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1.
J Photochem Photobiol B ; 241: 112670, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36841175

ABSTRACT

BACKGROUND: Antimicrobial blue light (aBL) kills a variety of bacteria, including Porphyromonas gingivalis. However, little is known about the transcriptomic response of P. gingivalis to aBL therapy. This study was designed to evaluate the selective cytotoxicity of aBL against P. gingivalis over human cells and to further investigate the genetic response of P. gingivalis to aBL at the transcriptome level. METHODS: Colony forming unit (CFU) testing, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM) were used to investigate the antimicrobial effectiveness of blue light against P. gingivalis. The temperatures of the irradiated targets were measured to prevent overheating. Multiple fluorescent probes were used to quantify reactive oxygen species (ROS) generation after blue-light irradiation. RNA sequencing (RNA-seq) was used to investigate the changes in global gene expression. Following the screening of target genes, real-time quantitative polymerase chain reaction (RT-qPCR) was performed to confirm the regulation of gene expression. RESULTS: A 405 nm aBL at 100 mW/cm2 significantly killed P. gingivalis within 5 min while sparing human gingival fibroblasts (HGFs). No obvious temperature changes were detected in the irradiated surface under our experimental conditions. RNA-seq showed that the transcription of multiple genes was regulated, and RT-qPCR revealed that the expression levels of the genes RgpA and RgpB, which may promote heme uptake, as well as the genes Ftn and FetB, which are related to iron homeostasis, were significantly upregulated. The expression levels of the FeoB-2 and HmuR genes, which are related to hydroxyl radical scavenging, were significantly downregulated. CONCLUSIONS: aBL strengthens the heme uptake and iron export gene pathways while reducing the ROS scavenging pathways in P. gingivalis, thus improving the accumulation of endogenous photosensitizers and enhancing oxidative damage to P. gingivalis.


Subject(s)
Color , Gene Expression Regulation, Bacterial , Genes, Bacterial , Iron , Light , Porphyrins , Porphyromonas gingivalis , Porphyrins/metabolism , Iron/metabolism , Porphyromonas gingivalis/cytology , Porphyromonas gingivalis/genetics , Porphyromonas gingivalis/metabolism , Porphyromonas gingivalis/radiation effects , Biological Transport/genetics , Biological Transport/radiation effects , Humans , Gingiva/cytology , Fibroblasts/cytology , Fibroblasts/radiation effects , Hydroxyl Radical/metabolism , Heme/metabolism , Up-Regulation/radiation effects , Homeostasis/radiation effects , Down-Regulation/radiation effects , Microbial Viability/radiation effects , Reactive Oxygen Species/metabolism , Aerobiosis , Genes, Bacterial/radiation effects , Gene Expression Regulation, Bacterial/genetics , Gene Expression Regulation, Bacterial/radiation effects
2.
Sci Rep ; 11(1): 15043, 2021 07 22.
Article in English | MEDLINE | ID: mdl-34294761

ABSTRACT

An emerging approach with potential in improving the treatment of neurodegenerative diseases and brain tumors is the use of focused ultrasound (FUS) to bypass the blood-brain barrier (BBB) in a non-invasive and localized manner. A large body of pre-clinical work has paved the way for the gradual clinical implementation of FUS-induced BBB opening. Even though the safety profile of FUS treatments in rodents has been extensively studied, the histological and behavioral effects of clinically relevant BBB opening in large animals are relatively understudied. Here, we examine the histological and behavioral safety profile following localized BBB opening in non-human primates (NHPs), using a neuronavigation-guided clinical system prototype. We show that FUS treatment triggers a short-lived immune response within the targeted region without exacerbating the touch accuracy or reaction time in visual-motor cognitive tasks. Our experiments were designed using a multiple-case-study approach, in order to maximize the acquired data and support translation of the FUS system into human studies. Four NHPs underwent a single session of FUS-mediated BBB opening in the prefrontal cortex. Two NHPs were treated bilaterally at different pressures, sacrificed on day 2 and 18 post-FUS, respectively, and their brains were histologically processed. In separate experiments, two NHPs that were earlier trained in a behavioral task were exposed to FUS unilaterally, and their performance was tracked for at least 3 weeks after BBB opening. An increased microglia density around blood vessels was detected on day 2, but was resolved by day 18. We also detected signs of enhanced immature neuron presence within areas that underwent BBB opening, compared to regions with an intact BBB, confirming previous rodent studies. Logistic regression analysis showed that the NHP cognitive performance did not deteriorate following BBB opening. These preliminary results demonstrate that neuronavigation-guided FUS with a single-element transducer is a non-invasive method capable of reversibly opening the BBB, without substantial histological or behavioral impact in an animal model closely resembling humans. Future work should confirm the observations of this multiple-case-study work across animals, species and tasks.


Subject(s)
Blood-Brain Barrier/metabolism , Blood-Brain Barrier/radiation effects , Neuronavigation/methods , Ultrasonic Waves , Animals , Behavior, Animal , Biological Transport/radiation effects , Biomarkers , Blood-Brain Barrier/diagnostic imaging , Cognition , Magnetic Resonance Imaging , Microbubbles , Models, Animal , Primates , Quantitative Trait, Heritable
3.
Neoplasia ; 23(7): 676-691, 2021 07.
Article in English | MEDLINE | ID: mdl-34139452

ABSTRACT

The blood-brain barrier (BBB) is a major anatomical and physiological barrier limiting the passage of drugs into brain. Central nervous system tumors can impair the BBB by changing the tumor microenvironment leading to the formation of a leaky barrier, known as the blood-tumor barrier (BTB). Despite the change in integrity, the BTB remains effective in preventing delivery of chemotherapy into brain tumors. Focused ultrasound is a unique noninvasive technique that can transiently disrupt the BBB and increase accumulation of drugs within targeted areas of the brain. Herein, we summarize the current understanding of different types of targeted ultrasound mediated BBB/BTB disruption techniques. We also discuss influence of the tumor microenvironment on BBB opening, as well as the role of immunological response following disruption. Lastly, we highlight the gaps between evaluation of the parameters governing opening of the BBB/BTB. A deeper understanding of physical opening of the BBB/BTB and the biological effects following disruption can potentially enhance treatment strategies for patients with brain tumors.


Subject(s)
Blood-Brain Barrier/metabolism , Blood-Brain Barrier/radiation effects , Brain Neoplasms/metabolism , Drug Delivery Systems , Tumor Microenvironment/radiation effects , Ultrasonic Waves , Animals , Biological Transport/radiation effects , Biological Variation, Population , Brain Neoplasms/drug therapy , Brain Neoplasms/etiology , Brain Neoplasms/pathology , Disease Models, Animal , High-Intensity Focused Ultrasound Ablation/adverse effects , High-Intensity Focused Ultrasound Ablation/methods , Humans , Neoplasm Metastasis , Permeability/radiation effects , Treatment Outcome , Tumor Microenvironment/drug effects , Tumor Microenvironment/immunology , Ultrasonic Therapy
4.
Article in English | MEDLINE | ID: mdl-33011226

ABSTRACT

The fluted giant clam, Tridacna squamosa, can perform light-enhanced shell formation, aided by its symbiotic dinoflagellates (Symbiodinium, Cladocopium, Durusdinium), which are able to donate organic nutrients to the host. During light-enhanced shell formation, increased Ca2+ transport from the hemolymph through the shell-facing epithelium of the inner mantle to the extrapallial fluid, where calcification occurs, is necessary. Additionally, there must be increased absorption of exogenous Ca2+ from the surrounding seawater, across the epithelial cells of the ctenidium (gill) into the hemolymph, to supply sufficient Ca2+ for light-enhanced shell formation. When Ca2+ moves across these epithelial cells, the low intracellular Ca2+ concentration must be maintained. Sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) regulates the intracellular Ca2+ concentration by pumping Ca2+ into the sarcoplasmic/endoplasmic reticulum (SR/ER) and Golgi apparatus. Indeed, the ctenidium and inner mantle of T. squamosa, expressed a homolog of SERCA (SERCA-like transporter) that consists of 3009 bp, encoding 1002 amino acids of 110.6 kDa. SERCA-like-immunolabeling was non-uniform in the cytoplasm of epithelial cells of ctenidial filaments, and that of the shell-facing epithelial cells of the inner mantle. Importantly, the protein abundance of SERCA-like increased significantly in the ctenidium and the inner mantle of T. squamosa after 12 h and 6 h, respectively, of light exposure. This would increase the capacity of pumping Ca2+ into the endoplasmic reticulum and avert a possible surge in the cytosolic Ca2+ concentration in epithelial cells of the ctenidial filaments during light-enhanced Ca2+ absorption, and in cells of the shell-facing epithelium of the inner mantle during light-enhanced shell formation.


Subject(s)
Animal Shells/metabolism , Bivalvia/metabolism , Calcium/metabolism , Membrane Transport Proteins/metabolism , Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism , Amino Acid Sequence , Animal Shells/radiation effects , Animals , Biological Transport/radiation effects , Bivalvia/genetics , Bivalvia/radiation effects , Blotting, Western , Gene Expression Regulation/radiation effects , Light , Lighting , Membrane Transport Proteins/genetics , Microscopy, Fluorescence , Reverse Transcriptase Polymerase Chain Reaction , Sarcoplasmic Reticulum Calcium-Transporting ATPases/genetics , Sequence Homology, Amino Acid
5.
Nat Commun ; 11(1): 4907, 2020 09 30.
Article in English | MEDLINE | ID: mdl-32999289

ABSTRACT

Global alterations in the metabolic network provide substances and energy to support tumor progression. To fuel these metabolic processes, extracellular matrix (ECM) plays a dominant role in supporting the mass transport and providing essential nutrients. Here, we report a fibrinogen and thrombin based coagulation system to construct an artificial ECM (aECM) for selectively cutting-off the tumor metabolic flux. Once a micro-wound is induced, a cascaded gelation of aECM can be triggered to besiege the tumor. Studies on cell behaviors and metabolomics reveal that aECM cuts off the mass transport and leads to a tumor specific starvation to inhibit tumor growth. In orthotopic and spontaneous murine tumor models, this physical barrier also hinders cancer cells from distant metastasis. The in vivo gelation provides an efficient approach to selectively alter the tumor mass transport. This strategy results in a 77% suppression of tumor growth. Most importantly, the gelation of aECM can be induced by clinical operations such as ultrasonic treatment, surgery or radiotherapy, implying this strategy is potential to be translated into a clinical combination regimen.


Subject(s)
Biomimetic Materials/administration & dosage , Extracellular Matrix/chemistry , Neoplasms/therapy , Animals , Biological Transport/drug effects , Biological Transport/radiation effects , Biomimetic Materials/chemistry , Biomimetic Materials/radiation effects , Cell Line, Tumor/transplantation , Cell Proliferation/drug effects , Cell Proliferation/radiation effects , Chemoradiotherapy/methods , Disease Models, Animal , Extracellular Matrix/metabolism , Extracellular Matrix/radiation effects , Female , Fibrinogen/administration & dosage , Fibrinogen/chemistry , Fibrinogen/radiation effects , Gels , Humans , Injections, Intravenous , Metabolomics , Mice , Neoplasms/metabolism , Thrombin/administration & dosage , Thrombin/chemistry , Thrombin/radiation effects , Ultrasonic Therapy/methods , Ultrasonic Waves
6.
Mol Plant ; 13(12): 1802-1815, 2020 12 07.
Article in English | MEDLINE | ID: mdl-33075506

ABSTRACT

Several photorespiratory bypasses have been introduced into plants and shown to improve photosynthesis by increasing chloroplastic CO2 concentrations or optimizing energy balance. We recently reported that an engineered GOC bypass could increase photosynthesis and productivity in rice. However, the grain yield of GOC plants was unstable, fluctuating in different cultivation seasons because of varying seed setting rates. In this study, we designed a synthetic photorespiratory shortcut (the GCGT bypass) consisting of genes encoding Oryza sativa glycolate oxidase and Escherichia coli catalase, glyoxylate carboligase, and tartronic semialdehyde reductase. The GCGT bypass was guided by an optimized chloroplast transit peptide that targeted rice chloroplasts and redirected 75% of carbon from glycolate metabolism to the Calvin cycle, identical to the native photorespiration pathway. GCGT transgenic plants exhibited significantly increased biomass production and grain yield, which were mainly attributed to enhanced photosynthesis due to increased chloroplastic CO2 concentrations. Despite the increases in biomass production and grain yield, GCGT transgenic plants showed a reduced seed setting rate, a phenotype previously reported for the GOC plants. Integrative transcriptomic, physiological, and biochemical assays revealed that photosynthetic carbohydrates were not transported to grains in an efficient manner, thereby reducing the seed setting rate. Taken together, our results demonstrate that the GCGT photorespiratory shortcut confers higher yield by promoting photosynthesis in rice, mainly through increasing chloroplastic CO2 concentrations.


Subject(s)
Biomass , Light , Oryza/growth & development , Oryza/radiation effects , Photosynthesis/radiation effects , Seeds/growth & development , Biological Transport/radiation effects , Carbohydrate Metabolism/radiation effects , Carbon Dioxide/metabolism , Cell Respiration/radiation effects , Chloroplasts/metabolism , Chloroplasts/radiation effects , Chloroplasts/ultrastructure , Gene Expression Regulation, Plant/radiation effects , Metabolome/radiation effects , Oryza/genetics , Plant Leaves/metabolism , Plant Leaves/radiation effects , Plant Leaves/ultrastructure , Plants, Genetically Modified , Seeds/radiation effects , Transcriptome/genetics
7.
Mol Neurobiol ; 57(11): 4530-4548, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32748371

ABSTRACT

Aquaporin 9 (AQP9) is an aquaglyceroporin that can transport lactate. Accumulating evidence suggests that astrocyte-to-neuron lactate shuttle (ANLS) plays a critical role in energy metabolism in neurons, including retinal ganglion cells (RGCs). To test the hypothesis that AQP9, in concert with monocarboxylate transporters (MCTs), participates in ANLS to maintain function and survival of RGCs, Aqp9-null mice and wild-type (WT) littermates were subjected to optic nerve crush (ONC) with or without intravitreal injection of an MCT2 inhibitor. RGC density was similar between the Aqp9-null mice and WT mice without ONC, while ONC resulted in significantly more RGC density reduction in the Aqp9-null mice than in the WT mice at day 7. Positive scotopic threshold response (pSTR) amplitude values were similar between the two groups without ONC, but were significantly more reduced in the Aqp9-null mice than in the WT mice 7days after ONC. MCT2 inhibitor injection accelerated RGC death and pSTR amplitude reduction only in the WT mice with ONC. Immunolabeling revealed that both RGCs and astrocytes expressed AQP9, that ONC predominantly reduced astrocytic AQP9 expression, and that MCTs 1, 2, and 4 were co-localized with AQP9 at the ganglion cell layer. These retinal MCTs were also co-immunoprecipitated with AQP9 in the WT mice. ONC decreased the co-immunoprecipitation of MCTs 1 and 4, but did not impact co-immunoprecipitation of MCT2. Retinal glucose transporter 1 expression was increased in Aqp9-null mice. Aqp9 gene deletion reduced and increased the intraretinal L-lactate and D-glucose concentrations, respectively. Results suggest that AQP9 acts as the ANLS to maintain function and survival of RGCs.


Subject(s)
Aquaporins/genetics , Astrocytes/metabolism , Gene Deletion , Lactic Acid/metabolism , Monocarboxylic Acid Transporters/metabolism , Neurons/metabolism , Optic Nerve/pathology , Retinal Ganglion Cells/pathology , Animals , Aquaporins/metabolism , Aquaporins/radiation effects , Astrocytes/radiation effects , Biological Transport/radiation effects , Cell Death , Cell Survival/radiation effects , Dark Adaptation/radiation effects , Electroretinography , Energy Metabolism/radiation effects , Glucose/metabolism , Glucose Transporter Type 1/metabolism , Glucose Transporter Type 3/metabolism , Light , Mice, Inbred C57BL , Mice, Knockout , Monocarboxylic Acid Transporters/antagonists & inhibitors , Nerve Crush , Neurons/radiation effects , Night Vision/radiation effects , Optic Nerve/physiopathology , Optic Nerve/radiation effects , Retinal Ganglion Cells/metabolism , Retinal Ganglion Cells/radiation effects , Sensory Thresholds/radiation effects
8.
Biochim Biophys Acta Bioenerg ; 1861(10): 148257, 2020 10 01.
Article in English | MEDLINE | ID: mdl-32621805

ABSTRACT

Export of reducing power from chloroplasts to cytoplasm serves to balance the NADPH/ATP ratio that is optimal for CO2 assimilation. Rapid cytoplasmic streaming in characean algae conveys the exported metabolites downstream towards the shaded plastids where envelope transporters may operate for the import of reducing power in accordance with the direction of concentration gradients. Import of reducing equivalents by chloroplasts in the analyzed area transiently enhances the pulse-modulated chlorophyll fluorescence F' controlled by the redox state of photosystem II acceptor QA. When the microfluidic pathway was transferred to darkness while the analyzed cell area remained in dim background light, the amplitude of cyclosis-mediated F' changes dropped sharply and then recovered within 5-10 min. The suppression of long-distance signaling indicates temporal depletion of transmitted metabolites in the streaming cytoplasm. The return to overall background illumination induced an exceptionally large F' response to the first local light pulse admitted to a remote cell region. This indicates the appearance of excess reductants in the streaming cytoplasm at a certain stage of photosynthetic induction. The results suggest highly dynamic exchange of metabolites between stationary chloroplasts lining the microfluidic pathway and the streaming cytoplasm upon light-dark and dark-light transitions. Evidence is obtained that slow stages of chlorophyll fluorescence induction in algae with rapid cytoplasmic streaming directly depend on cyclosis-mediated long-distance delivery of metabolites produced far beyond the analyzed cell area.


Subject(s)
Chara/cytology , Cytoplasm/metabolism , Biological Transport/radiation effects , Chara/metabolism , Chara/radiation effects , Darkness , Kinetics
9.
Biochim Biophys Acta Bioenerg ; 1861(10): 148258, 2020 10 01.
Article in English | MEDLINE | ID: mdl-32619428

ABSTRACT

Upon a dark-to-light transition, multiple species of cyanobacteria release a certain amount of H+ from the inside to the outside of their cells. Previous studies revealed the plasma membrane-localizing Proton exchange A (PxcA) is involved in the light-induced H+ extrusion in the cyanobacterium Synechocystis sp. PCC6803. Among oxygenic phototrophs, two PxcA homologs are conserved; they are the nuclear-encoded Day-length-dependent delayed-greening1 (DLDG1) and the plastid-encoded Ycf10 in Arabidopsis thaliana. We previously identified the putative DLDG1/Ycf10-interacting protein, Fluctuating-light acclimation protein1 (FLAP1), required for pH regulation in Arabidopsis chloroplasts. Synechocystis has PxcA and FLAP1 homologs designated here as PxcA like (PxcL) and FLAP1 homolog A (FlpA). Synechocystis mutants lacking pxcA, pxcL, and flpA were constructed and characterized to gain more insight into regulatory mechanisms of light-induced H+ extrusion in cyanobacteria. pH change kinetics of the extracellular solvent after shifting Synechocystis cells from dark to light indicated that PxcA is essential for the light-induced H+ extrusion, and both PxcA and PxcL were involved in H+ uptake. Mutational loss of flpA resulted in altered PxcA- and PxcL-dependent H+ efflux/influx activities, and the flpA-null mutant showed inhibited growth under dark-light cycles, indicating the importance of FlpA function for photosynthetic growth under fluctuating light. Collectively, these data suggest that PxcA is involved in H+ efflux immediately after light irradiation for the rapid formation of the H+ concentration gradient across the thylakoid membranes, PxcL is involved in H+ influx for activation of the Calvin-Benson-Bassham cycle, and FlpA controls the H+ transport under fluctuating light.


Subject(s)
Bacterial Proteins/metabolism , Light , Plastids/metabolism , Protons , Sequence Homology, Amino Acid , Synechocystis/metabolism , Synechocystis/radiation effects , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Biological Transport/radiation effects , Hydrogen-Ion Concentration , Mutation , Synechocystis/genetics
10.
Plant Physiol ; 183(3): 1268-1280, 2020 07.
Article in English | MEDLINE | ID: mdl-32430463

ABSTRACT

Exquisitely regulated plastid-to-nucleus communication by retrograde signaling pathways is essential for fine-tuning of responses to the prevailing environmental conditions. The plastidial retrograde signaling metabolite methylerythritol cyclodiphosphate (MEcPP) has emerged as a stress signal transduced into a diverse ensemble of response outputs. Here, we demonstrate enhanced phytochrome B protein abundance in red light-grown MEcPP-accumulating ceh1 mutant Arabidopsis (Arabidopsis thaliana) plants relative to wild-type seedlings. We further establish MEcPP-mediated coordination of phytochrome B with auxin and ethylene signaling pathways and uncover differential hypocotyl growth of red light-grown seedlings in response to these phytohormones. Genetic and pharmacological interference with ethylene and auxin pathways outlines the hierarchy of responses, placing ethylene epistatic to the auxin signaling pathway. Collectively, our findings establish a key role of a plastidial retrograde metabolite in orchestrating the transduction of a repertoire of signaling cascades. This work positions plastids at the zenith of relaying information coordinating external signals and internal regulatory circuitry to secure organismal integrity.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/growth & development , Arabidopsis/metabolism , Ethylenes/metabolism , Indoleacetic Acids/metabolism , Phytochrome B/metabolism , Adaptation, Physiological/drug effects , Adaptation, Physiological/radiation effects , Arabidopsis/drug effects , Arabidopsis/radiation effects , Arabidopsis Proteins/genetics , Biological Transport/drug effects , Biological Transport/radiation effects , Biosynthetic Pathways/drug effects , Biosynthetic Pathways/genetics , Biosynthetic Pathways/radiation effects , Epistasis, Genetic/drug effects , Epistasis, Genetic/radiation effects , Gene Expression Regulation, Plant/drug effects , Gene Expression Regulation, Plant/radiation effects , Genes, Plant , Hypocotyl/drug effects , Hypocotyl/growth & development , Hypocotyl/radiation effects , Indoleacetic Acids/pharmacology , Light , Mutation/genetics , Phytochrome B/genetics , Signal Transduction/drug effects , Signal Transduction/radiation effects
12.
Plant Sci ; 290: 110299, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31779902

ABSTRACT

Forest understory species have to acclimatize to highly heterogeneous light conditions inside forest canopies in order to utilize available resources efficiently. Light sensitivity and response speed of hydraulic conductance (KL) of common hazel (Corylus avellana L.) to fast changes in irradiance was studied in leaves from three different growth light conditions-sun-exposed, moderate shade, and deep shade. The KL of sun-exposed leaves was approximately 3-fold higher when compared to deep-shade leaves, indicating a strong dependence of leaf hydraulic capacity on light conditions. The KL of sun-exposed leaves increased by a factor of nearly four from minimal values recorded in darkness to maximal values in high light compared to deep-shade leaves. Reaction speed of KL to reach maximum values in response to light was nearly five times higher for sun-exposed vs deep-shade leaves. Plasticity indices of KL for sun-exposed and deep-shade leaves were 0.44 and 0.27, respectively. Higher light sensitivity enables a faster and more plastic response of KL to variable light conditions in sun leaves and enhances the ability of plants to maximize resource utilization under more beneficial environmental conditions.


Subject(s)
Adaptation, Physiological , Corylus/physiology , Plant Leaves/physiology , Sunlight , Biological Transport/physiology , Biological Transport/radiation effects , Corylus/radiation effects , Plant Leaves/radiation effects
13.
Electromagn Biol Med ; 39(1): 1-8, 2020.
Article in English | MEDLINE | ID: mdl-31884821

ABSTRACT

Measurement of cell transmembrane potential (TMP) is a complex methodology involving patch-clamp methods or fluorescence-based potentiometric markers, which have limited to no applicability during ultrafast charging and relaxation phenomena. In such a case, analytical methods are applied for evaluation of the voltage potential changes in biological cells. In this work, the TMP-based electrotransfer mechanism during ultra-high frequency (≥1 MHz) electric fields is studied and the phenomenon of rapid membrane charge accumulation, which is non-occurrent during conventional low-frequency electroporation is simulated using finite element method (FEM). The influence of extracellular medium conductivity (0.1, 1.5 S/m) and pulse rise/fall times (10-50 ns) TMP generation are presented. It is shown that the medium conductivity has a dramatic influence on the electroporation process in the high-frequency range of applied pulsed electric fields (PEF). The applied model allowed to grasp the differences in polarization between 100 and 900 ns PEF and enabled successful prediction of the experimental outcome of propidium iodide electrotransfer into CHO-K1 cells and the conductivity-dependent patterns of MHz range PEF-triggered electroporation were determined. The results of this study form recommendations for development and pre-evaluation of future PEF protocols and generators based on ultra-high frequency electroporation for anticancer and gene therapies.


Subject(s)
Electroporation , Finite Element Analysis , Microwaves , Animals , Biological Transport/radiation effects , CHO Cells , Cricetulus , Extracellular Space/metabolism , Extracellular Space/radiation effects , Membrane Potentials/radiation effects , Propidium/metabolism
14.
Radiat Res ; 193(2): 171-185, 2020 02.
Article in English | MEDLINE | ID: mdl-31877256

ABSTRACT

Radiation-induced lung injury (RILI) is a common and severe side effect of thoracic radiotherapy, which compromises patients' quality of life. Recent studies revealed that early vascular injury, especially microvascular damage, played a central role in the development of RILI. For this reason, early vascular protection is essential for RILI therapy. The ATP-sensitive K+ (KATP) channel is an ATP-dependent K+ channel with multiple subunits. The protective role of the KATP channel in vascular injury has been demonstrated in some published studies. In this work, we investigated the effect of KATP channel on RILI. Our findings confirmed that the KATP channel blocker glibenclamide, rather than the KATP channel opener pinacidil, remitted RILI, and in particular, provided protection against radiation-induced vascular injury. Cytology experiments verified that glibenclamide enhanced cell viability, increased the potential of proliferation after irradiation and attenuated radiation-induced apoptosis. Involved mechanisms included increased Ca2+ influx and PKC activation, which were induced by glibenclamide pretreatment. In conclusion, the KATP channel blocker glibenclamide remitted RILI and inhibited the radiation-induced apoptosis of vascular endothelial cells by increased Ca2+ influx and subsequent PKC activation.


Subject(s)
Apoptosis/drug effects , Calcium/metabolism , Glyburide/pharmacology , KATP Channels/antagonists & inhibitors , Lung Injury/prevention & control , Protein Kinase C/metabolism , Radiation Injuries, Experimental/prevention & control , Animals , Apoptosis/radiation effects , Biological Transport/drug effects , Biological Transport/radiation effects , Enzyme Activation/drug effects , Enzyme Activation/radiation effects , Gene Expression Regulation/drug effects , Gene Expression Regulation/radiation effects , Human Umbilical Vein Endothelial Cells/cytology , Human Umbilical Vein Endothelial Cells/drug effects , Human Umbilical Vein Endothelial Cells/radiation effects , Humans , Lung Injury/etiology , Lung Injury/metabolism , Lung Injury/pathology , Male , Mice , Mice, Inbred C57BL , Phosphorylation/drug effects , Phosphorylation/radiation effects , Potassium Channel Blockers/pharmacology , Radiation Injuries, Experimental/metabolism , Radiation Injuries, Experimental/pathology , Radiation Pneumonitis/prevention & control
15.
Plant Physiol ; 181(4): 1459-1467, 2019 12.
Article in English | MEDLINE | ID: mdl-31601643

ABSTRACT

Plasmodesmata (PD) are essential for plant development, but little is known about their regulation. Several studies have linked PD transport to chloroplast-centered signaling networks, but the physiological significance of this connection remains unclear. Here, we show that PD transport is strongly regulated by light and the circadian clock. Light promotes PD transport during the day, but light is not sufficient to increase rates of PD transport at night, suggesting a circadian gating mechanism. Silencing expression of the core circadian clock gene, LHY/CCA1, allows light to strongly promote PD transport during subjective night, confirming that the canonical plant circadian clock controls the PD transport light response. We conclude that PD transport is dynamically regulated during the day/night cycle. Due to the many roles of PD in plant biology, this discovery has strong implications for plant development, physiology, and pathogenesis.


Subject(s)
Arabidopsis/physiology , Circadian Clocks/radiation effects , Light , Nicotiana/physiology , Plant Cells/metabolism , Plant Cells/radiation effects , Plasmodesmata/metabolism , Plasmodesmata/radiation effects , Arabidopsis/radiation effects , Biological Transport/radiation effects , Photoperiod , Plant Leaves/growth & development , Plant Leaves/radiation effects , Nicotiana/radiation effects
16.
Gen Physiol Biophys ; 38(5): 379-388, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31411574

ABSTRACT

With the rapidly increasing use of mobile phones and their close-contact usage to the brain, there are some concerns about the possible neuronal effects induced by exposure to excessive electromagnetic radiation. Exposure to a radiofrequency electromagnetic field (RF-EMF) of 835 MHz (4.0 W/kg specific absorption rate (SAR) 5 h/day for 12 weeks) may affect hypothalamic presynaptic neurons in C57BL/6 mice. The number and size of the synaptic vesicles (SVs) in the hypothalamic presynaptic terminals were significantly decreased after RF-EMF exposure. Further, the density (SVs numbers/µm) of docking and fusing SVs in the active zones of the presynaptic terminal membrane was significantly decreased in hypothalamic neurons. The expression levels of synapsin I/II and synaptotagmin 1, two regulators of SV trafficking in neurons, were also significantly decreased in the hypothalamus. In parallel, the expression of calcium channel was significantly decreased. These changes in SVs in the active zones may directly decrease the release of neurotransmitters in hypothalamic presynaptic terminals. Therefore, we further studied the possible changes in hypothalamic function by testing the core body temperature and body weight and performed the buried pellet test. The trafficking of SVs was changed by RF-EMF; however, we could not find any significant phenotypical changes in our experimental condition.


Subject(s)
Hypothalamus/metabolism , Hypothalamus/radiation effects , Radio Waves , Synaptic Vesicles/metabolism , Synaptic Vesicles/radiation effects , Animals , Biological Transport/radiation effects , Male , Mice , Mice, Inbred C57BL
17.
New Phytol ; 223(3): 1241-1252, 2019 08.
Article in English | MEDLINE | ID: mdl-31077397

ABSTRACT

High concentrations of dissolved inorganic carbon in stems of herbaceous and woody C3 plants exit leaves in the dark. In the light, C3 species use a small portion of xylem-transported CO2 for leaf photosynthesis. However, it is not known if xylem-transported CO2 will exit leaves in the dark or be used for photosynthesis in the light in Kranz-type C4 plants. Cut leaves of Amaranthus hypochondriacus were placed in one of three solutions of [NaH13 CO3 ] dissolved in KCl water to measure the efflux of xylem-transported CO2 exiting the leaf in the dark or rates of assimilation of xylem-transported CO2 * in the light, in real-time, using a tunable diode laser absorption spectroscope. In the dark, the efflux of xylem-transported CO2 increased with increasing rates of transpiration and [13 CO2 *]; however, rates of 13 Cefflux in A. hypochondriacus were lower compared to C3 species. In the light, A. hypochondriacus fixed nearly 75% of the xylem-transported CO2 supplied to the leaf. Kranz anatomy and biochemistry likely influence the efflux of xylem-transported CO2 out of cut leaves of A. hypochondriacus in the dark, as well as the use of xylem-transported CO2 * for photosynthesis in the light. Thus increasing the carbon use efficiency of Kranz-type C4 species over C3 species.


Subject(s)
Carbon Dioxide/metabolism , Carbon/metabolism , Xylem/metabolism , Biological Transport/radiation effects , Carbon Cycle/radiation effects , Cell Respiration/radiation effects , Darkness , Light , Photosynthesis/radiation effects , Plant Leaves/physiology , Plant Leaves/radiation effects , Plant Transpiration/physiology , Plant Transpiration/radiation effects , Xylem/radiation effects
18.
New Phytol ; 223(3): 1230-1240, 2019 08.
Article in English | MEDLINE | ID: mdl-31081546

ABSTRACT

Traditionally, leaves were thought to be supplied with CO2 for photosynthesis by the atmosphere and respiration. Recent studies, however, have shown that the xylem also transports a significant amount of inorganic carbon into leaves through the bulk flow of water. However, little is known about the dynamics and proportion of xylem-transported CO2 that is assimilated, vs simply lost to transpiration. Cut leaves of Populus deltoides and Brassica napus were placed in either KCl or one of three [NaH13 CO3 ] solutions dissolved in water to simultaneously measure the assimilation and the efflux of xylem-transported CO2 exiting the leaf across light and CO2 response curves in real-time using a tunable diode laser absorption spectroscope. The rates of assimilation and efflux of xylem-transported CO2 increased with increasing xylem [13 CO2 *] and transpiration. Under saturating irradiance, rates of assimilation using xylem-transported CO2 accounted for c. 2.5% of the total assimilation in both species in the highest [13 CO2 *]. The majority of xylem-transported CO2 is assimilated, and efflux is small compared to respiration. Assimilation of xylem-transported CO2 comprises a small portion of total photosynthesis, but may be more important when CO2 is limiting.


Subject(s)
Carbon Dioxide/metabolism , Carbon/metabolism , Xylem/metabolism , Biological Transport/radiation effects , Light , Photosynthesis/radiation effects , Plant Leaves/metabolism , Plant Leaves/radiation effects , Xylem/radiation effects
19.
Plant Physiol ; 180(2): 1185-1197, 2019 06.
Article in English | MEDLINE | ID: mdl-30948554

ABSTRACT

Plants exhibit diverse polar behaviors in response to directional and nondirectional environmental signals, termed tropic and nastic movements, respectively. The ways in which plants incorporate directional information into tropic behaviors is well understood, but it is less well understood how nondirectional stimuli, such as ambient temperatures, specify the polarity of nastic behaviors. Here, we demonstrate that a developmentally programmed polarity of auxin flow underlies thermo-induced leaf hyponasty in Arabidopsis (Arabidopsis thaliana). In warm environments, PHYTOCHROME-INTERACTING FACTOR4 (PIF4) stimulates auxin production in the leaf. This results in the accumulation of auxin in leaf petioles, where PIF4 directly activates a gene encoding the PINOID (PID) protein kinase. PID is involved in polarization of the auxin transporter PIN-FORMED3 to the outer membranes of petiole cells. Notably, the leaf polarity-determining ASYMMETRIC LEAVES1 (AS1) directs the induction of PID to occur predominantly in the abaxial petiole region. These observations indicate that the integration of PIF4-mediated auxin biosynthesis and polar transport, and the AS1-mediated developmental shaping of polar auxin flow, coordinate leaf thermonasty, which facilitates leaf cooling in warm environments. We believe that leaf thermonasty is a suitable model system for studying the developmental programming of environmental adaptation in plants.


Subject(s)
Arabidopsis/growth & development , Movement , Plant Leaves/physiology , Arabidopsis/genetics , Arabidopsis/radiation effects , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Biological Transport/radiation effects , Gene Expression Regulation, Plant/radiation effects , Genes, Plant , Gravitation , Indoleacetic Acids/metabolism , Light , Models, Biological , Plant Leaves/radiation effects , Temperature , Transcription, Genetic/radiation effects
20.
Nanoscale ; 11(13): 5974-5981, 2019 Mar 28.
Article in English | MEDLINE | ID: mdl-30892307

ABSTRACT

The biological identity of nanoparticles (NPs) is defined by a protein layer formed on their surface, called protein corona (PC), once they meet the biological milieu. Any change in the PC composition may influence the biological fate of NPs. The PC composition is strongly dependent on several parameters including the physicochemical properties of NPs, and biological and environmental factors. As one of the main features of plasmonic NPs is their capacity to induce local heating by laser irradiation, we hypothesized that laser irradiation may change the biological identity of NPs and therefore alter their biological fate. To test this hypothesis, here we investigated the effects of either simultaneous or sequential laser irradiation on the conformations of a few proteins selected from two main categories of plasma proteins (i.e. human serum albumin and human fibrinogen) on the surfaces of gold nanorods (AuNRs). The outcomes revealed a significant role of laser irradiation on conformational changes of fibrinogen compared to albumin. Moreover, the effects of plasmonic heating - at various times - on the achieved corona composition from interactions of AuNRs and human plasma with various concentrations were monitored. Consequently, the cellular uptake of the corona coated AuNRs was measured in two cell types: malignant (MCF-7) and normal (MCF-10A) breast cell lines. The results demonstrated a substantial reduction in the cellular uptake of AuNRs in response to an increase in the laser irradiation time, especially in MCF-10A. Our results may pave the way for a mechanistic understanding of the biological identity of plasmonic NPs which in turn can help their safe and efficient clinical translations.


Subject(s)
Fibrinogen/chemistry , Lasers , Nanotubes/chemistry , Serum Albumin/chemistry , Biological Transport/radiation effects , Cell Line, Tumor , Circular Dichroism , Fibrinogen/metabolism , Gold/chemistry , Humans , Lysosomes/metabolism , Protein Corona/chemistry , Protein Structure, Tertiary , Serum Albumin/metabolism
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