QUANTITATION OF HUMAN RED BLOOD CELL FIXATION BY GLUTARALDEHYDE

The uptake of glutaraldehyde by human red blood cells has been measured as a function of time by a freezing point osmometer. The rate of attachment of glutaraldehyde to the cell proteins is high over the first hour, declining to zero over a period of a few days. The number of glutaraldehyde molecules cross-linking with each hemoglobin molecule is of the order of 200, in reasonable agreement with the calculated number of attachment sites. The cell membrane is immediately highly permeable to glutaraldehyde. Selective permeability to ions is lost during fixation. Ionic equilibrium is obtained only after a few hours. An optimum fixation technique for shape preservation is suggested

Nitrite oxidation exceeds reduction and fixed nitrogen loss in anoxic Pacific waters

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Babbin, A. R., Buchwald, C., Morel, F. M. M., Wankel, S. D., & Ward, B. B. Nitrite oxidation exceeds reduction and fixed nitrogen loss in anoxic Pacific waters. Marine Chemistry, 224, (2020): 103814, doi:10.1016/j.marchem.2020.103814.The diversity of nitrogen-based dissimilatory metabolisms in anoxic waters continues to increase with additional studies to the marine oxygen deficient zones (ODZs). Although the microbial oxidation of nitrite (NO2–) has been known for over a century, studies of the pathways and microbes involved have generally proceeded under the assumption that nitrite oxidation to nitrate requires dioxygen (O2). Anaerobic NO2– oxidation until now has been conclusively shown only for anammox bacteria, albeit only as a limited sink for NO2– in their metabolism compared to the NO2– reduced to N2. Here, using direct experimental techniques optimized for replicating in situ anoxic conditions, we show that NO2– oxidation is substantial, widespread, and consistent across the ODZs of the eastern tropical Pacific Ocean. Regardless of the specific oxidant, NO2– oxidation rates are up to an order of magnitude larger than simultaneous N2 production rates for which these zones are known, and cannot be explained by anammox rates alone. Higher rates of NO2– oxidation over reduction in anoxic waters are paradoxical but help to explain how anammox rates can be enhanced over denitrification in shallow anoxic waters (σθ < 26.4) at the edge of the ODZs but not within the ODZ core. Furthermore, nitrite oxidation may be the key to reconciliation of the perceived imbalance of the global fixed nitrogen loss budget.This work was funded by National Science Foundation grants OCE–1029951 to B.B.W, BIO–1402109 to A.R.B., and OCE-1260373 to S.D.W. Additional financial support to A.R.B. was provided by Simons Foundation grant 622065 and the generous contributions of Dr. Bruce L. Heflinger

Gross and net production during the spring bloom along the Western Antarctic Peninsula

Author Posting. © The Author(s), 2014. This is the author's version of the work. It is posted here by permission of New Phytologist Trust for personal use, not for redistribution. The definitive version was published in New Phytologist 205 (2015): 182-191, doi:10.1111/nph.13125.This study explores some of the physiological mechanisms responsible for high productivity near the shelf in the Western Antarctic Peninsula despite a short growing season and cold temperature. We measured gross and net primary production at Palmer Station during the summer 2012/2013 via three different techniques: 1) incubation with H218O; 2) incubation with 14CO2; and 3) in situ measurements of O2/Ar and triple oxygen isotope. Additional laboratory experiments were performed with the psychrophilic diatom Fragilariopsis cylindrus. During the spring bloom, which accounted for more than half of the seasonal gross production at Palmer Station, the ratio of net to gross production reached a maximum greater than ~60%, among the highest ever reported. The use of multiple-techniques showed that these high ratios resulted from low heterotrophic respiration and very low daylight autotrophic respiration. Laboratory experiments revealed a similar ratio of net to gross O2 production in F.cylindrus and provided the first experimental evidence for an important level of cyclic electron flow (CEF) in this organism. The low ratio of community respiration to gross primary production observed during the bloom at Palmer Station may be characteristic of high latitude coastal ecosystems and partially supported by a very active CEF in psychrophilic phytoplankton.This study was supported by funds from the US National Science Foundation (Award numbers 1040965 and 1043593). Funding to PDT was provided by the Natural Science and Engineering Research Council of Canada

Metabolic balance of coastal Antarctic waters revealed by autonomous pCO2 and ΔO2/Ar measurements

Author Posting. © American Geophysical Union, 2014. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geophysical Research Letters 41 (2014): 6803–6810, doi:10.1002/2014GL061266.We use autonomous gas measurements to examine the metabolic balance (photosynthesis minus respiration) of coastal Antarctic waters during the spring/summer growth season. Our observations capture the development of a massive phytoplankton bloom and reveal striking variability in pCO2 and biological oxygen saturation (ΔO2/Ar) resulting from large shifts in community metabolism on time scales ranging from hours to weeks. Diel oscillations in surface gases are used to derive a high-resolution time series of net community production (NCP) that is consistent with 14C-based primary productivity estimates and with the observed seasonal evolution of phytoplankton biomass. A combination of physical mixing, grazing, and light availability appears to drive variability in coastal Antarctic NCP, leading to strong shifts between net autotrophy and heterotrophy on various time scales. Our approach provides insight into the metabolic responses of polar ocean ecosystems to environmental forcing and could be employed to autonomously detect climate-dependent changes in marine primary productivity.This study was supported by funds from the U.S. National Science Foundation (OPP awards ANT-0823101, ANT-1043559, ANT-1043593, and ANT-1043532) as well as support for PDT and ECA from the National Science and Engineering Research Council of Canada.2015-04-0

Cadmium isotopic composition in the ocean

Author Posting. © Elsevier B.V., 2006. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Geochimica et Cosmochimica Acta 70 (2006): 5104-5118, doi:10.1016/j.gca.2006.07.036.The oceanic cycle of cadmium is still poorly understood, despite its importance for phytoplankton growth and paleoceanographic applications. As for other elements that are biologically recycled, variations in isotopic composition may bring unique insights. This article presents i) a protocol for the measurement of cadmium isotopic composition (Cd IC) in seawater and in phytoplankton cells; ii) the first Cd IC data in seawater, from two full depth stations, in the northwest Pacific and the northwest Mediterranean Sea; iii) the first Cd IC data in phytoplankton cells, cultured in vitro. The Cd IC variation range in seawater found at these stations is not greater than 1.5 eCd/amu units, only slightly larger than the mean uncertainty of measurement (0.8 eCd/amu). Nevertheless, systematic variations of the Cd IC and concentration in the upper 300m of the northwest Pacific suggest the occurrence of Cd isotopic fractionation by phytoplankton uptake, with a fractionation factor of 1.6±1.4 eCd/amu units. This result is supported by the culture experiment data suggesting that freshwater phytoplankton (Chlamydomonas reinhardtii and Chlorella sp.) preferentially take up light Cd isotopes, with a fractionation factor of 3.4±1.4 eCd/amu units. Systematic variations of the Cd IC and hydrographic data between 300 and 700m in the northwest Pacific have been tentatively attributed to the mixing of the mesothermal (temperature maximum) water (eCd/amu=-0.9±0.8) with the North Pacific Intermediate Water (eCd/amu=0.5±0.8). In contrast, no significant Cd IC variation is found in the northwest Mediterranean Sea. This observation was attributed to the small surface Cd depletion by phytoplankton uptake and the similar Cd IC of the different water masses found at this site. Overall, these data suggest that i) phytoplankton uptake fractionates Cd isotopic composition to a measurable degree (fractionation factors of 1.6 and 3.4 eCd/amu units, for the in situ and culture experiment data, respectively), ii) an open ocean profile of Cd IC shows upper water column variations consistent with preferential uptake and regeneration of light Cd isotopes, and iii) different water masses may have different Cd IC. This isotopic system could therefore provide information on phytoplankton Cd uptake and on water mass trajectories and mixing in some areas of the ocean. However, the very small Cd IC variations found in this study indicate that applications of Cd isotopic composition to reveal aspects of the present or past Cd oceanic cycle will be very challenging and may require further analytical improvements. Better precision could possibly be obtained with larger seawater samples, a better chemical separation of tin and a more accurate mass bias correction through the use of the double spiking technique

Human pallidothalamic and cerebellothalamic tracts: anatomical basis for functional stereotactic neurosurgery

Anatomical knowledge of the structures to be targeted and of the circuitry involved is crucial in stereotactic functional neurosurgery. The present study was undertaken in the context of surgical treatment of motor disorders such as essential tremor (ET) and Parkinson’s disease (PD) to precisely determine the course and three-dimensional stereotactic localisation of the cerebellothalamic and pallidothalamic tracts in the human brain. The course of the fibre tracts to the thalamus was traced in the subthalamic region using multiple staining procedures and their entrance into the thalamus determined according to our atlas of the human thalamus and basal ganglia [Morel (2007) Stereotactic atlas of the human thalamus and basal ganglia. Informa Healthcare Inc., New York]. Stereotactic three-dimensional coordinates were determined by sectioning thalamic and basal ganglia blocks parallel to stereotactic planes and, in two cases, by correlation with magnetic resonance images (MRI) from the same brains prior to sectioning. The major contributions of this study are to provide: (1) evidence that the bulks of the cerebellothalamic and pallidothalamic tracts are clearly separated up to their thalamic entrance, (2) stereotactic maps of the two tracts in the subthalamic region, (3) the possibility to discriminate between different subthalamic fibre tracts on the basis of immunohistochemical stainings, (4) correlations of histologically identified fibre tracts with high-resolution MRI, and (5) evaluation of the interindividual variability of the fibre systems in the subthalamic region. This study should provide an important basis for accurate stereotactic neurosurgical targeting of the subthalamic region in motor disorders such as PD and ET

Transit Timing Observations from Kepler: III. Confirmation of 4 Multiple Planet Systems by a Fourier-Domain Study of Anti-correlated Transit Timing Variations

We present a method to confirm the planetary nature of objects in systems with multiple transiting exoplanet candidates. This method involves a Fourier-Domain analysis of the deviations in the transit times from a constant period that result from dynamical interactions within the system. The combination of observed anti-correlations in the transit times and mass constraints from dynamical stability allow us to claim the discovery of four planetary systems Kepler-25, Kepler-26, Kepler-27, and Kepler-28, containing eight planets and one additional planet candidate.Comment: Accepted to MNRA

Chalk-Ex—fate of CaCO3 particles in the mixed layer : evolution of patch optical properties

Author Posting. © American Geophysical Union, 2009. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research 114 (2009): C07020, doi:10.1029/2008JC004902.The fate of particles in the mixed layer is of great relevance to the global carbon cycle as well as to the propagation of light in the sea. We conducted four manipulative field experiments called “Chalk-Ex” in which known quantities of uniform, calcium carbonate particles were injected into the surface mixed layer. Since the production term for these patches was known to high precision, the experimental design allowed us to focus on terms associated with particle loss. The mass of chalk in the patches was evaluated using the well-calibrated light-scattering properties of the chalk plus measurements from a variety of optical measurements and platforms. Patches were surveyed with a temporal resolution of hours over spatial scales of tens of kilometers. Our results demonstrated exponential loss of the chalk particles with time from the patches. There was little evidence for rapid sinking of the chalk. Instead, horizontal eddy diffusion appeared to be the major factor affecting the dispersion of the chalk to concentrations below the limits of detection. There was unequivocal evidence of subduction of the chalk along isopycnals and subsequent formation of thin layers. Shear dispersion is the most likely mechanism to explain these results. Calculations of horizontal eddy diffusivity were consistent with other mixed layer patch experiments. Our results provide insight into the importance of physics in the formation of subsurface particle maxima in the sea, as well as the importance of rapid coccolith production and critical patch size for maintenance of natural coccolithophore blooms in nature.We would like to thank the Office of Naval Research/Optical and Biological Oceanography Program for their support of Chalk-Ex with awards N000140110042 (WMB) and N00014-01-1-0141 (AJP). Additional funding for this work came from ONR (N00014-05-1- 0111) and NASA (NNG04Gl11G, NNX08AC27G, NNG04HZ25C) to W.M.B