Development and initial validation of a theory of planned behavior questionnaire: Assessment of purchase intentions towards products associated with CRM campaigns

AbstractThis research develops and validates scales based on the theory of planned behavior (TPB) to measure purchase intention towards products associated with cause-related marketing (CRM) campaigns in the South Asian context. Despite few studies using global measures of TPB in specific contexts to predict behavioral intention towards CRM campaigns, this study develops and uses belief-based formative indicators that can be used as an intervention to bring about behavioral changes for positive campaign outcomes. A mixed methods approach was used, including focus group discussions and open-ended questionnaires, to collect qualitative data from 62 participants, resulting in the development of the formative indicators of the measurement instrument. The scales were then combined with global measures of reflective indicators and validated using data collected from 1035 respondents in a quantitative study. The results support the TPB theory and show that the scales have strong internal consistency, reliability, and validity. The findings indicate that behavioral beliefs (β = 0.834, p < 0.001), normative beliefs (β = 0.631, p < 0.001), and control beliefs (β = 0.725, p < 0.001) significantly impact attitude, subjective norm, and perceived behavioral control respectively. Attitude (β = 0.374, p < 0.001), subjective norms (β = 0.218, p < 0.001), and perceived behavioral control (β = 0.320, p < 0.001) significantly influence purchase intentions, with attitude having the most significant impact. The study also found that purchase intention significantly affects purchase behavior (β = 0030.530, p < 0.001). And therefore, this study strengthens the theory of planned behavior in the context of CRM campaigns, aligning with the broader field of ethical consumption

Centripetal nuclear shape fluctuations associate with chromatin condensation in early prophase.

The nucleus plays a central role in several key cellular processes, including chromosome organisation, DNA replication and gene transcription. Recent work suggests an association between nuclear mechanics and cell-cycle progression, but many aspects of this connection remain unexplored. Here, by monitoring nuclear shape fluctuations at different cell cycle stages, we uncover increasing inward fluctuations in late G2 and in early prophase, which are initially transient, but develop into instabilities when approaching the nuclear-envelope breakdown. We demonstrate that such deformations correlate with chromatin condensation by perturbing both the chromatin and the cytoskeletal structures. We propose that the contrasting forces between an extensile stress and centripetal pulling from chromatin condensation could mechanically link chromosome condensation with nuclear-envelope breakdown, two main nuclear processes occurring during mitosis

Role of the nuclear membrane protein Emerin in front-rear polarity of the nucleus

Cell polarity refers to the intrinsic asymmetry of cells, including the orientation of the cytoskeleton. It affects cell shape and structure as well as the distribution of proteins and organelles. In migratory cells, front-rear polarity is essential and dictates movement direction. While the link between the cytoskeleton and nucleus is well-studied, we aim to investigate if front-rear polarity can be transmitted to the nucleus. We show that the knock-down of emerin, an integral protein of the nuclear envelope, abolishes preferential localization of several nuclear proteins. We propose that the frontally biased localization of the endoplasmic reticulum, through which emerin reaches the nuclear envelope, is sufficient to generate its observed bias. In primary emerin-deficient myoblasts, its expression partially rescues the polarity of the nucleus. Our results demonstrate that front-rear cell polarity is transmitted to the nucleus and that emerin is an important determinant of nuclear polarity.ISSN:2041-172

nucGEMs probe the biophysical properties of the nucleoplasm

Abstract The cell interior is highly crowded and far from thermodynamic equilibrium. This environment can dramatically impact molecular motion and assembly, and therefore influence subcellular organization and biochemical reaction rates. These effects depend strongly on length-scale, with the least information available at the important mesoscale (10-100 nanometers), which corresponds to the size of crucial regulatory molecules such as RNA polymerase II. It has been challenging to study the mesoscale physical properties of the nucleoplasm because previous methods were labor-intensive and perturbative. Here, we report nuclear Genetically Encoded Multimeric nanoparticles (nucGEMs). Introduction of a single gene leads to continuous production and assembly of protein-based bright fluorescent nanoparticles of 40 nm diameter. We implemented nucGEMs in budding and fission yeast and in mammalian cell lines. We found differences in particle motility between the nucleus and the cytosol at the mesoscale, that mitotic chromosome condensation ejects nucGEMs from the nucleus, and that nucGEMs are excluded from heterochromatin and the nucleolus. nucGEMs enable hundreds of nuclear rheology experiments per hour, and allow evolutionary comparison of the physical properties of the cytosol and nucleoplasm

The ESCRT machinery counteracts Nesprin-2G-mediated mechanical forces during nuclear envelope repair

Transient nuclear envelope ruptures during interphase (NERDI) occur due to cytoskeletal compressive forces at sites of weakened lamina, and delayed NERDI repair results in genomic instability. Nuclear envelope (NE) sealing is completed by endosomal sorting complex required for transport (ESCRT) machinery. A key unanswered question is how local compressive forces are counteracted to allow efficient membrane resealing. Here, we identify the ESCRT-associated protein BROX as a crucial factor required to accelerate repair of the NE. Critically, BROX binds Nesprin-2G, a component of the linker of nucleoskeleton and cytoskeleton complex (LINC). This interaction promotes Nesprin-2G ubiquitination and facilitates the relaxation of mechanical stress imposed by compressive actin fibers at the rupture site. Thus, BROX rebalances excessive cytoskeletal forces in cells experiencing NE instability to promote effective NERDI repair. Our results demonstrate that BROX coordinates mechanoregulation with membrane remodeling to ensure the maintenance of nuclear-cytoplasmic compartmentalization and genomic stability