Bacterial Isolate Inhabiting Spitsbergen Soil Modifies the Physiological Response of Phaseolus coccineus in Control Conditions and under Exogenous Application of Methyl Jasmonate and Copper Excess

The aim of the study was to demonstrate the potential of the promotion and regulation of plant physiology and growth under control and copper stress conditions, and the impact of the exogenous application of methyl jasmonate on this potential. Runner bean plants were treated with methyl jasmonate (1 or 10 µM) (J; J1 or J10) and Cu (50 µM), and inoculated with a bacterial isolate (S17) originating from Spitsbergen soil, and identified as Pseudomonas luteola using the analytical profile index (API) test. Above- and under-ground plant parts were analyzed. The growth parameters; the concentration of the photosynthetic pigments, elements, flavonoids (FLAVO), phenolics (TPC), allantoin (ALLA), and low molecular weight organic acids (LMWOAs); the activity of antioxidant enzymes and enzymes of resistance induction pathways (e.g., superoxide dismutase (SOD), catalase (CAT), ascorbate (APX) and guaiacol (GPX) peroxidase, glucanase (GLU), and phenylalanine (PAL) and tyrosine ammonia-lyase (TAL)), and the antioxidant capacity (AC) were studied. The leaves exhibited substantially higher ALLA and LMWOA concentrations as well as PAL and TAL activities, whereas the roots mostly had higher activities for a majority of the enzymes tested (i.e., SOD, CAT, APX, GPX, and GLU). The inoculation with S17 mitigated the effect of the Cu stress. Under the Cu stress and in the presence of J10, isolate S17 caused an elevation of the shoot fresh weight, K concentration, and TAL activity in the leaves, and APX and GPX (also at J1) activities in the roots. In the absence of Cu, isolate S17 increased the root length and the shoot-to-root ratio, but without statistical significance. In these conditions, S17 contributed to a 236% and 34% enhancement of P and Mn, respectively, in the roots, and a 19% rise of N in the leaves. Under the Cu stress, S17 caused a significant increase in FLAVO and TPC in the leaves. Similarly, the levels of FLAVO, TPC, and AC were enhanced after inoculation with Cu and J1. Regardless of the presence of J, inoculation at Cu excess caused a reduction of SOD and CAT activities, and an elevation of GPX. The effects of inoculation were associated with the application of Cu and J, which modified plant response mainly in a concentration-dependent manner (e.g., PAL, TAL, and LMWOA levels). The conducted studies demonstrated the potential for isolate S17 in the promotion of plant growth.

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Forced Expression of Keratin 16 Alters the Adhesion, Differentiation, and Migration of Mouse Skin Keratinocytes

Molecular Biology of the Cell ◽

10.1091/mbc.11.10.3315 ◽

2000 ◽

Vol 11 (10) ◽

pp. 3315-3327 ◽

Cited By ~ 39

Author(s):

Matthew Wawersik ◽

Pierre A. Coulombe

Keyword(s):

Mouse Skin ◽

Cellular Level ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Keratin 16 ◽

Skin Keratinocytes ◽

Steady State Levels ◽

And Migration ◽

Mouse Skin Keratinocytes ◽

The Impact

Injury to the skin results in an induction of keratins K6, K16, and K17 concomitant with activation of keratinocytes for reepithelialization. Forced expression of human K16 in skin epithelia of transgenic mice causes a phenotype that mimics several aspects of keratinocyte activation. Two types of transgenic keratinocytes, with forced expression of either human K16 or a K16-C14 chimeric cDNA, were analyzed in primary culture to assess the impact of K16 expression at a cellular level. High K16-C14-expressing and low K16-expressing transgenic keratinocytes behave similar to wild type in all aspects tested. In contrast, high K16-expressing transgenic keratinocytes show alterations in plating efficiency and calcium-induced differentiation, but proliferate normally. Migration of keratinocytes is reduced in K16 transgenic skin explants compared with controls. Finally, a subset of high K16-expressing transgenic keratinocytes develops major changes in the organization of keratin filaments in a time- and calcium concentration-dependent manner. These changes coincide with alterations in keratin content while the steady-state levels of K16 protein remain stable. We conclude that forced expression of K16 in progenitor skin keratinocytes directly impacts properties such as adhesion, differentiation, and migration, and that these effects depend upon determinants contained within its carboxy terminus.

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The cytoprotective effects of bilirubin and biliverdin on rat hepatocytes and human erythrocytes and the impact of albumin

Biochemistry and Cell Biology ◽

10.1139/o91-123 ◽

1991 ◽

Vol 69 (12) ◽

pp. 828-834 ◽

Cited By ~ 41

Author(s):

Tai-Wing Wu ◽

Doug Carey ◽

Jun Wu ◽

Hiroshi Sugiyama

Keyword(s):

Rat Hepatocytes ◽

Human Erythrocytes ◽

Red Cells ◽

Peroxyl Radicals ◽

Blood Levels ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Hepatocyte Necrosis ◽

The Impact ◽

First Time

The hypothesis that unconjugated bilirubin and biliverdin are cytoprotective antioxidants has been examined for the first time in systems containing cells. In primary rat hepatocytes exposed to xanthine oxidase and hypoxanthine, bilirubin (0–60 μM) failed to prolong cell survival. In contrast, biliverdin (20–100 μM) markedly delayed hepatocyte necrosis in a concentration-dependent manner. When 0.3 mM of albumin was present, bilirubin (0–50 μM) became protective of hepatocytes, while biliverdin was less dramatically enhanced in its cytoprotective effect. In human erythrocytes exposed to peroxyl radicals, bilirubin and biliverdin inhibited 50% cell lysis at lower concentrations than Trolox and ascorbate, respectively. Albumin alone appeared less cytoprotective in red cells than in hepatocytes, but its presence enhanced the effects of both pigments on erythrocytes. Of probable physiologic relevance, bilirubin with albumin present or biliverdin alone protected hepatocytes substantially (and to a lesser extent red cells) at the normal blood levels of bilirubin (3.4–26 μM). Moreover, the fact that the pigments are cytoprotective at higher bilirubin levels (e.g., 50–100 μM) tempts the speculation that they may be circulating cytoprotectors of overlooked importance in jaundice.Key words: cytoprotection, biliverdin, bilirubin, albumin.

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Cell-free Expression of Proton-Coupled Folate Transporter in the Presence of Nanodiscs

10.1101/2021.05.19.444806 ◽

2021 ◽

Author(s):

Hoa Quynh Do ◽

Carla M Bassil ◽

Elizabeth I Andersen ◽

Michaela Jansen

Keyword(s):

Tumor Cells ◽

Plasma Membranes ◽

In Vitro Translation ◽

Translation System ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

E Coli ◽

Membrane Scaffold ◽

The Impact

The Proton-Coupled Folate Transporter (PCFT) is a transmembrane transport protein that controls the absorption of dietary folates in the small intestine. PCFT also mediates uptake of chemotherapeutically used antifolates into tumor cells. PCFT has been identified within lipid rafts observed in phospholipid bilayers of plasma membranes, a micro environment that is altered in tumor cells. The present study aimed at investigating the impact of different lipids within Lipid-protein nanodiscs (LPNs), discoidal lipid structures stabilized by membrane scaffold proteins, to yield soluble PCFT expression in an E. coli lysate-based cell-free transcription/translation system. In the absence of detergents or lipids, we observed PCFT quantitatively as precipitate in this system. We then explored the ability of LPNs to support solubilized PCFT expression when present during in-vitro translation. LPNs consisted of either dimyristoyl phosphatidylcholine (DMPC), palmitoyl-oleoyl phosphatidylcholine (POPC), or dimyristoyl phosphatidylglycerol (DMPG). While POPC did not lead to soluble PCFT expression, both DMPG and DMPC supported PCFT translation directly into LPNs, the latter in a concentration dependent manner. The results obtained through this study provide insights into the lipid preferences of PCFT. Membrane-embedded or solubilized PCFT will enable further studies with diverse biophysical approaches to enhance the understanding of the structure and molecular mechanism of folate transport through PCFT.

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(R,S)-Ketamine Metabolites (R,S)-norketamine and (2S,6S)-hydroxynorketamine Increase the Mammalian Target of Rapamycin Function

Anesthesiology ◽

10.1097/aln.0000000000000285 ◽

2014 ◽

Vol 121 (1) ◽

pp. 149-159 ◽

Cited By ~ 76

Author(s):

Rajib K. Paul ◽

Nagendra S. Singh ◽

Mohammed Khadeer ◽

Ruin Moaddel ◽

Mitesh Sanghvi ◽

...

Keyword(s):

Parent Compound ◽

Mammalian Target Of Rapamycin ◽

Mtor Pathway ◽

Target Of Rapamycin ◽

Dependent Manner ◽

Α7 Nicotinic Acetylcholine Receptor ◽

Concentration Dependent Manner ◽

Pheochromocytoma Cells ◽

The Impact

Abstract Background: Subanesthetic doses of (R,S)-ketamine are used in the treatment of neuropathic pain and depression. In the rat, the antidepressant effects of (R,S)-ketamine are associated with increased activity and function of mammalian target of rapamycin (mTOR); however, (R,S)-ketamine is extensively metabolized and the contribution of its metabolites to increased mTOR signaling is unknown. Methods: Rats (n = 3 per time point) were given (R,S)-ketamine, (R,S)-norketamine, and (2S,6S)-hydroxynorketamine and their effect on the mTOR pathway determined after 20, 30, and 60 min. PC-12 pheochromocytoma cells (n = 3 per experiment) were treated with escalating concentrations of each compound and the impact on the mTOR pathway was determined. Results: The phosphorylation of mTOR and its downstream targets was significantly increased in rat prefrontal cortex tissue by more than ~2.5-, ~25-, and ~2-fold, respectively, in response to a 60-min postadministration of (R,S)-ketamine, (R,S)-norketamine, and (2S,6S)-hydroxynorketamine (P < 0.05, ANOVA analysis). In PC-12 pheochromocytoma cells, the test compounds activated the mTOR pathway in a concentration-dependent manner, which resulted in a significantly higher expression of serine racemase with ~2-fold increases at 0.05 nM (2S,6S)-hydroxynorketamine, 10 nM (R,S)-norketamine, and 1,000 nM (R,S)-ketamine. The potency of the effect reflected antagonistic activity of the test compounds at the α7-nicotinic acetylcholine receptor. Conclusions: The data demonstrate that (R,S)-norketamine and (2S,6S)-hydroxynorketamine have potent pharmacological activity both in vitro and in vivo and contribute to the molecular effects produced by subanesthetic doses of (R,S)-ketamine. The results suggest that the determination of the mechanisms underlying the antidepressant and analgesic effects of (R,S)-ketamine requires a full study of the parent compound and its metabolites.

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Interaction of Graphene Oxide Modified with Linear and Branched PEG with Monocytes Isolated from Human Blood

Nanomaterials ◽

10.3390/nano12010126 ◽

2021 ◽

Vol 12 (1) ◽

pp. 126

Author(s):

Pavel Khramtsov ◽

Maria Bochkova ◽

Valeria Timganova ◽

Anton Nechaev ◽

Sofya Uzhviyuk ◽

...

Keyword(s):

Graphene Oxide ◽

Immune Cells ◽

Human Peripheral Blood ◽

Oxide Surface ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Composition And Structure ◽

Luminescent Signal ◽

Ros Formation ◽

The Impact

Multiple graphene-based therapeutics have recently been developed, however potential risks related to the interaction between nanomaterials and immune cells are still poorly understood. Therefore, studying the impact of graphene oxide on various populations of immune cells is of importance. In this work, we aimed to investigate the effects of PEGylated graphene oxide on monocytes isolated from human peripheral blood. Graphene oxide nanoparticles with lateral sizes of 100–200 nm and 1–5 μm were modified with linear and branched PEG (GO-PEG). Size, elemental composition, and structure of the resulting nanoparticles were characterized. We confirmed that PEG was successfully attached to the graphene oxide surface. The influence of GO-PEG on the production of reactive oxygen species (ROS), cytokines, phagocytosis, and viability of monocytes was studied. Uptake of GO-PEG by monocytes depends on PEG structure (linear or branched). Branched PEG decreased the number of GO-PEG nanoparticles per monocyte. The viability of monocytes was not altered by co-cultivation with GO-PEG. GO-PEG decreased the phagocytosis of Escherichia coli in a concentration-dependent manner. ROS formation by monocytes was determined by measuring luminol-, lucigenin-, and dichlorodihydrofluorescein-dependent luminescence. GO-PEG decreased luminescent signal probably due to inactivation of ROS, such as hydroxyl and superoxide radicals. Some types of GO-PEG stimulated secretion of IL-10 by monocytes, but this effect did not correlate with their size or PEG structure.

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Size of Cells and Physicochemical Properties of Membranes are Related to Flavor Production during Sake Brewing in the Yeast Saccharomyces cerevisiae

Membranes ◽

10.3390/membranes10120440 ◽

2020 ◽

Vol 10 (12) ◽

pp. 440

Author(s):

Tsuyoshi Yoda ◽

Tomoaki Saito

Keyword(s):

Isoamyl Alcohol ◽

Lipid Vesicles ◽

Caproic Acid ◽

Yeast Cells ◽

Dependent Manner ◽

Yeast Saccharomyces Cerevisiae ◽

Concentration Dependent Manner ◽

Generalized Polarization ◽

Flavor Components ◽

The Impact

Ethyl caproate (EC) and isoamyl acetate (IA) are key flavor components of sake. Recently, attempts have been made to increase the content of good flavor components, such as EC and IA, in sake brewing. However, the functions of EC and IA in yeast cells remain poorly understood. Therefore, we investigated the effects of EC and IA using cell-sized lipid vesicles. We also investigated lipid vesicles containing EC and/or caproic acid (CA) as well as IA and/or isoamyl alcohol (IAA). CA and IAA are precursors of EC and IA, respectively, and are important flavors in sake brewing. The size of a vesicle is influenced by flavor compounds and their precursors in a concentration-dependent manner. We aimed to establish the conditions in which the vesicles contained more flavors simultaneously and with different ratios. Interestingly, vesicles were largest in a mixture of 50% of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) with 25% EC and 25% CA or a mixture of 50% DOPC with 25% IA and 25% IAA. The impact of flavor additives on membrane fluidity was also studied using Laurdan generalized polarization. During the production process, flavors may regulate the fluidity of lipid membranes.

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Evaluation of Physiologic and Pathologic Alterations of Coagulation and Fibrinolysis Using a New Global Assay.

Blood ◽

10.1182/blood.v104.11.2985.2985 ◽

2004 ◽

Vol 104 (11) ◽

pp. 2985-2985

Author(s):

Neil A. Goldenberg ◽

William E. Hathaway ◽

Linda Jacobson ◽

Marilyn J. Manco-Johnson

Keyword(s):

Pregnant Women ◽

Statistical Significance ◽

Healthy Adults ◽

Clot Formation ◽

Healthy Children ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Platelet Poor Plasma ◽

Coagulation And Fibrinolysis

Abstract BACKGROUND: The Clot Formation and Lysis (CloFAL) assay, modified from prior global assay methods by He et al., 1999, and Smith et al., 2003, involves activation of coagulation and fibrinolysis in platelet-poor plasma via addition of a physiologic reactant solution in a multi-well microassay plate. Continuous spectropohotometric data analysis allows measurement of kinetic absorbance changes of the plasma sample over three hours, which yields a unique clot formation and lysis curve. Using parameters of the curve, coagulation and fibrinolytic indices (CI and FI) are calculated relative to a simultaneously run pooled normal plasma standard. METHODS: Platelet-poor plasma obtained from pregnant women at term (n=24), neonatal cord blood (n=29), and healthy children (n=22) were analyzed using the CloFAL assay. Healthy adult (n=22) plasma samples, as well as those of individuals with factor deficiencies, were obtained commercially. Fibrinolytic alterations in vitro were also investigated. Intra-assay coefficients of variation (CVs) for normal controls ranged from 3–12% for all assay parameters, with inter-assay CVs of 5–15%. RESULTS: Representative CloFAL curves for healthy adults and children, pregnant women, and newborn infants are shown in Figure 1. Coagulation potential (measured by median CI) was significantly increased, while fibrinolytic capacity (measured by median FI) was markedly decreased, in pregnant women as compared to healthy adults (CI: 239% vs. 115%, FI: 59% vs. 95%; P<0.001 for each). By contrast, CI was decreased, and FI notably increased, in neonatal cords versus children, although the former comparison did not achieve statistical significance (CI: 58% vs. 69%, P=0.09; FI: 210% vs. 142%, P<0.001). The influence of deficiencies of coagulation factors and fibrinolytic regulators upon CloFAL parameters was also investigated. The greatest impact upon CI occurred with severe deficiency of fibrinogen or factors II, V, VII, VIII, IX, or X. Furthermore, CI was sensitive to deficiencies of factor VIII and fibrinogen in a concentration-dependent manner. In addition, FI was increased by PAI-1 deficiency and by inhibition of thrombin-activatable fibrinolysis inhibitor (TAFI) activation, and was zero in the setting of aminocaproic acid treatment. Further studies revealed that the influence of heparin concentrations of up to 2 U/mL in plasma was completely reversible by heparinase treatment of samples prior to assay. CONCLUSION: These results indicate that the CLoFAL assay is reproducible and analytically sensitive to known physiologic and pathologic alterations in coagulation and fibrinolysis. The application of this global assay to patients with a variety of disorders of thrombosis and hemostasis is currently ongoing. Figure Figure

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Physiological and Biochemical Changes in Moth Bean (Vigna aconitifolia L.) under Cadmium Stress

Journal of Botany ◽

10.1155/2016/6403938 ◽

2016 ◽

Vol 2016 ◽

pp. 1-13 ◽

Cited By ~ 5

Author(s):

Poornima D. Vijendra ◽

Kavitha M. Huchappa ◽

Roopa Lingappa ◽

Giridhara Basappa ◽

Sathisha G. Jayanna ◽

...

Keyword(s):

Growth Parameters ◽

Cadmium Stress ◽

Protein Carbonyls ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Vigna Aconitifolia ◽

Moth Bean ◽

Aerial Parts ◽

Drought Resistant ◽

Physiological And Biochemical

Moth bean (Vigna aconitifolia L.), a drought resistant legume, possesses high nutritional value. Cadmium (Cd) is a nonessential and the most toxic heavy metal in plants. The present study was to test the hypothesis of whether moth bean being a drought resistant legume can withstand the cadmium stress. Ten-day-old moth bean seedlings were subjected to cadmium stress and investigated for a period of 15 days every 3-day intervals. Cadmium quantification in moth bean tissues suggests root accumulation and translocation to aerial parts in a concentration dependent manner. Results of physiological and biochemical studies revealed that cadmium has affected the growth parameters like shoot and root lengths and tissue dry weights. Significant alternations in relative water content and cell membrane stability were observed in stressed seedlings. Similarly superoxide radical, lipoxygenase activity, membrane lipid peroxidation products, protein carbonyls, and reduced glutathione and nonprotein thiols were found increased in stressed seedlings compared to controls. However, hydrogen peroxide and ascorbic acid levels were not altered significantly in both stressed and control seedlings. Cadmium translocation ability from roots to aerial parts and elevated levels of nonenzymatic antioxidants in stressed seedlings suggest the cadmium stress withstanding ability of moth bean.

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Polyethylene glycol exerted toxicity to growth of Bacillus subtilis NRS-762

10.7287/peerj.preprints.26817 ◽

2018 ◽

Author(s):

Wenfa Ng

Keyword(s):

Bacillus Subtilis ◽

Polyethylene Glycol ◽

Optical Density ◽

Culture Broth ◽

Threshold Concentration ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Ph Variation ◽

The One ◽

The Impact

Polyethylene glycol is commonly used in fermentations as an anti-foam for preventing the rise of foam to the top plate of the bioreactor, which increases contamination risk. However, its potential toxicity to growth of various microorganisms is not well understood at the strain and species level. Hence, the objective of this study was to understand the impact of different concentrations of polyethylene glycol (0, 1, 5 and 10 g/L) on the aerobic growth of Bacillus subtilis NRS-762 (ATCC 8473) in LB Lennox medium in shake flasks at 30 oC and 230 rpm rotational shaking. Experiment results indicated that polyethylene glycol (PEG) (molecular weight ~8000 Da), at all concentrations tested, exerted some toxicity towards the growth of B. subtilis NRS-762 in LB Lennox medium. Specifically, maximal optical density obtained declined with greater exposure to PEG in a concentration-dependent manner, up to a threshold concentration of 5 g/L PEG. For example, maximal optical density obtained in B. subtilis NRS-762 without addition of PEG was 4.4, but the value obtained on exposure to 1 g/L of the anti-foam decreased to 4.1 and a further 3.8 on exposure of cells to 5 g/L and 10 g/L PEG. Similarly, growth rates of B. subtilis NRS-762 also decreased in a concentration-dependent manner with PEG concentration up to a threshold concentration of 5 g/L PEG. pH variation in culture broth, however, revealed that the pH profiles for exposure to PEG at all concentrations overlapped each other and was similar to the one of cells without exposure to the anti-foam; thereby, highlighting that metabolic processes in B. subtilis NRS-762 were not significantly affected by exposure to PEG. Collectively, polyethylene glycol anti-foam exerted toxicity effect on B. subtilis NRS-762 biomass formation, and possibly metabolism. The latter may not be sufficiently significant to affect the types of metabolites secreted by the bacterium, and thus, could not be detected by measurement of culture broth pH. Overall, the results should inform the choice and concentration of PEG for culturing B. subtilis in biotechnological applications.

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Up-regulation of autophagy by low concentration of salicylic acid delays methyl jasmonate-induced leaf senescence

10.1101/814608 ◽

2019 ◽

Author(s):

Runzhu Yin ◽

Jingfang Yu ◽

Yingbin Ji ◽

Jian Liu ◽

Lixin Cheng ◽

...

Keyword(s):

Salicylic Acid ◽

Methyl Jasmonate ◽

Leaf Senescence ◽

High Throughput Sequencing ◽

Autophagic Flux ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Regulatory Pathways ◽

Low Concentration ◽

Protein Protein Interaction

ABSTRACTCrosstalk between salicylic acid (SA) and jasmonic acid (JA) signaling plays an important role in molecular regulation of plant senescence. Our previous works found that SA could delay methyl jasmonate (MeJA)-induced leaf senescence in a concentration-dependent manner. Here, the effect of low concentration of SA (LCSA) application on MeJA-induced leaf senescence was further assessed. High-throughput sequencing (RNA-Seq) results showed that LCSA did not have dominant effects on the genetic regulatory pathways of basal metabolism like nitrogen metabolism, photosynthesis and glycolysis. The ClusterONE was applied to identify discrete gene modules based on protein-protein interaction (PPI) network. Interestingly, an autophagy-related (ATG) module was identified in the differentially expressed genes (DEGs) that exclusively induced by MeJA together with LCSA. RT-qPCR confirmed that the expression of most of the determined ATG genes were upregulated by LCSA. Remarkably, in contrast to wild type (Col-0), LCSA cannot alleviate the leaf yellowing phenotype in autophagy defective mutants (atg5-1 and atg7-2) upon MeJA treatment. Confocal and western blot results showed that LCSA increased the number of autophagic bodies and autophagic flux during MeJA-induced leaf senescence. Collectively, our work revealed up-regulation of autophagy by LCSA as a key regulator to alleviate MeJA-induced leaf senescence.

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