Excluding indigenous bioethical concerns when regulating frozen embryo storage: An Aotearoa New Zealand case study.

This article undertakes a close reading of the parliamentary debates associated with the topic of embryo cryopreservation in Aotearoa New Zealand. From our critical readings, we argue that there is a lack of transparency over the ethical reasons for enforcing a maximum storage limit. We demonstrate that arguments for the retention of this limit are associated (in New Zealand) with arguments based upon 'build-up avoidance' and 'conflict avoidance' as social goods based on Pakeha [New Zealander of European descent] cultural world views rather than identifiable universal ethical principles. We illustrate that the avoidance of embryo accumulation and related conflict was only achieved by the denial of indigenous spiritual and cultural concerns, while also shifting the ethical burdens of disposition on to clinic staff and those members of the public who protested against enforced cryopreserved embryo disposal. The Pakeha cultural concept of 'tidy housekeeping' emerges as a presumed ethical and social good in the New Zealand situation. This is despite abundant literature documenting the suffering created through forced decision-making upon disposition.

Plasma fatty acids as markers for desaturase and elongase activities in spinal cord injured males.

OBJECTIVE: To investigate the use of surrogate plasma fatty acid analysis to provide further insights into the underlying adiposity and the development of metabolic syndrome in men with spinal cord injury (SCI). DESIGN: Case-control, cross-sectional study. SETTING: Community-based individuals with spinal cord injury and healthy controls. PARTICIPANTS: Twenty men with SCI age, height and weight matched with 20 able-bodied controls. OUTCOME MEASURES: Lean tissue (LTM) and fat mass (FM) were determined using dual energy X-ray absorptiometry. Fasting blood samples were taken for analysis of fatty acids, adiponectin, insulin, glucose and leptin. Enzymatic indices were calculated using relevant fatty acids. RESULTS: Total FM, leptin, stearoyl coenzyme A desaturase (SCD) Δ9 (SCD-16, 16:1/16:0, and SCD-18, 18:1/18:0) indices and Δ6 desaturase index were significantly higher (P < 0.05) in the SCI group than the controls. Significant differences between the groups was observed for several individual fatty acids. Correlational analysis revealed a different pattern between blood biomarkers and indices of SCDs, de novo lipogenesis and elongase. Associations between the desaturase and elongase indices and biomarkers in the controls followed those reported elsewhere for able bodied participants; the same associations were not observed in the SCI group. CONCLUSION: We have identified disturbances in fatty acid biosynthesis in SCI individuals likely associated with the development of adipose tissue below the lesion and a decrease in LTM. Loss of LTM may disturb the normal skeletal muscle-fatty acid metabolic processes leading to the disruption of metabolic homeostasis, previously identified in persons with SCI.

The futility of fertility research? Barriers to embryo research in New Zealand.

AIMS: Successive New Zealand Health Ministers have failed to approve guidelines for research using viable human embryos, which effectively places a blanket ban on all research that "uses" viable human embryos in this country. This includes research that aims to improve currently available reproductive technologies, illustrated by a failed application to ministerial ethics committees for a clinical research project investigating the efficacy of in vitro fertilisation procedures. However, no data currently exists describing the degree to which these restrictions are inhibiting reproductive research in this country. METHODS: We have conducted a qualitative survey of New Zealand researchers from 20 major academic, clinical and governmental institutes to qualify the impact these restrictions are having on New Zealand's research outputs. RESULTS: The results suggest dissatisfaction with the current guidelines, and the lack of guidance from the Ministry of Health and associated ethics committees regarding what constitutes embryo research and therefore what research can be performed. CONCLUSIONS: The lack of current guidelines regarding the use of embryos for research is restricting improvements to established reproductive technologies, and any future research. We suggest that the Minister of Health instructs ministerial advisory and ethics committees to review the current guidelines and to define the term "use of embryos".

Proteomic Analysis of Liver from Human Lipoprotein(a) Transgenic Mice Shows an Oxidative Stress and Lipid Export Response.

BACKGROUND: Mouse models of hypercholesterolaemia have been used to identify arterial proteins involved in atherosclerosis. As the liver is extremely sensitive to dyslipidemia, one might expect major changes in the abundance of liver proteins in these models even before atherosclerosis develops. METHODS: Lipid levels were measured and a proteomic approach was used to quantify proteins in the livers of mice with an elevated low-density lipoprotein (LDL) and the presence of lipoprotein(a) [Lp(a)] but no atherosclerosis. RESULTS: The livers of Lp(a) mice showed an increased triglyceride but reduced phospholipid and oxidised lipid content. Two-dimensional gel electrophoresis and mass spectrometry analysis identified 24 liver proteins with significantly increased abundance in Lp(a) mice (P<0.05). A bioinformatic analysis of the 24 proteins showed the major effect was that of an enhanced antioxidant and lipid efflux response with significant increases in antioxidant (Park7, Gpx1, Prdx6, and Sod1) and lipid metabolism proteins (Fabp4, Acaa2, apoA4, and ApoA1). Interestingly, human liver cells treated with Lp(a) showed significant increases in Gpx1 and Prdx6 but not Sod1 or Park7. CONCLUSIONS: The presence of human LDL and Lp(a) in mice promotes an enhanced flux of lipids into the liver which elicits an antioxidant and lipid export response before the onset of atherosclerosis. The antioxidant response can be reproduced in human liver cells treated with Lp(a).

Cyclic Variation of Cellular Clock Proteins in the Mouse Estrous Ovary.

BACKGROUND: The mammalian ovary is controlled by a number of biological rhythms, which regulate the recruitment and release of mature oocytes. The main objective of this study was to investigate the role of cellular clock proteins during follicle maturation in the mouse estrous ovary. METHODS: Immunohistochemical (IHC) studies were performed on ovaries from 50 estrous staged mice culled at two time points of 09:00 [day] and 01:00 [mid-point of the dark cycle]. Six antibodies were used to identify the expression of core cellular clock proteins (BMAL1, CLOCK, CRY1, CRY2, PER1 and PER2) within the ovary and four follicle stages, primordial, primary, antral and corpus lutea. IHC data was scored using the Allred protocol and significance determined by Mann-Whitney tests. Differences were considered significant at p<0.05. RESULTS: All four follicle stages presented greater BMAL1 and CLOCK protein scores during the day and up regulation of CRY1-2 and PER1-2 at night. In primordial follicles, BMAL1 and CLOCK increases were significant (p<0.05) and CRY-1 and PER-1 were highly significant (p<0.001), and CRY-2 did not reach significance. Primary follicles demonstrated a similar response with BMAL1 and CLOCK, and CRY-1, PER-1-2 all reaching significant expression (p<0.05; p<0.001; p<0.001 respectively). CRY-2 expression was not significant. Antral follicles did not show significant BMAL1 or CLOCK expression, CRY-1 and PER-1 were highly significant (p<0.001) and CRY-2 had a small but significant increase (p<0.05). Corpus lutea demonstrated significant BMAL1 increase but CLOCK had no significant variation. CRY-1, PER1-2 increases were highly significant (p<0.001) and CRY-2 was up regulated but failed to reach significance. CONCLUSION: The ovary demonstrated a cellular clock response to the light: dark cycle and in addition, as the ovarian follicles mature changes in the positive and negative arms of both clock responsive proteins were observed.

Valuing embryos as both commodities and singularities.

An argument put forward against gamete and embryo donation, sale and research, is that to do so would treat the gametes or embryos as objects with no intrinsic value as human. Instead, gametes and embryos created and used for donation, sale or research, can be considered more like a commodity created and traded for economic exchange--something that is valuable only for the amount of money or other goods and services that others are willing to exchange. While Kant asserts that humans have dignity rather than object worth, the provision of human gametes and embryos are progressively becoming utilities for resolving childlessness and for certain research investigations. In this paper we discuss the commodity market and the relationship to human reproduction material.

Choice, Rights, and Virtue: Prenatal Testing and Styles of Moral Reasoning in Aotearoa/New Zealand.

Using a Foucauldian biopower analytic, this article combines insights from several ethnographic research projects around the moral reasoning styles underpinning debates over selective reproductive technologies in Aotearoa/New Zealand. We show that divergent or shared public, private, state, individual, and community moral reasoning styles become highly politicized truth discourses that have the potential to, and at times do, affect one another, modifying a dominant, state-supported, principal-based bioethics framework. The styles of moral reasoning that we identify pivot on an aspirational cultural ideal of the provision of choice to citizens, which is taken as an appropriate position from which to regulate selective reproductive technologies.

Ethics of mitochondrial therapy for deafness.

Mitochondrial therapy may provide the relief to many families with inherited mitochondrial diseases. However, it also has the potential for use in non-fatal disorders such as inherited mitochondrial deafness, providing an option for correction of the deafness using assisted reproductive technology. In this paper we discuss the potential for use in correcting mitochondrial deafness and consider some of the issues for the deaf community.

Mannose-binding lectin 2 gene polymorphism and susceptibility to upper respiratory tract infection among endurance athletes.

The aim of this study was to investigate the influence of mannose-binding lectin 2 (MBL2)-exon-1 gene polymorphisms on upper respiratory tract infection (URTI) incidence among endurance athletes. To this end, 100 healthy elite male athletes participating in the study were classified as either healthy or prone to frequent URTI. Blood samples, DNA isolation, multiplex polymerase chain reaction (PCR) and conventional PCR-RFLP were performed. Genomic DNA was extracted from peripheral leukocytes of whole blood samples using the QIAmp DNA Blood Mini Kit. For comparison of the distribution of genotypes between two groups and for estimating odds ratios (OR) for URTI susceptibility in relation to the MBL2-exon-1 polymorphism, Pearson's chi-square and logistic regression method were used, respectively. The MBL2-exon-1 genotype distribution differed between athletes with URTI and healthy athletes (χ(2) = 7.81, p = 0.02). The AO and AO + OO genotypes of MBL2 were observed at a greater frequency in the illness-prone group compared with the healthy group (34.04% vs. 11.32%). In conclusion, findings from this study have identified a potential role of genetic variation in influencing the risk for URTI in athletic populations and single-nucleotide polymorphisms (SNPs) in the MBL2-exon-1 genes were associated with an altered risk profile. These measures may have a predictive value in the identification of individuals who are more likely to experience recurrent infections when exposed to high physical stress in the areas of athletic endeavour.

Numerical identity: the creation of tri-parental embryos to correct inherited mitochondrial disease.

Inherited mitochondrial disorders affect between 1 in 5000 to 1 in 8000 people. These are a heterogeneous group of maternally-inherited disorders, with an array of outcomes such as heart and liver failure, defects in energy metabolism, blindness, deafness, loss of motor skills and premature death. Recently the Human Fertilisation and Embryology Authority provided advice to the UK Government to permit the use of enucleated donated oocytes with normal (wild-type) mitochondria (a currently prohibited IVF technique) to be used as recipients of nuclear DNA from intending mothers to overcome transmission of mitochondrial disorders. In this short communication we present the basis for this radical new IVF technology, and discuss the implications for its use both in the context of treating a group of inherited disorders and the current New Zealand IVF legislation.

Screening criteria: the need to deal with new developments and ethical issues in newborn metabolic screening.

Newborn metabolic screening is the most widespread application of screening technology and provides the most comprehensive application of genetics in health services, where the Guthrie blood spot cards allow screening for metabolic diseases in close to 100 % of all newborn babies. Despite over 40 years of use and significant benefits to well in excess of 100,000 children worldwide, there is remarkably little consensus in what conditions should be screened for and response to new advances in medicine relating to programme expansion. In this article, the international criteria for newborn metabolic screening are considered, and we propose that these criteria are poorly developed in relation to the baby, its family and society as a whole. Additionally, the ethical issues that should inform the application of screening criteria are often not developed to a level where a consensus might easily be achieved. We also consider that when family interests are factored in to the decision-making process, they have a significant influence in determining the list of diseases in the panel, with countries or states incorporating family and societal values being the most responsive. Based on our analysis, we propose that decision criteria for metabolic screening in the newborn period should be adapted to specifically include parent and family interests, community values, patients' rights, duties of government and healthcare providers, and ethical arguments for action in the face of uncertainty.

Somatic reactivation of expression of the silent maternal Mest allele and acquisition of normal reproductive behaviour in a colony of Peg1/Mest mutant mice.

Genomic imprinting confers allele-specific expression in less than 1% of genes, in a parent-of-origin specific fashion. In humans and mice the Peg1/Mest gene (Mest) is maternally repressed, and paternally expressed. Mest is expressed in embryogenic mesoderm-derived tissues and in adult brain, and paternal mutations in Mest lead to growth retardation and defective maternal behaviour. Despite our current understanding of mechanisms associated with the establishment of imprinting of Mest and other imprinted genes, it is unclear to what extent Mest imprinting needs to be maintained in adult tissues. Aberrations of imprinting are known to occur in certain rare syndromes, and involve either inherited mutations, or constitutive epigenetic alterations occurring soon after fertilization. Imprinting abnormalities may also occur in the aging somatic tissues of adult individuals. Here we report an occurrence of post-embryonic somatic variability of Mest allelic expression in a colony of mice where heterozygotes at the imprinted Mest locus for a mutation inherited from the father spontaneously expressed the normally silenced allele from the mother. In addition, a newly acquired ability to overcome the deficit in maternal reproductive behaviour had occurred in the mutant mice, but this appeared not to be directly linked to the Mest mutation. Our results suggest that at least one allele of Mest expression is required in the somatic tissues of adult individuals and that under certain conditions (such as in the presence of a Mest insertional mutation or in an altered genetic background), somatically acquired alterations of allelic expression at the Mest locus may occur.

Varying amounts of different aldehydes present in the cryoprotectants dimethyl sulphoxide and 1,2-propanediol.

Using high-pressure liquid chromatography two cryoprotectant solvents, dimethyl sulphoxide (four manufacturers) and 1,2-propanediol (one manufacturer) were investigated for aldehyde content. Fractionation of the aldehydes by high pressure liquid chromatography identified up to 11 aldehydes and two ketones in both cryoprotectant solvents in varying concentrations, which differed between manufacturer and container type. Of the 11 aldehydes identified, formaldehyde and acetylaldehyde were consistently in the greatest concentrations. As the low molecular weight aldehydes identified contain reactive polarised carbonyl groups they represent a potential source of intracellular damage when used in oocyte cryopreservation.

Proteomic analysis of aortae from human lipoprotein(a) transgenic mice shows an early metabolic response independent of atherosclerosis.

BACKGROUND: Elevated low density lipoprotein (LDL) and lipoprotein(a) are independent risk factors for the development of atherosclerosis. Using a proteomic approach we aimed to determine early changes in arterial protein expression in transgenic mice containing both human LDL and lipoprotein(a) in circulation. METHODS AND RESULTS: Plasma lipid analyses showed the lipoprotein(a) transgenic mice had significantly higher lipid levels than wildtype, including a much increased LDL and high density lipoprotein (HDL) cholesterol. Analysis of aortae from lipoprotein(a) mice showed lipoprotein(a) accumulation but no lipid accumulation or foam cells, leaving the arteries essentially atherosclerosis free. Using two-dimensional gel electrophoresis and mass spectrometry, we identified 34 arterial proteins with significantly altered abundance (P<0.05) in lipoprotein(a) transgenic mice compared to wildtype including 17 that showed a ≥2 fold difference. Some proteins of interest showed a similarly altered abundance at the transcript level. These changes collectively indicated an initial metabolic response that included a down regulation in energy, redox and lipid metabolism proteins and changes in structural proteins at a stage when atherosclerosis had not yet developed. CONCLUSIONS: Our study shows that human LDL and lipoprotein(a) promote changes in the expression of a unique set of arterial proteins which may be early indicators of the metabolic disturbances preceding atherosclerosis.

Amino acid transport system L activity in developing mouse ovarian follicles.

BACKGROUND: Little is known about metabolic processes in the developing ovarian follicle. Using mouse ovarian follicles, we investigated uptake of L-leucine by follicles at varying stages of maturity in the presence of insulin-like growth factor (IGF)-1. METHODS Mouse ovarian follicles were cultured in vitro for 5 days in increasing concentrations of IGF-1, and follicle diameter and atresia measured as endpoints for growth. Uptake of (3)H-leucine was measured in follicles at different stages of development. In optimal IGF-1-mediated growth conditions, competitive inhibition of (3)H-leucine uptake by 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH), a non-metabolizable substrate analogue of L-leucine, was performed to demonstrate specificity of influx, via system L transporters. To test whether uptake rates were dependent on intracellular amino acid availability, follicles from in vitro cultures were pre-treated with L-phenylalanine prior to (3)H-leucine uptake. RESULTS: Follicle development (P< 0.001) and survival (P< 0.001) increased with IGF-1 treatment. As pre-antral follicles progressed to late antral stage, we observed an increase in L-leucine uptake, which was reduced in pre-ovulatory follicles. BCH decreased L-leucine uptake rates in early antral (P< 0.05), antral (P< 0.001) and pre-ovulatory follicles (P< 0.01). L-leucine influx increased in follicles preloaded with phenylalanine (trans-stimulation). In follicles lacking free intracellular amino acids (zero-trans suppression), uptake rate was reduced (P< 0.05). CONCLUSIONS: These results demonstrate, for the first time, evidence of specific system L amino acid transport in intact, mouse ovarian follicles and profile L-leucine uptake during folliculogenesis. A better understanding of ovarian follicle metabolic pathways is necessary for improved in vitro maturation as well as determining the impact of altered metabolism on fertility.

Upright water-based exercise to improve cardiovascular and metabolic health: a qualitative review.

Research regarding the benefits of exercise for cardiovascular and metabolic health is extensive and well-documented. However, weight-bearing exercise may not be suitable for individuals with orthopaedic or musculoskeletal limitations, excess adiposity or other medical conditions. Water-based exercise may provide an attractive alternative to land-based exercise for achieving improved health and fitness in these populations. Although swimming is a popular form of water-based exercise it requires specific skills and is often undertaken at intensities that may not be safely prescribed in patient populations. Therefore upright, water-based exercise has been suggested as a viable water-based alternative. However, surprisingly little is known about the effects of upright water-based exercise on improvements in cardiovascular and metabolic health. Limited evidence from water-based studies indicate that regular deep or shallow water exercise can exert beneficial effects on cardiorespiratory fitness, strength, and body fat distribution. However, the impacts of water-based exercise on lipid profile, bodyweight, and carbohydrate metabolism are still unclear. Further studies are warranted to establish the effects of non-swimming, water-based exercise on cardiometabolic risks in humans.

Thiols stabilize cobblestone morphology of cultured mesothelial cells.

Cellular thiols including GSH (glutathione) and L-Cys (L-cysteine) are essential for cell signalling, growth and differentiation. L-Cys is derived from the extracellular thiol pool and is the rate-limiting compound for intracellular GSH biosynthesis. The present study investigated the effect of thiol-supplemented medium on cell growth, phenotype and total GSH of cultured hPMCs (human peritoneal mesothelial cells). Cells were cultured in medium M199 supplemented with 2% serum, with 'plus' or without 'minus' L-Cys and compared with medium supplemented with either ß-ME (ß-mercaptoethanol) (0.25 mmol/l) or the receptor tyrosine kinase ligand EGF (epidermal growth factor, 100 ng/ml). ß-ME produced a disproportionate increase in total GSH compared with L-Cys and other thiols tested [(procysteine (2-oxothiazolidine-4-carboxylic acid) or NAC (N-acetyl-L-cysteine)], while growth and morphology were identical. Cell behaviour of primary hPMCs is characterized by the transition of fibroblastoid to cobblestone morphology during early passage. L-Cys and ß-ME promoted a rapid MET (mesenchymal-to-epithelial transition) within 3 days of culture, confirmed by the presence of cobblestone cells, intact organelles, abundant microvilli, primary cilia and cortical actin. In contrast, EGF produced a biphasic response consisting of delayed growth and retention of a fibroblastoid morphology. During a rapid log phase of growth, MET was accompanied by rapid catch-up growth. Thiols may stabilize the epithelial phenotype by engaging redox-sensitive receptors and transcription factors that modulate differentiation. These data may benefit researchers working on thiol-mediated cell differentiation and strategies to regenerate damage to serosal membranes.

Stereological assessment of developing mouse ovarian follicles in an in vitro culture system.

The aim of this project was to validate the use of ovarian follicular diameter measurement as an indicator of growth using stereology as an independent measure. A strong linear correlation was observed between follicular volume and diameter, and between follicular volume and the total number of granulosa cells. These results have provided a useful validation of a widely used technique for determining follicular growth in vitro.

'Going-to-have-cancerness': a study of living with increased risk of BRCA1 and BRCA2 mutations for six South Island women.

INTRODUCTION: Mutations in the BRCA (breast cancer) 1 and 2 genes are thought to lead to 5-10% of breast cancers. AIM: A qualitative study to explore six New Zealand women's experiences of living with increased risk for a genetic susceptibility to breast cancer. METHODS: Six women were interviewed using semi-structured interviews, to explore their experiences of living at high risk for developing breast cancer due to familial and/or individual genetic susceptibilities. Results were analysed using thematic coding. After a three-year interval, interviewees were contacted again to discuss their experiences (although two were lost to follow-up). FINDINGS: The women held fatalistic views on developing cancer and drew on family experience as much as biomedical research to assess their situation. They became increasingly immersed in biomedical screening and prophylaxis without accompanying improvement to their peace of mind and with unrealistic ideas of it 'preventing' cancer. The biomedical management options and advice they reported receiving was factually inconsistent and a discrepancy emerged between women's expectations of breast cancer health services (including genetic testing) and the delivered support and services. CONCLUSION: This small sample group cannot be used to draw implications on the views of the wider group of higher risk patients, but for these six women, genetic testing, screening and prophylaxis have not provided peace of mind; rather the reverse has occurred. The findings are provocative as they challenge the biomedical idea of patients' experience of managing their genetic risk information as routinely positive.

Cryosolvent interaction with cellular actin using 3T3-L1cells as a model system.

Previous immunolocalisation studies using intact cells have identified modification of the cytoskeleton by cryoprotectants. In the present study we have used a proteomics approach to directly resolve the interactive effects of 3T3-L1cells exposed to two cryoprotectants, dimethyl sulphoxide (Me(2)SO) and 1,2-propanediol (PROH) in 5,10, 20 and 50(v/v) percent solutions, respectively. Two-dimensional protein electrophoresis and Western blot analysis of the cell extracts identified a range of immunoreactive actin fragments with varying molecular weights and isoelectric points at all cryoprotectant concentrations. The addition of either 10mM l-cysteine or reduced glutathione to the cells prior to cryprotectant exposure modified the actin fragmentation. In this preliminary report, we have provided direct evidence of actin fragmentation when exposed to cryoprotectants and have demonstrated that the use of redox agents can modify the cryprotectant action.