Enhanced Thermal Stability and Reduced Aggregation in an Antibody Fab Fragment at Elevated Concentrations

The aggregation of protein therapeutics such as antibodies remains a major challenge in the biopharmaceutical industry. The present study aimed to characterize the impact of the protein concentration on the mechanisms and potential pathways for aggregation, using the antibody Fab fragment A33 as the model protein. Aggregation kinetics were determined for 0.05 to 100 mg/mL Fab A33, at 65 °C. A surprising trend was observed whereby increasing the concentration decreased the relative aggregation rate, ln(v) (% day-1), from 8.5 at 0.05 mg/mL to 4.4 at 100 mg/mL. The absolute aggregation rate (mol L-1 h-1) increased with the concentration following a rate order of approximately 1 up to a concentration of 25 mg/mL. Above this concentration, there was a transition to an apparently negative rate order of -1.1 up to 100 mg/mL. Several potential mechanisms were examined as possible explanations. A greater apparent conformational stability at 100 mg/mL was observed from an increase in the thermal transition midpoint (Tm) by 7-9 °C, relative to those at 1-4 mg/mL. The associated change in unfolding entropy (△Svh) also increased by 14-18% at 25-100 mg/mL, relative to those at 1-4 mg/mL, indicating reduced conformational flexibility in the native ensemble. Addition of Tween or the crowding agents Ficoll and dextran, showed that neither surface adsorption, diffusion limitations nor simple volume crowding affected the aggregation rate. Fitting of kinetic data to a wide range of mechanistic models implied a reversible two-state conformational switch mechanism from aggregation-prone monomers (N*) into non-aggregating native forms (N) at higher concentrations. kD measurements from DLS data also suggested a weak self-attraction while remaining colloidally stable, consistent with macromolecular self-crowding within weakly associated reversible oligomers. Such a model is also consistent with compaction of the native ensemble observed through changes in Tm and △Svh

Association of plasma and CSF cytochrome P450, soluble epoxide hydrolase and ethanolamides metabolism with Alzheimer’s disease

Abstract: Alzheimer’s disease shares inflammatory origin with cardiometabolic disorders. Lipid mediators, including oxylipins, endocannabinoids, bile acids and steroids are potent regulators of inflammation, energy metabolism and cell proliferation with well-established involvement in cardiometabolic diseases. However, their role in Alzheimer’s disease is poorly understood. In the current study we provide comprehensive analysis of plasma and CSF lipid mediators in a case-control comparison of patients with Alzheimer’s disease, utilizing a targeted quantitative mass spectrometry approach. In both plasma and CSF, we observed Alzheimer’s disease patients to have elevated components of cytochrome P450/soluble epoxide hydrolase pathway and lower levels of fatty acids ethanolamides, when compared to the healthy controls. Multivariate analysis revealed that circulating metabolites of soluble epoxide hydrolase together with ethanolamides are strong and independent predictors for Alzheimer’s disease. Both metabolic pathways are potent regulators of inflammation with soluble epoxide hydrolase being reported to be upregulated in the brains of Alzheimer’s disease patients. This study provides further evidence for the involvement of inflammation in Alzheimer’s disease and argues for further research into the role of the cytochrome P450/soluble epoxide hydrolase pathway and fatty acid ethanolamides in this disorder. Further, these findings suggest that a combined pharmacological intervention targeting both metabolic pathways may have therapeutic benefits for Alzheimer’s disease

The Changing Carbon Cycle in the Southern Ocean

Various human activities, including fossil fuel combustion and forest clearing, emit about eight petagrams (or billion tons) of carbon in the form of CO2 into the atmosphere annually. The global ocean absorbs about two petagrams of CO2, and about a half of that amount is absorbed by the Southern Ocean south of 30°S, thus slowing the rapid accumulation of CO2 in the atmosphere. Partial pressure of CO2 (pCO2) is a measure of the chemical driving force for the CO2 exchange between the ocean and the atmosphere. This paper discusses its space and time distribution over the Southern Ocean. The major sink zone for atmospheric CO2 is located in a latitude belt between 30°S and 50°S, where the biological utilization of CO2 and cooling of warm subtropical waters flowing southward produce low seawater pCO2. Strong winds in this zone also enhance the ocean's uptake. Although the source-sink conditions vary over a wide range through the seasons in the areas south of 50°S, this zone is a small sink on an annual average. Winter observations show that surface water pCO2 values in the source region for Antarctic Intermediate Water have increased at a rate faster than the atmospheric increase rate, suggesting that the ocean CO2 sink intensity has been weakening for several decades and has changed from a net sink to a net source since 2005. The results of ocean general circulation-biogeochemistry model studies are found to be consistent with the observations

The changing carbon cycle in the Southern Ocean

Various human activities, including fossil fuel combustion and forest clearing, emit about eight petagrams (or billion tons) of carbon in the form of CO2 into the atmosphere annually. The global ocean absorbs about two petagrams of CO2, and about a half of that amount is absorbed by the Southern Ocean south of 30 S, thus slowing the rapid accumulation of CO2 in the atmosphere. Partial pressure of CO, (pCO(2)) is a measure of the chemical driving force for the CO2 exchange between the ocean and the atmosphere. This paper discusses its space and time distribution over the Southern Ocean. The major sink zone for atmospheric CO2 is located in a latitude belt between 30 S and 50 S, where the biological utilization of CO2 and cooling of warm subtropical waters flowing southward produce low seawater pCO(2). Strong winds in this zone also enhance the ocean's uptake. Although the source-sink conditions vary over a wide range through the seasons in the areas south of 50 S, this zone is a small sink on an annual average. Winter observations show that surface water pCO(2) values in the source region for Antarctic Intermediate Water have increased at a rate faster than the atmospheric increase rate, suggesting that the ocean CO2 sink intensity has been weakening for several decades and has changed from a net sink to a net source since 2005. The results of ocean general circulation-biogeochemistry model studies are found to be consistent with the observations.This is the publisher’s final pdf. The published article is copyrighted by the Oceanography Society and can be found at: www.tos.org/.Keywords: Dioxide, exchange, Antarctic circumpolar current, Surface water

Autonomous Seawater \u3ci\u3ep\u3c/i\u3eCO\u3csub\u3e2\u3c/sub\u3e and pH Time Series From 40 Surface Buoys and the Emergence of Anthropogenic Trends

Ship-based time series, some now approaching over 3 decades long, are critical climate records that have dramatically improved our ability to characterize natural and anthropogenic drivers of ocean carbon dioxide (CO2) uptake and biogeochemical processes. Advancements in autonomous marine carbon sensors and technologies over the last 2 decades have led to the expansion of observations at fixed time series sites, thereby improving the capability of characterizing sub-seasonal variability in the ocean. Here , we present a data product of 40 individual autonomous moored surface ocean pCO2 (partial pressure of CO2) time series established between 2004 and 2013, 17 also include autonomous pH measurements. These time series characterie a wide range of surface ocean carbonate conditions in diffferent oceanic (17 sites), coastal (13 sites), and coral reef (10 sites) regimes. A time of trend emergence (ToE) methodology applied ot the time series that exhibit well-constrained daily to interannual variability and an estimate of decadal variability indicates that the length of sustained observations necessary to detect statistically significant anthropogenic trends varies by marine environment. The ToE estisites, and 9 to 22 years at the coral reef sites. Only two open ocean pCO2 and pH range from 8 to 15 years at the open ocean sites, 16 to 41 years at the coastal sites, and 9 to 22 years at the coral reef sites. Only two open ocean pCO2 time series, Woods Hole Oceanographic Institution Hawaii Ocean Time-series Station (WHOTS) in the subtropical North Pacific and Stratus n the South Pacific gyre, have been deployed longer than the estimated trend detection time and, for these, deseasoned monthly means show estimated anthropogenic trends of 1.9 ± 0.3 and 1.6 ± 0.3 μatm yr-1, respectively. In the future, it is possible that updates to this product will allow for the estimation of anthropogenic trends at more sites; however, the product currently provides a valuable tool in an accessible format for evaluating climatology and natural variability of surface ocean carbonate chemistry in a variety of regions. Data are available at https://doi.org/10.7289/V5DB8043 and https://www.nodc.noaa.gov/ocads/oceans/Moorings/ndp097.html (Sutton et al., 2018)

CATS II long-term anthropometric and metabolic effects of maternal sub-optimal thyroid function in offspring and mothers

Context and Objectives The Controlled Antenatal Thyroid Screening Study I (CATS-I) was a randomized controlled trial investigating the effects of levothyroxine therapy for suboptimal gestational thyroid function (SGTF), comparing outcomes in children of treated (SGTF-T) with untreated (SGTF-U) women during pregnancy. This follow-up study, CATS-II, reports the long-term effects on anthropometric, bone, and cardiometabolic outcomes in mothers and offspring and includes a group with normal gestational thyroid function (NGTF). Design & Participants 332 mothers (197 NGTF, 56 SGTF-U, 79 SGTF-T) aged 41.2±5.3 years (mean±SD) and 326 paired children assessed 9.3±1.0 years after birth for (i) body mass index (BMI); (ii) lean, fat, and bone mass by dual-energy X-ray absorptiometry; (iii) blood pressure, augmentation index, and aortic pulse-wave-velocity; and (iv) thyroid function, lipids, insulin, and adiponectin. The difference between group means was compared using linear regression. Results Offspring’s measurements were similar between groups. Although maternal BMI was similar between groups at CATS-I, after 9 years (at CATS-II) SGTF-U mothers showed higher BMI (median [interquartile ratio] 28.3 [24.6-32.6] kg/m2) compared with NGTF (25.8 [22.9-30.0] kg/m2; P = 0.029), driven by fat mass increase. At CATS-II SGTF-U mothers also had higher thyroid-stimulating hormone (TSH) values (2.45 [1.43-3.50] mU/L) than NGTF (1.54 [1.12-2.07] mU/L; P = 0.015), since 64% had never received levothyroxine. At CATS-II, SGTF-T mothers had BMI (25.8 [23.1-29.8] kg/m2, P = 0.672) and TSH (1.68 [0.89-2.96] mU/L; P = 0.474) values similar to NGTF mothers. Conclusions Levothyroxine supplementation of women with SGTF did not affect long-term offspring anthropometric, bone, and cardiometabolic measurements. However, absence of treatment was associated with sustained long-term increase in BMI and fat mass in women with SGTF