title: Quiescence induces epigenetic changes in bovine fibroblasts and improves their reprogramming into cloned embryos

authors: Kallingappa, Prasanna Kumar

subjects: Quiescence, Epigenetics, Reprogramming, Somatic cell nuclear transfer (SCNT), Histone Methylation, Histone Acetylation, Histone varients, DNA methylation, Polycomb group (PcG) proteins

abstract: Cloning by somatic cell nuclear transfer (SCNT) forces cells to lose their lineage-specific epigenetic marks and become totipotent again. This reprogramming process often results in epigenetic and transcriptional aberrations that compromise development. Development rates after SCNT can thus serve as a functional assay for genome-wide epigenetic reprogramming. Dolly the sheep, the first mammalian SCNT clone, was derived from a donor cell that was induced into quiescence by serum starvation. We hypothesized that quiescence alters the epigenetic status of donor cells and elevates their reprogrammability. In order to test this idea, we compared chromatin composition and cloning efficiency of serum-starved, quiescent (G₀), bovine fibroblasts vs non-starved, diploid G₁ controls. Mechanically synchronized G₁ cells were generated by mitotic shake-off and harvested within 3 h post-mitosis. Based on morphological assessment and EdU incorporation during continuous labeling, >93% of cells were captured in G₁.
Using quantitative confocal immunofluorescence microscopy and fluorometric ELISA, we show that G₀ fibroblasts were significantly hypomethylated at lysines (K) of histone 3 (H3), specifically H3K4me3, H3K9me2, H3K9me3 and H3K27me3, but not H3K9me1. Histone acetylation was reduced at H3K9 and H4K5, increased at H3K12 and remained unchanged at H3K16. G₀ cells also significantly reduced DNAme. In addition, they significantly down-regulated the nuclear abundance of RNA polymerase II, histone variant H2A.Z, as well as Polycomb group (PcG) proteins EED, SUZ12, PHC1 and RING2. Histone variant H3.3, PcG proteins EZH2 and histone deacetylase HDAC1 did not change compared to the G₁ controls. Following NT into metaphase-arrested oocytes, G₀ DNA condensed slower than that of G₁ cells, indicating a more relaxed chromatin configuration. After seven days of in vitro culture, H3K9me3, but not H3K4me3, H3K27me3, SUZ12 and RING2, remained hypomethylated in G₀- vs G₁-derived NT blastocysts, both in the inner cell mass and trophectoderm. Furthermore, G₀ donors significantly improved development into cloned blastocysts. In conclusion, quiescence induced long-term epigenetic changes, specifically H3K9me3 hypomethylation, that correlated with increased donor cell reprogrammability.

text: Quiescence induces epigenetic changes in bovine fibroblasts and improves their reprogramming into cloned embryos Cloning by somatic cell nuclear transfer (SCNT) forces cells to lose their lineage-specific epigenetic marks and become totipotent again. This reprogramming process often results in epigenetic and transcriptional aberrations that compromise development. Development rates after SCNT can thus serve as a functional assay for genome-wide epigenetic reprogramming. Dolly the sheep, the first mammalian SCNT clone, was derived from a donor cell that was induced into quiescence by serum starvation. We hypothesized that quiescence alters the epigenetic status of donor cells and elevates their reprogrammability. In order to test this idea, we compared chromatin composition and cloning efficiency of serum-starved, quiescent (G₀), bovine fibroblasts vs non-starved, diploid G₁ controls. Mechanically synchronized G₁ cells were generated by mitotic shake-off and harvested within 3 h post-mitosis. Based on morphological assessment and EdU incorporation during continuous labeling, >93% of cells were captured in G₁.
Using quantitative confocal immunofluorescence microscopy and fluorometric ELISA, we show that G₀ fibroblasts were significantly hypomethylated at lysines (K) of histone 3 (H3), specifically H3K4me3, H3K9me2, H3K9me3 and H3K27me3, but not H3K9me1. Histone acetylation was reduced at H3K9 and H4K5, increased at H3K12 and remained unchanged at H3K16. G₀ cells also significantly reduced DNAme. In addition, they significantly down-regulated the nuclear abundance of RNA polymerase II, histone variant H2A.Z, as well as Polycomb group (PcG) proteins EED, SUZ12, PHC1 and RING2. Histone variant H3.3, PcG proteins EZH2 and histone deacetylase HDAC1 did not change compared to the G₁ controls. Following NT into metaphase-arrested oocytes, G₀ DNA condensed slower than that of G₁ cells, indicating a more relaxed chromatin configuration. After seven days of in vitro culture, H3K9me3, but not H3K4me3, H3K27me3, SUZ12 and RING2, remained hypomethylated in G₀- vs G₁-derived NT blastocysts, both in the inner cell mass and trophectoderm. Furthermore, G₀ donors significantly improved development into cloned blastocysts. In conclusion, quiescence induced long-term epigenetic changes, specifically H3K9me3 hypomethylation, that correlated with increased donor cell reprogrammability.

year: 2013

title: The Impacts of Social Media and Electronic Mediating: Improving Educational Outcomes for Saudi Women within an Electronic Social Networking Environment

authors: Alkahtani, Norah

subjects: Social Networking

abstract: This dissertation reports on a study about the effects of social media, such as social networks used by female university students in the context of a gender segregated system in Saudi Arabia. 

The result clearly reflects that social media has a positive effect on the collaborations of students, as they are able to keep their cultural and religious values without physically intermingling with the opposite sex, which is a requirement of religion and culture.

This research was carried out through two stages. The first stage was an exploratory study carried out for post graduate Saudi students at The University of Waikato. Students from both genders were granted scholarships from the King of Saudi Arabia, “King Abdullah”. They have had the experience of studying within two different cultures, in Saudi Arabia and New Zealand. They have had good experiences within the mixed gender environment at The University of Waikato.  

The second stage was action research, to diagnose the importance of social networks in the education system for Saudi females at Saudi universities. The segregation between genders in Saudi Arabia is a big issue at universities but with the appearance of social networks such as Facebook genders can interact easily without being physically in the same place. 

The action research took place through three cycles at three different universities; King Saud University and Imam University, which are public universities and the last cycle in private Universities, in Riyadh. 

The results collecting data revealed that electronic social networks could help promote collaboration between both genders in Saudi Arabia. Both genders felt more comfortable dealing with the opposite gender under the umbrella of education without harming their culture or religion.

text: The Impacts of Social Media and Electronic Mediating: Improving Educational Outcomes for Saudi Women within an Electronic Social Networking Environment This dissertation reports on a study about the effects of social media, such as social networks used by female university students in the context of a gender segregated system in Saudi Arabia. 

The result clearly reflects that social media has a positive effect on the collaborations of students, as they are able to keep their cultural and religious values without physically intermingling with the opposite sex, which is a requirement of religion and culture.

This research was carried out through two stages. The first stage was an exploratory study carried out for post graduate Saudi students at The University of Waikato. Students from both genders were granted scholarships from the King of Saudi Arabia, “King Abdullah”. They have had the experience of studying within two different cultures, in Saudi Arabia and New Zealand. They have had good experiences within the mixed gender environment at The University of Waikato.  

The second stage was action research, to diagnose the importance of social networks in the education system for Saudi females at Saudi universities. The segregation between genders in Saudi Arabia is a big issue at universities but with the appearance of social networks such as Facebook genders can interact easily without being physically in the same place. 

The action research took place through three cycles at three different universities; King Saud University and Imam University, which are public universities and the last cycle in private Universities, in Riyadh. 

The results collecting data revealed that electronic social networks could help promote collaboration between both genders in Saudi Arabia. Both genders felt more comfortable dealing with the opposite gender under the umbrella of education without harming their culture or religion.

year: 2012

title: The southern initiative: How indigenous values inspire social innovation and impact

authors: Niu, Xiaoliang

abstract: Indigenous values are increasingly recognised in helping organisations contribute to wellbeing within and beyond the workplace. Adopting the theoretical lens of Māori economies of wellbeing, this case study examines how The Southern Initiative (TSI), a unit within Auckland Council, incorporates Māori values to co-create place-based solutions and foster whānau (family) wellbeing. Through kōrero (conversations) with three people, a wānanga (collaborative discussion) with TSI members, and analysis of organisational literature, we identified how TSI's organising approach synthesises social innovation and bureaucracy. We found that indigeneity-embedded intrapreneurship, distributed leadership, and whānau-centred design support TSI's innovations. Mana (prestige) emerged as a primary organising principle, sustaining TSI's approach to achieving systemic change. By bridging Indigenous paradigms and conventional managerial practice, this case study demonstrates how Māori values can transform public sector management, elevate social justice, and encourage community resilience. These findings highlight culturally grounded frameworks for delivering social impact and shaping equitable outcomes.

text: The southern initiative: How indigenous values inspire social innovation and impact Indigenous values are increasingly recognised in helping organisations contribute to wellbeing within and beyond the workplace. Adopting the theoretical lens of Māori economies of wellbeing, this case study examines how The Southern Initiative (TSI), a unit within Auckland Council, incorporates Māori values to co-create place-based solutions and foster whānau (family) wellbeing. Through kōrero (conversations) with three people, a wānanga (collaborative discussion) with TSI members, and analysis of organisational literature, we identified how TSI's organising approach synthesises social innovation and bureaucracy. We found that indigeneity-embedded intrapreneurship, distributed leadership, and whānau-centred design support TSI's innovations. Mana (prestige) emerged as a primary organising principle, sustaining TSI's approach to achieving systemic change. By bridging Indigenous paradigms and conventional managerial practice, this case study demonstrates how Māori values can transform public sector management, elevate social justice, and encourage community resilience. These findings highlight culturally grounded frameworks for delivering social impact and shaping equitable outcomes.

year: 2025