title: Hyperglycaemia and oxidative stress upregulate HSP60 & HSP70 expression in HeLa cells

authors: Hall, Luke

subjects: HSP60

abstract: Heat Shock Proteins 60 & 70 (HSP60 & HSP70) are intracellular protein that has been shown to be present at elevated levels in systemic circulation in Type 2 Diabetes mellitus (T2DM) patients. Conditions that lead to its secretion, and the mechanism of its translocation from cells, have not yet been defined. The aim of this study was to determine if specific cell stressors associated with T2DM, namely hyperglycaemia and oxidative stress, result in the upregulation of HSP60 in human cells in vitro. Human HeLa cells were grown in media supplemented with 100 mM glucose, 200 μM hydrogen peroxide (H₂O₂), and 50 μM sodium azide. Initially, the effect of these treatments on cell growth rate was examined, with each treatment significantly inhibiting growth rate. LDH and MTT assays were also used to successfully demonstrate that these treatments do not significantly increase cell lysis, but do significantly impair mitochondrial dehydrogenase activity. To confirm this mitochondria specific form of inhibition, DCFDA assay were used to investigate any increases in intracellular reactive oxygen species (ROS) generation. All three treatments resulted in significantly increased ROS generation, with greater ROS production occurring with a greater exposure time. Interestingly, when the protein levels of HSP60 and HSP70 were measured after 3 and 7 days of exposure of the HeLa cells to 100 mM glucose, 200 μM H₂O₂, and 50 μM sodium azide significant induction of these two molecular stress proteins were observed ranging from 2.43-5.08 fold compared to untreated control cells.

text: Hyperglycaemia and oxidative stress upregulate HSP60 & HSP70 expression in HeLa cells Heat Shock Proteins 60 & 70 (HSP60 & HSP70) are intracellular protein that has been shown to be present at elevated levels in systemic circulation in Type 2 Diabetes mellitus (T2DM) patients. Conditions that lead to its secretion, and the mechanism of its translocation from cells, have not yet been defined. The aim of this study was to determine if specific cell stressors associated with T2DM, namely hyperglycaemia and oxidative stress, result in the upregulation of HSP60 in human cells in vitro. Human HeLa cells were grown in media supplemented with 100 mM glucose, 200 μM hydrogen peroxide (H₂O₂), and 50 μM sodium azide. Initially, the effect of these treatments on cell growth rate was examined, with each treatment significantly inhibiting growth rate. LDH and MTT assays were also used to successfully demonstrate that these treatments do not significantly increase cell lysis, but do significantly impair mitochondrial dehydrogenase activity. To confirm this mitochondria specific form of inhibition, DCFDA assay were used to investigate any increases in intracellular reactive oxygen species (ROS) generation. All three treatments resulted in significantly increased ROS generation, with greater ROS production occurring with a greater exposure time. Interestingly, when the protein levels of HSP60 and HSP70 were measured after 3 and 7 days of exposure of the HeLa cells to 100 mM glucose, 200 μM H₂O₂, and 50 μM sodium azide significant induction of these two molecular stress proteins were observed ranging from 2.43-5.08 fold compared to untreated control cells.

year: 2013

title: Timing of initiation of reverse displacement on the Taranaki Fault, northern Taranaki Basin: Constraints from the on land record (Oligocene Te Kuiti Group)

authors: Tripathi, Anand Ratnakar Prasad

subjects: Sequence Stratigraphy

abstract: Structures associated with the wedge of basement overthrust into Taranaki Basin along the Taranaki Fault, are regarded as hydrocarbon plays and have been tested by drilling through the tip of the overthrust. The timing of initiation of reverse displace ment on Taranaki Fault is difficult to interpret from available seismic reflection data across it because the evidence has been masked by later movements. The record from the basin, as summarised in King & Thrasher (1996), suggests that the fault evolved from normal to reverse character during the mid-Oligocene. This was inferred from formation of a foredeep parallel to, and west of, Taranaki Fault and a marked increase in its paleo-water depth, as indicated by foraminiferal assemblages of Late Oligocene age.  
A comprehensive re-assessment of the lithostratigraphy and sequence stratigraphy of the Late Eocene-Oligocene Te Kuiti Group exposed on land east of Taranaki Fault in central-western North Island, between Port Waikato and Awakino, provides new constraints on the early history of Taranaki Fault displacement. New age control has been achieved by a review of existing foraminiferal biostratigraphy combined with determination of Sr isotope ages from macrofossil samples. Six unconformity-bound sequences have been identified and mapped within the Te Kuiti Group. A major subaerial unconformity between sequences TK3 and TK4 combined with a basinward shift in the position of onlap for sequence TK4 indicate a dramatic change in stratigraphic development and basin dynamics during the mid-upper Whaingaroan at c. 29 Ma, corresponding to the change from mild extension (sag basin) to shortening across the Taranaki Fault Zone. We consider sequences TK4 – TK6 to each represent tectonic cycles of subsidence and basin inversion and we attribute the origin of these cycles to periodic locking of the Taranaki Fault décollement in underlying Murihiku basement, the accumulating strain causing uplift in the basin east of the fault zone, followed by free displacement, relaxation in the upper crust and subsidence. A 1st order model is presented of the Late Oligocene to earliest Miocene vertical and horizontal displacement of basement on the Taranaki Fault Zone for a west –east transect through Awakino. It implies that the mid- to Late Oligocene displace¬ment on the fault zone in the vicinity of Awakino was episodic, and that the thrust belt was narrow (c. 15 km). North of Kawhia Harbour there will have been a different displacement history with most of the total displacement occurring during the devel opment of the c. 29 Ma unconformity at the base of Sequence TK4, whereas to the south between Awakino and Kawhia Harbour the majority of the total displacement occurred during the Otaian and at the end of it. The model also shows that the start of reverse/thrust displacement on Taranaki Fault must have involved the development of a completely new fault trace(s), rather than involving a change of sense of movement on the pre-existing normal fault. The Manganui Fault is part of the Taranaki Fault Zone and probably became active at c. 27 Ma during development of the unconformity between sequences TK4 & TK5. The model presented here has been validated against the subsurface Oligocene stratigraphy in Taranaki Basin.

text: Timing of initiation of reverse displacement on the Taranaki Fault, northern Taranaki Basin: Constraints from the on land record (Oligocene Te Kuiti Group) Structures associated with the wedge of basement overthrust into Taranaki Basin along the Taranaki Fault, are regarded as hydrocarbon plays and have been tested by drilling through the tip of the overthrust. The timing of initiation of reverse displace ment on Taranaki Fault is difficult to interpret from available seismic reflection data across it because the evidence has been masked by later movements. The record from the basin, as summarised in King & Thrasher (1996), suggests that the fault evolved from normal to reverse character during the mid-Oligocene. This was inferred from formation of a foredeep parallel to, and west of, Taranaki Fault and a marked increase in its paleo-water depth, as indicated by foraminiferal assemblages of Late Oligocene age.  
A comprehensive re-assessment of the lithostratigraphy and sequence stratigraphy of the Late Eocene-Oligocene Te Kuiti Group exposed on land east of Taranaki Fault in central-western North Island, between Port Waikato and Awakino, provides new constraints on the early history of Taranaki Fault displacement. New age control has been achieved by a review of existing foraminiferal biostratigraphy combined with determination of Sr isotope ages from macrofossil samples. Six unconformity-bound sequences have been identified and mapped within the Te Kuiti Group. A major subaerial unconformity between sequences TK3 and TK4 combined with a basinward shift in the position of onlap for sequence TK4 indicate a dramatic change in stratigraphic development and basin dynamics during the mid-upper Whaingaroan at c. 29 Ma, corresponding to the change from mild extension (sag basin) to shortening across the Taranaki Fault Zone. We consider sequences TK4 – TK6 to each represent tectonic cycles of subsidence and basin inversion and we attribute the origin of these cycles to periodic locking of the Taranaki Fault décollement in underlying Murihiku basement, the accumulating strain causing uplift in the basin east of the fault zone, followed by free displacement, relaxation in the upper crust and subsidence. A 1st order model is presented of the Late Oligocene to earliest Miocene vertical and horizontal displacement of basement on the Taranaki Fault Zone for a west –east transect through Awakino. It implies that the mid- to Late Oligocene displace¬ment on the fault zone in the vicinity of Awakino was episodic, and that the thrust belt was narrow (c. 15 km). North of Kawhia Harbour there will have been a different displacement history with most of the total displacement occurring during the devel opment of the c. 29 Ma unconformity at the base of Sequence TK4, whereas to the south between Awakino and Kawhia Harbour the majority of the total displacement occurred during the Otaian and at the end of it. The model also shows that the start of reverse/thrust displacement on Taranaki Fault must have involved the development of a completely new fault trace(s), rather than involving a change of sense of movement on the pre-existing normal fault. The Manganui Fault is part of the Taranaki Fault Zone and probably became active at c. 27 Ma during development of the unconformity between sequences TK4 & TK5. The model presented here has been validated against the subsurface Oligocene stratigraphy in Taranaki Basin.

year: 2008

title: Evaluation of Rongo Atea: Alcohol and Other Drug Treatment Centre for Adolescents

authors: Paki, Helen Mary

subjects: Alcohol

abstract: Abstract Rongo Ātea is a residential abstinence-based Kaupapa Māori alcohol and other drug programme accommodating youth between the ages of 13 and 17. This evaluation investigates the role that staff play at Rongo Ātea along with a number of other factors including the physical environment; stages of change; programme implementation and aftercare. As a Kaupapa Māori programme, the role of how culture and identity can influence positive change is also explored along with the development of a youth-focused approach. I utilised a three phase framework borrowed from the work of Mason Durie (2008) to organise themes. These three phases, which Durie (2008) based on marae encounters, include Whakapiri (Engagement); Whakamarama (Enlightenment); and Whakamana (Empowerment) as they aptly reflect a three phase intervention approach: detoxification and early programme engagement; learning and development through ongoing programme commitment; and post treatment outcomes and aftercare. The data collection phase of this evaluation took place in 2006 and was initiated by the manager of Rongo Ātea who requested an evaluation to identify programme strengths and limitations from the perspectives of staff and students, and to make recommendations to Rongo Ātea that would assist with further programme developments. I utilised a collaborative and participatory approach (Bishop, 1996; Patton, 1990). Kaupapa Māori research principles were reflected in the use of ‘kanohi kitea’— face to face contact (Smith, 1999). With appropriate training and management support, staff could have a greater influence on programme outcomes. Evaluation findings suggest that drug and alcohol intervention and treatment for young people in New Zealand is significantly under-resourced, particularly in the areas of detoxification and aftercare. To be effective, residential treatment programmes should consist of a three stage programme covering detoxification; treatment; and aftercare incorporating an integrated approach. A greater emphasis on working with whānau alongside the young person is recommended.

text: Evaluation of Rongo Atea: Alcohol and Other Drug Treatment Centre for Adolescents Abstract Rongo Ātea is a residential abstinence-based Kaupapa Māori alcohol and other drug programme accommodating youth between the ages of 13 and 17. This evaluation investigates the role that staff play at Rongo Ātea along with a number of other factors including the physical environment; stages of change; programme implementation and aftercare. As a Kaupapa Māori programme, the role of how culture and identity can influence positive change is also explored along with the development of a youth-focused approach. I utilised a three phase framework borrowed from the work of Mason Durie (2008) to organise themes. These three phases, which Durie (2008) based on marae encounters, include Whakapiri (Engagement); Whakamarama (Enlightenment); and Whakamana (Empowerment) as they aptly reflect a three phase intervention approach: detoxification and early programme engagement; learning and development through ongoing programme commitment; and post treatment outcomes and aftercare. The data collection phase of this evaluation took place in 2006 and was initiated by the manager of Rongo Ātea who requested an evaluation to identify programme strengths and limitations from the perspectives of staff and students, and to make recommendations to Rongo Ātea that would assist with further programme developments. I utilised a collaborative and participatory approach (Bishop, 1996; Patton, 1990). Kaupapa Māori research principles were reflected in the use of ‘kanohi kitea’— face to face contact (Smith, 1999). With appropriate training and management support, staff could have a greater influence on programme outcomes. Evaluation findings suggest that drug and alcohol intervention and treatment for young people in New Zealand is significantly under-resourced, particularly in the areas of detoxification and aftercare. To be effective, residential treatment programmes should consist of a three stage programme covering detoxification; treatment; and aftercare incorporating an integrated approach. A greater emphasis on working with whānau alongside the young person is recommended.

year: 2010