Photosynthesis and light-dependent proton pumps increase boundary layer pH in tropical macroalgae: A proposed mechanism to sustain calcification under ocean acidification

Ocean acidification (OA) projections predict ocean pH to decline between 0.2 and 0.4 by 2100 with potential negative consequences for marine calcifiers without acclimation or adaption strategies to accomodate greater H+ in seawater. Biotic control of calcified reef macroalgae thalli surface diffusive boundary layer (DBL) chemistry may overcome low pH in seawater as one strategy to accommodate OA conditions. To investigate this strategy, we examined surface DBL O2 and pH dynamics in five calcifying macroalgae (Halimeda, Udotea, Jania, Neogoniolithon, crustose coralline algae CCA) from the Florida Reef Tract under ambient (8.1) and low (7.65) pH using microsensors (100�μm) at the thalli surface in a flow-through flume. The role of photosynthesis and photosystem II (PSII)-independent proton pumps in controlling DBL pH were examined. Four of the five macroalgae exhibited a strong positive linear relationship between O2 production and increasing pH in the first 15�30�s of irradiance. Once a quasi-steady-state O2 concentration was reached (300�s), all species had DBL pH that were higher (0.02�0.32) than bulk seawater. The DBL pH increase was greatest at low pH and dependent on PSII. Some evidence was found for a light-dependent, but PSII-independent, proton pump. High DBL � pH upon illumination was likely in response to carbon concentrating mechanisms (CCMs) for photosynthesis. CCMs may be a HCO3��H+ symport, OH� antiport or other DIC transport system, accompanied by proton efflux. HCO3� dehydration by external carbonic anhydrase (CAext) also produces OH� that can neutralize H+ in the DBL. CO2 or HCO3� uptake for photosynthesis may also engage H+/OH� fluxes as part of intracellular acid-base regulation changing DBL pH. A higher � pH within the DBL at low pH could be accounted for by greater CO2 diffusion and/or lower efficiencies in exporting cellular H+ across a lower concentration gradient, and/or a more efficient removal of H+ by CAext-driven dehydration of HCO3�. In the dark, � pH was less than in the light as these dynamics were primarily due to photosynthesis. We present a conceptual model of inorganic carbon uptake and ion transport pathways, as well as other processes associated with photosynthesis that drive DBL � pH and sustain tropical macroalgal calcification in the light under OA. In the dark, unless PSII-independent proton pumps are present, which do not appear to be ubiquitous amongst species, acidification processes likely dominate, resulting in CaCO3 net dissolution, particularly under OA conditions

Lines Missing Every Random Point

We prove that there is, in every direction in Euclidean space, a line that misses every computably random point. We also prove that there exist, in every direction in Euclidean space, arbitrarily long line segments missing every double exponential time random point.Comment: Added a section: "Betting in Doubly Exponential Time.

Identifying Host Genetic Risk Factors in the Context of Public Health Surveillance for Invasive Pneumococcal Disease

Host genetic factors that modify risk of pneumococcal disease may help target future public health interventions to individuals at highest risk of disease. We linked data from population-based surveillance for invasive pneumococcal disease (IPD) with state-based newborn dried bloodspot repositories to identify biological samples from individuals who developed invasive pneumococcal disease. Genomic DNA was extracted from 366 case and 732 anonymous control samples. TagSNPs were selected in 34 candidate genes thought to be associated with host response to invasive pneumococcal disease, and a total of 326 variants were successfully genotyped. Among 543 European Americans (EA) (182 cases and 361 controls), and 166 African Americans (AA) (53 cases and 113 controls), common variants in surfactant protein D (SFTPD) are consistently underrepresented in IPD. SFTPD variants with the strongest association for IPD are intronic rs17886286 (allelic OR 0.45, 95% confidence interval (CI) [0.25, 0.82], with p = 0.007) in EA and 5′ flanking rs12219080 (allelic OR 0.32, 95%CI [0.13, 0.78], with p = 0.009) in AA. Variants in CD46 and IL1R1 are also associated with IPD in both EA and AA, but with effects in different directions; FAS, IL1B, IL4, IL10, IL12B, SFTPA1, SFTPB, and PTAFR variants are associated (p≤0.05) with IPD in EA or AA. We conclude that variants in SFTPD may protect against IPD in EA and AA and genetic variation in other host response pathways may also contribute to risk of IPD. While our associations are not corrected for multiple comparisons and therefore must be replicated in additional cohorts, this pilot study underscores the feasibility of integrating public health surveillance with existing, prospectively collected, newborn dried blood spot repositories to identify host genetic factors associated with infectious diseases

Microwave Spectroscopy

Contains reports on nine research projects.United States Department of Energy (Contract DA36-039-sc-73014

Laser Photochemistry and Transient Raman Spectroscopy of Silyl-Substituted Fischer-Type Carbene Complexes

Pulsed laser irradiation of the silyl-substituted carbene complexes (CO)5W=C(XR)SiPh3 (XR = NC4H8 (1); = OEt (2)) in various solvents has been investigated using transient absorbance and time-resolved resonance Raman scattering as monitoring techniques. Irradiation of (1) in noncoordinating or weakly-coordinating solvents at 355 nm within the ligand field absorption band results in the rapid formation, within the laser pulse duration, of a permanent photoproduct. Saturation of the irradiated solution with CO results in regeneration of the starting complex 1. IR and Raman spectral data suggest that the photoproduct is the internally stabilized 16- electron species (CO)4W=C(NC,iH8)SiPh3. The observations are discussed in relation to the previously reported formation of the same 16-electron species by thermolysis of 1. When the irradiation is carried out in CH3CN as solvent, UV-visible evidence suggests formation of the photosubstituted species (CO)4(CH3CN) W=C(NC4Hg)SiPh3. No photoactivity, either transient or permanent, is seen in any solvent when the irradiation is carried out at 416 nm, a wavelength which falls within the MLCT absorption region of 1. When the ethoxy-substituted carbene complex 2 is irradiated in either the LF or MLCT absorption regions a transient species forms rapidly, within the laser pulse duration, and decays on a time scale of several μß, with a lifetime dependent on solvent polarity but independent of CO concentration in solution. Time-resolved resonance Raman studies in which the sample is photolyzed at 355 nm and probed by means of a delayed pulse at 406 nm show the formation and decay of a transient consistent with the flash photolysis results. The data are interpreted in terms of photoinduced anti-syn isomerization of 2 about the Ccarbene~0 bond

Microwave Spectroscopy

Contains research objectives and reports on five research projects.Signal Corps Contract DA36-039-sc-73014Signal Corps Contract DA36-039-sc-7489

Development of novel methods for non-canonical myeloma protein analysis with an innovative adaptation of immunofixation electrophoresis, native top-down mass spectrometry, and middle-down de novo sequencing

OBJECTIVES: Multiple myeloma (MM) is a malignant plasma cell neoplasm, requiring the integration of clinical examination, laboratory and radiological investigations for diagnosis. Detection and isotypic identification of the monoclonal protein(s) and measurement of other relevant biomarkers in serum and urine are pivotal analyses. However, occasionally this approach fails to characterize complex protein signatures. Here we describe the development and application of next generation mass spectrometry (MS) techniques, and a novel adaptation of immunofixation, to interrogate non-canonical monoclonal immunoproteins. METHODS: Immunoprecipitation immunofixation (IP-IFE) was performed on a Sebia Hydrasys Scan2. Middle-down de novo sequencing and native MS were performed with multiple instruments (21T FT-ICR, Q Exactive HF, Orbitrap Fusion Lumos, and Orbitrap Eclipse). Post-acquisition data analysis was performed using Xcalibur Qual Browser, ProSight Lite, and TDValidator. RESULTS: We adapted a novel variation of immunofixation electrophoresis (IFE) with an antibody-specific immunosubtraction step, providing insight into the clonal signature of gamma-zone monoclonal immunoglobulin (M-protein) species. We developed and applied advanced mass spectrometric techniques such as middle-down de novo sequencing to attain in-depth characterization of the primary sequence of an M-protein. Quaternary structures of M-proteins were elucidated by native MS, revealing a previously unprecedented non-covalently associated hetero-tetrameric immunoglobulin. CONCLUSIONS: Next generation proteomic solutions offer great potential for characterizing complex protein structures and may eventually replace current electrophoretic approaches for the identification and quantification of M-proteins. They can also contribute to greater understanding of MM pathogenesis, enabling classification of patients into new subtypes, improved risk stratification and the potential to inform decisions on future personalized treatment modalities

NIOX VERO: Individualized Asthma Management in Clinical Practice

As we move toward an era of precision medicine, novel biomarkers of disease will enable the identification and personalized treatment of new endotypes. In asthma, fractional exhaled nitric oxide (FeNO) serves as a surrogate marker of airway inflammation that often correlates with the presence of sputum eosinophils. The increase in FeNO is driven by an upregulation of inducible nitric oxide synthase (iNOS) by cytokines, which are released as a result of type-2 airway inflammation. Scientific evidence supports using FeNO in routine clinical practice. In steroid-naive patients and in patients with mild asthma, FeNO levels decrease within days after corticosteroid treatment in a dose-dependent fashion and increase after steroid withdrawal. In difficult asthma, FeNO testing correlates with anti-inflammatory therapy compliance. Assessing adherence by FeNO testing can remove the confrontational aspect of questioning a patient about compliance and change the conversation to one of goal setting and ways to improve disease management. However, the most important aspect of incorporating FeNO in asthma management is the reduction in the risk of exacerbations. In a recent primary care study, reduction of exacerbation rates and improved symptom control without increasing overall inhaled corticosteroid (ICS) use were demonstrated when a FeNO-guided anti-inflammatory treatment algorithm was assessed and compared to the standard care. A truly personalized asthma management approach—showing reduction of exacerbation rates, overall use of ICS and neonatal hospitalizations—was demonstrated when FeNO testing was applied as part of the treatment algorithm that managed asthma during pregnancy. The aim of this article is to describe how FeNO and the NIOX VERO® analyzer can help to optimize diagnosis and treatment choices and to aid in the monitoring and improvement of clinical asthma outcomes in children and adults