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A DNA-based fluorescent reporter maps HOCl production in the maturing phagosome
摘要： Phagocytes destroy pathogens by trapping them in a transient organelle called the phagosome, where they are bombarded with reactive oxygen species (ROS) and reactive nitrogen species (RNS). Imaging reactive species within the phagosome would directly reveal the chemical dynamics underlying pathogen destruction. Here we introduce a fluorescent, DNA-based combination reporter, cHOClate, which simultaneously images hypochlorous acid (HOCl) and pH quantitatively. Using cHOClate targeted to phagosomes in live cells, we successfully map phagosomal production of a specific ROS, HOCl, as a function of phagosome maturation. We found that phagosomal acidification was gradual in macrophages and upon completion, HOCl was released in a burst. This revealed that phagosome–lysosome fusion was essential not only for phagosome acidification, but also for providing the chloride necessary for myeloperoxidase activity. This method can be expanded to image several kinds of ROS and RNS and be readily applied to identify how resistant pathogens evade phagosomal killing.
关键词： hypochlorous acid,phagosome,reactive oxygen species,DNA-based reporter,myeloperoxidase
更新于2025-09-04 15:30:14
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Variability of Multispectral Lidar 3D and Intensity Features with Individual Tree Height and Its Influence on Needleleaf Tree Species Identification
摘要： Tree species identification is important in forest management. The multispectral lidar Titan of Teledyne Optech Inc. can improve tree species separation by providing classification features computed from the three-channel intensities, ratios and normalized differences. However, the value of features used in classification algorithms (e.g., random forest, RF) may vary with tree size. The focus of the present study is to show how tree height influences the 3D and intensity features, how this relationship may affect the species classification accuracy, and how different classification strategies may circumvent this problem. Six needleleaf species (Pinus resinosa, Pinus strobus, Pinus sylvestris, Larix laricina, Picea abies and Picea glauca), found in plantations of different ages, were sampled to train classifiers. Some features yielded a good discriminatory power for species identification, despite their relation to tree height (r2 up to 0.6). Two classification strategies—a) using only size-invariant features (SIF) and b) training separate classifiers per tree height strata (HSC)—were compared to a standard classification (STD: all features, without height stratification). The accuracy of the SIF approach was lowest, useful variables being removed due to their relationship to tree height. The HSC provided only a minor improvement over the STD results.
关键词： tree species identification,Teledyne Optech Inc.,Titan,random forest,3D features,intensity features,multispectral lidar
更新于2025-09-04 15:30:14
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Global model of an atmospheric-pressure capacitive discharge in helium with air impurities from 100 to 10000 ppm
摘要： Helium is a common working gas for cold atmospheric plasmas (CAPs) and this is often mixed with other gases, such as oxygen and nitrogen, to increase its reactivity. Air is often found in these plasmas and it can be either introduced deliberately as a precursor or entrapped in systems that operate in open atmosphere. In either case, the presence of small traces of air can cause a profound change on the composition of the plasma and consequently its application efficacy. In this paper, a global model for He+Air CAPs is developed, in which 59 species and 866 volume reactions are incorporated, and a new boundary condition is used for the mass transport at the interface between the plasma and its surrounding air gas. The densities of reactive species and the power dissipation characteristics are obtained as a function of air concentrations spanning from 100 to 10000 ppm. As the air concentration increases, the dominant cation changes from O2+ to NO+ and then to NO2+, the dominant anion changes from O2- to NO2- and then to NO3-, the dominant ground state reactive oxygen species changes from O to O3, and the dominant ground state reactive nitrogen species changes from NO to HNO2. O2(a) is the most abundant metastable species and its density is orders of magnitude larger than other metastable species for all air concentrations considered in the study. Ion Joule heating is found important due to the electronegative nature of the plasma, which leads to the fast decrease of electron density when the air concentration is larger than 1000 ppm. The generation and loss pathways of important biologically relevant reactive species such as O, O2-, O3, OH, H2O2, NO, HNO2, HNO3 are discussed and differences with the pathways observed in He+O2, He+H2O, Ar+Air and pure air plasmas are highlighted. Based on the simulation results, a simplified chemistry set with 47 species and 109 volume reactions is proposed. This simplified model greatly reduces the computational load while maintaining the accuracy of the simulation results within a factor of 2. The simplified chemistry model is computationally much less intensive, facilitating its integration into multidimensional fluid models for the study of the spatio-temporal evolution of He+Air CAPs.
关键词： helium,simplified chemistry model,reactive species,global model,cold atmospheric plasmas,power dissipation,air impurities
更新于2025-09-04 15:30:14
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Photobiomodulation at Multiple Wavelengths Differentially Modulates Oxidative Stress <i>In Vitro</i> and <i>In Vivo</i>
摘要： Photobiomodulation (PBM) is emerging as an effective strategy for the management of multiple inflammatory conditions, including oral mucositis (OM) in cancer patients who receive chemotherapy or radiotherapy. Still, the poor understanding of the mechanisms by which the light interacts with biological tissues and the heterogeneity of light sources and protocols employed worldwide significantly limits its applicability. Reactive oxygen species (ROS) are massively generated during the early phases of OM and play a major role in the pathogenesis of inflammation in general. Here, we report the results of a clinical and experimental study, aimed at evaluating the effect of laser light at different wavelengths on oxidative stress in vivo in oncologic patients suffering from OM and in vitro in two cell types abundantly present within the inflamed oral mucosa, neutrophil polymorphonuclear (PMN) granulocytes, and keratinocytes. In addition to standard ROS detection methods, we exploited a roGFP2-Orp1 genetically encoded sensor, allowing specific, quantitative, and dynamic imaging of redox events in living cells in response to oxidative stress and PBM. We found that the various wavelengths differentially modulate ROS production. In particular, the 660 nm laser light increases ROS production when applied either before or after an oxidative stimulus. In contrast, the 970 nm laser light exerted a moderate antioxidant activity both in the saliva of OM patients and in both cell types. The most marked reduction in the levels of ROS was detected in cells exposed either to the 800 nm laser light or to the combination of the three wavelengths. Overall, our study demonstrates that PBM exerts different effects on the redox state of both PMNs and keratinocytes depending on the used wavelength and prompts the validation of a multiwavelength protocol in the clinical settings.
关键词： Reactive oxygen species,Laser therapy,Photobiomodulation,Oxidative stress,Oral mucositis
更新于2025-09-04 15:30:14
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[Methods in Molecular Biology] Microfluidic Electrophoresis Volume 1906 (Methods and Protocols) || Microchip Electrophoresis Containing Electrodes for Integrated Electrochemical Detection
摘要： Microchip electrophoresis is a versatile separation technique. Electrochemical detection is suitable to apply to microdevices due to its easy integration to the fabrication process and good sensitivity and selectivity. Here we describe the procedures to prepare Pt band electrodes deposited on glass to couple to poly-dimethylsiloxane (PDMS) microchips aiming the separation and detection of nitrite using an isolated potentiostat.
关键词： Gated injection,Polydimethylsiloxane,Reactive nitrogen species,Amperometric detection,Isolated potentiostat
更新于2025-09-04 15:30:14
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A versatile dual-species Zeeman slower for caesium and ytterbium
摘要： We describe the design, construction, and operation of a versatile dual-species Zeeman slower for both Cs and Yb, which is easily adaptable for use with other alkali metals and alkaline earths. With the aid of analytic models and numerical simulation of decelerator action, we highlight several real-world problems affecting the performance of a slower and discuss effective solutions. To capture Yb into a magneto-optical trap (MOT), we use the broad 1S0 to 1P1 transition at 399 nm for the slower and the narrow 1S0 to 3P1 intercombination line at 556 nm for the MOT. The Cs MOT and slower both use the D2 line (62S1/2 to 62P3/2) at 852 nm. The slower can be switched between loading Yb or Cs in under 0.1 s. We demonstrate that within a few seconds the Zeeman slower loads more than 109 Yb atoms and 108 Cs atoms into their respective MOTs. These are ideal starting numbers for further experiments on ultracold mixtures and molecules.
关键词： caesium,Zeeman slower,dual-species,ytterbium,ultracold atoms,magneto-optical trap
更新于2025-09-04 15:30:14