| 1. |
- Cutas, Daniela, 1978, et al.
(författare)
-
Legal imperialism in the regulation of stem cell research and therapy: the problem of extraterritorial jurisdiction
- 2010
-
Ingår i: Capps BJ & Campbell AV (eds.). CONTESTED CELLS: Global Perspectives on the Stem Cell Debate. - London : Imperial College Press. - 9781848164376 ; , s. 95-119
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- Countries worldwide have very different national regulations on human embryonic stem (ES) cell research, informed by a range of ethical values. Some countries find reason to extend the applicability of their regulations on such research to its citizens when they visit other countries. Extraterritorial jurisdiction has recently been identified as a potential challenge towards global regulation of ES cell research. This chapter explores the implications and impact of extraterritorial jurisdiction and global regulation of ES cell research on researchers, clinicians and national health systems, and how this may affect patients. The authors argue that it would make ethical sense for ES cell restrictive countries to extend its regulations on ES cell research beyond its borders, because, if these countries really consider embryo destruction to be objectionable on the basis on the status of the embryo, then they ought to count it morally on par with murder (and thus have a moral imperative to protect embryos from the actions of its own citizens). However, doing so could lead to a legal situation that would result in substantial harm to central values in areas besides research, such as health care, the job market, basic freedom of movement, and strategic international finance and politics. Thus, it seems that restrictive extraterritorial jurisdiction in respect to ES cell research would be deeply problematic, given that the ethical permissibility of ES cell research is characterised by deep and wide disagreement.
|
|
| 2. |
- Porosk, Ly, et al.
(författare)
-
Endpoint and Kinetic Approaches for Assessing Transfection Efficacy in Mammalian Cell Culture
- 2022. - 3
-
Ingår i: Cell Penetrating Peptides. - New York : Humana Press. - 9781071617519 - 9781071617526 ; , s. 529-545
-
Bokkapitel (refereegranskat)abstract
- The efficacy of transfection reagents and nanoparticles is often assessed by measuring levels of expressed reporter protein. Fluorescence and luminescence based assays provide sensitive, quantifiable and repeatable approaches. The genes expressing reporter protein can be integrated into the cells to create stable reporter cell lines or can be expressed from a transfected plasmid. Green fluorescent protein, luciferase, and secreted alkaline phosphatase are well-established reporters with versatile applications. Monitoring changes in live cells during and after transfection offer opportunities to reveal related mechanisms, efficacy, and bottlenecks of transfection. In this chapter, we describe the experimental setup and considerations for in vitro screening of delivery vectors. This can further be extended to measurements in reporter cell lines.
|
|
| 3. |
- Gustafson, Deborah R.
(författare)
-
Adipose Tissue Complexities in Dyslipidemias
- 2019
-
Ingår i: Dyslipidemia. - London : IntechOpen. - 9781839680045 - 9781839680038 - 9781839680052 ; , s. 1-22
-
Bokkapitel (refereegranskat)abstract
- Adipose tissue is the largest organ in the human body and, in excess, contributes to dyslipidemias and the dysregulation of other vascular and metabolic processes. Adipose tissue is heterogeneous, comprised of several cell types based on morphology, cellular age, and endocrine and paracrine function. Adipose tissue depots are also regional, primarily due to sex differences and genetic variation. Adipose tissue is also characterized as subcutaneous vs. visceral. In addition, fatty deposits exist outside of adipose tissue, such as those surrounding the heart, or as infiltration of skeletal muscle. This review focuses on adipose tissue and its contribution to dyslipidemias. Dyslipidemias are defined as circulating blood lipid levels that are too high or altered. Lipids include both traditional and nontraditional species. Leaving aside traditional definitions, adipose tissue contributes to dyslipidemias in a myriad of ways. To address a small portion of this topic, we reviewed (a) adipose tissue location and cell types, (b) body composition, (c) endocrine adipose, (d) the fat-brain axis, and (e) genetic susceptibility. The influence of these complex aspects of adipose tissue on dyslipidemias and human health, illustrating that, once again, that adipose tissue is a quintessential, multifunctional tissue of the human body, will be summarized.
|
|
| 4. |
- Guterstam, Peter, et al.
(författare)
-
Characterization of cellular internalization pathways for CPP-mediated oligonucleotide delivery
- 2011
-
Ingår i: Cell-penetrating peptides. - New York : Humana Press. - 9781607619185 ; , s. 219-230
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- The methods for evaluating internalization pathways of cellular CPP-mediated ON delivery utilizing a pre-mRNA splice correction assay and fluorescence-based quantification are described. Examples for characterization of CPP uptake routes, employing various endocytosis inhibitors, and special treatment conditions are demonstrated. The methods are developed to characterize cellular delivery of pre-mRNA splice switching peptide nucleic acids conjugated to CPPs by disulfide bond.
|
|
| 5. |
- Lammi, Mikko, 1961-, et al.
(författare)
-
Responses of mammalian cells to mechanical forces
- 2001. - Vol. 1
-
Ingår i: Recent Research Developments in Biophysics and Biochemistry. - Trivandrum, India : Research Signpost. - 8177360574 ; , s. 77-89
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- All cells and tissues of our body are continuously subject to various mechanical stresses. These forces include, e.g., compression, shear stress, hydrostatic pressure and osmotic pressure. The range of forces vary from few pascals to several megapascals in magnitude. In many cases, mechanical forces are required for the tissues to maintain their normal functional structure and composition. However, excessive forces in the end may lead to adverse responses. In this paper, we review the data available from many different tissues in order to compare the signaling mechanisms involved in cellular mechanotransduction, and how the cells respond to forces that are too strenuous for them to withstand. The possible stress reactions caused by excessive forces are also discussed.
|
|
| 6. |
- Wang, Yuanmo, 1986, et al.
(författare)
-
Artificial Cells for Dissecting Exocytosis
- 2023
-
Ingår i: Methods in Molecular Biology. - New York, NY : Springer US. - 1940-6029 .- 1064-3745. ; 2565, s. 261-279
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- The fusion of vesicles and exocytosis release of neurotransmitters into the extracellular space for detection and chemical signal decoding by neighboring cells is the key process in neuronal communication. It is important to understand what regulates exocytosis because the amount of neurotransmitters released into the synaptic cleft has a direct impact on brain function such as cognition learning and memory as well as on brain malfunctions. Much success in molecular biology can be credited for the existence of simplified model systems. Therefore, for gaining deeper insights into the details of exocytosis and what controls vesicle-mediated neurotransmission, functional artificial cells for exocytosis have been developed that can be used for studying various biophysical aspects and roles of molecules affecting exocytosis, which is difficult to study in living cells. Here, we describe the design and fabrication of specific artificial cell models and how chemical measurements at these cells can be implemented for probing dynamics of the exocytosis fusion pore and its effect on the regulation of neurochemical release. We introduce bottom-up synthetic methods for constructing model cells using protein-free giant unilamellar vesicles (GUV) as starting material, which allows further tuning of molecular complexity in a manner that is not possible in living cells and therefore can be used for dissecting the role of essential molecular components affecting the exocytosis process. The experimental setup uses microscopy video recording, micromanipulation and microelectroinjection techniques, and amperometry detection to study neurotransmitter release from these cells mimicking exocytosis.
|
|
| 7. |
- EL Andaloussi, Samir, et al.
(författare)
-
Application of PepFect peptides for the delivery of splice-correcting oligonucleotides
- 2011
-
Ingår i: Cell-penetrating peptides. - New York : Humana Press. - 9781607619185 ; , s. 361-373
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- One oligonucleotide-based approach that appear very promising for the treatment of different genetic disorders are based on so-called splice-correcting oligonucleotides (SCOs) that are exploited to manipulate splicing patterns. In order to increase the bioavailability, cell-penetrating peptides (CPPs) have readily been covalently conjugated to SCOs to facilitate cellular internalization. While being a successful strategy for the delivery of uncharged oligonucleotides (ONs), it is extremely difficult to generate covalent conjugates between commonly used negatively charged ON analogs and cationic CPPs. Furthermore, high concentrations of ONs in the micromolar range are often needed to obtain biological responses, most likely as a result of endosomal entrapment of material. Therefore, exploring other vectorization methods using CPPs with endosomolytic properties are highly desired. A method of using stearyl modified CPP (i.e., TP10) analogs, named PepFect3 and PepFect4, are being described for the transfection of antisense SCOs using a simple one-step co-incubation procedure. These peptides form complexes with SCOs and efficiently promote cellular uptake by facilitating endosomal escape. This chapter describes the methods of how to form and characterize these nanoparticles and the cellular assay used to address the delivery.
|
|
| 8. |
|
|
| 9. |
- EL Andaloussi, Samir, et al.
(författare)
-
Cell-penetrating peptides-based strategies for the delivery of splice redirecting antisense oligonucleotides
- 2011
-
Ingår i: Therapeutic Oligonucleotides. - New York : Humana Press. ; 764, s. 75-89
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- Progress in our understanding of the molecular pathogenesis of human malignancies has provided therapeutic targets amenable to oligonucleotide (ON)-based strategies. Antisense ON-mediated splicing regulation in particular offers promising prospects since the majority of human genes undergo alternative splicing and since splicing defects have been found in many diseases. However, their implementation has been hampered so far by the poor bioavailability of nucleic acids-based drugs. Cell-penetrating peptides (CPPs) now appear as promising non-viral delivery vector for non-permeant biomolecules. We describe here new CPPs allowing the delivery of splice redirecting steric-block ON using either chemical conjugation or non-covalent complexation. We also describe a convenient and robust splice redirecting assay which allows the quantitative assessment of ON nuclear delivery.
|
|
| 10. |
- Roomans, Godfried M., 1951-, et al.
(författare)
-
X-Ray Microanalysis in the Scanning Electron Microscope
- 2014. - 3
-
Ingår i: Electron microscopy. - New York, USA : Humana Press. - 9781627037761 - 9781627037754 ; , s. 639-661
-
Bokkapitel (refereegranskat)abstract
- X-ray microanalysis conducted using the scanning electron microscope is a technique that allows the determination of chemical elements in bulk or semi-thick specimens. The lowest concentration of an element that can be detected is in the order of a few mmol/kg or a few hundred parts per million, and the smallest amount is in the order of 10(-18) g. The spatial resolution of the analysis depends on the thickness of the specimen. For biological specimen analysis, care must be taken to prevent displacement/loss of the element of interest (usually ions). Protocols are presented for the processing of frozen-hydrated and freeze-dried specimens, as well as for the analysis of small volumes of fluid, cell cultures, and other specimens. Aspects of qualitative and quantitative analysis are covered, including limitations of the technique.
|
|
