title: Postantibiotic effect of purified melittin from honeybee (Apis mellifera) venom against Escherichia coli and Staphylococcus aureus

authors: Han, Sangmi

subjects: bee venom

abstract: Since the ancient times, the antibacterial application of honeybee venom (BV) has been practised and persisted. We investigated the antibacterial activity of whole BV and purified melittin against Escherichia coli and Staphylococcus aureus by the minimum inhibitory concentrations (MICs) and the postantibiotic effects (PAEs). The in vitro PAEs of whole BV and isolated melittin were determined using E. coli and S. aureus. The PAEs of whole BV against E. coli and S. aureus were 0.15 h (1   MIC), 2.4 h (5   MIC), and 3.45 h (1   MIC), respectively. The PAEs of melittin against E. coli and S. aureus were 0.1 h (1   MIC), 3.2 h (5   MIC), and 4.35 h (1   MIC), respectively. These results suggest that whole BV and melittin will be developed as a novel antibacterial drug.

text: Postantibiotic effect of purified melittin from honeybee (Apis mellifera) venom against Escherichia coli and Staphylococcus aureus Since the ancient times, the antibacterial application of honeybee venom (BV) has been practised and persisted. We investigated the antibacterial activity of whole BV and purified melittin against Escherichia coli and Staphylococcus aureus by the minimum inhibitory concentrations (MICs) and the postantibiotic effects (PAEs). The in vitro PAEs of whole BV and isolated melittin were determined using E. coli and S. aureus. The PAEs of whole BV against E. coli and S. aureus were 0.15 h (1   MIC), 2.4 h (5   MIC), and 3.45 h (1   MIC), respectively. The PAEs of melittin against E. coli and S. aureus were 0.1 h (1   MIC), 3.2 h (5   MIC), and 4.35 h (1   MIC), respectively. These results suggest that whole BV and melittin will be developed as a novel antibacterial drug.

year: 2009

title: A promoter characterisation of the Ets transcription factor gene, Ese-2/Elf-5

authors: Speed, Kathy Ann

abstract: E26 transformation specific (Ets) proteins are a versatile class of transcription factors. They act both up and downstream of signal transduction cascades, are causative in cancer generation and are able to control the expression of other transcription factors. Thus they play an important role in cellular growth, proliferation and differentiation. The ets gene family is characterised by an 85 amino acid DNA binding domain and currently consists of around 30 members. They have been shown to be proto-oncogenes and it is this feature that has elicited much research into their regulation and function in both normal and neoplasic tissue. The high level of conservation within the ETS domain of the family identifies all Ets proteins as sequence-specific DNA-binding proteins. Many have been identified as having a role in the regulation of gene expression. The retention of many ets genes throughout evolution implies there would be few redundant genes (Thomas, 1993) and therefore each gene would have a unique biological role. Because of the common DNA binding domain, the regulation of the ets genes must be complex to provide specificity of action throughout the family (Laudet et al., 1999).

A cDNA sequence was isolated from mammary gland tissue and characterised as belonging to the Ets family of transcription factors. The gene was designated BrETS for Breast ETS. Recent publications have described the gene as Elf-5 (Zhou et al., 1998) and ESE-2 (Oettgen et al., 1999). BrEts showed a restricted pattern of spatial expression, being strongly expressed in the mammary gland. Its expression in the mammary gland also exhibits a tightly controlled temporal pattern, which implicates it in mammary processes such as development and differentiation. As BrETS is highly expressed in the mammary gland (where intense extracellular remodelling occurs), and has been localised to the epithelial tissue, it may play a role in the regulation of matrix metalloproteinases or their inhibitors.

Genomic DNA spanning the BrETS locus was isolated in this study. It was found to encompass approximately 3kb of upstream sequence, the first two exons and intron 1. Multiple transcription start sites have been identified, indicative of a TATA-less, tissue- specific gene. Deletion constructs of the proximal promoter (850bp) were examined in cell culture. The mammary gland derived cell lines HC11 and Comma1D were used, in conjunction with a chloramphenicol acetyltransferase reporter system. A region of sequence between -70 and -200bp was identified as having up-regulated transcriptional activity above an empty expression vector in cell culture. Analysis of more distal promoter constructs resulted in decreased activity, to a level representative of basal activity. The first intron (6kb) was also examined in vitro as a complete construct and as smaller fragments. Low levels of reporter activity were seen with these constructs in cell culture. DNasel hypersensitive analysis on the immediate promoter region revealed the presence of a hypersensitive site within the first intron. This implies a regulatory element of potential importance is located here, though further research is required to determine the element(s) and significance.

The identification of regulatory regions may allow for the determination of tissue-specific elements and regions that control the expression of the gene. This information may be later extrapolated and manipulated in vivo as a means of controlling the expression of the BrETS gene.

text: A promoter characterisation of the Ets transcription factor gene, Ese-2/Elf-5 E26 transformation specific (Ets) proteins are a versatile class of transcription factors. They act both up and downstream of signal transduction cascades, are causative in cancer generation and are able to control the expression of other transcription factors. Thus they play an important role in cellular growth, proliferation and differentiation. The ets gene family is characterised by an 85 amino acid DNA binding domain and currently consists of around 30 members. They have been shown to be proto-oncogenes and it is this feature that has elicited much research into their regulation and function in both normal and neoplasic tissue. The high level of conservation within the ETS domain of the family identifies all Ets proteins as sequence-specific DNA-binding proteins. Many have been identified as having a role in the regulation of gene expression. The retention of many ets genes throughout evolution implies there would be few redundant genes (Thomas, 1993) and therefore each gene would have a unique biological role. Because of the common DNA binding domain, the regulation of the ets genes must be complex to provide specificity of action throughout the family (Laudet et al., 1999).

A cDNA sequence was isolated from mammary gland tissue and characterised as belonging to the Ets family of transcription factors. The gene was designated BrETS for Breast ETS. Recent publications have described the gene as Elf-5 (Zhou et al., 1998) and ESE-2 (Oettgen et al., 1999). BrEts showed a restricted pattern of spatial expression, being strongly expressed in the mammary gland. Its expression in the mammary gland also exhibits a tightly controlled temporal pattern, which implicates it in mammary processes such as development and differentiation. As BrETS is highly expressed in the mammary gland (where intense extracellular remodelling occurs), and has been localised to the epithelial tissue, it may play a role in the regulation of matrix metalloproteinases or their inhibitors.

Genomic DNA spanning the BrETS locus was isolated in this study. It was found to encompass approximately 3kb of upstream sequence, the first two exons and intron 1. Multiple transcription start sites have been identified, indicative of a TATA-less, tissue- specific gene. Deletion constructs of the proximal promoter (850bp) were examined in cell culture. The mammary gland derived cell lines HC11 and Comma1D were used, in conjunction with a chloramphenicol acetyltransferase reporter system. A region of sequence between -70 and -200bp was identified as having up-regulated transcriptional activity above an empty expression vector in cell culture. Analysis of more distal promoter constructs resulted in decreased activity, to a level representative of basal activity. The first intron (6kb) was also examined in vitro as a complete construct and as smaller fragments. Low levels of reporter activity were seen with these constructs in cell culture. DNasel hypersensitive analysis on the immediate promoter region revealed the presence of a hypersensitive site within the first intron. This implies a regulatory element of potential importance is located here, though further research is required to determine the element(s) and significance.

The identification of regulatory regions may allow for the determination of tissue-specific elements and regions that control the expression of the gene. This information may be later extrapolated and manipulated in vivo as a means of controlling the expression of the BrETS gene.

year: 2002

title: Te Whatu Kakahu: Assessment in Kaupapa Maori Early Childhood Practice

authors: Rameka, Lesley Kay

subjects: Kaupapa Maori

abstract: Through the exploration of Kaupapa Māori assessment approaches, I examine the reclaiming and reframing of Māori ways of knowing and being within early childhood practice. Assessment is the vehicle for reclaiming and reframing while Kaupapa Māori theory is the fuel that ignites and drives the vehicle.

The effects of successive education policies remain today with Māori children, and their families continuing to disengage from education and consistently receiving disproportionately lower outcomes, opportunities and benefits. Reclaiming and reframing Māori ways of knowing and being within early childhood assessment thinking and practice is a means of addressing the cultural and educational disparities faced by Māori children within an education system that upholds western cultural and educational superiority, privilege and truths.  Key questions in this process of reclaiming and reframing are: Who has the power to define?  Whose truths are being reflected and how are these truths constructed? 

The metaphor of whatu kākahu or weaving of clothing has been used to frame this thesis. The process has involved weaving the Kaupapa Māori theory elements of conscientisation, resistance, transformative praxis and Māori ways of knowing and being, across and within historical, cultural and educational paradigms and understandings, to fashion assessment kākahu that afford comfort, warmth and flexibility in a contemporary early childhood context. This research case studies the progress of three Māori early childhood services and kōhanga reo towards the development of Kaupapa Māori early childhood assessment understandings and framings (kākahu), that reflect their particular ways of knowing and being, context and aspirations for children. This thesis has been about their assessment journeys. These journeys are a work in progress and that work continues.

A qualitative, Kaupapa Māori research methodology was used to gather, collate and analyse data in this research.  In accordance with Kaupapa Māori research aspirations and expectations, this research focuses on areas of importance and concern for Māori, and involved retrieving space for Māori voices to be heard. This research can be seen as a means of privileging Māori approaches, perspectives and ways of knowing and being in early childhood assessment practice.

Kaupapa Māori assessment is an important agenda for early childhood.   It builds upon Māori philosophical and epistemological understandings that express Māori ways of knowing and being. Kaupapa Māori assessment is able to contribute significantly to children’s learning and potential growth and is an important tool in constructing educational outcomes for Māori children. It is therefore an important agenda for early childhood.

text: Te Whatu Kakahu: Assessment in Kaupapa Maori Early Childhood Practice Through the exploration of Kaupapa Māori assessment approaches, I examine the reclaiming and reframing of Māori ways of knowing and being within early childhood practice. Assessment is the vehicle for reclaiming and reframing while Kaupapa Māori theory is the fuel that ignites and drives the vehicle.

The effects of successive education policies remain today with Māori children, and their families continuing to disengage from education and consistently receiving disproportionately lower outcomes, opportunities and benefits. Reclaiming and reframing Māori ways of knowing and being within early childhood assessment thinking and practice is a means of addressing the cultural and educational disparities faced by Māori children within an education system that upholds western cultural and educational superiority, privilege and truths.  Key questions in this process of reclaiming and reframing are: Who has the power to define?  Whose truths are being reflected and how are these truths constructed? 

The metaphor of whatu kākahu or weaving of clothing has been used to frame this thesis. The process has involved weaving the Kaupapa Māori theory elements of conscientisation, resistance, transformative praxis and Māori ways of knowing and being, across and within historical, cultural and educational paradigms and understandings, to fashion assessment kākahu that afford comfort, warmth and flexibility in a contemporary early childhood context. This research case studies the progress of three Māori early childhood services and kōhanga reo towards the development of Kaupapa Māori early childhood assessment understandings and framings (kākahu), that reflect their particular ways of knowing and being, context and aspirations for children. This thesis has been about their assessment journeys. These journeys are a work in progress and that work continues.

A qualitative, Kaupapa Māori research methodology was used to gather, collate and analyse data in this research.  In accordance with Kaupapa Māori research aspirations and expectations, this research focuses on areas of importance and concern for Māori, and involved retrieving space for Māori voices to be heard. This research can be seen as a means of privileging Māori approaches, perspectives and ways of knowing and being in early childhood assessment practice.

Kaupapa Māori assessment is an important agenda for early childhood.   It builds upon Māori philosophical and epistemological understandings that express Māori ways of knowing and being. Kaupapa Māori assessment is able to contribute significantly to children’s learning and potential growth and is an important tool in constructing educational outcomes for Māori children. It is therefore an important agenda for early childhood.

year: 2012