fish

  • 文章类型: Journal Article
    最近,关于蚂蚁染色体中核糖体基因进化模式的假设一直在讨论中。这些假设之一提出了染色体位置和rDNA位点数量之间的关系,表明末端位置通过减数分裂期间的异位重组促进rDNA簇的分散,而染色体内位置将它们限制为单个染色体对。另一个假设表明,由于染色体分裂,rDNA位点的增殖可能与膜翅目中染色体数量的增加有关。在这项研究中,我们在15个新的蚂蚁物种中对rDNA位点进行了物理映射,并回顾了自Teixeira等人修订以来可用的rDNA数据。(2021a)。我们的目标是调查新数据是否证实了染色体位置和rDNA位点数量之间的关系,以及染色体数目的增加是否显著影响蚂蚁核型rDNA簇的分散。将我们的新数据与2021年后发布的有关蚂蚁细胞遗传学的所有信息相结合,组装了40个新物种和9个新属。大多数物种在单个染色体对上表现出染色体内rDNA位点,而三个物种在多个染色体对的末端区域显示了这些基因。一方面,假设rDNA簇的染色体位置可能促进rDNA位点在蚂蚁基因组中的分散,如前所述,加强了,但是,另一方面,染色体裂变是核糖体基因在蚂蚁体内分散的主要机制的假说可能会被驳斥。此外,在某些属中,rDNA位点的位置在所研究的物种中保持相似,而在其他人中,这些基因的分布显示出物种之间的显著差异,表明染色体进化更加动态.
    Recently, hypotheses regarding the evolutionary patterns of ribosomal genes in ant chromosomes have been under discussion. One of these hypotheses proposes a relationship between chromosomal location and the number of rDNA sites, suggesting that terminal locations facilitate the dispersion of rDNA clusters through ectopic recombination during meiosis, while intrachromosomal locations restrict them to a single chromosome pair. Another hypothesis suggests that the multiplication of rDNA sites could be associated with an increase in the chromosome number in Hymenoptera due to chromosomal fissions. In this study, we physically mapped rDNA sites in 15 new ant species and also reviewed data on rDNA available since the revision by Teixeira et al. (2021a). Our objectives were to investigate whether the new data confirm the relationship between chromosomal location and the number of rDNA sites, and whether the increase in the chromosome number is significant in the dispersion of rDNA clusters in ant karyotypes. Combining our new data with all information on ant cytogenetics published after 2021, 40 new species and nine new genera were assembled. Most species exhibited intrachromosomal rDNA sites on a single chromosome pair, while three species showed these genes in terminal regions of multiple chromosome pairs. On one hand, the hypothesis that the chromosomal location of rDNA clusters may facilitate the dispersion of rDNA sites in the ant genome, as previously discussed, was strengthened, but, on the other hand, the hypothesis of chromosomal fission as the main mechanism for dispersion of ribosomal genes in ants is likely to be refuted. Furthermore, in certain genera, the location of rDNA sites remained similar among the species studied, whereas in others, the distribution of these genes showed significant variation between species, suggesting a more dynamic chromosomal evolution.
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  • 文章类型: Journal Article
    脊椎动物的体细胞生长主要受生长激素(GH)/胰岛素样生长因子I(IGF-I)轴的控制。表观遗传机制在鱼类中调节该轴的作用尚不清楚。这项工作旨在优化和评估使用短期培养的垂体和肝脏外植体从养殖鱼,金头seabreamSparusaurata,用于研究与GH/IGF-I轴调节有关的表观遗传机制。我们关于生存能力的结果,结构,扩散,和外植体的功能支持它们在短期测定中的使用。垂体外植体暴露于DNA甲基化抑制剂地西他滨(5-Aza-2'-脱氧胞苷;DAC)后,gh表达无变化,尽管通过改变dnmt3bb和tet1表达来响应DAC,和TET活动,产生整体DNA羟甲基化的增加。相反,在肝脏外植体中,DAC对dnmt和tet的表达或活性没有影响,而是改变了GH-IGF-I轴基因的表达。特别是,DAC和染料木素使igfbp2a的表达增加,而igfbp4,ghri和ghrii的表达减少,这表明增长受损。虽然肝脏外植体与S-腺苷甲硫氨酸(SAM)的孵育没有产生明显的影响,建议营养素必须确保鱼类肝脏内的甲基化环境以维持适当的生长,这需要进一步的体内验证。如本文所述,来自S.aurata的垂体和肝脏外植体可以进一步用于筛选表观遗传调节剂的抑制剂或激活剂,以及评估养殖鱼类GH-IGF-I变异背后的表观遗传机制。
    Somatic growth in vertebrates is mainly controlled by the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis. The role of epigenetic mechanisms in regulating this axis in fish is far from being understood. This work aimed to optimize and evaluate the use of short-term culture of pituitary and liver explants from a farmed fish, the gilthead seabream Sparus aurata, for studying epigenetic mechanisms involved in GH/IGF-I axis regulation. Our results on viability, structure, proliferation, and functionality of explants support their use in short-term assays. Pituitary explants showed no variation in gh expression after exposure to the DNA methylation inhibitor decitabine (5-Aza-2\'-deoxycytidine; DAC), despite responding to DAC by changing dnmt3bb and tet1 expression, and TET activity, producing an increase in overall DNA hydroxymethylation. Conversely, in liver explants, DAC had no effects on dnmt s and tet s expression or activity, but modified the expression of genes from the GH-IGF-I axis. In particular, the expression of igfbp2a was increased and that of igfbp4, ghri and ghrii was decreased by DAC as well as by genistein, which is suggestive of impaired growth. While incubation of liver explants with S-adenosylmethionine (SAM) produced no clear effects, it is proposed that nutrients must ensure the methylation milieu within the liver in the fish to sustain proper growth, which need further in vivo verification. Pituitary and liver explants from S. aurata can be further used as described herein for the screening of inhibitors or activators of epigenetic regulators, as well as for assessing epigenetic mechanisms behind GH-IGF-I variation in farmed fish.
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  • 文章类型: Journal Article
    人类表皮生长因子受体2(HER2)-低乳腺癌(BC)是BC的一种亚型,最近被认为是具有独特临床和分子特征的独立临床实体。它被定义为低水平的HER2蛋白表达,这将其与其他更具侵略性的BC亚型区分开来。早期研究表明,它可能比HER2阳性BC具有更有利的预后,因为它不太可能扩散到身体的其他部位,并且可能对标准的BC治疗(如化疗)更敏感,放射治疗,和激素治疗。鉴于HER2低BC的相对新出现,关于该亚型仍有很多需要了解;正在进行的研究集中在确定导致HER2低BC的潜在基因突变,以及开发可以改善该疾病患者预后的靶向治疗.这篇综述旨在总结目前关于HER2低BC的临床知识。目的是更好地了解这一实体,并为潜在的干预措施和新的护理标准铺平道路。
    Human epidermal growth factor receptor 2 (HER2)-low breast cancer (BC) is a subtype of BC that has been recently recognized as a separate clinical entity with distinct clinical and molecular characteristics. It is defined by a low level of HER2 protein expression, which distinguishes it from other more aggressive BC subtypes. Early studies suggest that it may have a more favorable prognosis than HER2-positive BC, as it is less likely to spread to other parts of the body and may be more responsive to standard BC treatments such as chemotherapy, radiation therapy, and hormone therapy. Given the relative new emergence of HER2-low BC, there is still much to be learned about this subtype; ongoing research is focused on identifying the underlying genetic mutations that contribute to HER2-low BC as well as developing targeted therapies that can improve outcomes for patients with this disease. This review is aimed at summarizing the current clinical knowledge on HER2-low BC, with the aim of creating a better understanding of this entity and paving the way for potential interventions and a new standard of care.
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  • 文章类型: Case Reports
    背景:脊索瘤,一种罕见的由脊索组织引起的恶性肿瘤,通常发生在脊柱轴。只有少数已发表的原发性肺脊索瘤的报道。在这里,我们介绍了一例原发性肺脊索瘤,并讨论了诊断罕见脊索瘤的重要考虑因素。
    方法:我们报告一例39岁男性原发性肺脊索瘤,有睾丸混合生殖细胞卵黄囊肿瘤和畸胎瘤病史。计算机断层扫描显示左下叶缓慢生长的实性病变。我们对可疑生殖细胞肿瘤肺转移进行了楔形切除术。组织学上,大的圆形或卵圆形细胞与嗜酸性细胞的细胞质被大的细胞包围,颗粒,轻度嗜酸性细胞浆。肿瘤细胞为生唾液。免疫组织化学检查对短尾畸形呈阳性,S-100蛋白,上皮膜抗原,波形蛋白,和细胞角蛋白AE1/AE3,提示肺脊索瘤。睾丸混合生殖细胞肿瘤的重新检查未发现脊索元素。尽管某些区域的短尾染色呈阳性,苏木精和伊红(HE)染色未显示脊索瘤的典型形态特征。肺肿瘤的互补荧光原位杂交(FISH)证实不存在同染色体12p和12p扩增。因此,确定了原发性肺脊索瘤的最终诊断.
    结论:在有睾丸混合生殖细胞肿瘤病史的患者中,肺和睾丸肿瘤的HE和Brachyury染色的组织形态学比较,使用FISH分析肺肿瘤中的12p和12p扩增对于诊断罕见肺脊索瘤至关重要。
    BACKGROUND: Chordoma, a rare malignant tumor arising from notochordal tissue, usually occurs along the spinal axis. Only a few published reports of primary lung chordomas exist. Herein, we present a case of primary lung chordoma and discuss important considerations for diagnosing rare chordomas.
    METHODS: We report a case of primary lung chordoma in a 39-year-old male with a history of testicular mixed germ-cell tumor of yolk sac and teratoma. Computed tomography revealed slow-growing solid lesions in the left lower lobe. We performed wedge resection for suspected germ-cell tumor lung metastasis. Histologically, large round or oval cells with eosinophilic cytoplasm were surrounded by large cells with granular, lightly eosinophilic cytoplasm. Tumor cells were physaliphorous. Immunohistochemistry was positive for brachyury, S-100 protein, epithelial membrane antigen, vimentin, and cytokeratin AE1/AE3, suggesting pulmonary chordoma. Re-examination of the testicular mixed germ-cell tumor revealed no notochordal elements. Although some areas were positive for brachyury staining, hematoxylin and eosin (HE) staining did not show morphological features typical of chordoma. Complementary fluorescence in situ hybridization (FISH) of the lung tumor confirmed the absence of isochromosome 12p and 12p amplification. Thus, a final diagnosis of primary lung chordoma was established.
    CONCLUSIONS: In patients with a history of testicular mixed germ cell tumors, comparison of histomorphology using HE and Brachyury staining of lung and testicular tumors, and analyzing isochromosome 12p and 12p amplification in lung tumors using FISH is pivotal for the diagnosis of rare lung chordomas.
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  • 文章类型: Journal Article
    高分辨率质谱(HRMS)已成为环境和食品安全分析的重要工具。这篇综述重点介绍了HRMS如何用于分析鱼类中的化学污染物。测量和记录鱼类中的化学污染物不仅可以作为环境条件的指标,还可以监测这些动物的健康状况,并有助于保护人类食物的重要来源。包括兽药在内的污染物的发生率和意义,人类药物和个人护理产品,杀虫剂,持久性有机污染物,全氟化和多氟化物质,和海洋毒素将被审查。HRMS相对于传统MS的优势在于其能够扩展可检测和鉴定的化合物的数量。HRMS是否用于靶向分析物,或更广泛地用于可疑筛查和非靶向分析。化合物的种类,种类的鱼或海鲜,数据采集和分析的选项,并总结了最近HMRS方法对鱼类化学污染物的意外发现的报道。
    High resolution mass spectrometry (HRMS) has become an important tool in environmental and food safety analysis. This review highlights how HRMS has been used to analyze chemical contaminants in fish. Measuring and documenting chemical contaminants in fish serves not only as an indicator of environmental conditions but can also monitor the health of these animals and help protect an important source of human food. The incidence and significance of contaminants including veterinary drugs, human drugs and personal care products, pesticides, persistent organic pollutants, per- and poly fluorinated substances, and marine toxins will be reviewed. The advantage of HRMS over traditional MS is its ability to expand the number of compounds that can be detected and identified. This is true whether HRMS is used for targeted analytes, or more broadly for suspect screening and nontargeted analyses. The classes of compounds, types of fish or seafood, options for data acquisition and analysis, and reports of unexpected findings from recent HMRS methods for chemical contaminants in fish are summarized.
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  • 文章类型: Journal Article
    在海洋环境中,对许多分类群来说,生物多样性的知识仍然不完整,要求进行评估以了解和监测生物多样性的丧失。环境DNA(eDNA)元转录编码是监测海洋生物多样性的有力工具,因为它可以在单个样品中同时表征多个分类单元。然而,环境DNA元编码产生的数据通常不容易重用。为eDNA衍生数据实施FAIR原则和标准可以促进科学界的数据共享。
    这项研究的重点是在瓜德罗普岛的背风海岸使用eDNA元编码检测海洋脊椎动物的生物多样性,法属西印度群岛的海洋生物多样性热点。此处使用DarwinCore标准与MIMARKS标准相结合来共享事件和DNA衍生数据。
    UNASSIGNED: In the marine environment, knowledge of biodiversity remains incomplete for many taxa, requiring assessments to understand and monitor biodiversity loss. Environmental DNA (eDNA) metabarcoding is a powerful tool for monitoring marine biodiversity, as it enables several taxa to be characterised simultaneously in a single sample. However, the data generated by environmental DNA metabarcoding are often not easily reusable. Implementing FAIR principles and standards for eDNA-derived data can facilitate data-sharing within the scientific community.
    UNASSIGNED: This study focuses on the detection of marine vertebrate biodiversity using eDNA metabarcoding on the leeward coast of Guadeloupe, a known hotspot for marine biodiversity in the French West Indies. Occurrences and DNA-derived data are shared here using DarwinCore standards combined with MIMARKS standards.
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  • 文章类型: Journal Article
    最近的全球变暖趋势对全球生态系统构成了重大威胁。这种全球气候变化也影响了水生生态系统的污染水平,从而影响人类健康。为了解决这些问题,进行了一项实验,以研究铁纳米颗粒(Fe-NPs)对砷和氨毒性以及高温胁迫(AsNH3T)的缓解作用。Fe-NP是使用鱼废物生物合成的,并以10、15和20mgkg-1的饮食掺入饲料配方中。在涉及540条鱼的完全随机设计后,共设计了12种处理,一式三份。15mgkg-1饮食中的Fe-NPs显着降低了暴露于多种应激源的鱼中的皮质醇水平。HSP70、DNA损伤诱导蛋白(DDIP)、DNA损伤被应激源(AsNH3T)上调,被Fe-NP下调。凋亡基因(Cas3a和3b)和解毒基因(CYP450),金属硫蛋白(MT),在遭受AsNH3T胁迫的鱼中,Fe-NP在15mgkg-1饮食下下调了诱导型一氧化氮合酶(iNOS)。免疫相关基因,如肿瘤坏死因子(TNFα),免疫球蛋白(Ig),白细胞介素(IL)被Fe-NPs上调,表明鱼在As+NH3+T胁迫下的免疫力增强。相反,在AsNH3T胁迫下,鱼类在15mgkg-1饮食下的Fe-NP明显下调了Toll样受体(TLR)的表达。免疫学属性,如氯化硝基蓝四唑,总蛋白质,白蛋白,球蛋白,A:G比,和髓过氧化物酶(MPO)通过饮食Fe-NPs在15mgkg-1饮食的鱼,不管压力因素。抗氧化基因(CAT,SOD,和GPx)也被鱼类中的Fe-NPs增强。与生长性能相关的基因,如生长激素调节剂(GHR1和GHRβ),生长激素(GH),和胰岛素样生长因子(IGF1X和IGF2X),被上调,在压力下促进鱼类生长,而SMT和MYST在饮食中被Fe-NPs下调。在15mgkg-1饮食下,饮食Fe-NPs改善了各种生长性能指标。值得注意的是,Fe-NP还增强了砷的解毒作用,并降低了细菌感染后的累积死亡率。总之,这项研究强调,饮食中的Fe-NPs可以通过调节鱼类的基因表达来有效减轻砷和氨的毒性以及高温胁迫。
    The recent trend of global warming poses a significant threat to ecosystems worldwide. This global climate change has also impacted the pollution levels in aquatic ecosystems, subsequently affecting human health. To address these issues, an experiment was conducted to investigate the mitigating effects of iron nanoparticles (Fe-NPs) on arsenic and ammonia toxicity as well as high temperature stress (As+NH3+T). Fe-NPs were biologically synthesized using fish waste and incorporated into feed formulations at 10, 15, and 20 mg kg-1 diet. A total of 12 treatments were designed in triplicate following a completely randomized design involving 540 fish. Fe-NPs at 15 mg kg-1 diet notably reduced the cortisol levels in fish exposed to multiple stressors. The gene expressions of HSP 70, DNA damage-inducible protein (DDIP), and DNA damage were upregulated by stressors (As+NH3+T) and downregulated by Fe-NPs. Apoptotic genes (Cas 3a and 3b) and detoxifying genes (CYP 450), metallothionein (MT), and inducible nitric oxide synthase (iNOS) were downregulated by Fe-NPs at 15 mg kg-1 diet in fish subjected to As+NH3+T stress. Immune-related genes such as tumor necrosis factor (TNFα), immunoglobulin (Ig), and interleukin (IL) were upregulated by Fe-NPs, indicating enhanced immunity in fish under As+NH3+T stress. Conversely, Toll-like receptor (TLR) expression was notably downregulated by Fe-NPs at 15 mg kg-1 diet in fish under As+NH3+T stress. Immunological attributes such as nitro blue tetrazolium chloride, total protein, albumin, globulin, A:G ratio, and myeloperoxidase (MPO) were improved by dietary Fe-NPs at 15 mg kg-1 diet in fish, regardless of stressors. The antioxidant genes (CAT, SOD, and GPx) were also strengthened by Fe-NPs in fish. Genes associated with growth performance, such as growth hormone regulator (GHR1 and GHRβ), growth hormone (GH), and insulin-like growth factor (IGF 1X and IGF 2X), were upregulated, enhancing fish growth under stress, while SMT and MYST were downregulated by Fe-NPs in the diet. Various growth performance indicators were improved by dietary Fe-NPs at 15 mg kg-1 diet. Notably, Fe-NPs also enhanced arsenic detoxification and reduced the cumulative mortality after a bacterial infection. In conclusion, this study highlights that dietary Fe-NPs can effectively mitigate arsenic and ammonia toxicity as well as high temperature stress by modulating gene expression in fish.
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  • 文章类型: Journal Article
    嗜冷黄杆菌(Fp)引起沙门氏菌的细菌性冷水病。在宿主-病原体相互作用期间,革兰氏阴性菌,比如Fp,释放外膜囊泡(OMV)窝载货物,比如DNA,RNA和毒力因子。这项研究旨在表征OMV小RNA(sRNA)在Fp-虹鳟鱼宿主-病原体相互作用中的潜在作用。从Fp分离OMV内携带的sRNA。来自全细胞Fp及其分离的OMV的RNA-Seq数据集表明与亲本细胞相比,OMV中特定sRNA的大量富集。许多OMV包装的sRNA位于Fp的致病性岛。报道了在具有不同程度毒力的65个菌株中sRNA的保守性。Fp抗性和易感虹鳟鱼遗传系感染后第5天,宿主和病原体转录组的双重RNA-Seq揭示了OMV包装的sRNA及其预测的宿主免疫基因靶标的相关表达。体外,用OMV处理虹鳟鱼上皮细胞系RTgill-W1显示出细胞毒性的迹象,并伴随着宿主基因表达的动态变化。OMV处理的细胞,类似于抗Fp的鱼,显示细胞因子信号抑制因子1(SOCS1)基因的表达下调,提示吞噬体成熟的诱导。OMV处理后调节宿主基因表达的其他迹象包括来自吞噬细胞的有利元件,内吞和抗原呈递途径,除了HSP70,HSP90和伴侣蛋白,这为OMV在增强宿主免疫反应中的潜在作用提供了证据。总之,该研究确定了新型微生物靶标和OMV的固有特性,可以开辟治疗和预防鱼类感染的新途径。
    Flavobacterium psychrophilum (Fp) causes Bacterial Cold Water Disease in salmonids. During host-pathogen interactions, gram-negative bacteria, such as Fp, release external membrane vesicles (OMVs) harbouring cargos, such as DNA, RNA and virulence factors. This study aimed to characterise the potential role of the OMVs\' small RNAs (sRNAs) in the Fp-rainbow trout host-pathogen interactions. sRNAs carried within OMVs were isolated from Fp. RNA-Seq datasets from whole-cell Fp and their isolated OMVs indicated substantial enrichment of specific sRNAs in the OMVs compared to the parent cell. Many of the OMV-packaged sRNAs were located in the pathogenicity islands of Fp. Conservation of sRNAs in 65 strains with variable degrees of virulence was reported. Dual RNA-Seq of host and pathogen transcriptomes on day 5 post-infection of Fp -resistant and -susceptible rainbow trout genetic lines revealed correlated expression of OMV-packaged sRNAs and their predicted host\'s immune gene targets. In vitro, treatment of the rainbow trout epithelial cell line RTgill-W1 with OMVs showed signs of cytotoxicity accompanied by dynamic changes in the expression of host genes when profiled 24 h following treatment. The OMV-treated cells, similar to the Fp -resistant fish, showed downregulated expression of the suppressor of cytokine signalling 1 (SOCS1) gene, suggesting induction of phagosomal maturation. Other signs of modulating the host gene expression following OMV-treatment include favouring elements from the phagocytic, endocytic and antigen presentation pathways in addition to HSP70, HSP90 and cochaperone proteins, which provide evidence for a potential role of OMVs in boosting the host immune response. In conclusion, the study identified novel microbial targets and inherent characteristics of OMVs that could open up new avenues of treatment and prevention of fish infections.
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  • 文章类型: Journal Article
    随着科学的进步和新基因组技术(NGTs)的发展,为各种目的而修饰的生物范围正在迅速扩大,包括广泛的分类群。决策者特别感兴趣的是,如何更好地了解哪些新开发的产品可以在不久和更远的将来进入市场并释放到环境中。监管机构,和风险评估员。为了满足这些信息需求,我们对四组生物的潜在环境应用进行了地平线扫描(HS):陆生动物(不包括昆虫和具有基因驱动的应用),鱼,藻类和微生物。我们采用了正式的范围审查方法,包括对科学文献进行结构化搜索,然后进行资格筛选,辅以对灰色文学的调查,以及监管网站和数据库。在所有四类生物中,我们确定了基础和高级研究阶段的广泛潜在应用,以及数量有限的应用程序,或者准备被放在上面,市场。对转基因动物包括鱼类的研究集中在养殖动物,主要针对提高性能的性状,影响繁殖,或传达对疾病的抵抗力。在HS中鉴定的转基因藻类都是单细胞的,超过一半的文章涉及生物燃料生产。转基因藻类在环境中的应用包括生物防治和生物修复,这也是转基因微生物的主要应用。从风险评估者的角度来看,这些潜在的应用需要多种可能的危害途径。目前经验水平有限,现有科学资料数量有限,风险评估人员和主管部门迫切需要为此做好准备。
    With scientific progress and the development of new genomic techniques (NGTs), the spectrum of organisms modified for various purposes is rapidly expanding and includes a wide range of taxonomic groups. An improved understanding of which newly developed products may be introduced into the market and released into the environment in the near and more distant future is of particular interest for policymakers, regulatory authorities, and risk assessors. To address this information need, we conducted a horizon scanning (HS) of potential environmental applications in four groups of organisms: terrestrial animals (excluding insects and applications with gene drives), fish, algae and microorganisms. We applied a formal scoping review methodology comprising a structured search of the scientific literature followed by eligibility screening, complemented by a survey of grey literature, and regulatory websites and databases. In all four groups of organisms we identified a broad range of potential applications in stages of basic as well as advanced research, and a limited number of applications which are on, or ready to be placed on, the market. Research on GM animals including fish is focused on farmed animals and primarily targets traits which increase performance, influence reproduction, or convey resistance against diseases. GM algae identified in the HS were all unicellular, with more than half of the articles concerning biofuel production. GM algae applications for use in the environment include biocontrol and bioremediation, which are also the main applications identified for GM microorganisms. From a risk assessor\'s perspective these potential applications entail a multitude of possible pathways to harm. The current limited level of experience and limited amount of available scientific information could constitute a significant challenge in the near future, for which risk assessors and competent authorities urgently need to prepare.
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  • 文章类型: Journal Article
    这项研究引入了流式细胞术和荧光原位杂交(Flow-FISH)的优化整合,作为益生菌产品中革兰氏阳性细菌的特异性计数方法。克服了传统方法的局限性。增强的Flow-FISH技术将流式细胞术的快速和自动化能力与FISH的高特异性相结合,促进益生菌混合物中物种水平的活细胞分化。通过分析鼠李糖乳杆菌的冻干样品,植物乳杆菌,和动物双歧杆菌亚种。乳酸,和商业产品,该研究突出了优化的Flow-FISH协议的优势,包括减少杂交时间至1.5小时和消除离心步骤。用广泛接受的计数方法进行板计数和活/死(L/D)染色的比较评价。研究表明,Flow-FISH产生的活细胞计数高于平板计数,从而通过强调其低估实际可行细胞数量的倾向来挑战传统的“黄金标准”。针对L/D染色,Flow-FISH取得了可比的结果,which,尽管每种技术的基础前提不同,证实了我们方法的准确性和可靠性。总之,优化的Flow-FISH方案代表了益生菌研究和质量控制的重大飞跃。这种方法提供了一种快速、健壮,以及益生菌计数的高度特异性替代方法,超越传统方法。它能够对益生菌产品进行更详细和可靠的分析,为精确的质量控制和研究见解铺平了道路。强调其显著改善该领域的潜力。
    This study introduces an optimized integration of flow cytometry and fluorescence in situ hybridization (Flow-FISH) as an approach for the specific enumeration of gram-positive bacteria in probiotic products, overcoming the limitations of conventional methods. The enhanced Flow-FISH technique synergizes the rapid and automated capabilities of flow cytometry with the high specificity of FISH, facilitating the differentiation of viable cells at the species level within probiotic blends. By analyzing lyophilized samples of Lacticaseibacillus rhamnosus, Lactiplantibacillus plantarum, and Bifidobacterium animalis subsp. lactis, and a commercial product, the study highlights the optimized Flow-FISH protocol\'s advantages, including reduced hybridization times to 1.5 h and elimination of centrifugation steps. Comparative evaluations with the widely accepted enumeration methods plate count and Live/Dead (L/D) staining were conducted. The study revealed that Flow-FISH produces higher viable cell counts than plate count, thereby challenging the traditional \"gold standard\" by highlighting its predisposition to underestimate actual viable cell numbers. Against L/D staining, Flow-FISH achieved comparable results, which, despite the different foundational premises of each technique, confirms the accuracy and reliability of our method. In conclusion, the optimized Flow-FISH protocol represents a significant leap forward in probiotic research and quality control. This method provides a rapid, robust, and highly specific alternative for the enumeration of probiotic bacteria, surpassing traditional methodologies. Its ability to enable a more detailed and reliable analysis of probiotic products paves the way for precise quality control and research insights, underscoring its potential to improve the field significantly.
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