scholarly journals Definition of a Novel Plasmid-Based Gene Transfection Protocol of Mammalian Skeletal Muscles by Means of In Vivo Electroporation

2020 ◽  
Vol 21 (18) ◽  
pp. 6494
Author(s):  
Enrico P. Spugnini ◽  
Manuel Scimeca ◽  
Bruno Amadio ◽  
Giancarlo Cortese ◽  
Maurizio Fanciulli ◽  
...  
Keyword(s):  
Gene Transfection ◽  
Dna Transfection ◽  
In Vivo Electroporation ◽  
Electric Pulses ◽  
Hind Limbs ◽  
Transfection Protocol ◽  
Gait Abnormalities ◽  
Definition Of ◽  
The Right

We describe an original electroporation protocol for in vivo plasmid DNA transfection. The right hind limbs of C57 mice are exposed to a specifically designed train of permeabilizing electric pulses by transcutaneous application of tailored needle electrodes, immediately after the injection of pEGFP-C1 plasmid encoding GFP (Green Fluorescente Protein). The electroporated rodents show a greater GFP expression than the controls at three different time points (4, 10, and 15 days). The electroporated muscles display only mild interstitial myositis, with a significant increase in inflammatory cell infiltrates. Finally, mild gait abnormalities are registered in electroporated mice only in the first 48 h after the treatment. This protocol has proven to be highly efficient in terms of expression levels of the construct, is easy to apply since it does not require surgical exposure of the muscle and is well tolerated by the animals because it does not cause evident morphological and functional damage to the electroporated muscle.

Delivery of Plasmid DNA Through Intratumoral Infusion and Electroporation

2000 ◽  
Author(s):  
Fan Yuan ◽  
David Zaharoff ◽  
Xiao-Yu Zhang ◽  
Frank Lohr ◽  
Mark W. Dewhirst ◽  
...  
Keyword(s):  
Gene Expression ◽  
Solid Tumors ◽  
Plasmid Dna ◽  
Ex Vivo ◽  
Electric Pulse ◽  
Interleukin 12 ◽  
Growth Delay ◽  
In Vivo Electroporation ◽  
Electric Pulses

Abstract We investigated DNA transport in the interstitial space and across cell membrane facilitated by intratumoral infusion and in vivo electroporation, respectively. In the study, a rat fibrosarcoma was perfused ex vivo, and apparent hydraulic conductivity (Kapp) was quantified under different perfusion conditions. In addition, three plasmid DNA vectors were infused into solid tumors. Immediately after infusion, tumors were treated with or without electric pulses. Gene expression and tumor growth delay were determined at different time points after electroporation. We found that Kapp was very sensitive to the perfusion pressure, presumably due to perfusion-induced tissue deformation. Treatment of tumors with electric pulse facilitated gene expression in vivo. The growth of tumors treated with plasmid DNA encoding interleukin 12 (IL-12) and electric pulses was slower than those treated with IL-12 or electric pulses alone. These data suggest that gene delivery in solid tumors could be improved significantly through combination of intratumoral infusion and in vivo electroporation.

Delivery of Plasmid DNA Through Intratumoral Infusion and Electroporation

2000 ◽  
Author(s):  
Fan Yuan ◽  
David Zaharoff ◽  
Xiao-Yu Zhang ◽  
Frank Lohr ◽  
Mark W. Dewhirst ◽  
...  
Keyword(s):  
Gene Expression ◽  
Solid Tumors ◽  
Plasmid Dna ◽  
Ex Vivo ◽  
Electric Pulse ◽  
Interleukin 12 ◽  
Growth Delay ◽  
In Vivo Electroporation ◽  
Electric Pulses

Abstract We investigated DNA transport in the interstitial space and across cell membrane facilitated by intratumoral infusion and in vivo electroporation, respectively. In the study, a rat fibrosarcoma was perfused ex vivo, and apparent hydraulic conductivity (Kapp) was quantified under different perfusion conditions. In addition, three plasmid DNA vectors were infused into solid tumors. Immediately after infusion, tumors were treated with or without electric pulses. Gene expression and tumor growth delay were determined at different time points after electroporation. We found that Kapp was very sensitive to the perfusion pressure, presumably due to perfusion-induced tissue deformation. Treatment of tumors with electric pulse facilitated gene expression in vivo. The growth of tumors treated with plasmid DNA encoding interleukin 12 (IL-12) and electric pulses was slower than those treated with IL-12 or electric pulses alone. These data suggest that gene delivery in solid tumors could be improved significantly through combination of intratumoral infusion and in vivo electroporation.

scholarly journals Radiation-Guided Peptide Delivery in a Mouse Model of Nasopharyngeal Carcinoma

2016 ◽  
Vol 2016 ◽  
pp. 1-7
Author(s):  
Pei-cheng Lin ◽  
Jun-yan He ◽  
Yu-yin Le ◽  
Kai-xin Du ◽  
Wei-feng Zhu ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Mouse Model ◽  
Fluorescence Intensity ◽  
Imaging System ◽  
Control Group ◽  
Recombinant Peptide ◽  
Hind Limbs ◽  
The Right ◽  
Experimental Group

Purpose.This study aimed to evaluate the characteristics of the HVGGSSV peptide, exploring radiation-guided delivery in a mouse model of nasopharyngeal carcinoma.Methods.Mice with CNE-1 nasopharyngeal carcinoma were assigned to two different groups treated with Cy7-NHS and Cy7-HVGGSSV, respectively. Meanwhile, each mouse received a single dose of 3 Gy radiation. Biological distribution of the recombinant peptide was assessed on anin vivosmall animal imaging system.Results.The experimental group showed maximum fluorescence intensity in irradiated tumors treated with Cy7-labeled HVGGSSV, while untreated (0 Gy) control tumors showed lower intensity levels. Fluorescence intensities of tumors in the right hind limbs of experimental animals were7.84×107±1.13×107,1.35×108±2.66×107,4.05×108±1.75×107,5.57×108±3.47×107, and9.26×107±1.73×107photons/s/cm2higher compared with left hind limb values at 1, 2, 15, 24, and 48 h, respectively. Fluorescence intensities of tumor in the right hind limbs of the experimental group were1.66×108±1.71×107,1.51×108±3.23×107,5.38×108±1.96×107,5.89×108±3.57×107, and1.62×108±1.69×107photons/s/cm2higher compared with control group values at 1, 2, 15, 24, and 48 h, respectively. Fluorescence was not specifically distributed in the control group. Compared with low fluorescence intensity in the heart, lungs, and tumors, high fluorescence distribution was found in the liver and kidney at 48 h.Conclusions.HVGGSSV was selectively bound to irradiated nasopharyngeal carcinoma, acting as a targeting transport carrier for radiation-guided drugs that are mainly metabolized in the kidney and liver.

Evaluation of 99mTc-Label led Vitamin K4 as an Agent for Testicular Imaging

1991 ◽  
Vol 30 (01) ◽  
pp. 35-39 ◽  
Author(s):  
H. S. Durak ◽  
M. Kitapgi ◽  
B. E. Caner ◽  
R. Senekowitsch ◽  
M. T. Ercan
Keyword(s):  
Soft Tissue ◽  
Room Temperature ◽  
Normal Subjects ◽  
Left Testis ◽  
Wide Range ◽  
Activity Ratios ◽  
The Right ◽  
Radioactivity Levels

Vitamin K4 was labelled with 99mTc with an efficiency higher than 97%. The compound was stable up to 24 h at room temperature, and its biodistribution in NMRI mice indicated its in vivo stability. Blood radioactivity levels were high over a wide range. 10% of the injected activity remained in blood after 24 h. Excretion was mostly via kidneys. Only the liver and kidneys concentrated appreciable amounts of radioactivity. Testis/soft tissue ratios were 1.4 and 1.57 at 6 and 24 h, respectively. Testis/blood ratios were lower than 1. In vitro studies with mouse blood indicated that 33.9 ±9.6% of the radioactivity was associated with RBCs; it was washed out almost completely with saline. Protein binding was 28.7 ±6.3% as determined by TCA precipitation. Blood clearance of 99mTc-l<4 in normal subjects showed a slow decrease of radioactivity, reaching a plateau after 16 h at 20% of the injected activity. In scintigraphic images in men the testes could be well visualized. The right/left testis ratio was 1.08 ±0.13. Testis/soft tissue and testis/blood activity ratios were highest at 3 h. These ratios were higher than those obtained with pertechnetate at 20 min post injection.99mTc-l<4 appears to be a promising radiopharmaceutical for the scintigraphic visualization of testes.

Enhanced Increases in Cytosolic Ca2+ in ADP-Stimulated Platelets from Patients with Delta-Storage Pool Deficiency – A Possible Indicator of Interactions between Granule-Bound ADP and the Membrane ADP Receptor

1997 ◽  
Vol 77 (02) ◽  
pp. 376-382 ◽  
Author(s):  
Bruce Lages ◽  
Harvey J Weiss
Keyword(s):  
Platelet Rich Plasma ◽  
Limited Range ◽  
Dense Granule ◽  
Receptor Desensitization ◽  
Fura 2 ◽  
Storage Pool ◽  
Low Levels ◽  
The Right

SummaryThe possible involvement of secreted platelet substances in agonist- induced [Ca2+]i increases was investigated by comparing these increases in aspirin-treated, fura-2-loaded normal platelets and platelets from patients with storage pool deficiencies (SPD). In the presence and absence of extracellular calcium, the [Ca2+]i response induced by 10 µM ADP, but not those induced by 0.1 unit/ml thrombin, 3.3 µM U46619, or 20 µM serotonin, was significantly greater in SPD platelets than in normal platelets, and was increased to the greatest extent in SPD patients with Hermansky-Pudlak syndrome (HPS), in whom the dense granule deficiencies are the most severe. Pre-incubation of SPD-HPS and normal platelets with 0.005-5 µM ADP produced a dose-dependent inhibition of the [Ca2+]i response induced by 10 µ M ADP, but did not alter the [Ca2+]i increases induced by thrombin or U46619. Within a limited range of ADP concentrations, the dose-inhibition curve of the [Ca2+]i response to 10 µM ADP was significantly shifted to the right in SPD-HPS platelets, indicating that pre-incubation with greater amounts of ADP were required to achieve the same extent of inhibition as in normal platelets. These results are consistent with a hypothesis that the smaller ADP-induced [Ca2+]i increases seen in normal platelets may result from prior interactions of dense granule ADP, released via leakage or low levels of activation, with membrane ADP receptors, causing receptor desensitization. Addition of apyrase to platelet-rich plasma prior to fura-2 loading increased the ADP-induced [Ca2+]i response in both normal and SPD-HPS platelets, suggesting that some release of ADP derived from both dense granule and non-granular sources occurs during in vitro fura-2 loading and platelet washing procedures. However, this [Ca2+]i response was also greater in SPD-HPS platelets when blood was collected with minimal manipulation directly into anticoagulant containing apyrase, raising the possibility that release of dense granule ADP resulting in receptor desensitization may also occur in vivo. Thus, in addition to enhancing platelet activation, dense granule ADP could also act to limit the ADP-mediated reactivity of platelets exposed in vivo to low levels of stimulation.

THE CONCEPT OF CONSTITUTIONAL AND LEGAL GUARANTEES OF ELECTORAL RIGHTS

2020 ◽  
Vol 10 (4) ◽  
pp. 85-90
Author(s):  
VLADIMIR TROYAN ◽  
Keyword(s):  
Scientific Research ◽  
Formal Aspect ◽  
Universal Approach ◽  
Right To Vote ◽  
Definition Of ◽  
The Right

The relevance of the interpretation of constitutional and legal guarantees of the right to vote is mediated by isolated scientific research in this area, as well as the lack of a universal approach to legal guarantees. In this regard, the purpose of the article is to argue and disclose the author’s definitive aspect of the claimed guarantees. In the work, the author named and characterized the normative (based exclusively on legal means) with the perspective of a branch of legal and technical; regulatory and institutional (combines the formal aspect with the activities of authorized entities) and associated legal (including a set of legal and other aspects) approaches to the definition of legal guarantees. Based on the second approach, as well as combining the guarantees of the right to vote directly guarantees of the subjective right itself and guarantees of its implementation, the author offers a definition of constitutional and legal guarantees of the right to vote.

administrasi ketatausahaan sekolah

2020 ◽  
Author(s):  
Isra Revenia
Keyword(s):  
Information Needs ◽  
Control Functions ◽  
Definition Of ◽  
The Right ◽  
And Control

This article is made to know the destinantion and the administrasi functions of the school in order to assist the leader of an organazation in making decisions and doing the right thing, recording of such statements in addition to the information needs also pertains to the function of accountabilitty and control functions. Administrative administration is the activity of recording for everything that happens in the organization to be used as information for leaders. While the definition of administration is all processing activities that start from collecting (receiving), recording, processing, duplicating, minimizing and storing all the information of correspondence needed by the organization. Administration is as an activity to determine everything that happens in the organization, to be used as material for information by the leadership, which includes all activities ranging from manufacturing, managing, structuring to all the preparation of information needed by the organization.

SELF ASSESSMENT TO KNOW UNDERSTANDING MATHEMATIC CONCEPT

2018 ◽  
Vol 3 (1) ◽  
pp. 14-21
Author(s):  
Deni Iriyadi
Keyword(s):  
Qualitative Study ◽  
Qualitative Analysis ◽  
Resolution Limit ◽  
Self Assessment ◽  
Limit Concept ◽  
Research Instrument ◽  
Definition Of ◽  
The Right

This research is a qualitative study aimed to determine the students' understanding of the concept of matter limit. The subjects were students of class XI IPA 1 SMA Negeri 1 Watampone. The concept includes the definition of the limit. Data obtained using a research instrument in the form of self-assessment and then proceed with the interview subjects were selected based on the results of self-assessment has been done before. Analysis using qualitative analysis of students' understanding of the concept of the limit concept. The results of this study indicate that students' understanding of concepts some of which are not / do not understand especially regarding definitions limit. In addition students are also wrong about the resolution limit. Students who understand the concept of limit dinyakatakan them restate concepts, including examples and classify the sample to non-completion of function and limit the right results.

DIRECTIONS FOR PROVISION OF PUBLIC BODIES’ TRANSPARENCY AND ACCOUNTABILITY IN CONDITIONS OF PARTICIPATORY DEMOCRACY DEVELOPMENT

2018 ◽  
Vol 1 (11) ◽  
pp. 144-154
Author(s):  
Olga Mykhailоvna Ivanitskaya
Keyword(s):  
Public Administration ◽  
Participatory Democracy ◽  
Main Task ◽  
The Public ◽  
State Governance ◽  
Legislative Initiative ◽  
The Government ◽  
Definition Of ◽  
The Right ◽  
Transparency And Accountability

The article is devoted to issues of ensuring transparency and ac- countability of authorities in the conditions of participatory democracy (democ- racy of participation). It is argued that the public should be guaranteed not only the right for access to information but also the prerequisites for expanding its par- ticipation in state governance. These prerequisites include: the adoption of clearly measurable macroeconomic and social goals and the provision of control of the processes of their compliance with the government by citizens of the country; ex- tension of the circle of subjects of legislative initiative due to realization of such rights by citizens and their groups; legislative definition of the forms of citizens’ participation in making publicly significant decisions, design of relevant orders and procedures, in particular participation in local referendum; outlining methods and procedures for taking into account social thought when making socially im- portant decisions. The need to disclose information about resources that are used by authorities to realize the goals is proved as well as key performance indicators that can be monitored by every citizen; the efforts made by governments of coun- tries to achieve these goals. It was noted that transparency in the conditions of representative democracy in its worst forms in a society where ignorance of the thought of society and its individual members is ignored does not in fact fulfill its main task — to establish an effective dialogue between the authorities and so- ciety. There is a distortion of the essence of transparency: instead of being heard, society is being asked to be informed — and passively accept the facts presented as due. In fact, transparency and accountability in this case are not instruments for the achievement of democracy in public administration, but by the form of a tacit agreement between the subjects of power and people, where the latter passes the participation of an “informed observer”.

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