LC-MS/MS Method for Measurement of Thiopurine Nucleotides (TN) in Erythrocytes and Association of TN Concentrations With TPMT Enzyme Activity.

Monitoring concentrations of thiopurine metabolites is used clinically to prevent adverse effects in patients on thiopurine drug therapy. We developed a LC-MS/MS method for the quantification of 6-thioguanine (6-TG) and 6-methylmercaptopurine (6-MMP) in red blood cells (RBCs). This method utilizes an automated cell washer for RBC separation from whole blood samples and washing of the separated RBCs. The lower limit of quantification of the method was 0.2 µmol/L for 6-TG (∼50 pmol/8 × 108 RBC) and 4 µmol/L for 6-MMP (∼1,000 pmol/8 × 108 RBC). The total imprecision of the assay was <3.0%. The upper limit of linearity for 6-TG and 6-MMP was 7.5 µmol/L and 150 µmol/L, respectively. The stability of the thiopurine metabolites under pre- and post-analytically relevant conditions was also evaluated. A good agreement was observed between this method and validated LC-MS/MS methods from three laboratories, except for ∼40% low bias for 6-MMP observed in one of the methods. The assessment of the association between 6-TG and 6-MMP concentrations with thiopurine S-methyltransferase (TPMT) phenotype and genotype demonstrated a statistically significant difference in the thiopurine metabolite concentrations between the TPMT groups with normal and intermediate activity of 6-MMP (p < 0.0001), while the difference in 6-TG concentrations was statistically not significant (p = 0.096). Among the samples with normal TPMT activity, higher concentrations of 6-MMP (p = 0.015) were observed in pediatric samples than in the samples of adults. No statistically significant differences were observed in the distributions of 6-TG and 6-MMP concentrations among the evaluated genotypes.

The impact of the COVID-19 pandemic on economic security and pregnancy intentions among people at risk of pregnancy.

OBJECTIVE: To understand how the COVID-19 pandemic affected women of reproductive age, specifically their economic conditions, desire for pregnancy, and access to contraceptive services during the pandemic. STUDY DESIGNS: A total of 554 women respondents age 18 to 49 and reside in the United States were recruited using social media between May 16, 2020 and June 16, 2020. Logistic regression models assessed predictors of reporting pandemic-related changes in economic conditions, desire for pregnancy, and contraceptive access. RESULTS: Compared to White/Caucasian respondents, Hispanics/Latinx and Black/African Americans have 4 times the odds of experiencing inability to afford food, transportation, and/or housing (p < 0.01) during the pandemic; Hispanics/Latinx have twice the odds of experiencing food insecurity (p < 0.05). Inability to afford food, transportation, and/or housing was associated with drop in desire to be pregnant (p < 0.01). Despite the 25% of participants who reported a drop in desire for pregnancy, 1 in 6 reported difficulty accessing contraceptives, particularly those who experienced reduced income (p < 0.01). CONCLUSIONS: In our sample, the pandemic unevenly affected people from different socioeconomic groups. Many simultaneously experienced reduced income, difficulties in accessing contraception, and a greater desire to avoid a pregnancy. This combination of factors increases the chance that people will experience unintended pregnancies. IMPLICATIONS: The pandemic caused economic hardship and an increased desire to postpone or prevent pregnancy at the same time that it created new barriers to contraceptive services. This pattern may lead to a potential net effect of an increase in unintended pregnancy, particularly among people who had difficulty affording food, transportation, and/or housing during the pandemic.

State of the science in reconciling top-down and bottom-up approaches for terrestrial CO2 budget.

Robust estimates of CO2 budget, CO2 exchanged between the atmosphere and terrestrial biosphere, are necessary to better understand the role of the terrestrial biosphere in mitigating anthropogenic CO2 emissions. Over the past decade, this field of research has advanced through understanding of the differences and similarities of two fundamentally different approaches: "top-down" atmospheric inversions and "bottom-up" biosphere models. Since the first studies were undertaken, these approaches have shown an increasing level of agreement, but disagreements in some regions still persist, in part because they do not estimate the same quantity of atmosphere-biosphere CO2 exchange. Here, we conducted a thorough comparison of CO2 budgets at multiple scales and from multiple methods to assess the current state of the science in estimating CO2 budgets. Our set of atmospheric inversions and biosphere models, which were adjusted for a consistent flux definition, showed a high level of agreement for global and hemispheric CO2 budgets in the 2000s. Regionally, improved agreement in CO2 budgets was notable for North America and Southeast Asia. However, large gaps between the two methods remained in East Asia and South America. In other regions, Europe, boreal Asia, Africa, South Asia, and Oceania, it was difficult to determine whether those regions act as a net sink or source because of the large spread in estimates from atmospheric inversions. These results highlight two research directions to improve the robustness of CO2 budgets: (a) to increase representation of processes in biosphere models that could contribute to fill the budget gaps, such as forest regrowth and forest degradation; and (b) to reduce sink-source compensation between regions (dipoles) in atmospheric inversion so that their estimates become more comparable. Advancements on both research areas will increase the level of agreement between the top-down and bottom-up approaches and yield more robust knowledge of regional CO2 budgets.

Comment on "Saturation of the southern ocean CO2 sink due to recent climate change".

Unlike Le Quéré et al. (Reports, 22 June 2007, p. 1735), we do not find a saturating Southern Ocean carbon sink due to recent climate change. In our ocean model, observed wind forcing causes reduced carbon uptake, but heat and freshwater flux forcing cause increased uptake. Our inversions of atmospheric carbon dioxide show that the Southern Ocean sink trend is dependent on network choice.

Towards robust regional estimates of CO2 sources and sinks using atmospheric transport models.

Information about regional carbon sources and sinks can be derived from variations in observed atmospheric CO2 concentrations via inverse modelling with atmospheric tracer transport models. A consensus has not yet been reached regarding the size and distribution of regional carbon fluxes obtained using this approach, partly owing to the use of several different atmospheric transport models. Here we report estimates of surface-atmosphere CO2 fluxes from an intercomparison of atmospheric CO2 inversion models (the TransCom 3 project), which includes 16 transport models and model variants. We find an uptake of CO2 in the southern extratropical ocean less than that estimated from ocean measurements, a result that is not sensitive to transport models or methodological approaches. We also find a northern land carbon sink that is distributed relatively evenly among the continents of the Northern Hemisphere, but these results show some sensitivity to transport differences among models, especially in how they respond to seasonal terrestrial exchange of CO2. Overall, carbon fluxes integrated over latitudinal zones are strongly constrained by observations in the middle to high latitudes. Further significant constraints to our understanding of regional carbon fluxes will therefore require improvements in transport models and expansion of the CO2 observation network within the tropics.