Actins

肌动蛋白
  • 文章类型: Journal Article
    目的:本研究的目的是研究α平滑肌肌动蛋白和转化生长因子β的免疫组织化学表达,并比较它们在口腔乳头状鳞状细胞癌中的表达和在不同组织学分级的口腔鳞状细胞癌中的表达。然后进行这些免疫组织化学表达和组织学发现之间的相关性。研究的问题是“OPSCC中α-SMA和TGF-β免疫表达的百分比是否与常规OSCC中的不同?”。
    方法:这将通过收集口腔乳头状鳞状细胞癌和不同等级的口腔鳞状细胞癌的档案块来实现,用转化生长因子β和α平滑肌肌动蛋白染色标本,然后测量各组表达的平均染色指数和两种标记物的面积百分比。
    结果:结果显示,在高分化鳞状细胞癌的所有病例中,上皮中转化生长因子β的表达都很高,大多数口腔乳头状鳞状细胞癌,和低分化口腔鳞状细胞癌。另一方面,不同级别的口腔鳞状细胞癌显示间质中α-平滑肌肌动蛋白的高染色指数表达。而口腔乳头状鳞状细胞癌的病例为中度或低染。
    结论:与不同组织学分级相比,口腔乳头状鳞状细胞癌具有良好的预后,预后不仅取决于组织学分级,还取决于其他预后因素。
    OBJECTIVE: The aim of the study is to investigate the immunohistochemical expression of both Alpha smooth muscle actin and Transforming Growth Factor beta and compare their expression in oral papillary squamous cell carcinoma with their expression in different histological grades of oral squamous cell carcinoma. A correlation between these immuno-histochemical expressions and histological findings will then be performed. The research question is \"Do the percentages of α-SMA and TGF-β immune-expression in OPSCC differ from that in the conventional OSCC?\".
    METHODS: This will be achieved by collecting archival blocks of oral papillary squamous cell carcinoma and different grades of oral squamous cell carcinoma, staining the specimens with Transforming Growth Factor beta and alpha smooth muscle actin, then measuring the mean staining index of expression in each group and the area percent of both markers.
    RESULTS: Results revealed that transforming growth factor beta expression in the epithelium was high in all cases of well-differentiated squamous cell carcinoma, most oral papillary squamous cell carcinoma, and poorly differentiated oral squamous cell carcinoma. On the other hand, different grades of oral squamous cell carcinoma showed a high staining index of alpha smooth muscle actin expression in the stroma. While cases of oral papillary squamous cell carcinoma were either moderate or low-staining.
    CONCLUSIONS: Oral papillary squamous cell carcinoma has a favourable prognosis compared to different histological grades, and the prognosis does not depend only on histological grade but also on other prognostic factors.
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  • 文章类型: Journal Article
    背景:口腔粘膜下纤维化(OSF)是一种癌前病变,口腔鳞状细胞癌(OSCC)是影响口腔粘膜的最常见恶性肿瘤。OSF向OSCC的恶性转化估计发生在7-13%的病例中。肌成纤维细胞(MFs)在生理和病理过程中发挥关键作用,比如伤口愈合和肿瘤发生,分别。本研究旨在探讨MFs在OSF及其恶性转化过程中的作用。
    方法:总共,收集94个福尔马林固定石蜡包埋的组织块,包括正常口腔粘膜(NOM;n=10),早期中度OSF(EMOSF;n=29),高级OSF(AOSF;n=29),癌旁OSF(POSF;n=21),和OSCC(n=5)样本。α-平滑肌肌动蛋白用于MFs的免疫组织化学鉴定。
    结果:NOM表现出不常见的MFs表达。在AOSF中发现了较高的MFs染色指数,其次是EMOSF和NOM。此外,从EMOSF到POSF和OSCC,MF的染色指数显着增加。MOM中MFs的染色指数,EMOSF,AOSF,POSF,OSCC分别为0.14±0.2、1.69±1.4、2.47±1.2、3.57±2.6和8.86±1.4。所有结果均有统计学意义(P<0.05)。
    结论:随着疾病从轻度转化到恶性转化,MFs的表达逐渐增加,表明MFs在与OSF相关的纤维化和潜在肿瘤发生中的作用。
    BACKGROUND: Oral submucous fibrosis (OSF) is a precancerous lesion, with oral squamous cell carcinoma (OSCC) being the most prevalent malignancy affecting the oral mucosa. The malignant transformation of OSF into OSCC is estimated to occur in 7-13% of cases. Myofibroblasts (MFs) play pivotal roles in both physiological and pathological processes, such as wound healing and tumorigenesis, respectively. This study aimed to explore the involvement of MFs in the progression of OSF and its malignant transformation.
    METHODS: In total, 94 formalin-fixed paraffin-embedded tissue blocks were collected, including normal oral mucosa (NOM; n = 10), early-moderate OSF (EMOSF; n = 29), advanced OSF (AOSF; n = 29), paracancerous OSF (POSF; n = 21), and OSCC (n = 5) samples. Alpha-smooth muscle actin was used for the immunohistochemical identification of MFs.
    RESULTS: NOM exhibited infrequent expression of MFs. A higher staining index of MFs was found in AOSF, followed by EMOSF and NOM. Additionally, a significant increase in the staining index of MFs was found from EMOSF to POSF and OSCC. The staining index of MFs in NOM, EMOSF, AOSF, POSF, and OSCC was 0.14 ± 0.2, 1.69 ± 1.4, 2.47 ± 1.2, 3.57 ± 2.6, and 8.86 ± 1.4, respectively. All results were statistically significant (P < 0.05).
    CONCLUSIONS: The expression of MFs exhibited a gradual increase as the disease progressed from mild to malignant transformation, indicating the contributory role of MFs in the fibrogenesis and potential tumorigenesis associated with OSF.
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  • 文章类型: Journal Article
    背景和目的:伤口愈合包含多种因素,并且需要建立基底膜成分之间的相互作用。颗粒浓度的定量可以用作评估生物力学肌肉特性的有价值的生物标志物。这项研究的目的是检查VI型肌层胶原的免疫表达和免疫浓度,弹性蛋白,α-平滑肌肌动蛋白,和平滑肌肌球蛋白重链,以及剖宫产(CS)后子宫瘢痕中血小板和分化簇31的表达。材料和方法:从健康的孕妇队列中获得总共177例活检,特别是在CS的外科手术过程中。参与者被分为七个不同的组。第1组由初产妇组成,共有52人。随后的组是根据自上一次CS以来所经过的持续时间来组织的。分析集中于所列出的颗粒的免疫表达和免疫浓度。结果:VI型胶原蛋白的肌层免疫浓度未观察到显着变化,弹性蛋白,平滑肌肌球蛋白,和内皮细胞分化簇31在分析组中。发现子宫肌层中α-平滑肌肌动蛋白的浓度在自先前CS以来不到2年内接受CS的患者中明显更高。与那些手术间隔较长的人相比。结论:我们的发现表明子宫肌层瘢痕VI型胶原的免疫浓度,弹性蛋白,平滑肌肌球蛋白重链,α-平滑肌肌动蛋白,和分化的内皮细胞标记物簇31保持一致,而不管自先前CS以来经过的持续时间。研究结果表明,CS后13个月后,子宫肌肉的生物力学特性没有显着变化。
    Background and Objectives: Wound healing encompasses a multitude of factors and entails the establishment of interactions among components of the basement membrane. The quantification of particle concentrations can serve as valuable biomarkers for assessing biomechanical muscle properties. The objective of this study was to examine the immunoexpression and immunoconcentration of myometrial collagen type VI, elastin, alpha-smooth muscle actin, and smooth muscle myosin heavy chain, as well as the expression of platelets and clusters of differentiation 31 in the uterine scar following a cesarean section (CS). Materials and Methods: A total of 177 biopsies were procured from a cohort of pregnant women who were healthy, specifically during the surgical procedure of CS. The participants were categorized into seven distinct groups. Group 1 consisted of primiparas, with a total of 52 individuals. The subsequent groups were organized based on the duration of time that had elapsed since their previous CS. The analysis focused on the immunoexpression and immunoconcentration of the particles listed. Results: No significant variations were observed in the myometrial immunoconcentration of collagen type VI, elastin, smooth muscle myosin, and endothelial cell cluster of differentiation 31 among the analyzed groups. The concentration of alpha-smooth muscle actin in the myometrium was found to be significantly higher in patients who underwent CS within a period of less than 2 years since their previous CS, compared to those with a longer interval between procedures. Conclusions: Our findings indicate that the immunoconcentration of uterine myometrial scar collagen type VI, elastin, smooth muscle myosin heavy chain, alpha-smooth muscle actin, and endothelial cell marker cluster of differentiation 31 remains consistent regardless of the duration elapsed since the previous CS. The findings indicate that there are no significant alterations in the biomechanical properties of the uterine muscle beyond a period of 13 months following a CS.
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    文章类型: Journal Article
    肌动蛋白成核因子主要有三类:Arp2/3复合物,Spire和Formin.尖顶通过使稳定的纵向四聚体成核并将肌动蛋白结合到微丝的生长端来组装微丝。早在1999年,惠灵顿等人。确定Spire是一种肌动蛋白成核剂,然而,多年来,大多数研究都集中在Arp2/3和Formin蛋白上;关于Spire作为肌动蛋白成核因子的成员的研究相对较少。最近的研究表明,Spire通过肌动蛋白的合成参与囊泡运输,在神经发育中起重要作用。在本文中,我们回顾了结构,Spire的表达和功能,及其与疾病的关联,以便为Spire的研究确定有意义的潜在方向。
    There are three main classes of actin nucleation factors: Arp2/3 complexes, Spire and Formin. Spire assembles microfilaments by nucleating stable longitudinal tetramers and binding actin to the growing end of the microfilament. As early as 1999, Wellington et al. identified Spire as an actin nucleating agent, however, over the years, most studies have focused on Arp2/3 and Formin proteins; there has been relatively less research on Spire as a member of the actin nucleating factors. Recent studies have shown that Spire is involved in the vesicular transport through the synthesis of actin and plays an important role in neural development. In this paper, we reviewed the structure, expression and function of Spire, and its association with disease in order to identify meaningful potential directions for studies on Spire.
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  • 文章类型: Journal Article
    哺乳动物具有6种高度保守的肌动蛋白亚型,具有非冗余的生物学功能。同工型特异性的分子基础,然而,由于缺乏工具,仍然难以捉摸。这里,我们描述了IntAct的发展,一种内部标记策略,用于研究固定细胞和活细胞中的肌动蛋白同工型。我们在β-肌动蛋白中鉴定了允许标签整合的残基对,并使用了敲入细胞系来证明IntActβ-肌动蛋白表达和丝掺入与野生型没有区别。此外,IntActβ-肌动蛋白仍然与常见的肌动蛋白结合蛋白(ABP)相关,并且可以在活细胞中靶向。通过显示肌动蛋白同工型特异性分布在人细胞中保持,我们证明了IntAct用于肌动蛋白同工型研究的可用性。最后,我们观察到标记的肌动蛋白变体在酵母肌动蛋白斑块中的变体依赖性掺入,电缆,和细胞动力学环证明了跨物种的适用性。一起,我们的数据表明,IntAct是研究肌动蛋白同工型定位的通用工具,动力学,和分子相互作用。
    Mammals have 6 highly conserved actin isoforms with nonredundant biological functions. The molecular basis of isoform specificity, however, remains elusive due to a lack of tools. Here, we describe the development of IntAct, an internal tagging strategy to study actin isoforms in fixed and living cells. We identified a residue pair in β-actin that permits tag integration and used knock-in cell lines to demonstrate that IntAct β-actin expression and filament incorporation is indistinguishable from wild type. Furthermore, IntAct β-actin remains associated with common actin-binding proteins (ABPs) and can be targeted in living cells. We demonstrate the usability of IntAct for actin isoform investigations by showing that actin isoform-specific distribution is maintained in human cells. Lastly, we observed a variant-dependent incorporation of tagged actin variants into yeast actin patches, cables, and cytokinetic rings demonstrating cross species applicability. Together, our data indicate that IntAct is a versatile tool to study actin isoform localization, dynamics, and molecular interactions.
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  • 文章类型: Journal Article
    目的探讨细胞外基质蛋白ABI3BP对水泡性口炎病毒(VSV)基因组复制及固有免疫信号通路的影响.方法用小干扰RNA(siRNA)转染人皮肤成纤维细胞BJ-5ta细胞中ABI3BP基因。建立VSV-绿色荧光蛋白(VSV-GFP)感染的细胞模型。采用phalloidin免疫荧光染色法检测ABI3BP敲低细胞的形态变化和F-肌动蛋白应激纤维形成。通过RT-qPCR检测VSV-GFP感染后BJ-5ta细胞中病毒复制的mRNA水平;进行蛋白质印迹检测干扰素调节因子3(IRF3)和TANK结合激酶1(TBK1)磷酸化水平的变化。结果成功建立了VSV-GFP感染的BJ-5ta细胞模型。实现了ABI3BP对BJ-5ta细胞的有效敲低。Phalloidin免疫荧光染色显示ABI3BP基因敲低后细胞内F-肌动蛋白的结构重排。与对照组相比,当感染复数0.1和1的VSV时,ABI3BP敲低细胞中VSV-GFP的基因拷贝数分别增加2.2-3.5倍(P<0.01)和2.2-4.0倍(P<0.01)。病毒感染后,ABI3BP敲低细胞中病毒蛋白的表达显着增加。I型干扰素途径的激活,如磷酸化IRF3和磷酸化TBK1所确定,在VSV-GFP感染后ABI3BP敲低细胞中显著降低。结论细胞外基质蛋白ABI3BP在维持肌动蛋白结构的形成和重排中起重要作用。ABI3BP基因缺失促进RNA病毒复制,ABI3BP是维持I型干扰素通路完整性的重要分子。
    Objective To explore the influence of extracellular matrix protein ABI-interactor 3-binding protein (ABI3BP) on vesicular stomatitis virus (VSV) genome replication and innate immune signaling pathway.Methods The small interfering RNA (siRNA) was transfected to knock down ABI3BP gene in human skin fibroblast BJ-5ta cells. VSV-green fluorescent protein (VSV-GFP)-infected cell model was established. The morphological changes and F-actin stress fiber formation were detected on ABI3BP knockdown cells by phalloidin immunofluorescence staining. The mRNA level of virus replication was detected by RT-qPCR in BJ-5ta cells after VSV-GFP infection; western blotting was performed to detect the changes in interferon regulatory factor 3 (IRF3) and TANK-binding kinase 1 (TBK1) phosphorylation levels.Results The VSV-GFP-infected BJ-5ta cell model was successfully established. Efficient knockdown of ABI3BP in BJ-5ta cells was achieved. Phalloidin immunofluorescence staining revealed structural rearrangement of intracellular F-actin after ABI3BP gene knockdown. Compared with the control group, the gene copy number of VSV-GFP in ABI3BP knockdown cells increased by 2.2 - 3.5 times (P<0.01) and 2.2 - 4.0 times (P<0.01) respectively when infected with VSV of multiplicity of infection 0.1 and 1. The expression of viral protein significantly increased in ABI3BP knockdown cells after virus infection. The activation of type-I interferon pathway, as determined by phosphorylated IRF3 and phosphorylated TBK1, was significantly decreased in ABI3BP knockdown cells after VSV-GFP infection.Conclusions Extracellular matrix protein ABI3BP plays an important role in maintaining the formation and rearrangement of actin structure. ABI3BP gene deletion promotes RNA virus replication, and ABI3BP is an important molecule that maintains the integrity of type I interferon pathway.
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  • 文章类型: Journal Article
    骨重建过程中过度的骨吸收破骨细胞活性是骨疾病的主要特征。比如骨质疏松症。因此,抑制破骨细胞形成和骨吸收可以成为各种骨疾病的有效治疗靶点。Gryllusbiomaculatus(GB)由于其高营养价值和环境可持续性,最近被批准为替代食品来源。传统上,已知GB具有各种药理特性,包括解热和降血压活性,最近有报道称它具有各种生物活性,包括对炎症的保护作用,氧化应激,胰岛素抵抗,和酒精引起的肝损伤。然而,GB对破骨细胞分化和骨代谢的影响尚未得到证实。在这项研究中,我们证实了GB提取物(GBE)对核因子κB受体活化因子配体(RANKL)诱导的破骨细胞形成的抑制作用。探讨GBE对RANKL诱导的破骨细胞分化及功能的影响,我们进行了TRAP和F-肌动蛋白染色,以及骨吸收试验。通过Western印迹分析和定量实时聚合酶链反应揭示了负责破骨细胞生成调节的GBE的细胞内机制。我们研究了GBE与破骨细胞特异性分子表达之间的关系,以进一步阐明潜在的机制。发现GBE通过降低Akt的磷酸化显著抑制破骨细胞生成,p38,JNK,和ERK,以及Btk-PLCγ2信号,在参与早期破骨细胞生成的途径中以及通过随后的c-Fos抑制,NFATc1和破骨细胞生成特异性标记基因。此外,GBE抑制F-肌动蛋白环阳性破骨细胞的形成和成熟破骨细胞的骨吸收活性。我们的发现表明,GBE是涉及破骨细胞的骨骼疾病的潜在功能性食物和治疗候选药物。
    Excessive bone-resorbing osteoclast activity during bone remodeling is a major feature of bone diseases, such as osteoporosis. Therefore, the inhibition of osteoclast formation and bone resorption can be an effective therapeutic target for various bone diseases. Gryllus biomaculatus (GB) has recently been approved as an alternative food source because of its high nutritional value and environmental sustainability. Traditionally, GB has been known to have various pharmacological properties, including antipyretic and blood pressure-lowering activity, and it has recently been reported to have various biological activities, including protective effects against inflammation, oxidative stress, insulin resistance, and alcohol-induced liver injury. However, the effect of GB on osteoclast differentiation and bone metabolism has not yet been demonstrated. In this study, we confirmed the inhibitory effect of GB extract (GBE) on the receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast formation. To determine the effect of GBE on RANKL-induced osteoclast differentiation and function, we performed TRAP and F-actin staining, as well as a bone-resorbing assay. The intracellular mechanisms of GBE responsible for the regulation of osteoclastogenesis were revealed by Western blot analysis and quantitative real-time polymerase chain reaction. We investigated the relationship between GBE and expression of osteoclast-specific molecules to further elucidate the underlying mechanisms. It was found that GBE significantly suppressed osteoclastogenesis by decreasing the phosphorylation of Akt, p38, JNK, and ERK, as well as Btk-PLCγ2 signaling, in pathways involved in early osteoclastogenesis as well as through the subsequent suppression of c-Fos, NFATc1, and osteoclastogenesis-specific marker genes. Additionally, GBE inhibited the formation of F-actin ring-positive osteoclasts and bone resorption activity of mature osteoclasts. Our findings suggest that GBE is a potential functional food and therapeutic candidate for bone diseases involving osteoclasts.
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  • 文章类型: Journal Article
    壁虎脚趾垫表面的分层设计及其可逆粘附性多年来一直启发材料科学家。已经开发出具有令人印象深刻的粘合性能的微米和纳米图案表面来模仿壁虎的性能。虽然在一些示例中实现的粘合性能已经超过了生活同行,制造表面的耐久性是有限的,自我更新和恢复生物系统固有功能的能力是不可想象的。在这里,使用Bibron壁虎(Chondrodactylusbibronii)的皮肤样本研究了壁虎刚毛的形态发生。壁虎刚毛在皮肤表皮内不同的细胞-细胞层界面处发育为专门的顶端分化结构。作为模板结构元件的F-肌动蛋白和微管的主要作用对于刚毛的分层形态的发展是必要的。并确定了角蛋白和角膜β蛋白的稳定作用。落叶从底层的单个细胞生长,突出到上层的四个相邻细胞中。通过促进细胞-细胞界面的断裂和高纵横比刚毛的释放,所得的多细胞连接可以在脱落过程中发挥作用。结果有助于理解刚毛再生,并可能激发未来的概念,以生物工程自我再生图案化的粘合剂表面。
    The hierarchical design of the toe pad surface in geckos and its reversible adhesiveness have inspired material scientists for many years. Micro- and nano-patterned surfaces with impressive adhesive performance have been developed to mimic gecko\'s properties. While the adhesive performance achieved in some examples has surpassed living counterparts, the durability of the fabricated surfaces is limited and the capability to self-renew and restore function-inherent to biological systems-is unimaginable. Here the morphogenesis of gecko setae using skin samples from the Bibron´s gecko (Chondrodactylus bibronii) is studied. Gecko setae develop as specialized apical differentiation structures at a distinct cell-cell layer interface within the skin epidermis. A primary role for F-actin and microtubules as templating structural elements is necessary for the development of setae\'s hierarchical morphology, and a stabilization role of keratins and corneus beta proteins is identified. Setae grow from single cells in a bottom layer protruding into four neighboring cells in the upper layer. The resulting multicellular junction can play a role during shedding by facilitating fracture of the cell-cell interface and release of the high aspect ratio setae. The results contribute to the understanding of setae regeneration and may inspire future concepts to bioengineer self-renewable patterned adhesive surfaces.
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  • 文章类型: Journal Article
    基底细胞癌(BCC)和鳞状细胞癌(SCC)是非黑色素瘤皮肤肿瘤的两种最常见类型。有时会遇到临床或组织病理学诊断挑战。CD56和平滑肌肌动蛋白(SMA)在BCC中高度表达。我们的目的是调查这些标记的频率,B细胞淋巴瘤2(Bcl-2)和Ki67。进行这项研究是为了提出一种可能具有诊断价值的方法。本研究包括128例BCC和39例SCC病例。CD56、SMA、应用Bcl-2和Ki67免疫组织化学染色。99例(77.3%)BCC和6例(15.4%)SCC显示CD56免疫反应性。在78.1%的BCC和5.1%的SCC病例中检测到SMA表达。CD56、SMA、Bcl-2表达在BCC病例中明显增高。在SCC病例中发现Ki67增殖指数明显更高。当基底鳞癌病例与SCC病例进行比较时,肿瘤与CD56、SMA、获得Bcl-2。CD56和SMA,除了Bcl-2,更喜欢BCC。Ki67也应包括在该组中以证明增殖活性。
    UNASSIGNED: Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are the 2 most common types of nonmelanoma skin tumors. Clinical or histopathological diagnostic challenges are encountered on occasion. CD56 and smooth muscle actin (SMA) are highly expressed in BCCs. We aimed to investigate the frequency of these markers, along with B-cell lymphoma 2 (Bcl-2) and Ki67. This study was conducted to propose a method that could possibly be of diagnostic value. One hundred twenty-eight BCC and 39 SCC cases were included in this study. CD56, SMA, Bcl-2, and Ki67 immunohistochemical stains were applied. Ninety-nine (77.3%) BCC and 6 (15.4%) SCC cases showed CD56 immunoreactivity. SMA expression was detected in 78.1% of BCC and 5.1% of SCC cases. CD56, SMA, and Bcl-2 expressions were significantly higher in BCC cases. The Ki67 proliferation index was found significantly higher in SCC cases. When basosquamous carcinoma cases were compared with SCC cases, a significant correlation between tumors and expression of CD56, SMA, and Bcl-2 were obtained. CD56 and SMA, in addition to Bcl-2, favor BCC. Ki67 should also be included in the panel to demonstrate the proliferative activity.
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  • 文章类型: Comparative Study
    背景:因子VIII(FVIII)产物的结构和化学修饰可影响其在FVIII活性测定中的行为。因此,评估FVIII产品在这些测定中的性能是重要的。Efanesoccogalfa是一类新的FVIII替代疗法,旨在提供高持续因子活性水平和延长的血浆半衰期。
    目的:评估在一阶段凝血测定(OSA)和显色底物测定(CSA)中测量efanesoctocogalfaFVIII活性的准确性。
    方法:将未检测到FVIII活性的人血浆掺入fanesoctocogalfa或全长重组FVIII产品比较物,octocogalfa,在标称浓度为0.80IU/mL时,0.20IU/mL,或0.05IU/mL,基于标记的效力。临床止血实验室(N=35)使用内部测定测试了盲化样品。分析了来自51种OSA(14种活化的部分凝血活酶时间[aPTT]试剂)和42种CSA(8种试剂盒)的数据。
    结果:使用具有肌动蛋白FSL和多种其他aPTT试剂的OSA在所有浓度下可靠地测量了Efanesoccogalfa活性(标称活性的±25%)。一些试剂发生FVIII活性的低估和高估。对于任何种类的aPTT激活剂都没有观察到具体的趋势。使用CSA和OSA以肌动蛋白FS作为aPTT试剂,在评估的浓度上始终观察到2至3倍的高估。
    结论:某些特定的OSA和大多数CSA发生低估或高估,这在以前的其他改性FVIII替代产品中已经观察到。使用几种OSA方法和商业血浆标准以可接受的准确性和可靠性测量了FVIII活性。
    BACKGROUND: Structural and chemical modifications of factor VIII (FVIII) products may influence their behaviour in FVIII activity assays. Hence, it is important to assess the performance of FVIII products in these assays. Efanesoctocog alfa is a new class of FVIII replacement therapy designed to provide both high sustained factor activity levels and prolonged plasma half-life.
    OBJECTIVE: Evaluate the accuracy of measuring efanesoctocog alfa FVIII activity in one-stage clotting assays (OSAs) and chromogenic substrate assays (CSAs).
    METHODS: Human plasma with no detectable FVIII activity was spiked with efanesoctocog alfa or a full-length recombinant FVIII product comparator, octocog alfa, at nominal concentrations of 0.80 IU/mL, 0.20 IU/mL, or 0.05 IU/mL, based on labelled potency. Clinical haemostasis laboratories (N = 35) tested blinded samples using in-house assays. Data from 51 OSAs (14 activated partial thromboplastin time [aPTT] reagents) and 42 CSAs (eight kits) were analyzed.
    RESULTS: Efanesoctocog alfa activity was reliably (±25% of nominal activity) measured across all concentrations using OSAs with Actin FSL and multiple other aPTT reagents. Under- and overestimation of FVIII activity occurred with some reagents. No specific trend was observed for any class of aPTT activators. A two- to three-fold overestimation was consistently observed using CSAs and the OSA with Actin FS as the aPTT reagent across evaluated concentrations.
    CONCLUSIONS: Under- or overestimation occurred with some specific OSAs and most CSAs, which has been previously observed with other modified FVIII replacement products. Efanesoctocog alfa FVIII activity was measured with acceptable accuracy and reliability using several OSA methods and commercial plasma standards.
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