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Micro Flow-Through Thermocycler with Simple Meandering Channel with Symmetric Temperature Zones for Disposable PCR-Devices in Microscope Slide Format

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Journal of Bionic Engineering 2008年第4期5卷 291-298页

作者： Anett Reichert Jana Felbel Mark Kielpinski Matthias Urban Benno Steinbrecht thomas henkel Microfluidics Department Institute of Photonic Technology Arvato Digital Services Manufacturing EMEA

Chip-based flow-through PCR implements the PCR as a continuous process for nucleic acid analytics. The sample is transported in a winding channel through temperature zones required for denaturation, annealing and extension. Main fields of application are the monitoring of continuous processes for rapid identification of contaminants and quality control as well as high throughput screening of cells or microorganisms. A modular arrangement with five heating zones for flow-through PCR is discussed and evaluated. The special heater arrangement allows the implementation of up to 40 cycles on the footprint of a microscope slide, which is placed on top ofa 5 zones heating plate. Liquid/liquid two phase flow of PCR reaction mixture and mineral oil have been applied to create a segmented flow process scheme. In that way, the developed system may provide flow-through PCR as a unit operation for the droplet based microfluidics platform. The single use of disposable devices is commonly preferred due to the sensitivity of the PCR process to contaminations. All-glass microfluidic chips and disposable chip devices, made from polycarbonate as a replication with identically geometry, have been fabricated and tested. For the first time, microchannel geometries with nearly circular profile developed by all-glass technology have been transferred to mass fabrication by injection compression molding. Both devices have been successfully applied for the detection of the tumor suppressor gene p53. Although product yield and selectivity of the amplification process do not depend on the chip material, a well defined, reliable segmented flow regime could only be realized in the all-glass chip.

关键词： lab-on-a-chip PCR microfluidics two-phase-flow injection molding point-of-care biomedical

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专用性芯片实验室的开发

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实验与分析 2009年第5期 55-56页

作者： thomas henkel 德国图像技术研究所微流控制装置部

采用一种新工艺，可以比以前更为快捷地在最小的空间内为化学和生物分析量身定做芯片实验室。通过引入建立在液滴基础上的微流控制装置，可以大大地缩短分析时间。

关键词：芯片实验室专用性开发生物分析控制装置

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从天然提取物或分离部位中以碳酸酐酶II为靶点的抗骨质疏松活性筛选(英文)

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云南植物研究 2003年第6期25卷 724-729页

作者：付祥谭宁华姜立花贾锐锐嵇长久 LANDRO James FEKETE Alexander MUELLER Hartwig henkel thomas 中国科学院昆明植物研究所植物化学与西部植物资源持续利用国家重点实验室云南昆明650204 Bayer Corporation IRT Bayer AG PH-R-EU

目的是以碳酸酐酶II (CAII)为靶点筛选其抑制剂 ,以期寻找抗骨质疏松活性样品。实验是在 96孔酶标板上对来源于 1 78个科、 60 8个属、 1 0 2 0种动植物 2 91 9个提取物或分离部位样品在CA II模型上进行了批量筛选。结果表明在 1 0 μg... 详细信息

目的是以碳酸酐酶II (CAII)为靶点筛选其抑制剂 ,以期寻找抗骨质疏松活性样品。实验是在 96孔酶标板上对来源于 1 78个科、 60 8个属、 1 0 2 0种动植物 2 91 9个提取物或分离部位样品在CA II模型上进行了批量筛选。结果表明在 1 0 μg ml浓度下发现了来源于 40个科、61个属、72个种的 1 0 0个样品有活性 ,其中 5个样品的IC50 小于 2 50 μg ml,2 2个样品的IC50 在 2 51～ 5 0 0 μg ml范围 ,73个样品的IC50 在 5 0 1～1 0 0 0 μg ml范围。通过以上工作我们认为以碳酸酐酶II为分子靶点的体外筛选方法稳定、方便、快速、微量、有效 ,特别适用于天然产物的抗骨质疏松活性筛选。

关键词：抗骨质疏松活性筛选碳酸酐酶Ⅱ 天然产物提取物分离部位

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从天然提取物或分离部位中以烯醇式丙酮酸转移酶为靶点的抗细菌活性筛选(英文)

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云南植物研究 2003年第1期25卷 90-94页

作者：姜立花谭宁华杨亚滨汪俊松付祥 Lars JOHANNSEN Hartwig MUELLER thomas henkel 中国科学院昆明植物研究所植物化学与西部植物资源持续利用国家重点实验室云南昆明650204 Bayer AG PH-R-EU

目的是以烯醇式丙酮酸转移酶 (EPT)为靶点筛选其抑制剂 ,以期寻找抗细菌活性样品。实验是在 96孔酶标板上对来源于 16 9个科、 5 6 0个属、 916种动植物 2 4 90个提取物或分离部位样品在EPT模型上进行了批量筛选。结果表明在 96 15 μg... 详细信息

目的是以烯醇式丙酮酸转移酶 (EPT)为靶点筛选其抑制剂 ,以期寻找抗细菌活性样品。实验是在 96孔酶标板上对来源于 16 9个科、 5 6 0个属、 916种动植物 2 4 90个提取物或分离部位样品在EPT模型上进行了批量筛选。结果表明在 96 15 μg/ml浓度下发现了来源于 80个科、 16 9个属、 2 18个种的 30 9个样品有活性 ,其中 14个样品的IC50 小于 10 0 0 μg/ml,4 0个样品的IC50 在 10 0 1～ 30 0 0 μg/ml范围 ,83个样品的IC50 在 30 0 1～ 5 0 0 0 μg/ml范围 ,172个样品的IC50 在 5 0 0 1～ 96 15 μg/ml范围。通过以上工作我们认为以烯醇式丙酮酸转移酶为分子靶点的体外筛选方法稳定、方便、快速、微量、有效。

关键词：烯醇式丙酮酸转移酶抑制剂抗细菌活性天然产物天然提取物分离部位体外筛选

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Tuning beneficial calcineurin phosphatase activation to counterα-synuclein toxicity in a yeast model of Parkinson’s disease

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Neural Regeneration Research 2025年第1期20卷 199-200页

作者： Srishti Chawla Mikael Molin thomas Nystrom Institute for Biomedicine Sahlgrenska AcademyCentre for Ageing and Health-AgeCapUniversity of GothenburgGothenburgSweden Department of Life Sciences Chalmers University of TechnologyGothenburgSweden

Calcineurin(CN)is a calcium-and calmodulindependent serine/threonine that has been studied in many model organisms including yeast,filamentous fungi,plants,and *** biological functions range from ion homeostasis and virulence in lower eukaryotes to T-cell activation in humans by human nuclear factors of activated *** is a heterodimeric protein consisting of a catalytic subunit,calcineurin A(Cna1p),which contains an active site with a dinuclear metal center,and a regulatory Ca^(2+) binding subunit called calcineurin B(Cnb1p)required to activate *** calcineurin B subunit has been highly conserved through evolution:For example,the mammalian calcineurin B shows 54%identity with calcineurin B from Saccharomyces cerevisiae.

关键词： activation dinuclear conserved

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Sperm preparation： state-of-the-art--physiological aspects and application of advanced sperm preparation methods

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Asian Journal of Andrology 2012年第2期14卷 260-269页

作者： Ralf henkel Department of Medical Biosciences University of the Western Cape Bellville 7535 South Africa

For assisted reproduction technologies （ART）, numerous techniques were developed to isolate spermatozoa capable of fertilizing oocytes. While early methodologies only focused on isolating viable, motile spermatozoa, with progress of ART, particularly intracytoplasmic sperm injection （ICSI）, it became clear that these parameters are insufficient for the identification of the most suitable spermatozoon for fertilization. Conventional sperm preparation techniques, namely, swim-up, density gradient centrifugation and glass wool filtration, are not efficient enough to produce sperm populations free of DNA damage, because these techniques are not physiological and not modeled on the stringent sperm selection processes taking place in the female genital tract. These processes only allow one male germ cell out of tens of millions to fuse with the oocyte. Sites of sperm selection in the female genital tract are the cervix, uterus, uterotubal junction, oviduct, cumulus oophorus and the zona pellucida. Newer strategies of sperm preparation are founded oni （i） morphological assessment by means of＇motile sperm organelle morphological examination （MSOM E）＇; （ii） electrical charge; and （ili） molecular binding characteristics of the sperm cell. Whereas separation methods based on electrical charge take advantage of the sperm＇s adherence to a test tube surface or separate in an electrophoresis, molecular binding techniques use Annexin V or hyaluronic acid （HA） as substrates. Techniques in this category are magnet-activated cell sorting, Annexin V-activated glass wool filtration, flow cytometry and picked spermatozoa for ICSI （PICSI） from HA-coated dishes and HA-containing media. Future developments may include Raman microspectrometry, confocal light absorption and scattering spectroscopic microscopy and polarization microscopy.

关键词： advanced sperm selection biomimetics conventional sperm selection female genital tract physiological sperm selection

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Microglia:a promising therapeutic target in spinal cord injury

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Neural Regeneration Research 2025年第2期20卷 454-463页

作者： Xiaowei Zha Guoli Zheng thomas Skutella Karl Kiening Andreas Unterberg Alexander Younsi Department of Neurosurgery Heidelberg University HospitalHeidelbergGermany Department of Neuroanatomy Institute for Anatomy and Cell BiologyHeidelberg UniversityHeidelbergGermany

Microglia are present throughout the central nervous system and are vital in neural repair,nutrition,phagocytosis,immunological regulation,and maintaining neuronal *** a healthy spinal cord,microglia are accountable for immune surveillance,however,when a spinal cord injury occurs,the microenvironment drastically changes,leading to glial scars and failed axonal *** this context,microglia vary their gene and protein expression during activation,and proliferation in reaction to the injury,influencing injury responses both favorably and unfavorably.A dynamic and multifaceted injury response is mediated by microglia,which interact directly with neurons,astrocytes,oligodendrocytes,and neural stem/progenitor *** a clear understanding of their essential nature and origin,the mechanisms of action and new functions of microglia in spinal cord injury require extensive *** review summarizes current studies on microglial genesis,physiological function,and pathological state,highlights their crucial roles in spinal cord injury,and proposes microglia as a therapeutic target.

关键词： astrocytes cytokines functional recovery immune regulation M1/M2 activation macrophages microglia neuroinflammation spinal cord injury therapy

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Leukocytes and oxidative stress： dilemma for sperm Function and male fertility

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Asian Journal of Andrology 2011年第1期13卷 43-52页

作者： Ralf R henkel Department of Medical Biosciences University of the Western Cape Bellville 7535 South Africa

Spermatozoa are constantly exposed to the interphase between oxidation through high amounts of reactive oxygen species （ROS） and leukocytes, and reduction by means of scavengers and antioxidants. Considering the very special functions as being the only cells with such high polarization and exerting their functions outside the body, even in a different individual, the female genital tract, the membranes of these cells are chemically composed of an extraordinary high amount of polyunsaturated fatty acids. This in turn, renders them very susceptible to oxidative stress, which is defined as an imbalance between oxidation and reduction towards the oxidative status. As a result, ROS deriving from both leukocytes and the male germ cells themselves cause a process called ＇lipid peroxidation＇ and other damages to the sperm cell. On the other hand, a certain limited amount of ROS iS essential in order to trigger vital physiological reactions in cells, including capacitation or the acrosome reaction in sperm. The treatment of patients with antioxidants to compensate the oxidative status caused by oxidative stress is highly debated as uncontrolled antioxidative treatment might derail the system towards the reduced status, which is also unphysiological and can even induce cancer. This paradox is called the ＇antioxidant paradox＇. Therefore, a proper andrological diagnostic work-up, including the evaluation of ROS levels and the antioxidant capacity of the semen, has to he carried out beforehand, aimed at keeping the fine balance between oxidation and scavenging of vital amounts of ROS.

关键词： antioxidants balance beneficial effects detrimental effects leukocytes male infertility reactive oxygen species sperm functions

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Sleep as a window to understand and regulate Alzheimer's disease:emerging roles of thalamic reticular nucleus

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Neural Regeneration Research 2025年第6期20卷 1711-1712页

作者： Haoqi Sun Shiqian Shen Robert J.thomas M.Brandon Westover Can Zhang Department of Neurology Beth Israel Deaconess Medical CenterHarvard Medical SchoolBostonMAUSA Center for Translational Pain Research Department of AnesthesiaCritical Care and Pain MedicineMassachusetis General HospitalHarvard Medical SchoolBostonMAUSA Division of Pulmonary Critical Care and Sleep MedicineDepartment of MedicineBeth Israel Deaconess Medical CenterHarvard Medical SchoolBostonMAUSA Genetics and Aging Research Unit McCance Center for Brain HealthMass General Institute for Neurodegenerative DiseaseDepartment of NeurologyMassachusetts General HospitalHarvard Medical SchoolBostonMAUSA

Introduction:Alzheimer 's disease(AD) is a common neurodegenerative disorder and the primary cause of dementia. Considerable evidence supports the “amyloid hypothesis,” stating that the pathogenesis of AD is primarily caused by the deposition of amyloid-β(Aβ), which drives tau phosphorylation, neuroinflammation, and neurodegeneration in the brain. The amyloid hypothesis is strengthened by the significant and moderate benefit of lecanemab, a humanized antibody through an anti-amyloid mechanism,showing slowed clinical decline(van Dyck et al.,2023). The recent positive results of anti-amyloid trials have brought back focus on the amyloid hypothesis through biochemical, genetic, and pharmacological approaches(Zhang, 2023).

关键词： Alzheimer inflammation hypothesis

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Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation:an appeal for performance verification

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Asian Journal of Andrology 2019年第5期21卷 438-444页

作者： Rakesh Sharma Sajal Gupta Ralf henkel Ashok Agarwal American Center for Reproductive Medicine Cleveland ClinicClevelandOH 44195USA Department of Medical Bioscience University of the Western CapeBellville 7535South Africa

Lack of standardized,reproducible protocols and referencO values is among the challenges faced when using new or upgraded versions of instruments in reproductive laboratories and flow *** deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assay combined with flow cytometry routinely used for diagnostic measurement of sperm DNA fragmentation(SDF)is a unique *** change in the setting of the standard instrument,including upgrades of hardware or software,can lead to different results and may affect clinicians'decision for ***,we compared TUNEL results of SDF obtained from a standard(C6)flow cytometer with a newer version of the same instrument(C6 Plus)and examined the cutoff,sensitivity,and specificity without calibration(adjustment)and after *** sperm preparation and matched acquisition settings were used to examine the performance of two flow *** strength of agreement of the results between the two observers was also *** adjustment of the settings,overall concordance became high and the two cytometers showed 100%positive and negative predictive value with 100%area under the *** overall correlation coefficient observed between C6 and C6 Plus was highly significant(P99%.The in terobserver agreeme nt on C6 flow cytometer for the two observers was 0.801±0.062 and 0.746±0.044 for C6 *** demonstrated a strong agreement between the samples tested on the two flow cytometers after calibration and established the robustness of both instruments.

关键词： DNA fragmentation flow cytometer oxidative stress sperm TUNEL assay

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