RESUMO
In 2018, data from a surveillance study in Botswana evaluating adverse birth outcomes raised concerns that women on antiretroviral therapy (ART) containing dolutegravir (DTG) may be at increased risk for neural tube defects (NTDs). The mechanism of action for DTG involves chelation of Mg2+ ions in the active site of the viral integrase. Plasma Mg2+ homeostasis is maintained primarily through dietary intake and reabsorption in the kidneys. Inadequate dietary Mg2+ intake over several months results in slow depletion of plasma Mg2+ and chronic latent hypomagnesemia, a condition prevalent in women of reproductive age worldwide. Mg2+ is critical for normal embryonic development and neural tube closure. We hypothesized that DTG therapy might slowly deplete plasma Mg2+ and reduce the amount available to the embryo, and that mice with pre-existing hypomagnesemia due to genetic variation and/or dietary Mg2+ insufficiency at the time of conception and initiation of DTG treatment would be at increased risk for NTDs. We used two different approaches to test our hypothesis: 1) we selected mouse strains that had inherently different basal plasma Mg2+ levels and 2) placed mice on diets with different concentrations of Mg2+. Plasma and urine Mg2+ were determined prior to timed mating. Pregnant mice were treated daily with vehicle or DTG beginning on the day of conception and embryos examined for NTDs on gestational day 9.5. Plasma DTG was measured for pharmacokinetic analysis. Our results demonstrate that hypomagnesemia prior to conception, due to genetic variation and/or insufficient dietary Mg2+ intake, increases the risk for NTDs in mice exposed to DTG. We also analyzed whole-exome sequencing data from inbred mouse strains and identified 9 predicted deleterious missense variants in Fam111a that were unique to the LM/Bc strain. Human FAM111A variants are associated with hypomagnesemia and renal Mg2+ wasting. The LM/Bc strain exhibits this same phenotype and was the strain most susceptible to DTG-NTDs. Our results suggest that monitoring plasma Mg2+ levels in patients on ART regimens that include DTG, identifying other risk factors that impact Mg2+ homeostasis, and correcting deficiencies in this micronutrient might provide an effective strategy for mitigating NTD risk.
RESUMO
Visual assessments of disease severity are currently the industry standard for quantification of the necrosis caused by Cercospora beticola on sugar beet (Beta vulgaris) leaves. We compared the precision, reproducibility, and sensitivity of a multispectral radiometer to visual disease assessments. Individual wavebands from the radiometer, as well as vegetative indices calculated from the individual wavebands, were compared with visual disease estimates from two raters at each of two research sites. Visual assessments and radiometric measurements were partially repeated immediately after the first assessment at each site. Precision, as measured by reduced coefficients of variation, was improved for all vegetative indices and individual waveband radiometric measures compared with visual assessments. Visual assessments, near-infrared singlewaveband reflectance values, and four of the six vegetative indices had high treatment F values, suggesting greater sensitivity at discriminating disease severity levels. Reproducibility, as measured by a test-retest method, was high for visual assessments, single-waveband reflectance at 810 nm, and several of the vegetative indices. The use of radiometric methods has the potential to increase the precision of assessments of Cercospora leaf spot foliar symptoms of sugar beet while eliminating potential bias. We recommend this method be used in conjunction with visual disease assessments to improve precision of assessments and guard against potential bias in evaluations.