Novel application of confocal laser scanning microscopy and 3D volume rendering toward improving the resolution of the fossil record of charcoal.

Published onlineHistorical ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, Non-P.H.S.This is the final version of the article. It first appeared from PLoS via http://dx.doi.org/10.1371/journal.pone.0072265Variations in the abundance of fossil charcoals between rocks and sediments are assumed to reflect changes in fire activity in Earth's past. These variations in fire activity are often considered to be in response to environmental, ecological or climatic changes. The role that fire plays in feedbacks to such changes is becoming increasingly important to understand and highlights the need to create robust estimates of variations in fossil charcoal abundance. The majority of charcoal based fire reconstructions quantify the abundance of charcoal particles and do not consider the changes in the morphology of the individual particles that may have occurred due to fragmentation as part of their transport history. We have developed a novel application of confocal laser scanning microscopy coupled to image processing that enables the 3-dimensional reconstruction of individual charcoal particles. This method is able to measure the volume of both microfossil and mesofossil charcoal particles and allows the abundance of charcoal in a sample to be expressed as total volume of charcoal. The method further measures particle surface area and shape allowing both relationships between different size and shape metrics to be analysed and full consideration of variations in particle size and size sorting between different samples to be studied. We believe application of this new imaging approach could allow significant improvement in our ability to estimate variations in past fire activity using fossil charcoals.This research was supported by funding from a European Union Marie Curie Intra-European Fellowship (FILE-PIEF-GA-2009-25378 to CMB), a Marie Curie Career Integration Grant (PyroMap PCIG10-GA-2011-303610 to CMB), a University of Exeter Outward Mobility Academic Fellowship (to CMB) and the US National Science Foundation (DBI-1052997 to SWP). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Correction factors for δ 18 O-derived global sea surface temperature reconstructions from diagenetically altered intervals of coral skeletal density banding

Reconstruction of sea surface temperature (SST) from the δ18O and Sr/Ca composition of coral skeletal density banding (CSDB), identified with x-ray diffraction and micro computed tomography, provides invaluable centuries-long records of ocean circulation and climate change. Comparison with age-equivalent instrument measurements of SST over the last 125 years has proven these δ18O-derived SST reconstructions to be generally reliable. However, notable exceptions occur within discrete CSDB stratigraphic intervals that yield δ18O-derived SST underestimates of as much as 9°C with respect to instrument measured SST. Here we combine high-resolution optical and electron microscopy with geochemical modeling to establish correction factors for the impact of marine seafloor physical, chemical, and biological alteration (diagenesis) within these altered intervals of CSDB stratigraphy. Four cores were collected from Porites coral heads across a 4-24 m water depth bathymetric transect at Myrmidon Reef, Great Barrier Reef, Australia. Precise mapping of diagenetic aragonite cementation was completed within CSDB patterns digitally overlaid on 35 petrographic thin sections fully covering 2.1 m of core. The vast majority of core skeletal material exhibited little to no diagenetic aragonite cementation. However, extensive diagenetic alteration was observed within discrete CSDB intervals near the base of the two deeper water Porites heads. This diagenesis serves to modify skeletal density and CSDB stratigraphy in these intervals, as well as structurally reinforce the coral skeleton. Reliable δ18O-based SST correction factors for these diagenetically altered CSDB intervals are established here by applying the percent mixing of diagenetic aragonite cement to a binary mixing model. This approach, with quantitative extents of mixing established with both microscopy and existing globally distributed coral δ18O and Sr/Ca data sets, accurately restores modern and fossil coral δ18O-derived SST records. Results indicate that as little as 5% mixing of diagenetic aragonite cement with original coral skeleton will cause δ18O-based SST anomalies of 0.9°C

Corals regulate the distribution and abundance of Symbiodiniaceae and biomolecules in response to changing water depth and sea surface temperature

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Sivaguru, M., Todorov, L. G., Fouke, C. E., Munro, C. M. O., Fouke, K. W., Fouke, K. E., Baughman, M. E., & Fouke, B. W. Corals regulate the distribution and abundance of Symbiodiniaceae and biomolecules in response to changing water depth and sea surface temperature. Scientific Reports, 11(1), (2021): 2230, https://doi.org/10.1038/s41598-021-81520-0.The Scleractinian corals Orbicella annularis and O. faveolata have survived by acclimatizing to environmental changes in water depth and sea surface temperature (SST). However, the complex physiological mechanisms by which this is achieved remain only partially understood, limiting the accurate prediction of coral response to future climate change. This study quantitatively tracks spatial and temporal changes in Symbiodiniaceae and biomolecule (chromatophores, calmodulin, carbonic anhydrase and mucus) abundance that are essential to the processes of acclimatization and biomineralization. Decalcified tissues from intact healthy Orbicella biopsies, collected across water depths and seasonal SST changes on Curaçao, were analyzed with novel autofluorescence and immunofluorescence histology techniques that included the use of custom antibodies. O. annularis at 5 m water depth exhibited decreased Symbiodiniaceae and increased chromatophore abundances, while O. faveolata at 12 m water depth exhibited inverse relationships. Analysis of seasonal acclimatization of the O. faveolata holobiont in this study, combined with previous reports, suggests that biomolecules are differentially modulated during transition from cooler to warmer SST. Warmer SST was also accompanied by decreased mucus production and decreased Symbiodiniaceae abundance, which is compensated by increased photosynthetic activity enhanced calcification. These interacting processes have facilitated the remarkable resiliency of the corals through geological time.Financial support for this work was provided by the Office of Naval Research (N00014-00-1-0609), the Illinois Carl R. Woese Institute for Genomic Biology Undergraduate Summer Research Fellowship, the Illinois Carl R. Woese Institute for Genomic Biology Mark Tracy Fellowship for Translational Research, and the Illinois Department of Molecular and Cellular Biology Jenner Family Summer Research Fellowship and the Edward and Barbara Weil Research Fund provided to the University of Illinois Urbana-Champaign

Kinetic Models of Trymethylolpropane (TMP) and Castor Oil Methyl Ester Transesterification for engine oil synthesis

Non-digestible oil as renewable resources has grown a compulsive concern for engine oil synthesis. The synthesis of engine oil was conducted in a bath stirrer flask via two-steps catalytic transesterification. Modification of trimethylolpropane (TMP) and castor oil methyl ester (COME) transesterification was run under non- and impregnated Perna V. shells catalysts. Operation temperature effect on the castor trimethylol propane triester as engine lubricant was conditioned at range 90°C – 130°C. The optimal reaction temperature was detected at 110°C with 98.60 % TMP conversion and 71.29% triester composition. Excess dosage of COME was structured at 4:1 ratio to TMP for progressive reaction limitation. Kinetics of TMP and COME transesterifation were established for first- and second orders. The second order kinetic model best matchs the acquired data for an overall reaction rate constants of 0.0354 (% wt/wt min °C)-1. Synthesized engine oil resulted activation energy of 74.10 kJ/mol. Estimated pour and flash point were -17 °C and 221°C. Kinematic viscosity at 40°C was 64.31 mm2/s and density was 872 kg/m3 fulfilled the proposed lubricant standard. The kinetic models were proportionate to other oil seeds crops based engine oil, typically Jatropha seeds oil

Monoclonal antibodies for copper-64 PET dosimetry and radioimmunotherapy

BACKGROUND: We previously described a two-antibody model of (64)Cu radioimmunotherapy to evaluate low-dose, solid-tumor response. This model was designed to test the hypothesis that cellular internalization is critical in causing tumor cell death by mechanisms in addition to radiation damage. The purpose of the present study was to estimate radiation dosimetry for both antibodies (mAbs) using positron emission tomography (PET) imaging and evaluate the effect of internalization on tumor growth. RESULTS: Dosimetry was similar between therapy groups. Median time to tumor progression to 1 g ranged from 7–12 days for control groups and was 32 days for both treatment groups (p < 0.0001). No statistically significant difference existed between any control group or between the treatment groups. MATERIAL AND METHODS: In female nude mice bearing LS174T colon carcinoma xenografts, tumor dosimetry was calculated using serial PET images of three mice in each group of either internalizing (64)Cu-labeled DOTA-cBR96 (DOTA = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) or non-internalizing (64)Cu-labeled DOTA-cT84.66 from 3 to 48 h. For the therapy study, controls (n = 10) received saline, DOTA-cBR96 or DOTA-cT84.66. Treatment animals (n = 9) received 0.890 mCi of (64)Cu-labeled DOTA-cBR96 or 0.710 mCi of (64)Cu-labeled DOTA-cT84.66. Tumors were measured daily. CONCLUSIONS: PET imaging allows the use of (64)Cu for pre-therapy calculation of tumor dosimetry. In spite of highly similar tumor dosimetry, an internalizing antibody did not improve the outcome of (64)Cu radioimmunotherapy. Radio-resistance of this tumor cell line and copper efflux may have confounded the study. Further investigations of the therapeutic efficacy of (64)Cu-labeled mAbs will focus on interaction between (64)Cu and tumor suppressor genes and copper chaperones

Length of carotid stenosis predicts peri-procedural stroke or death and restenosis in patients randomized to endovascular treatment or endarterectomy.

BACKGROUND: The anatomy of carotid stenosis may influence the outcome of endovascular treatment or carotid endarterectomy. Whether anatomy favors one treatment over the other in terms of safety or efficacy has not been investigated in randomized trials. METHODS: In 414 patients with mostly symptomatic carotid stenosis randomized to endovascular treatment (angioplasty or stenting; n = 213) or carotid endarterectomy (n = 211) in the Carotid and Vertebral Artery Transluminal Angioplasty Study (CAVATAS), the degree and length of stenosis and plaque surface irregularity were assessed on baseline intraarterial angiography. Outcome measures were stroke or death occurring between randomization and 30 days after treatment, and ipsilateral stroke and restenosis ≥50% during follow-up. RESULTS: Carotid stenosis longer than 0.65 times the common carotid artery diameter was associated with increased risk of peri-procedural stroke or death after both endovascular treatment [odds ratio 2.79 (1.17-6.65), P = 0.02] and carotid endarterectomy [2.43 (1.03-5.73), P = 0.04], and with increased long-term risk of restenosis in endovascular treatment [hazard ratio 1.68 (1.12-2.53), P = 0.01]. The excess in restenosis after endovascular treatment compared with carotid endarterectomy was significantly greater in patients with long stenosis than with short stenosis at baseline (interaction P = 0.003). Results remained significant after multivariate adjustment. No associations were found for degree of stenosis and plaque surface. CONCLUSIONS: Increasing stenosis length is an independent risk factor for peri-procedural stroke or death in endovascular treatment and carotid endarterectomy, without favoring one treatment over the other. However, the excess restenosis rate after endovascular treatment compared with carotid endarterectomy increases with longer stenosis at baseline. Stenosis length merits further investigation in carotid revascularisation trials

Is callose a barrier for lead ions entering Lemna minor L. root cells?

Plants have developed a range of strategies for resisting environmental stresses. One of the most common is the synthesis and deposition of callose, which functions as a barrier against stress factor penetration. The aim of our study was to examine whether callose forms an efficient barrier against Pb penetration in the roots of Lemna minor L. exposed to this metal. The obtained results showed that Pb induced callose synthesis in L. minor roots, but it was not deposited regularly in all tissues and cells. Callose occurred mainly in the protoderm and in the centre of the root tip (procambial central cylinder). Moreover, continuous callose bands, which could form an efficient barrier for Pb penetration, were formed only in the newly formed and anticlinal cell walls (CWs); while in other CWs, callose formed only small clusters or incomplete bands. Such an arrangement of callose within root CWs inefficiently protected the protoplast from Pb penetration. As a result, Pb was commonly present inside the root cells. In the light of the results, the barrier role of callose against metal ion penetration appears to be less obvious than previously believed. It was indicated that induction of callose synthesis is not enough for a successful blockade of the stress factor penetration. Furthermore, it would appear that the pattern of callose distribution has an important role in this defence strategy

Oxidative Stress in Dilated Cardiomyopathy Caused by MYBPC3 Mutation

Cardiomyopathies can result from mutations in genes encoding sarcomere proteins including MYBPC3, which encodes cardiac myosin binding protein-C (cMyBP-C). However, whether oxidative stress is augmented due to contractile dysfunction and cardiomyocyte damage in MYBPC3-mutated cardiomyopathies has not been elucidated. To determine whether oxidative stress markers were elevated in MYBPC3-mutated cardiomyopathies, a previously characterized 3-month-old mouse model of dilated cardiomyopathy (DCM) expressing a homozygous MYBPC3 mutation (cMyBP-C(t/t)) was used, compared to wild-type (WT) mice. Echocardiography confirmed decreased percentage of fractional shortening in DCM versus WT hearts. Histopathological analysis indicated a significant increase in myocardial disarray and fibrosis while the second harmonic generation imaging revealed disorganized sarcomeric structure and my

Capturing the Surface Texture and Shape of Pollen: A Comparison of Microscopy Techniques

Research on the comparative morphology of pollen grains depends crucially on the application of appropriate microscopy techniques. Information on the performance of microscopy techniques can be used to inform that choice. We compared the ability of several microscopy techniques to provide information on the shape and surface texture of three pollen types with differing morphologies. These techniques are: widefield, apotome, confocal and two-photon microscopy (reflected light techniques), and brightfield and differential interference contrast microscopy (DIC) (transmitted light techniques). We also provide a first view of pollen using super-resolution microscopy. The three pollen types used to contrast the performance of each technique are: Croton hirtus (Euphorbiaceae), Mabea occidentalis (Euphorbiaceae) and Agropyron repens (Poaceae). No single microscopy technique provided an adequate picture of both the shape and surface texture of any of the three pollen types investigated here. The wavelength of incident light, photon-collection ability of the optical technique, signal-to-noise ratio, and the thickness and light absorption characteristics of the exine profoundly affect the recovery of morphological information by a given optical microscopy technique. Reflected light techniques, particularly confocal and two-photon microscopy, best capture pollen shape but provide limited information on very fine surface texture. In contrast, transmitted light techniques, particularly differential interference contrast microscopy, can resolve very fine surface texture but provide limited information on shape. Texture comprising sculptural elements that are spaced near the diffraction limit of light (∼250 nm; NDL) presents an acute challenge to optical microscopy. Super-resolution structured illumination microscopy provides data on the NDL texture of A. repens that is more comparable to textural data from scanning electron microscopy than any other optical microscopy technique investigated here. Maximizing the recovery of morphological information from pollen grains should lead to more robust classifications, and an increase in the taxonomic precision with which ancient vegetation can be reconstructed

Quantitative trait loci and candidate gene mapping of aluminum tolerance in diploid alfalfa

Aluminum (Al) toxicity in acid soils is a major limitation to the production of alfalfa (Medicago sativa subsp. sativa L.) in the USA. Developing Al-tolerant alfalfa cultivars is one approach to overcome this constraint. Accessions of wild diploid alfalfa (M. sativa subsp. coerulea) have been found to be a source of useful genes for Al tolerance. Previously, two genomic regions associated with Al tolerance were identified in this diploid species using restriction fragment length polymorphism (RFLP) markers and single marker analysis. This study was conducted to identify additional Al-tolerance quantitative trait loci (QTLs); to identify simple sequence repeat (SSR) markers that flank the previously identified QTLs; to map candidate genes associated with Al tolerance from other plant species; and to test for co-localization with mapped QTLs. A genetic linkage map was constructed using EST-SSR markers in a population of 130 BC(1)F(1) plants derived from the cross between Al-sensitive and Al-tolerant genotypes. Three putative QTLs on linkage groups LG I, LG II and LG III, explaining 38, 16 and 27% of the phenotypic variation, respectively, were identified. Six candidate gene markers designed from Medicago truncatula ESTs that showed homology to known Al-tolerance genes identified in other plant species were placed on the QTL map. A marker designed from a candidate gene involved in malic acid release mapped near a marginally significant QTL (LOD 2.83) on LG I. The SSR markers flanking these QTLs will be useful for transferring them to cultivated alfalfa via marker-assisted selection and for pyramiding Al tolerance QTLs