Cornelie Nienaber-Rousseau

Gene–environment and gene–gene interactions of specific MTHFR, MTR and CBS gene variants in relation to homocysteine in black South Africans

The methylenetetrahydrofolate reductase (MTHFR), cystathione-β-synthase (CBS) and methionine synthase (MTR) genes interact with each other and the environment. These interactions could influence homocysteine (Hcy) and diseases contingent thereon. We determined single nucleotide polymorphisms (SNPs) within these genes, their relationships and interactions with total Hcy concentrations within black South Africans to address the increased prevalence of diseases associated with Hcy. The MTHFR 677 TT and MTR 2756 AA genotypes were associated with higher Hcy concentrations (16.6 and 10.1 μmol/L; p<0.05) compared to subjects harboring the MTHFR 677 CT/CC and the MTR 2756 AG genotypes (10.5, 9.7 and 9.5 μmol/L, respectively). The investigated CBS genotypes did not influence Hcy. We demonstrated interactions between the area of residence and the CBS T833C/844ins68 genotypes (p=0.005) so that when harboring the wildtype allele, rural subjects had significantly higher Hcy than their urban counterparts, but when hosting the variant allele the environment made no difference to Hcy. Between the CBS T833C/844ins68 or G9276A and MTHFR C677T genotypes, there were two-way interactions (p=0.003 and=0.004, respectively), with regard to Hcy. Subjects harboring the MTHFR 677 TT genotype in combination with the CBS 833 TT/homozygous 844 non-insert or the MTHFR 677 TT genotype in combination with the CBS 9276 GA/GG displayed higher Hcy concentrations. Therefore, some of the investigated genotypes affected Hcy; residential area changed the way in which the CBS T833C/844ins68 SNPs influenced Hcy concentrations highlighting the importance of environmental factors; and gene-gene interactions allude to epistatic effects.

Interactions between C-Reactive Protein Genotypes with Markers of Nutritional Status in Relation to Inflammation

Inflammation, as indicated by C-reactive protein concentrations (CRP), is a risk factor for chronic diseases. Both genetic and environmental factors affect susceptibility to inflammation. As dietary interventions can influence inflammatory status, we hypothesized that dietary effects could be influenced by interactions with single nucleotide polymorphisms (SNPs) in the CRP gene. We determined 12 CRP SNPs, as well as various nutrition status markers in 2010 black South Africans and analyzed their effect on CRP. Interactions were observed for several genotypes with obesity in determining CRP. Lipid intake modulated the pro-inflammatory effects of some SNPs, i.e., an increase in both saturated fatty acid and monounsaturated fatty acid intake in those homozygous for the polymorphic allele at rs2808630 was associated with a larger increase in CRP. Those harboring the minor alleles at rs3093058 and rs3093062 presented with significantly higher CRP in the presence of increased triglyceride or cholesterol intake. When harboring the minor allele of these SNPs, a high omega-6 to -3 ratio was, however, found to be anti-inflammatory. Carbohydrate intake also modulated CRP SNPs, as HbA1C and fasting glucose levels interacted with some SNPs to influence the CRP. This investigation highlights the impact that nutritional status can have on reducing the inherent genetic susceptibility to a heightened systemic inflammatory state.

Genetic polymorphisms influencing total and γ' fibrinogen levels and fibrin clot properties in Africans

Inter‐ethnic variation in fibrinogen levels is hypothesized to be the result of differences in genetic background. No information is available regarding the contribution of genetics to fibrinogen γ′ in Africans. Only limited information is available regarding the interaction between genotypes and total and γ′ fibrinogen concentration in determining fibrin clot properties. Our aim was to investigate the effect of polymorphisms in the fibrinogen and Factor XIII genes on total and γ′ fibrinogen and clot properties (turbidimetry) in 2010 black Africans as well as to determine their interactions. Significant associations were observed between rs1049636 (FGG gene), with total fibrinogen levels and between rs2070011 (FGA promoter area) and fibrinogen γ′ levels. Significant associations were observed between single nucleotide polymorphisms (SNPs) in the FGA (rs2070011), FGB (rs1800787) and FGG (rs1049636) genes and fibre size. Significant interactions were found between total and/or γ′ fibrinogen levels and SNPs in the FGA (rs2070011), FGB (rs2227385, rs1800787, rs1800788, rs4220) and F13A1 genes (rs5985) in determining clot properties. The different SNPs influenced the relationships between total and γ′ fibrinogen levels with clot properties in opposing directions. Genetic influences may be ethnic‐specific and should not only focus on fibrinogen concentration, but also on functionality in determining its role in CVD.

Socio-Demographic and Lifestyle Factors Predict 5-Year Changes in Adiposity among a Group of Black South African Adults

The rising prevalence of obesity and excessive adiposity are global public health concerns. Understanding determinants of changes in adiposity over time is critical for informing effective evidence-based prevention or treatment. However, limited information is available to achieve this objective. Cultural, demographic, environmental, and behavioral factors including socio-economic status (SES) likely account for obesity development. To this end, we related these variables to anthropometric measures in 1058 black adult Tswana-speaking South Africans who were HIV negative in a prospective study over five years. Body mass index (BMI) and waist circumference increased in both sexes, whereas triceps skinfold thickness remained the same. Over the five years, women moved to higher BMI categories and more were diagnosed with central obesity. Age correlated negatively, whereas SES, physical activity, energy, and fat intake correlated positively with adiposity markers in women. In men, SES, marital status, physical activity, and being urban predicted increases in adiposity. For women, SES and urbanicity increased, whereas menopause and smoking decreased adiposity. Among men, smokers had less change in BMI than those that never smoked over five years. Our findings suggest that interventions, focusing on the urban living, the married and those with the highest SES—the high-risk groups identified herein—are of primary importance to contain morbidity and premature mortality due to obesity in black South Africans.

Degree of obesity influences the relationship of PAI-1 with body fat distribution and metabolic variables in African women.

INTRODUCTION Although the relationship of plasminogen activator inhibitor-1 (PAI-1) with obesity has been well established, the relationship of PAI-1 with different body fat distribution patterns is less clear particularly in non-white ethnicities. METHODS We investigated the cross-sectional association of PAI-1act with body fat % and two different body fat distribution patterns, namely sarcopenic obesity (SO) and visceral (VAT) compared to subcutaneous (SCAT) abdominal obesity, in 246 healthy African women by creating sub-groups according to different body fat distribution patterns. RESULTS The PAI-1act level of the SO group did not differ significantly from that of the excessive % body fat, non-sarcopenic group (p=0.8). The relationship of PAI-1act, with body fat %, insulin, triglycerides and appendicular skeletal mass (ASM) was influenced by body fat distribution patterns and degree of obesity. PAI-1act was higher (1.65 vs 0.16U/ml; p=0.001) in women with a proportionally higher abdominal VAT compared to higher abdominal SCAT compartment in the total study population, but not in the centrally obese sub-group (1.72 vs 0.83U/ml; p=0.5). Multiple regression models indicated that body fat % per se did not contribute significantly to PAI-1act variance in women with increased fat mass. CONCLUSION Fat distribution patterns and degree of obesity influenced the association of PAI-1act with insulin, triglycerides, ASM and body fat % in African women. In centrally obese women, abdominal VAT no longer contributed more to plasma PAI-1act, than abdominal SCAT. Inflammation and endothelial dysfunction contributed more to PAI-1act variance in obese African women than did body fat % per se.

The association between calf circumference and appendicular skeletal muscle mass index of black urban women in Tlokwe City

Background: Sarcopenia, the loss of muscle mass and strength, is associated with adverse health outcomes. Calf circumference (CC) has been proposed as a surrogate measure of muscle mass in the elderly; however, ethnic/sex specific cut-off values remain to be established. Objective: A study was undertaken to investigate the relationship between CC and appendicular skeletal muscle mass (ASM), and the ASM index (ASMI), as well as to determine whether CC could be used to diagnose sarcopenia. Methods: This was a cross-sectional study of 247 older black women living from Tlokwe, South Africa. ASM was measured using dual-energy X-ray absorptiometry, and the ASMI was calculated. Receiver operator characteristics curves and maximum Youden index were applied to identify a CC cut-off point for sarcopenia according to low gait speed (< 0.8 m/s), low hand-grip strength (< 16 kg) and low ASMI using a South African cut-off point for sarcopenia (ASMI < 4.94 kg/m2). Results: A strong positive correlation between CC and ASMI (r = 0.84, p < 0.001) was observed. The CC to predict low hand-grip strength was 34.3 cm and 37.8 cm for low gait speed. A CC of 29.9 cm was indicative of sarcopenia. The area under the curve for all outcomes was > 0.60. Conclusion: A CC of 30 cm is proposed as a simple and inexpensive way to predict, screen or diagnose sarcopenia in black women in low-resource health settings. An accessible, inexpensive screening or diagnostic tool could facilitate timely interventions and prevention.

CRP genotypes predict increased risk to co-present with low vitamin D and elevated CRP in a group of healthy black South African women

Low 25-hydroxyvitamin D (25(OH)D) and elevated C-reactive protein (CRP) concentrations are independently associated with adverse health outcomes, including cardiovascular disease (CVD). Although an inverse association between these factors has been described, the underlying mechanisms remain unknown. We postulate that environment–gene interactions, through which 25(OH)D interacts with single nucleotide polymorphisms (SNPs) within the CRP gene, modulate CRP; that certain CRP genotypes predispose individuals to a co-phenotype of low 25(OH)D and elevated CRP concentrations; and that this co-phenotype is associated with higher CVD risk. Twelve CRP SNPs were genotyped, and both 25(OH)D and CRP were quantified, in 505 black South African women. Alarmingly, 66% and 60% of the women presented with deficient/insufficient 25(OH)D and elevated CRP concentrations, respectively. CRP concentrations were higher in individuals with lower 25(OH)D concentrations. However, no 25(OH)D–CRP genotype interactions were evident. Several genotypes were associated with an altered risk of presenting with the co-phenotype, indicating a genetic predisposition. Women presenting with this co-phenotype had higher blood pressure and increased anthropometric measures, which may predispose them to develop CVD. We recommend increasing vitamin D fortification and supplementation efforts to reduce inflammation among black women with vitamin D deficiency, thereby possibly curbing diseases contingent on the co-phenotype described here.

Fibrinogen and clot-related phenotypes determined by fibrinogen polymorphisms: independent and IL-6-interactive associations

Interleukin-6 (IL-6) induces the expression of fibrinogen, and polymorphic variation within the fibrinogen genes is believed to alter the magnitude of this expression. The identification of the functional relevance of individual fibrinogen single nucleotide polymorphisms (SNPs) has been hindered by the high linkage disequilibrium (LD) reported in the European fibrinogen gene locus. This study investigated two novel and 12 known fibrinogen SNPs of potential functional relevance, in 2010 Tswana individuals known to have low LD. We aimed to identify functional polymorphisms that contribute to clot-related phenotypes and total and γ’ fibrinogen concentrations independently and through their interaction with IL-6, by taking advantage of the high fibrinogen and IL-6 concentrations and the low LD reported in black South Africans. Fibrinogen was significantly associated with IL-6, thereby mediating associations of IL-6 with clot formation and structure, although attenuating the association of IL-6 with clot lysis time. None of the common European fibrinogen haplotypes was present in this study population. Putative functional fibrinogen SNPs FGB–rs7439150, rs1800789 (–1420G/A) and rs1800787 (–148C/T) were significantly associated with fibrinogen concentration and altered clot properties, with several associations significantly influenced by IL-6 concentrations. The impact of harbouring several minor fibrinogen SNP alleles on the association of IL-6 and fibrinogen concentration was cumulative, with possession of each additional minor allele showing a stronger relationship of IL-6 with fibrinogen. This was also reflected in differences in clot properties, suggesting potential clinical relevance. Therefore, when investigating the effect of fibrinogen genetics on fibrinogen concentrations and CVD outcome, the possible interactions with modulating factors and the fact that SNP effects seem to be additive should be taken into account.

Dietary strategies to treat hyperhomocysteinaemia based on the biochemistry of homocysteine: a review

Abstract Hyperhomocysteinaemia is implicated in various diseases, including cardiovascular disease and cancer. Several conditions influence the concentration of homocysteine (Hcy), including demographic, genetic and lifestyle factors. With regard to the latter, dietary components may be manipulated as Hcy can be remethylated to methionine by folate, or metabolised by other one‑carbon nutrients, such as betaine and its precursor, choline. This metabolic interplay enables the nutritionist or dietitian to be able to lower Hcy concentrations cost‑effectively by tailoring an individual’s diet, or by food enrichment and fortification strategies. Evidence supports the safety and benefits of Hcy reduction by simple dietary intervention. B vitamins, and betaine and choline intake lower Hcy, whereas methionine and certain beverages (coffee, tea and alcohol) increase it. Therefore, dietary determinants of Hcy raise the prospect of a simple, inexpensive and safe means of treating and/or preventing diseases contingent on this sulphur‑containing protein.