, except for low CD8+ cell (percentage 10.51% and viable absolute count 12.37%). Comparison of results obtained with two different protocols for CD3+ cells determination shows no statistically significant difference. Statistically significant differences between results of the analysis of CD4+ cells in fresh samples and results obtained after storage at 4 °C (p = .004) and at room temperature (p = .018) were found. In conclusion, method for enumeration of T-lymphocyte subtypes can be used in routine work on BD FACS Canto II instrument for quality assessment of fresh cell products collected by leukapheresis procedure.

嵌合抗原受体-T(CAR-T)细胞疗法是目前最著名的免疫效应细胞疗法。对于 CAR T 细胞疗法来说,CD3+ T 细胞的测定对于作为起始材料的新鲜白细胞产品的质量控制是必要的。目的是在 BD FACS Canto II 流式细胞仪上使用单平台方法验证定量新鲜白细胞采集产物中 T 淋巴细胞亚型(CD3+、CD4+ 和 CD8+ 细胞)百分比和绝对计数的分析方法。验证研究包括测定精确度、真实度(偏差)、线性评估、携带、比较两种不同的流式细胞仪测定 CD3+ 细胞的方案所获得的结果以及稳定性研究。结果发现,运行间精密度(CVs)为 p = .004,室温下精密度(CVs)为 p = .018。总之,T淋巴细胞亚型的计数方法可用于 BD FACS Canto II 仪器的常规工作中,以评估通过白细胞清除程序收集的新鲜细胞产品的质量。
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引用次数: 0
Impact of breast cancer neoadjuvant chemotherapy on plasma and urine amino acid profile, plasma proteins and nitrogen metabolism. 乳腺癌新辅助化疗对血浆和尿液氨基酸谱、血浆蛋白和氮代谢的影响
IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Pub Date : 2024-07-01 Epub Date: 2024-06-27 DOI: 10.1080/00365513.2024.2369982
Milan Dastych, Miloš Holánek, Jana Gottwaldová, Zdenka Čermáková, Alena Mikušková

Neoadjuvant chemotherapy (NAC) is the preferred treatment option in locally advanced breast cancer (BC). The administration of NAC is associated with a wide range of adverse effects. This pilot observational prospective study examined the effect of NAC using anthracycline + cyclophosphamide (AC) followed by paclitaxel (PTx) on a portfolio of 22 plasma and urinary amino acids, plasma proteins (albumin, prealbumin, transferrin), and products of nitrogen metabolism (urea, creatinine, uric acid) in plasma and urine. Plasma and 24-h urine samples were obtained from ten patients with early breast cancer (N1-3 N0-2 M0), at the following time points: before the start of NAC and during the AC/PTx treatment period (a total of 8 measurements at three-weekly intervals). Amino acids were analyzed using ion exchange chromatography. There were no significant differences in the measured parameters in plasma and urine between pre-NAC and during AC- and PTx-treatment. No trend was detected. A significant difference in the portfolio of plasma and urinary amino acids was found only in the pre-treatment period compared to the control group. Levels of eight plasma amino acids (8/22) were significantly reduced and those of nine urine amino acids were increased (9/22). Nitrogenous catabolites in plasma and urine were not indicative of increased protein catabolism during the anthracycline and taxane treatment periods. A slightly positive nitrogen balance was accompanied by an average weight gain of 3.3 kg (range 0-6 kg). The AC/PTx treatment regimen did not cause significant changes in the monitored laboratory parameters.

新辅助化疗(NAC)是局部晚期乳腺癌(BC)的首选治疗方案。新辅助化疗与多种不良反应相关。这项试点观察性前瞻性研究考察了使用蒽环类+环磷酰胺(AC)和紫杉醇(PTx)的新辅助化疗对血浆和尿液中的 22 种氨基酸、血浆蛋白(白蛋白、前白蛋白、转铁蛋白)和氮代谢产物(尿素、肌酐、尿酸)的影响。十名早期乳腺癌患者(N1-3 N0-2 M0)在以下时间点采集了血浆和 24 小时尿液样本:开始接受 NAC 治疗前和 AC/PTx 治疗期间(共 8 次测量,每次间隔三周)。氨基酸采用离子交换色谱法进行分析。在 NAC 治疗前、AC 和 PTx 治疗期间,血浆和尿液中的测量参数没有明显差异。未发现任何趋势。只有在治疗前,血浆和尿液中的氨基酸组合与对照组相比有明显差异。八种血浆氨基酸(8/22)的水平显著降低,九种尿液氨基酸(9/22)的水平显著升高。在蒽环类和紫杉类药物治疗期间,血浆和尿液中的含氮代谢物并不表明蛋白质代谢增加。氮平衡略呈正值的同时,平均体重增加了 3.3 千克(范围为 0-6 千克)。AC/PTx 治疗方案并未导致监测到的实验室参数发生显著变化。
{"title":"Impact of breast cancer neoadjuvant chemotherapy on plasma and urine amino acid profile, plasma proteins and nitrogen metabolism.","authors":"Milan Dastych, Miloš Holánek, Jana Gottwaldová, Zdenka Čermáková, Alena Mikušková","doi":"10.1080/00365513.2024.2369982","DOIUrl":"10.1080/00365513.2024.2369982","url":null,"abstract":"<p><p>Neoadjuvant chemotherapy (NAC) is the preferred treatment option in locally advanced breast cancer (BC). The administration of NAC is associated with a wide range of adverse effects. This pilot observational prospective study examined the effect of NAC using anthracycline + cyclophosphamide (AC) followed by paclitaxel (PTx) on a portfolio of 22 plasma and urinary amino acids, plasma proteins (albumin, prealbumin, transferrin), and products of nitrogen metabolism (urea, creatinine, uric acid) in plasma and urine. Plasma and 24-h urine samples were obtained from ten patients with early breast cancer (N1-3 N0-2 M0), at the following time points: before the start of NAC and during the AC/PTx treatment period (a total of 8 measurements at three-weekly intervals). Amino acids were analyzed using ion exchange chromatography. There were no significant differences in the measured parameters in plasma and urine between pre-NAC and during AC- and PTx-treatment. No trend was detected. A significant difference in the portfolio of plasma and urinary amino acids was found only in the pre-treatment period compared to the control group. Levels of eight plasma amino acids (8/22) were significantly reduced and those of nine urine amino acids were increased (9/22). Nitrogenous catabolites in plasma and urine were not indicative of increased protein catabolism during the anthracycline and taxane treatment periods. A slightly positive nitrogen balance was accompanied by an average weight gain of 3.3 kg (range 0-6 kg). The AC/PTx treatment regimen did not cause significant changes in the monitored laboratory parameters.</p>","PeriodicalId":21474,"journal":{"name":"Scandinavian Journal of Clinical & Laboratory Investigation","volume":" ","pages":"237-244"},"PeriodicalIF":1.3,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141459060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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Scandinavian Journal of Clinical & Laboratory Investigation
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