| 1. |
- Tahmina, Qurratul-Ain
(författare)
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Gender Equality and Media Regulation Study : Bangladesh
- 2022
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Rapport (populärvet., debatt m.m.)abstract
- This study seeks to provide new knowledge and analysis about gender-equality related provisions in regulations, self-regulatory frameworks and policies concerning media in Bangladesh. It also explores their implementation and monitoring aspects.It seeks to provide clear recommendations and cite best practices that can assist stakeholders including law and policy-makers to promote gender equality in and through the media without compromising professional independence. For clarifying the contextual situations and ground realities, it also seeks to provide qualitative reflections accumulated through the research process.March through mid-October 2021, the study explored: to what extent and how have gender-equality, gender-sensitivity or gender-awareness issues been integrated into the regulation and self-regulation concerning both media structure and content. whether such integration and efforts could contribute to the overarching goal of increasing the freedom of expression for women and girls, for people of non-binary gender, for people belonging to sexual minorities, and also for men and boys in situations similarly relevant. if any such effort could lead to compromising or curtailing media freedom and independence.For these explorations, the study used mixed methodology, both quantitative and qualitative. A total of 43 Acts and Rules including the Constitution and 12 national-level media policies were selected for mapping and analysis. A questionnaire survey of 18 media houses were conducted, while seven regulators and self-regulators were interviewed.The study finds that provisions for gender equality and sensitivity in media regulations and national policies have two distinct features. The earlier provisions were concerned more about decency, obscenity and other such issues mainly from a moralistic perspective. The main concerns seemed to have been protecting the morality of society from effects of such contents, rather than protecting the rights of women.Then over the last couple of decades, gender-related provisions in media regulation and policies have been markedly influenced by development concerns. Media advocacy for women development along with other development goals started occupying a central space in regulatory and policy frameworks.10This era might have introduced a rights perspective but gender-equality in the media organisations has not been the dominant primary approach. Provisions on „gender-sensitivity‟ were more frequent. The laws and regulations lack a requirement for equal coverage of women or other gender minorities in media content as well. The policies may have some requirements of this from the perspective of development in various sectors.The laws varyingly provide eligibility criteria for ownership, which are basic, general and more or less common in nature, not specifying anything on gender-equality commitment. Such mentions would be more appropriate and necessary in policies. All policies, with the exception of one, are silent about gender-equality or gender-sensitivity commitments of owners. No law or policy, however, have any bar to a woman being an owner of a media outlet.Sector-wide self-regulatory frameworks are very rare, so are individual house-level ones. Written in- house policies on gender equality or sensitivity are very few.Regulatory and supervisory authority of all the media rests primarily with one central ministry, while gender-related supervision is the responsibility of another ministry. No implementation or monitoring mechanism could be located. Scarcity of data in this field is another major problem. Implementation and monitoring are generally weak in media organisations too.The study recommends more stress on having policies, especially within media organisations. While regulations are important, law alone cannot do much. Policy is more important, at the state level of course, but especially the self-regulatory ones. In the absence of an industry-level press commission, unions and professional organisations need to formulate codes of conducts. Monitoring should be strengthened both at regulatory and self-regulatory levels. Getting the editors and owners on board is important.The study particularly recommends, building on whatever gains have been made so far—building on the awareness amongst the media leaders about what is politically and ethically correct. Alongside looking for what gaps are there, one needs to examine how much and what have been achieved, and what factors made the changes happen. A crucially important task is liaising with the women‟s rights and gender rights movements and gain their support for attaining gender-equality and sensitivity in the media.
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| 2. |
- Bengtsson, Elina, et al.
(författare)
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Myosin-Induced Gliding Patterns at Varied [MgATP] Unveil a Dynamic Actin Filament
- 2016
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Ingår i: Biophysical Journal. - : Elsevier BV. - 0006-3495 .- 1542-0086. ; 111:7, s. 1465-1477
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Tidskriftsartikel (refereegranskat)abstract
- Actin filaments have key roles in cell motility but are generally claimed to be passive interaction partners in actin-myosin -based motion generation. Here, we present evidence against this static view based on an altered myosin-induced actin filament gliding pattern in an in vitro motility assay at varied [MgATP]. The statistics that characterize the degree of meandering of the actin filament paths suggest that for [MgATP] >= 0.25 mM, the flexural rigidity of heavy meromyosin (HMM)-propelled actin filaments is similar (without phalloidin) or slightly lower (with phalloidin) than that of HMM-free filaments observed in solution without surface tethering. When [MgATP] was reduced to <= 0.1 mM, the actin filament paths in the in vitro motility assay became appreciably more winding in both the presence and absence of phalloidin. This effect of lowered [MgATP] was qualitatively different from that seen when HMM was mixed with ATP-insensitive, N-ethylmaleimide-treated HMM (NEM-HMM; 25-30%). In particular, the addition of NEM-HMM increased a non-Gaussian tail in the path curvature distribution as well as the number of events in which different parts of an actin filament followed different paths. These effects were the opposite of those observed with reduced [MgATP]. Theoretical modeling suggests a 30-40% lowered flexural rigidity of the actin filaments at [MgATP] <= 0.1 mM and local bending of the filament front upon each myosin head attachment. Overall, the results fit with appreciable structural changes in the actin filament during actomyosin-based motion generation, and modulation of the actin filament mechanical properties by the dominating chemomechanical actomyosin state.
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| 3. |
- Lindberg, Frida W., et al.
(författare)
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Controlled Surface Silanization for Actin-Myosin and Biocompatibility of New Polymer Resists
- 2018
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Ingår i: Langmuir. - : American Chemical Society (ACS). - 0743-7463 .- 1520-5827. ; 34:30, s. 8777-8784
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Tidskriftsartikel (refereegranskat)abstract
- Molecular motor-based nanodevices require organized cytoskeletal filament guiding along motility-promoting tracks, confined by motility-inhibiting walls. One way to enhance motility quality on the tracks, particularly in terms of filament velocity but also the fraction of motile filaments, is to optimize the surface hydrophobicity. We have investigated the potential to achieve this for the actin myosin II motor system on trimethylchlorosilane (TMCS)-derivatized SiO2 surfaces to be used as channel floors in nanodevices. We have also investigated the ability to supress motility on two new polymer resists, TU7 (for nanoimprint lithography) and CSAR 62 (for electron beam and deep UV lithography), to be used as channel walls. We developed a chemical-vapor deposition tool for silanizing SiO2 surfaces in a controlled environment to achieve different surface hydrophobicities (measured by water contact angle). In contrast to previous work, we were able to fabricate a wide range of contact angles by varying the silanization time and chamber pressure using only one type of silane. This resulted in a significant improvement of the silanization procedure, producing a predictable contact angle on the surface and thereby predictable quality of the heavy meromyosin (HMM)-driven actin motility with regard to velocity. We observed a high degree of correlation between the filament sliding velocity and contact angle in the range 10-86 degrees, expanding the previously studied range. We found that the sliding velocity on TU7 surfaces was superior to that on CSAR 62 surfaces despite similar contact angles. In addition, we were able to suppress the motility on both TU7 and CSAR 62 by plasma oxygen treatment before silanization. These results are discussed in relation to previously proposed surface adsorption mechanisms of HMM and their relationship to the water contact angle. Additionally, the results are considered for the development of actin-myosin based nanodevices with superior performance with respect to actin-myosin functionality.
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| 4. |
- Lindberg, Frida W., et al.
(författare)
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Design and development of nanoimprint-enabled structures for molecular motor devices
- 2019
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Ingår i: Materials Research Express. - : IOP Publishing. - 2053-1591. ; 6:2
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Tidskriftsartikel (refereegranskat)abstract
- Devices based on molecular motor-driven cytoskeletal filaments, e.g., actin filaments, have been developed both for biosensing and biocomputational applications. Commonly, these devices require nanoscaled tracks for guidance of the actin filaments which has limited the patterning technique to electron beam lithography. Thus, large scale systems become intractable to fabricate at a high throughput within a reasonable time-frame. We have studied the possibility to fabricate molecular motor-based devices using the high throughput, high resolution technique of nanoimprint lithography. Molecular motor-based devices require wide open regions (loading zones) to allow filaments to land for later propulsion into the nanoscale tracks. Such open zones are challenging to fabricate using nanoimprint lithography due to the large amount of material displaced in the process. We found that this challenge can be overcome by introducing nanoscaled pillars inside the loading zones, into which material can be displaced during imprint. By optimising the resist thickness, we were able to decrease the amount of material displaced without suffering from insufficient filling of the stamp. Furthermore, simulations suggest that the shape and positioning of the pillars can be used to tailor the overall cytoskeletal filament transportation direction and behaviour. This is a potentially promising design feature for future applications that however, requires further optimisations before experimental realisation.
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| 5. |
- Rahman, Mohammad A.
(författare)
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Biophysical studies of the actin-myosin motor system and applications in nanoscience
- 2019
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- The actin-myosin motor system plays important roles in cellular processes. In addition, actin and myosin have been used for developments towards nanotechnological applications in recent years. Therefore, fundamental biophysical studies of actin and myosin and the actomyosin force generating cycle are important both in biology and for nanotechnology where the latter applications require methodological insights for optimization. This dual goal is central in the present thesis with major focus on factors that control the function (e.g. velocity) and the effectiveness of transport of filaments (e.g. filament flexural rigidity) through nanoscale channels with supplementation of methodological insights. The thesis thus provides evidence that actin is a dynamic filament whose flexural rigidity is different at different MgATP concentrations as well as in the presence or absence of myosin binding. Furthermore, probing the myosin ATPase cycle with the myosin inhibitor blebbistatin revealed that velocity is easily modified by this drug. Our detailed studies also suggest that actin-myosin force generation is preceded by Pi release and that blebbistatin changes the rate limiting transition in the cycle from the attachment step to a step between weakly attached states. The studies of actin dynamics and of the actomyosin force generating cycle were largely performed using in vitro motility assay (IVMA) where surface adsorbed myosin motor or its proteolytic fragments propel fluorescently labeled actin filaments. The IVMA is often taken as the basis for developments towards different nanotechnological applications. However, in the IVMA, actomyosin motility is often negatively affected by the presence of “dead”, non-functional myosin heads. Therefore, in this thesis, two popular methods, that are often used to remove dead myosin heads, are analyzed and compared. It was found that after affinity purification, the in vitro actin sliding velocity is reduced compared to the control conditions, something that was not seen with the use of blocking actin. Therefore, the effects of the affinity purification method should be considered when interpreting IVMA data. This is important while using IVMA both for fundamental studies and for nanotechnological applications. Another issue in the use of IVMAs in nanotechnological applications is the requirement for expensive and time-consuming fabrication of nanostructured devices. We therefore developed a suitable method for regenerating molecular motor based bionanodevices without a need to disassemble the flow cell. Evidence is presented that, use of proteinase K with a suitable detergent (SDS or Triton X100) lead to successful regeneration of devices where both actin-myosin and microtubule-kinesin motility are used. Lastly, this thesis presents efforts to immobilize engineered light sensitive myosin motors on trimethyl chlorosilane (TMCS) derivatized surfaces for light operated switching of myosin motor in order to control actin movement in nano-networks. This has potential for developing a programmable junction in a biocomputation network. In brief, the described results have contributed both to the fundamental understanding of actin and myosin properties and the actomyosin interaction mechanisms. They have also given technical insights for molecular motor based bionanotechnology.
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| 6. |
- Rahman, Mohammad A., et al.
(författare)
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Blebbistatin Effects Expose Hidden Secrets in the Force-Generating Cycle of Actin and Myosin
- 2018
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Ingår i: Biophysical Journal. - Rockville, MD : Biophysical Society. - 0006-3495 .- 1542-0086. ; 115:2, s. 386-397
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Tidskriftsartikel (refereegranskat)abstract
- Cyclic interactions between myosin II motors and actin filaments driven by ATP turnover underlie muscle contraction and have key roles in the motility of nonmuscle cells. A remaining enigma in the understanding of this interaction is the relationship between the force-generating structural change and the release of the ATP-hydrolysis product, inorganic phosphate (Pi), from the active site of myosin. Here, we use the small molecular compound blebbistatin to probe otherwise hidden states and transitions in this process. Different hypotheses for the Pi release mechanism are tested by interpreting experimental results from in vitro motility assays and isolated muscle fibers in terms of mechanokinetic actomyosin models. The data fit with ideas that actomyosin force generation is preceded by Pi release, which in turn is preceded by two serial transitions after/coincident with cross-bridge attachment. Blebbistatin changes the rate limitation of the cycle from the first to the second of these transitions, uncovering functional roles of an otherwise short-lived pre-power stroke state that has been implicated by structural data.
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| 7. |
- Rahman, Mohammad A., et al.
(författare)
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Comparative analysis of widely used methods to remove nonfunctional myosin heads for the in vitro motility assay
- 2018
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Ingår i: Journal of Muscle Research and Cell Motility. - : Springer. - 0142-4319 .- 1573-2657. ; 39:5-6, s. 175-187
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Tidskriftsartikel (refereegranskat)abstract
- The in vitro motility assay allows studies of muscle contraction through observation of actin filament propulsion by surface-adsorbed myosin motors or motor fragments isolated from muscle. A possible problem is that motility may be compromised by nonfunctional, "dead", motors, obtained in the isolation process. Here we investigate the effects on motile function of two approaches designed to eliminate the effects of these dead motors. We first tested the removal of heavy meromyosin (HMM) molecules with ATP-insensitive "dead" heads by pelleting them with actin filaments, using ultracentrifugation in the presence of 1 mM MgATP ("affinity purification"). Alternatively we incubated motility assay flow cells, after HMM surface adsorption, with non-fluorescent "blocking actin" (1 µM) to block the dead heads. Both affinity purification and use of blocking actin increased the fraction of motile filaments compared to control conditions. However, affinity purification significantly reduced the actin sliding speed in five out of seven experiments on silanized surfaces and in one out of four experiments on nitrocellulose surfaces. Similar effects on velocity were not observed with the use of blocking actin. However, a reduced speed was also seen (without affinity purification) if HMM or myosin subfragment 1 was mixed with 1 mM MgATP before and during surface adsorption. We conclude that affinity purification can produce unexpected effects that may complicate the interpretation of in vitro motility assays and other experiments with surface adsorbed HMM, e.g. single molecule mechanics experiments. The presence of MgATP during incubation with myosin motor fragments is critical for the complicating effects.
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| 8. |
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| 9. |
- Rahman, Mohammad A., et al.
(författare)
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Regeneration of Assembled, Molecular-Motor-Based Bionanodevices
- 2019
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Ingår i: Nano Letters. - : American Chemical Society (ACS). - 1530-6984 .- 1530-6992. ; 19:10, s. 7155-7163
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Tidskriftsartikel (refereegranskat)abstract
- The guided gliding of cytoskeletal filaments, driven by biomolecular motors on nano/microstructured chips, enables novel applications in biosensing and biocomputation. However, expensive and time-consuming chip production hampers the developments. It is therefore important to establish protocols to regenerate the chips, preferably without the need to dismantle the assembled microfluidic devices which contain the structured chips. We here describe a novel method toward this end. Specifically, we use the small, nonselective proteolytic enzyme, proteinase K to cleave all surface-adsorbed proteins, including myosin and kinesin motors. Subsequently, we apply a detergent (5% SDS or 0.05% Triton X100) to remove the protein remnants. After this procedure, fresh motor proteins and filaments can be added for new experiments. Both, silanized glass surfaces for actin-myosin motility and pure glass surfaces for microtubule-kinesin motility were repeatedly regenerated using this approach. Moreover, we demonstrate the applicability of the method for the regeneration of nano/microstructured silicon-based chips with selectively functionalized areas for supporting or suppressing gliding motility for both motor systems. The results substantiate the versatility and a promising broad use of the method for regenerating a wide range of protein-based nano/microdevices.
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| 10. |
- Salhotra, Aseem, et al.
(författare)
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Exploitation of Engineered Light-Switchable Myosin XI for Nanotechnological Applications
- 2023
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Ingår i: ACS Nano. - : American Chemical Society (ACS). - 1936-0851 .- 1936-086X. ; 17:17, s. 17233-17244
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Tidskriftsartikel (refereegranskat)abstract
- For certain nanotechnological applications of the contractile proteins actin and myosin, e.g., in biosensing and network-based biocomputation, it would be desirable to temporarily switch on/off motile function in parts of nanostructured devices, e.g., for sorting or programming. Myosin XI motor constructs, engineered with a light-switchable domain for switching actin motility between high and low velocities (light-sensitive motors (LSMs) below), are promising in this regard. However, they were not designed for use in nanotechnology, where longevity of operation, long shelf life, and selectivity of function in specific regions of a nanofabricated network are important. Here, we tested if these criteria can be fulfilled using existing LSM constructs or if additional developments will be required. We demonstrated extended shelf life as well as longevity of the actin-propelling function compared to those in previous studies. We also evaluated several approaches for selective immobilization with a maintained actin propelling function in dedicated nanochannels only. Whereas selectivity was feasible using certain nanopatterning combinations, the reproducibility was not satisfactory. In summary, the study demonstrates the feasibility of using engineered light-controlled myosin XI motors for myosin-driven actin transport in nanotechnological applications. Before use for, e.g., sorting or programming, additional work is however needed to achieve reproducibility of the nanofabrication and, further, optimize the motor properties.
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