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Functionality of a bicistronic construction containing HEXA and HEXB genes encoding β-hexosaminidase A for cell-mediated therapy of Gm2 gangliosidoses

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Neural Regeneration Research 2022年第1期17卷 122-129页

作者： Alisa A.Shaimardanova Daria S.Chulpanova Valeriya V.Solovyeva aleks.ndr m.aimaletdinov Albert A.Rizvanov Institute of Fundamental medicine and Biology Kazan Federal UniversityKazanRussia

Tay-Sachs disease and Sandhoff disease are severe hereditary neurodegenerative disorders caused by a deficiency ofβ-hexosaminidase A(HexA)enzyme,which results in the accumulation of Gm2 gangliosides in the nervous system *** this work,we analyzed the efficacy and safety of cell-mediated gene therapy for Sandhoff disease and Sandhoff disease using a bicistronic lentiviral vector encoding cDNA of HexAα-andβ-subunit genes separated by the nucleotide sequence of a P2A peptide(HEXA-HEXB).The functionality of the bicistronic construct containing the HEXA-HEXB genetic cassette was analyzed in a culture of HEK293T cells and human umbilical cord blood mononuclear cells(hUCBmCs).Our results showed that the enzymatic activity of HexA in the conditioned medium harvested from genetically modified HEK293T-HEXA-HEXB and hUCBmCs-HEXA-HEXB was increased by 23 and 8 times,respectively,compared with the conditioned medium of native *** blot analysis showed that hUCBmCs-HEXA-HEXB secreted both completely separated HEXA and HEXB proteins,and an uncleaved protein containing HEXA+HEXB linked by the P2A *** injection of genetically modified hUCBmCs-HEXA-HEXB to laboratory Wistar rats was carried out,and the HexA enzymatic activity in the blood plasma of experimental animals,as well as the number of live cells of immune system organs(spleen,thymus,bone marrow,lymph nodes)were determined.A significant increase in the enzymatic activity of HexA in the blood plasma of laboratory rats on days 6 and 9(by 2.5 and 3 times,respectively)after the administration of hUCBmCsHEXA-HEXB was *** the same time,the number of live cells in the studied organs remained ***,the functionality of the bicistronic genetic construct encoding cDNA of the HEXA and HEXB genes separated by the nucleotide sequence of the P2A peptide was shown in vitro and in *** hypothesize that due to the natural ability of hUCBmCs to overcome biological barriers,such a strategy can rest

关键词： bicistronic vector cell-mediated gene therapy Gm2 gangliosidosis P2A peptide Sandhoff disease Tay-Sachs disease umbilical cord blood mononuclear cells β-hexosaminidase

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Increasingβ-hexosaminidase A activity using genetically modified mesenchymal stem cells

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Neural Regeneration Research 2024年第1期19卷 212-219页

作者： Alisa A.Shaimardanova Daria S.Chulpanova Valeriya V.Solovуeva Shaza S.Issa Aysilu I.mullagulova Angelina A.Titova Yana O.mukhamedshina Anna V.Timofeeva Alexander m.aimaletdinov Islam R.Nigmetzyanov Albert A.Rizvanov Institute of Fundamental medicine and Biology Kazan Federal UniversityKazanRussia Department of Histology Cytology and EmbryologyKazan State Medical UniversityKazanRussia

Gm2 gangliosidoses are a group of autosomal-recessive lysosomal storage disorde *** diseases result from a deficiency of lysosomal enzymeβ-hexosaminidase A(HexA),which is responsible for Gm2 ganglioside *** deficiency causes the accumulation of Gm2-gangliosides mainly in the nervous system cells,leading to severe progressive neurodegeneration and *** date,there is no treatment for these ***-mediated gene therapy is considered a promising treatment for Gm2 *** study aimed to evaluate the ability of genetically modified mesenchymal stem cells(mSCs-HEXA-HEXB)to restore HexA deficiency in Tay-Sachs disease patient cells,as well as to analyze the functionality and biodistribution of mSCs in *** effectiveness of HexA deficiency cross-correction was shown in mutant mSCs upon intera ction with *** results also showed that the mSCs-HEXA-HEXB express the functionally active HexA enzyme,detectable in vivo,and intravenous injection of the cells does not cause an immune response in *** data suggest that genetically modified mesenchymal stem cells have the potentials to treat Gm2 gangliosidoses.

关键词： adeno-associated viral vectors cell therapy cell-mediated gene therapy gene therapy Gm2 gangliosidosis Sandhoff disease Tay-Sachs disease β-hexosaminidase

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超临界CO_2增强热卤水开采模型研究

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地质科技情报 2014年第6期33卷 233-240页

作者：杨国栋李义连马鑫 Atrens aleks 倪浩中国地质大学环境学院武汉430074 中国地质调查局水文地质环境地质调查中心河北保定071051 Queensland Geothermal Energy Centre of Excellence The University of Queensland

面对全球气候变化和能源紧缺的巨大压力,CO2地质利用技术成为研究的焦点。为了实现CO2减排及资源化利用,提出了利用超临界CO2增强卤水提取和地热能开采的CO2地质利用集成系统的概念。通过建立CO2注入热卤水储层的质量平衡模型,分析了不... 详细信息

面对全球气候变化和能源紧缺的巨大压力,CO2地质利用技术成为研究的焦点。为了实现CO2减排及资源化利用,提出了利用超临界CO2增强卤水提取和地热能开采的CO2地质利用集成系统的概念。通过建立CO2注入热卤水储层的质量平衡模型,分析了不同流量的卤水生产、CO2注入以及储层边界流对储层变化的影响,初步评估了该系统的CO2封存量、卤水提取量以及储层流体组成变化的时间尺度。研究表明,注入CO2提供了热卤水层的压力维持,促进卤水和地热资源的可持续开采,对于江陵凹陷研究区9×10^8 m^3的储层有效体积,注入9.95×10^6 t CO2可提取17.12×10^6 t卤水,时间尺度超过30a。对于50~1 000kg/s的卤水生产速度,可以产生0.9~18.8 mW电力。同时,该技术增加了CO2的封存容量和效率,有利于CO2大规模安全封存,经济和环境效益显著。

关键词： CO2地质利用卤水地热能开采 CCUS

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Target blood pressure in diabetes patients with hypertension——What is the accumulated evidence in 2011?

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2011年第8期12卷 611-623页

作者： Peter m.NILSSON Department of Clinical Sciences Lund UniversityUniversity Hospital

There is overwhelming evidence that hypertension is an important risk factor for both macrovascular and microvascular complications in patients with diabetes,but the problem remains to identify appropriate goals for preventive therapies.A number of guidelines (the European Society of Cardiology (ESC)/European Association for the Study of Diabetes (EASD) 2007,the Joint National Committee (JNC)-Ⅶ 2003,the American Diabetes Association (ADA) 2011) have for example advocated a blood pressure goal of less than 130/80 mmHg,but this suggestion has been challenged by findings in recent trials and meta-analyses (2011).The European Society of Hypertension (ESH) therefore recommends a systolic blood pressure goal of "well below" 140 *** on evidence from both randomized controlled trials (hypertension optimal treatment (HOT),action in diabetes and vascular disease:preterax and diamicron mR controlled evaluation (ADVANCE),action to control cardiovascular risk in diabetes (ACCORD)) and observational studies (ongoing telmisartan alone and in combination with ramipril global endpoint trial (ONTARGET),international verapamil-trandolapril study (INVEST),treat to new targets (TNT),and the National Diabetes Register (ndr)),it has been shown that the benefit for stroke reduction remains even at lower achieved blood pressure levels,but the risk of coronary events may be uninfluenced or even increased at lower systolic blood pressure *** a recent meta-analysis,it was therefore concluded that the new recommended goal should be 130-135 mmHg systolic blood pressure for most patients with type 2 *** risk factors should also be controlled with a more ambitious strategy applied in the younger patients with shorter diabetes duration,but a more cautious approach in the elderly and frail patients with a number of vascular or non-vascular *** patients from East Asia,such as China,the stroke risk is relatively higher than the risk of coronary *** must al

关键词： Blood pressure Cardiovascular Diabetes Goal Hypertension Treatment

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Optimized Deep Learning Approach for Efficient Diabetic Retinopathy Classification Combining VGG16-CNN

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Computers, materials & Continua 2023年第11期77卷 1855-1872页

作者： Heba m.El-Hoseny Heba F.Elsepae Wael A.mohamed Ayman S.Selmy Department of Computer Science the Higher Future Institute for Specialized Technological StudiesObour11828Egypt Department of Electrical Engineering Benha Faculty of EngineeringBenha UniversityBenha13511Egypt

Diabetic retinopathy is a critical eye condition that,if not treated,can lead to vision *** methods of diagnosing and treating the disease are time-consuming and ***,machine learning and deep transfer learning(DTL)techniques have shown promise in medical applications,including detecting,classifying,and segmenting diabetic *** advanced techniques offer higher accuracy and *** Diagnosis(CAD)is crucial in speeding up classification and providing accurate disease ***,these technological advancements hold great potential for improving the management of diabetic *** study’s objective was to differentiate between different classes of diabetes and verify the model’s capability to distinguish between these *** robustness of the model was evaluated using other metrics such as accuracy(ACC),precision(PRE),recall(REC),and area under the curve(AUC).In this particular study,the researchers utilized data cleansing techniques,transfer learning(TL),and convolutional neural network(CNN)methods to effectively identify and categorize the various diseases associated with diabetic retinopathy(DR).They employed the VGG-16CNN model,incorporating intelligent parameters that enhanced its *** outcomes surpassed the results obtained by the auto enhancement(AE)filter,which had an ACC of over 98%.The manuscript provides visual aids such as graphs,tables,and techniques and frameworks to enhance *** study highlights the significance of optimized deep TL in improving the metrics of the classification of the four separate classes of *** manuscript emphasizes the importance of using the VGG16CNN classification technique in this context.

关键词： No diabetic retinopathy(ndr) convolution layers(CNV layers) transfer learning data cleansing convolutional neural networks a visual geometry group(VGG16)

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A step-by-step protocol for formaldehyde-assisted isolation of regulatory elements from Arabidopsis thaliana

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Journal of Integrative Plant Biology 2014年第6期56卷 527-538页

作者： mohammad Amin Omidbakhshfard Flavia Vischi Winck Samuel Arvidsson Diego m.Ria?o-Pachón Bernd mueller-Roeber University of Potsdam Institute of Biochemistry and Biology Potsdam-Golm 14476 Germany max-Planck Institute of molecular Plant Physiology Potsdam-Golm 14476 Germany Brazilian Bioethanol Science and Technology Laboratory (CTBE) Brazilian Center of Research in Energy and Materials (CNPEM) Campinas Brazil

The control of gene expression by transcriptional regulators and other types of functional y relevant DNA transactions such as chromatin remodeling and replication underlie a vast spectrum of biological processes in al organisms. DNA transactions require the control ed interaction of proteins with DNA sequence motifs which are often located in nucleosome-depleted regions （ndrs） of the chromatin. Formaldehyde-assisted isolation of regulatory elements （FAIRE） has been established as an easy-to-implement method for the isolation of ndrs from a number of eukaryotic organisms, and it has been successful y employed for the discovery of new regulatory segments in genomic DNA from, for example, yeast, Drosophila, and humans. Until today, however, FAIRE has only rarely been employed in plant research and currently no detailed FAIRE protocol for plants has been published. Here, we provide a step-by-step FAIRE protocol for ndr discovery in Arabidopsis thaliana. We demonstrate that ndrs isolated from plant chromatin are readily amenable to quantitative polymerase chain reaction and next-generation sequencing. Only minor modification of the FAIRE protocol wil be needed to adapt it to other plants, thus facilitating the global inventory of regulatory regions across species.

关键词： Arabidopsis thaliana chromatin cis-regulatory elements epigenomics FAIRE-qPCR FAIRE-seq gene expression gene regulatory network transcription factor

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Analyzing the Effects of Key Design Factors of a Negative-Differential-Resistance(ndr)microfluidic Oscillator–an Equivalent-Circuit-model Approach

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Advances in Applied mathematics and mechanics 2022年第6期14卷 1381-1399页

作者： J.W.Wu H.m.Xia Z.P.Wang W.Wang H.J.Du School of mechanical Engineering Nanjing University of Science and TechnologyNanjingJiangsu 210094China Singapore Institute of manufacturing Technology 73 Nanyang DriveSingapore 637662Singapore School of mechanical and Aerospace Engineering Nanyang Technological University50 Nanyang AvenueSingapore 639798Singapore

Numerical study on dynamic hydroelastic problems is usually rather complex due to the coupling of fluid and solid ***,we demonstrate that the performance of a hydroelastic microfluidic oscillator can be analyzed using a simple equivalent circuit *** studies reveal that its transition from the steady state to the oscillation state follows the negative-differential-resistance(ndr)*** performance is mainly determined by a bias fluidic resistor,and a pressurevariant resistor which further relates to the bending stiffness of the elastic diaphragm and the depth of the oscillation *** this work,a numerical study is conducted to examine the effects of key design factors on the device robustness,the applicable fluid viscosity,the flow rate,and the transition *** underlying physics is interpreted,providing a new perspective on hydroelastic oscillation *** findings also provide design guidelines of the ndr fluidic oscillator.

关键词： microfluidic oscillator hydroelastics equivalent circuit model negative differential resistance

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北德意志广播电台的立体电视试验报告

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广播与电视技术 1983年第6期 57-60页

作者： m.文茨尔 ,杨玉泉

1982年2月北德意志广播电台(ndr)两次播出了立体电视节目,在观众中引起了很大的反响。对此得追溯到几乎100年前就为人所知的补色立体方法(Anaglyphen)原理。由两部组装在一起的电视摄象机摄取的两幅图象加上了红、绿两色。为了能看到立体效

关键词：立体图象德意志广播电台立体电视三维电视摄象机传感器立体眼镜

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Response of Salt-Tolerant Rice Varieties to Biocompost Application in Sodic Soil of Eastern Uttar Pradesh

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American Journal of Plant Sciences 2014年第1期5卷 7-13页

作者： Akhtar H. Khan Ashok K. Singh mubeen Sudhanshu Singh Najam W. Zaidi Uma S. Singh Stephan m. Haefele 1Department of Crop Physiology Narendra Deva University of Agriculture and Technology Kumarganj India Australian Centre for Plant Functional Genomics University of Adelaide Adelaide Australia Department of Crop Physiology Narendra Deva University of Agriculture and Technology Kumarganj India International Rice Research Institute New Delhi India

Sodic soils have immense productivity potential, if managed through proper technology interventions. Biocompost is prepared by composting pressmud (a sugar industry byproduct) received from cane juice filtration and spent wash received from distilleries through microbial aerobic decomposition and can be used to reclaim sodic soils. Field experiments were conducted during the wet season of 2011 and 2012 to study the effect of incorporation of biocompost in sodic soil with four treatments: T1—Control, T2—Biocompost at 2 t ha-1, T3—Biocompost at 4 t ha-1 and T4—Biocompost at 6 t ha-1. The two promising salt tolerant rice varieties preferred by farmers, Narendra usar 3 and ndr 359 were used as test crops, which can produce yields ranging between 2-4 t ha-1 in soil having a pH range of 9.2 to 10.5. Among the different doses of biocompost tested, application of biocompost at 6 t ha-1 registered highest yields, enabled by a higher biomass, ear bearing tiller (EBT), and grain fertility in both varieties. Narendra usar 3 was more responsive to treatments even at lower doses of biocompost than ndr 359, but ndr 359 yielded slightly higher than Narendra usar 3 in all treatments. Soil health was also improved evidently on better fertility and low soil pH and EC at harvest. Thus, biocompost can be considered as a commercially viable, environmentally acceptable and practically enforceable option for improving the crop productivity and soil fertility status.

关键词： Biocompost Narendra usar 3 ndr 359 Salt-Tolerant Varieties Sodic Soil

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常染色体显性遗传的单症状性肌强直持续状态

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世界核心医学期刊文摘(神经病学分册) 2006年第12期 20-21页

作者： Colding-Jφrgensen E. Duno m. Vissing J. 郭俊 Dr.Department of Clinical Neurophysiology 19 Glostrup HospitalUniversity of CopenhagenNdr.RingvejDK-2600 GlostrupDenmark

myotonia permanens is associated with a G1306E mutation in the SCN4A *** sporadic patients have been reported,but the clinical phenotype has not been fully *** authors report a family in which the disease is autosomal dominantly *** patients have severe myotonia,but the clinical picture is not qualitatively different from that seen in other nondystrophic myotonias.

关键词：肌强直神经肌肉疾病持续状态常染色体显性遗传

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