A high incidence of polymorphic CYP2C19 variants in archival blood samples from Papua New Guinea.

There is considerable inter-ethnic variability in the incidence of CYP2C19 genetic poor metabolisers ( var / var ). About 3 per cent of Caucasians are CYP2C19 var/var . By contrast, an extremely high incidence (70 per cent) is observed in the Melanesian island of Vanuatu. The colonisation of the Pacific Islands is believed to have involved migration through Papua New Guinea (PNG), and hence a high incidence may also be expected in this population. The reported incidence in PNG was only 36 per cent, however. PNG is a country of extensive ethnic diversity, and the incidence of the CYP2C19 var/var in other regional populations of PNG is currently not established. In this study, restriction fragment length polymorphism-polymerase chain reaction analysis of archival blood serum samples was used to determine the prevalence of the CYP2C19*2 and *3 variant alleles in three different ethnic and geographically isolated populations of PNG. In the largest population studied (Iruna), the frequency of both variant CYP2C19 alleles was high (0.37 and 0.34, respectively). Specifically, the frequency of the CYP2C19*3 allele was significantly higher than in the PNG (East Sepik) population reported previously (0.34 vs 0.16; p < 0.0001). In the Iruna population, 48.9 per cent of the samples were homozygous variants for CYP2C19*2 or *3 , which although higher was not statistically different from the East Sepik population (36 per cent). The results of this study indicated that other regional populations of PNG also have a relatively high incidence of the CYP2C19 genetic polymorphism compared with Caucasian populations. The high incidence reported in Vanuatu, however, may be due to genetic drift rather than a PNG founder population, as the Vanuatu population is dominated by the CYP2C19*2 allele, with a lower contribution from the *3 allelic variant.

A microsatellite map of white clover.

The white clover ( Trifolium repens) nuclear genome (n = 2x = 16) is an important yet under-characterised genetic environment. We have developed simple sequence repeat (SSR) genetic markers for the white clover genome by mining an expressed sequence tag (EST) database and by isolation from enriched genomic libraries. A total of 2,086 EST-derived SSRs (EST-SSRs) were identified among 26,480 database accessions. Evaluation of 792 EST-SSR primer pairs resulted in 566 usable EST-SSRs. Of these, 335 polymorphic EST-SSRs, used in concert with 30 genomic SSRs, detected 493 loci in the white clover genome using 92 F1 progeny from a pair cross between two highly heterozygous genotypes--364/7 and 6525/5. Map length, as estimated using the joinmap algorithm, was 1,144 cM and spanned all 16 homologues. The R (red leaf) locus was mapped to linkage group B1 and is tightly linked to the microsatellite locus prs318c. The eight homoeologous pairs of linkage groups within the white clover genome were identified using 96 homoeologous loci. Segregation distortion was detected in four areas (groups A1, D1, D2 and H2). Marker locus density varied among and within linkage groups. This is the first time EST-SSRs have been used to build a whole-genome functional map and to describe subgenome organisation in an allopolyploid species, and T. repens is the only Trifolieae species to date to be mapped exclusively with SSRs. This gene-based microsatellite map will enable the resolution of quantitative traits into Mendelian characters, the characterisation of syntenic relationships with other genomes and acceleration of white clover improvement programmes.

Advances in pasture plant breeding for animal productivity and health.

Plant breeding has had a substantial effect on the productivity and health of ruminant animals in New Zealand by improving the quantity, quality and reliability of grazed temperate pastures. Genetic changes have affected annual pasture productivity, seasonal growth, digestibility, protein/energy balance, level of rumen undegradable protein, leaf properties affecting intake, resistance to foliar diseases, and reductions in compounds that have an adverse impact on the health, welfare and reproductive fertility of ruminant animals. Most plant improvement programmes have achieved genetic gains in excess of 1% per year for a variety of target traits, and these gains are likely to continue given the high genetic variation available within forage plants. Significant heritable variation exists to improve forage quality, particularly for soluble carbohydrate and fibre fractions in grasses, and in the rate at which these change during the season. Deleterious animal health and welfare effects can be alleviated through the use of non-toxic endophytes in grasses, that do not produce lolitrem B and ergovaline. Use of improved cultivars, with the appropriate management, can add value to animal products.

The New Zealand Blood Donors' Health Study: baseline findings of a large prospective cohort study of injury.

INTRODUCTION: Cohort studies have contributed important scientific knowledge regarding the determinants of chronic diseases. Despite the need for etiologic investigations, this design has been infrequently used in injury prevention research. OBJECTIVES: To describe the baseline findings of the New Zealand Blood Donors' Health Study, a large prospective study designed to investigate relationships between lifestyle, psychosocial factors, and serious injury due to road crashes, falls, self harm, assault, work, sport, and recreation. METHODS: Participants were recruited from fixed and mobile collection sites of a voluntary non-profit blood donor program. Baseline exposure data (for example risk taking behaviors, alcohol and marijuana use, sleep habits, and depression) were collected using a self administered questionnaire. Outcome data regarding serious injury will be collected prospectively through computerized record linkage of participants' unique identifiers to national morbidity and mortality databases. RESULTS: In total, 22 389 participants enrolled in the study (81% response rate). The diverse study population included 36% aged 16-24 years, 20% rural residents, and large variability in exposures of interest. For example, in the 12 months before recruitment, 21% had driven a motor vehicle when they considered themselves over the legal limit for alcohol, and 11% had been convicted of traffic violations (excluding parking infringements). Twelve per cent had seriously considered attempting suicide sometime in their life. CONCLUSIONS: This is the first, large scale cohort study investigating determinants of serious injury in New Zealand and among the largest worldwide. Preliminary findings from prospective analyses that can inform injury prevention policy are expected within five years.

On techniques for the measurement of the mass fractal dimension of aggregates.

A review is presented of a number of techniques available for the characterisation of the structure of aggregates formed from suspensions of sub-micron particles. Amongst the experimental techniques that have been commonly used are scattering (light, X-ray or neutron), settling and imaging and these are the focus of this work. The theoretical basis for the application of fractal geometry to characterisation of flocs and aggregates is followed by a discussion of the strengths and limitations of the above techniques. Of the scattering techniques available, light scattering provides the greatest potential for use as a tool for structure characterisation even though interpretation of the scattered intensity pattern is complicated by the strong interaction of light and matter. Restructuring further complicates the analysis. Although settling has long been used to characterise particle behaviour, the absence of an accurate permeability model limits the technique as a means of determining the porosity of fractal aggregates. However, it can be argued that the determination of fractal dimension is relatively unaffected. The strength of image analysis lies in its ability to provide a great deal of information about particle morphology and the weaknesses lie in the difficulties with image processing and sample size as this is a particle counting technique. There are very few papers which compare the fractal dimension measured by more than one technique. Light scattering potentially provides a useful tool for checking settling results. However, further work is required to develop proper models for aggregate permeability and flow-through effects.

An improved permeability model for fractal aggregates settling in creeping flow.

Aggregates often have mass fractal properties and there is, therefore, considerable interest in the hydrodynamics of mass fractal aggregates. There is no appropriate model in the literature to account for the non-homogeneous permeability within mass fractal aggregates. The new model proposed treats an individual aggregate as a self-similar object made up of a settling swarm of permeable spheres which are themselves made up of a settling swarm of sub-spheres and so on to the level of primary particles. This staged approach makes appropriate use of the fractal description of the aggregate to account for non-homogeneous distribution of porosity and quantify the drag correction for flow through effects. It also allows the settling properties to be quantified for multiple level aggregate structures, such as those formed from microflocs or by restructuring.

The phenols and prodelphinidins of white clover flowers.

White clover flowers (Trifolium repens L.) contain an abundance of phenolics, namely cis- and trans-p-coumaric acid 4-O-beta-D-glucopyranoside, the 3-O-beta-D-galactopyranosides of myricetin, quercetin and kaempferol together with two new derivatives namely myricetin 3-O-(6"-acetyl)-beta-D-galactopyranoside and kaempferol 3-O-(6"-acetyl)-beta-D-galactopyranoside. Gallocatechin, epigallocatechin, gallocatechin-(4alpha-8)-epigallocatechin and their corresponding prodelphinidin polymers were also present. The 13C-NMR spectra showed that the polymers consisted of only gallocatechin and epigallocatechin monomeric units with the latter being about twice as abundant in the extenders but only slightly more than that in the terminating units. The average degree of polymerization was estimated by 13C-NMR and ES-MS, which gave a remarkably consistent result of about 5.8 flavanol units.

A prospective cohort study of blood donors: methodological issues in the investigation of injuries and chronic diseases.

Blood donors have made important contributions to research, most notably in cross-sectional seroprevalence studies. The proposed New Zealand Blood Donors Health Study is a prospective cohort study of 30,000 New Zealand donors designed to investigate the determinants of common injuries, cardiovascular disease and cancer. While robust from an analytic perspective, the execution of prospective cohort studies in many settings is impeded by methodological, economic and organisational barriers. We examined the operational considerations of implementing a large-scale cohort study at a transfusion centre and evaluated measures taken to optimise data collection procedures. A pilot study of 1,000 participants revealed donor motivation to participate in this research was high (91% response rate). Comprehensive exposure data on lifestyle, behavioural and psychosocial factors were obtained from 95% of participants. Substantial heterogeneity in levels of potential risk factors was noted among respondents. Detailed dietary habit information and a study blood sample were obtained from 67% and 100% of participants, respectively. Study recruitment and baseline data collection was feasible during routine donor visits with minimal interruption to donor centre staff and procedures. We conclude the study design and characteristics of the regional donor program enhance the efficiency and significance of the proposed research.

Clinical usage of intravenous immunoglobulins in Auckland.

AIMS: To review the indications for prescription of intravenous immunoglobulin (IVIg), in Auckland and the associated adverse effects. METHODS: All patients who received IVIg infusions in four major hospitals in Auckland in 1996 were identified from blood bank records. Clinical details were recorded from case notes. RESULTS: One hundred and thirty-five cases were identified and 131 records were available for review. Hypogammaglobulinaemia (n = 38) and thrombocytopenia (n = 38) were the two most common indications for 44% of Primary hypogammaglobulinaemia accounted for IVIg. the IVIg prescribed. IVIg was only used in one patient with hypogammaglobulinaemia secondary to chronic lymphocytic leukaemia and in one case of graft-versus-host disease. Adverse reactions were noted in 13 cases (10%). Headache (n = 4) and fever (n = 4) were the most commonly recorded side-effects. One patient developed acute renal failure after IVIg infusion. Written consent was documented in 7 patients (5%). CONCLUSIONS: The indications for IVIg prescription were generally consistent with the recommendations of the Australasian Society of Blood Transfusion. Use of IVIg in haematological conditions other than immune thrombocytopaenic purpura was infrequent. Most of the IVIg infusions were administered uneventfully but some minor side effects were recorded and one significant clinical event may have been causally related to IVIg infusion.

Clinical and molecular aspects of Gaucher disease in New Zealand.

AIM: To report on the clinical and molecular aspects of Gaucher disease in New Zealand. METHODS: Patients known to have Gaucher disease were contacted and clinical information was recorded by questionnaire. Blood samples from affected individuals and their families provided DNA material for mutation analysis of disease causing alleles. Patients were assayed for beta-glucocerebrosidase, the enzyme deficiency which causes Gaucher disease. RESULTS: Twelve of 14 patients and 10 carriers were confirmed by DNA analysis. One asymptomatic individual was diagnosed. Four known mutations (N370S, 1444p, R463c and RecNcIl) and one unknown mutation were found from the 34 disease producing alleles that were identified. Of these, the L444P and N370S alleles were the most common. Most patients exhibited a clinical disorder typical of type 1 Gaucher disease. Two recent patients with severe neuropathic Gaucher disease had died in childhood. All patients showed a deficiency in beta-glucocerebrosidase. CONCLUSION: Gaucher disease in New Zealand is represented in a small number of non Jewish individuals with varying severity. Identifiable mutations and clinical symptoms aid in expanding the Australasian picture of this well studied disease. Enzyme replacement therapy for these patients has recently commenced in New Zealand.

HLA-B27 polymorphism and worldwide susceptibility to ankylosing spondylitis.

HLA-B27 is strongly associated to ankylosing spondylitis (AS) and represents a family of eleven B27 alleles (B*2701-11). Our aim was to analyze the distribution of B27 subtypes by PCR/SSOP and genomic sequencing in a large group of populations (n = 17). 711 B27-positive samples from Caucasoid, Asian, African, Amerindian and Polynesian populations were selected to ascertain transracial gene mapping of the B27 subtypes. 476 of these were AS patients, chosen to investigate the contribution of B27 alleles to AS susceptibility. Some significant new findings have arisen from this study: 1) B*2705 was the predominant subtype in circumpolar and subarctic areas. B*2702 was found to be practically restricted to Caucasian populations, showing a higher frequency in Middle-East (Jews) and North Africa (Arabs/Berbers) groups. 2) B*2703 appears associated with AS in Western Africans. This is of remarkable interest since it was suggested that B*2703 would be negatively disease-associated. 3) Although B*2706 appears negatively associated with AS in Thais, we identified two patients from northern China carrying it. This may be a reflection of a disease heterogeneity and could indicate that more than one pathogenic agent can be involved in AS. B*2709 has been recently described as negatively associated with AS in Sardinians. The molecular changes His114Asp (B*2706) and Asp116His (B*2709) could modify the genetic susceptibility to AS.

Transfusion transmitted Yersinia enterocolitica infection in New Zealand.

AIMS: To update and summarise cases of transfusion-transmitted Yersinia enterocolitica infection in New Zealand and to evaluate critically suggested methods to reduce this rare but frequently fatal complication of blood transfusion. METHODS: Case reports of four recent transfusion-transmitted Y. enterocolitica infections in New Zealand are given and previous reports reviewed. Literature review and evaluation of proposed methods to decrease the incidence of transfusing yersinia contaminated blood. RESULTS: There have been eight cases of transfusion-transmitted Y. enterocolitica infection in New Zealand in the past five years. Four of the five deaths have been directly caused by the transfusion. This gives a transfusion incidence rate of one:65,000 and a fatality rate of one:104,000 units transfused. This fatality rate is more than 80 times higher than that reported in the United States. CONCLUSIONS: Why the incidence of transfusion-transmitted yersinia is so high is not clear, since we do not store blood as long as many other countries, particularly the United States. In Auckland, however, the cases came at a time when the number of yersinia isolates from the community is reported to be rising. Many suggestions for the prevention of this problem have been put forward reflecting the fact that there is as yet no perfect solution. Those which are easy to implement and cheap to perform are largely already in place and investigation is continuing into the other alternatives.

Donor work-up and transport of bone marrow--recommendations and requirements for a standardized practice throughout the world from the Donor Registries and Quality Assurance Working Groups of the World Marrow Donor Association (WMDA).

In October 1995 the World Marrow Donor Association (WMDA) was restructured in order to facilitate its primary function of establishing guidelines in relation to international bone marrow and blood stem cell transplants -- transplants in which the donor is in one country and the patient is in another country. Five new working groups were established -- Donor Registries, Ethics, Quality Assurance, Finances, and Stem Cells. This paper, prepared by members of the Donor Registries Working Group, in consultation with the Quality Assurance Working Group, provides recommendations for the 'donor work-up'. This term covers events that start when the definitive donor has been identified, includes the harvesting (collection) and transportation of the stem cell product and ends when the product reaches the transplant centre. The paper includes examples of the documentation intended to ensure compliance with the recommendations at all key points in the sequence.

Alpha-2-HS-glycoprotein polymorphism in New Zealand Europeans and New Zealand Maori.

The polymorphism of alpha-2-HS-glycoprotein (AHSG, alpha 2HS) was analysed in a sample of New Zealanders consisting of 194 New Zealand Europeans and 236 New Zealand Maori. Thin layer polyacrylamide gel isoelectric focusing followed by immunofixation revealed four different AHSG phenotypes in New Zealand Europeans and three different AHSG phenotypes in New Zealand Maori. The AHSG*2 frequency of 0.695 for the New Zealand Maori population was found to be one of the highest reported for any population. AHSG*2 appears to be a useful genetic marker for Maori in anthropological studies.

Entire nucleotide sequence and characterization of a hepatitis C virus of genotype V/3a.

The entire nucleotide sequence of a hepatitis C virus (HCV) genome (NZL1) of genotype V/3a was determined from overlapping cDNA clones obtained from a human carrier in New Zealand. It comprised 9425 nucleotides (nt) including a 5'-untranslated region of 339 nt, a single large open reading frame encoding a polyprotein of 3021 amino acids, a 3'-untranslated region of 23 nt, and 3'-terminal poly(U) stretches of variable lengths. The NZL1 genome was compared with 15 HCV isolates of other genotypes for which the full-length sequence has been determined. It differed from them by 31.1 to 34.3% in nucleotide sequence identity and by 24.5 to 29.1% in amino acid sequence identity, confirming the distinction of genotype V/3a from the other isolates.

The genetic affinity of Polynesians: evidence from Y chromosome polymorphisms.

Y-linked polymorphisms were studied in a sample of 60 Polynesians, and results were compared with findings from studies on other major population groups. Three previously unreported 49a/TaqI haplotypes were observed, two of which possess a new polymorphic fragment named I2. Frequency data for the 49a/TaqI, XY275, pDP31 and Y Alu polymorphisms indicate that Polynesians have greater affinity to Caucasoids than to African populations. Similar population frequency trends were not observed for the p21A1/TaqI polymorphism, supporting the hypothesis that this polymorphism has arisen more than once.