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Analysis of CA125 antigen in normal human seminal plasma highlights the molecular heterogeneity of underlying glycosylated species 正常人精浆中CA125抗原的分析突出了潜在糖基化物种的分子异质性
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-06-01 DOI: 10.3906/BIY-1610-32
N. Mitić, B. Milutinovic, M. Janković
Cancer antigen CA125 represents an extracellular part of transmembrane mucin MUC16 and may be expressed in tissues of the male reproductive tract. It reaches human seminal plasma (hSP) after undergoing the main processes involved in protein speciation: synthesis, posttranslational modifications, compartmentalization, and auto/proteolytic degradation. This study was aimed at profiling CA125-immunoreactive species in hSP from healthy subjects as a specific and unexplored source of this tumor-associated antigen expressed under normal physiological conditions. High molecular mass components from total hSP and corresponding acid-soluble hSP preparations were analyzed. The results indicated that antibodies recognizing two distinct immunogenic areas of CA125 antigen exhibited a common pattern of immunoreactive bands affected by low pH and reducing agents. They comprise a large, heavily glycosylated moiety and a mixture of low glycosylated species ranging from 100 to 150 kDa. High molecular mass CA125-immunoreactive smears overlapped core 2 O-glycans and the Lex glycotope, the latter also being abundant on distinct lower molecular mass immunoreactive bands. Within the bulk of normal hSP mucins, the CA125 antigen is present in low amounts and resides on heterogeneously glycosylated species that may be proteolysis-derived and sensitive to redox environments. Both factors influence the composition of hSP and therefore might affect speciation of mucin16 under normal and pathological conditions.
癌症抗原CA125代表跨膜粘蛋白MUC16的细胞外部分,并且可以在男性生殖道的组织中表达。它在经历蛋白质形态形成的主要过程后到达人类精浆(hSP):合成、翻译后修饰、区室化和自身/蛋白水解降解。本研究旨在分析健康受试者hSP中CA125免疫反应物质,作为在正常生理条件下表达的这种肿瘤相关抗原的特异性和未探索的来源。分析了总hSP和相应的酸溶性hSP制剂的高分子量组分。结果表明,识别CA125抗原两个不同免疫原性区域的抗体表现出受低pH和还原剂影响的免疫反应带的共同模式。它们包含大的、高度糖基化的部分和范围为100-150kDa的低糖基化物质的混合物。高分子量CA125免疫反应涂片与核心2 O-聚糖和Lex糖蛋白重叠,后者也在不同的低分子量免疫反应带上丰富。在大部分正常hSP粘蛋白中,CA125抗原以低量存在,并存在于可能由蛋白水解衍生并对氧化还原环境敏感的非均质糖基化物种上。这两个因素都会影响hSP的组成,因此可能会影响正常和病理条件下粘蛋白16的形态形成。
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引用次数: 0
Changes in apoptosis-related gene expression profiles in cancer cell lines exposed to usnic acid lichen secondary metabolite 枸橼酸地衣次生代谢物对癌细胞凋亡相关基因表达谱的影响
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-06-01 DOI: 10.3906/BIY-1609-40
A. B. Dinçsoy, Demet Duman
The presence of uninhibited side effects of cancer drugs often used in cancer treatment has stimulated the search for alternative therapeutic approaches. Therefore, anticarcinogenic effects of synthetic, herbal, and fungal drugs have been investigated for the treatment of various cancer types in recent studies. Lichens, symbiotic organisms of fungi and algae, synthesize metabolites with significant biological activities. The aim of the current study was to screen the anticancer potential of usnic acid on various types of nonmalignant cell lines (Vero, L929) and cancer cell lines (CaCo2, RD, Hep2C, HepG2, Wehi). The growth inhibitory effect of usnic acid was determined by MTT assay. Since this study was also designed to explore mRNA expression profiles, this paper is the first to look into the effects of usnic acid on apoptotic gene expression. The effects of usnic acid on the gene expression patterns of the tumor suppressor gene p53, proapoptotic gene Bcl-2, and Bax were studied with qRT-PCR. There was an approximately ninefold decrease in the p53 and Bcl-2 expression for usnic acid in the Wehi cancer cell line. Consequently, it is concluded that usnic acid has tumor inhibitory properties, and if indicated by further works like animal studies and clinical trials, it may be used therapeutically in the future.
经常用于癌症治疗的癌症药物的无抑制副作用的存在刺激了对替代治疗方法的探索。因此,在最近的研究中,合成药物、草药和真菌药物的抗癌作用已被研究用于治疗各种癌症类型。地衣是真菌和藻类的共生生物,合成具有重要生物活性的代谢产物。本研究的目的是筛选usnic酸对各种类型的非恶性细胞系(Vero,L929)和癌症细胞系(CaCo2,RD,Hep2C,HepG2,Wehi)的抗癌潜力。MTT法测定乌苏酸的生长抑制作用。由于这项研究也是为了探索信使核糖核酸的表达谱,因此本文首次研究了usnic酸对凋亡基因表达的影响。用qRT-PCR方法研究了usnic酸对抑癌基因p53、促凋亡基因Bcl-2和Bax基因表达模式的影响。在Wehi癌症细胞系中,usnic酸的p53和Bcl-2表达降低了约9倍。因此,可以得出结论,usnic酸具有肿瘤抑制特性,如果动物研究和临床试验等进一步工作表明,它可能在未来用于治疗。
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引用次数: 27
Histomorphometric evaluation of the testicular parenchyma of rats submittedto protein restriction during intrauterine and postnatal life 宫内和产后蛋白质限制大鼠睾丸实质的组织形态学评价
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-06-01 DOI: 10.3906/BIY-1610-20
J. M. D. Oliveira, Alluanan Adelson do Nascimento Silva, S. L. Souza, R. N. Morais, Elizabeth Neves de Melo, F. Maia, V. A. Júnior
The critical period of development is highly susceptible to disorders. Environmental contaminants, stress, and poor nutrition may permanently affect structurally and functionally an organism during adulthood. Protein restriction in intrauterine and neonatal periods may impair testicular cells and reduce steroidogenic activity. The current study investigated the effect of low protein diet during intrauterine and postnatal life on testicular function in immature and adult rats. Pregnant Wistar rats were fed either a normal protein diet or a low protein diet during pregnancy until birth and during lactation until weaning. The results showed that undernutrition during intrauterine life reduced epithelium height, tubular volume, Leydig cell volume, and serum testosterone levels, although it increased the support capacity of Sertoli cells. Tubular diameter, tubular volume, epithelium height, and Leydig cell population decreased when protein was restricted during lactation. Pregnant rats submitted to a low protein diet during the two periods generated male rats with a reduction in seminiferous tubule volume and length, Sertoli cell population, Leydig cell population and volume, and sperm production. Undernutrition during both the intrauterine and postnatal periods caused the most drastic testicular effects, as irreversible damage to Sertoli cell population and, consequently, to sperm production in the adult male.
发育的关键时期极易受到疾病的影响。环境污染物、压力和营养不良可能会在成年期对生物体的结构和功能产生永久性影响。宫内和新生儿期的蛋白质限制可能会损害睾丸细胞并降低类固醇生成活性。目前的研究调查了低蛋白饮食在宫内和产后对未成熟和成年大鼠睾丸功能的影响。怀孕的Wistar大鼠在怀孕至分娩期间和哺乳至断奶期间被喂食正常蛋白质饮食或低蛋白质饮食。结果表明,宫内营养不良降低了上皮高度、小管体积、Leydig细胞体积和血清睾酮水平,尽管它增加了支持细胞的支持能力。当蛋白质在哺乳期受到限制时,小管直径、小管体积、上皮高度和Leydig细胞数量减少。在这两个时期接受低蛋白饮食的怀孕大鼠产生的雄性大鼠生精小管体积和长度、支持细胞群、Leydig细胞群和体积以及精子产量都有所减少。宫内和产后营养不足对睾丸的影响最为严重,对支持细胞群造成不可逆转的损害,从而影响成年男性的精子生产。
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引用次数: 4
Induction of haploid plants in citrus through gamma-irradiated pollen and ascertainment of ovule age for maximum recovery of haploid plantlets 利用γ辐照花粉诱导柑橘单倍体植株及确定胚珠龄以最大限度恢复单倍体植株
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-06-01 DOI: 10.3906/BIY-1606-28
M. Kundu, A. Dubey, M. Srivastav, S. Malik
* Correspondence: manojhorti18@gmail.com
*通信:manojhorti18@gmail.com
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引用次数: 8
Morphology, development, and transplant potential of Prunus avium and Cornus sanguinea seedlings growing under different LED lights 不同LED灯下桃李和血茱萸幼苗的形态、发育和移植潜力
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-04-20 DOI: 10.3906/BIY-1607-19
F. Bantis, K. Radoglou
* Correspondence: kradoglo@fmenr.duth.gr
*通信:kradoglo@fmenr.duth.gr
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引用次数: 12
Improvement in androgenic response of borage (Borago officinalis L.) cultured anthers using antibrowning agents and picloram 抗褐变剂和picloram对琉璃苣培养花药雄激素反应的改善
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-04-20 DOI: 10.3906/BIY-1606-49
M. Abdollahi, Zahra Chardoli Eshaghi, M. Majdi
* Correspondence: m.abdollahi@basu.ac.ir
*通信:m.abdollahi@basu.ac.ir
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引用次数: 5
Optimization of monoclonal antibody expression in CHOcells by employing epigenetic gene regulation tools 应用表观遗传学基因调控工具优化CHO细胞单克隆抗体表达
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-04-02 DOI: 10.3906/biy-1702-18
Fatemeh Nematpour, F. Mahboudi, V. Khalaj, B. Vaziri, S. Ahmadi, M. Ahmadi, Saedeh Ebadat, Fatemeh Davami
Monoclonal antibodies (mAbs) are crucial in pharmaceutical biotechnology. Mammalian cell lines are the most preferred for their production. One of the significant challenging issues of mammalian expression systems is epigenetic gene silencing. Employing epigenetic gene regulation tools can increase the productivity of the mammalian cell lines. Sodium butyrate (NaBut) and valproic acid (VPA) regulate gene expression by inhibiting histone deacetylase. A ubiquitous chromatin-opening element (UCOE) can improve expression by reducing DNA methylation. Here, the separate and combined effects of NaBut and VPA histone deacetylase inhibitors (iHDACs) and UCOE on mAb synthesis were studied. Stable cell lines were generated by non-UCOE (CHO-HL) and UCOE-containing vectors (CHO-UHUL) and cultured in the presence and absence of NaBut or VPA. Expression analysis showed that CHO-UHUL gave a 4-fold greater yield than non-UCOE CHO-HL. Antibody production levels of the CHO-HL and CHO-UHUL cells increased 2-fold and 2.5-fold after NaBut and VPA treatment, respectively. These results indicate that UCOE has more impact on antibody expression than iHDACs. iHDAC treatment exhibited at least a 0.5-fold higher antibody yield in UCOE containing CHO-UHUL cells. Thus utilization of NaBut and VPA (iHDACs) and UCOE resulted in antibody expression improvement, and the combined use of them had a synergistic effect on antibody synthesis. Keywords: Monoclonal antibodies, Chinese hamster ovary, epigenetic gene silencing, chromatin-opening element, sodium butyrate, valproic acid
单克隆抗体(mab)在制药生物技术中至关重要。哺乳动物细胞系是它们生产的首选材料。表观遗传基因沉默是哺乳动物表达系统的重要挑战之一。利用表观遗传基因调控工具可以提高哺乳动物细胞系的生产力。丁酸钠(NaBut)和丙戊酸(VPA)通过抑制组蛋白去乙酰化酶调节基因表达。普遍存在的染色质打开元件(UCOE)可以通过减少DNA甲基化来改善表达。本研究研究了NaBut和VPA组蛋白去乙酰化酶抑制剂(ihdac)和UCOE对单克隆抗体合成的单独和联合作用。用不含ucoe (CHO-HL)和含ucoe (CHO-UHUL)的载体生成稳定的细胞系,并在NaBut或VPA存在和不存在的情况下培养。表达分析表明,CHO-UHUL的产量比非ucoe的CHO-HL高4倍。NaBut和VPA处理后,CHO-HL和CHO-UHUL细胞的抗体产生水平分别增加了2倍和2.5倍。这些结果表明UCOE比iHDACs对抗体表达的影响更大。在含有CHO-UHUL细胞的UCOE中,iHDAC处理的抗体产量至少提高了0.5倍。因此,利用NaBut和VPA (ihdac)和UCOE可提高抗体表达,两者联合使用对抗体合成具有协同作用。关键词:单克隆抗体,仓鼠卵巢,表观遗传基因沉默,染色质打开元件,丁酸钠,丙戊酸
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引用次数: 3
Cytotoxic and apoptotic effects of endemic Centaurea fenzlii Reichardt on the MCF-7 breast cancer cell line 地方性半毛茛对MCF-7乳腺癌细胞系的细胞毒性和凋亡作用
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-04-01 DOI: 10.3906/biy-1609-74
Ümit Yırtıcı, Fatih Göger, Mehmet Sarimahmut, A. Ergene
The main purpose of this study was to analyze the cytotoxic activity of an extract obtained from Centaurea fenzlii Reichardt, and the fractions eluted from this extract, in breast cancer cells. After isolation and structural analysis of the fractions were conducted, a meaningful cytotoxic effect was indicated. The goal of the analysis was to reveal the mechanism by which this effect occurs through researching the apoptotic side of these fractions and determining the amount of several proteins that are the products of the genes. Test substances were applied to breast cancer cells and the inhibitory concentration value 50 (IC50) that caused a cytotoxic effect was determined using MTT and ATP assays. The Centaurea fenzlii Reichardt dichloromethane extracts-ethyl acetate fractions (CFDCMEAF) exhibited a stronger growth-inhibitory effect on MCF-7 cells (45.771 μg/mL). The apoptotic effect was studied using double staining and flow cytometry. The death rate in the cells treated with the CFDCM-EAF IC50 dose was approximately 90%: 9.2% living cells, 22.8% necrotic cells, 62.3% late apoptotic cells, and 5.8% early apoptotic cells. Structural analysis of the CFDCM-EAF, which indicated significant cytotoxic effects, was performed using chromatographic methods. Hispidulin was the major component of the CFDCM-EAF by LC-APCI-MS/MS analysis.
本研究的主要目的是分析半人马提取物的细胞毒性活性,以及从该提取物中洗脱的部分,在乳腺癌细胞中的作用。经分离和结构分析,表明其具有明显的细胞毒作用。分析的目的是通过研究这些部分的凋亡方面和确定作为基因产物的几种蛋白质的数量来揭示这种作用发生的机制。将试验物质作用于乳腺癌细胞,采用MTT和ATP测定引起细胞毒性作用的抑制浓度50 (IC50)。半人马二氯甲烷提取物-乙酸乙酯组分(CFDCMEAF)对MCF-7细胞有较强的生长抑制作用(45.771 μg/mL)。采用双染色和流式细胞术观察其对细胞凋亡的影响。CFDCM-EAF IC50剂量处理的细胞死亡率约为90%,其中活细胞9.2%,坏死细胞22.8%,晚期凋亡细胞62.3%,早期凋亡细胞5.8%。采用色谱法对CFDCM-EAF进行结构分析,结果表明CFDCM-EAF具有明显的细胞毒作用。LC-APCI-MS/MS分析结果表明,CFDCM-EAF的主要成分为Hispidulin。
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引用次数: 11
Neuropeptide Y reduces migration capacity of human choriocarcinoma cell line by altering oxidative/antioxidative status 神经肽Y通过改变氧化/抗氧化状态来降低人绒毛膜癌细胞系的迁移能力
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-04-01 DOI: 10.3906/biy-1606-73
M. Matić, Milica G. Paunović, B. Ognjanović, Stajn As, Z. Saicic
Reduced migration capacity of trophoblast cells leads to poor placentation and correlates with severe pregnancy disorders such as intrauterine growth restriction and preeclampsia. Neuropeptide Y (NPY) is sympathetic cotransmitter involved in various physiological processes and its levels are significantly increased in preeclamptic pregnancy compared to healthy pregnancy. In this study the prooxidative role of NPY and its effects on migration capacity of human trophoblast cell line JEG-3 were investigated together with the effects of nitric oxide (NO) depletion, a molecule that was shown to play an important role in promoting cell migration. The cells were treated for 24 h (short-term stimulation) and 72 h (long-term stimulation) respectively with 1 nM NPY. Oxidative/antioxidative status and the migration index of cells were measured. The results showed increased concentrations of oxidative stress parameters (O2.-, H2O2) and molecules of the antioxidant defense system (reduced glutathione and oxidized glutathione), while the levels of intracellular nitrites (indicators of NO) and cell migration index were significantly decreased in trophoblast cells treated with NPY (both at 24 h and 72 h of exposure) compared to the control cells. These results suggest that NPY may significantly contribute to reduced migration capacity of trophoblast cells by generating oxidative stress and reducing the bioavailability of NO.
滋养细胞迁移能力降低导致胎盘发育不良,并与宫内生长受限和先兆子痫等严重妊娠疾病相关。神经肽Y (NPY)是参与多种生理过程的交感共递质,与正常妊娠相比,子痫前期妊娠的神经肽Y水平显著升高。本研究研究了NPY的促氧化作用及其对人滋养细胞JEG-3迁移能力的影响,并研究了一氧化氮(NO)耗竭的影响,一氧化氮在促进细胞迁移中起重要作用。1 nM NPY分别作用24 h(短期刺激)和72 h(长期刺激)。测定细胞的氧化/抗氧化状态和迁移指数。结果表明,氧化应激参数(O2;-, H2O2)和抗氧化防御系统分子(还原性谷胱甘肽和氧化性谷胱甘肽),而细胞内亚硝酸盐(NO指标)水平和细胞迁移指数在NPY处理的滋养细胞(暴露24 h和72 h)与对照细胞相比显著降低。这些结果表明,NPY可能通过产生氧化应激和降低NO的生物利用度而显著降低滋养细胞的迁移能力。
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引用次数: 1
Biocompatible polymeric coatings do not inherently reduce the cytotoxicity of iron oxide nanoparticles 生物相容性聚合物涂层本身并不会降低氧化铁纳米颗粒的细胞毒性
IF 2.2 4区 生物学 Q3 BIOLOGY Pub Date : 2017-04-01 DOI: 10.3906/BIY-1608-61
A. Ş. Ürkmez, Ece Bayir, E. Bilgi, M. Ozen
Nanotechnology in biomedical research is an emerging and promising tool for different purposes, like high-resolution medical imaging, diagnostics, and targeted drug delivery. Although some experimental efforts focused on determination of the cytotoxicity of nanoparticles (NPs) are present, there are many controversial results. In this study, chitosan (CS)- and poly(acrylic acid) (PAA)-coated iron oxide nanoparticles (IONPs) were used to investigate the possible cytotoxicity of coated IONPs on model mammalian cell line SaOs-2 by evaluating cellular viability and membrane integrity. Increasing concentrations of IONPs increased the cytotoxic effect on SaOs-2 cells with both CS- and PAA-coated IONPs. Cell viability on day 3 was as low as 40% and 48% at 1000 ?M concentration for PAAand CS-coated IONPs, respectively, in 1% FBS-supplemented media. Cytotoxicity determined by the released lactate dehydrogenase (LDH) was as high as 163% with 1000 ?M concentration of CS-IONPs, while there was no significant change in LDH release with PAAcoated IONPs at any concentration. This study reveals that IONPs coated with a biocompatible polymer, which are usually assumed to be nontoxic, show cytotoxicity with increasing concentration and incubation time. The cytotoxicity of nanoparticles intended for biomedical purposes must be evaluated using more than one approach.
生物医学研究中的纳米技术是一种新兴的、有前途的工具,可用于不同的目的,如高分辨率医学成像、诊断和靶向药物输送。虽然目前已经有一些实验致力于测定纳米颗粒(NPs)的细胞毒性,但仍有许多有争议的结果。本研究利用壳聚糖(CS)-和聚丙烯酸(PAA)-包被氧化铁纳米粒子(IONPs),通过评估细胞活力和膜完整性来研究包被的IONPs对哺乳动物模型细胞系SaOs-2可能的细胞毒性。增加的IONPs浓度增加了对CS和paa包被的SaOs-2细胞的细胞毒性作用。在添加1% fbs的培养基中,paa和cs包被的IONPs在1000 μ M浓度下,第3天的细胞存活率分别低至40%和48%。CS-IONPs浓度为1000 μ M时,乳酸脱氢酶(LDH)释放量高达163%,而PAAcoated IONPs在任何浓度下,LDH释放量均无显著变化。本研究表明,通常被认为是无毒的生物相容性聚合物包被的IONPs随着浓度和孵育时间的增加而显示出细胞毒性。用于生物医学目的的纳米颗粒的细胞毒性必须使用多种方法进行评估。
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引用次数: 4
期刊
Turkish Journal of Biology
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