• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1098-1105
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• 2017

Vesicular stomatitis virus G glycoprotein (VSV-G) has been widely used for pseudotyping retroviral, lentiviral, and artificial viral vectors. The objective of this study was to establish a potential approach for large-scale production of VSV-G. To this end, VSV-G was cloned with an N-terminal His-tag into Pichia pastoris expression vector pPIC3.5K. Three clones ($Mut^s$) containing the VSV-G expression cassette were identified by PCR. All clones proliferated normally in expansion medium, whereas the proliferation was reduced significantly under induction conditions. VSV-G protein was detected in cell lysates by western blot analysis, and the highest expression level was observed at 96 h post induction. VSV-G could also be obtained from the condition medium of yeast protoplasts. Furthermore, VSV-G could be incorporated into Ad293 cells and was able to induce cell fusion, leading to the transfer of cytoplasmic protein. Finally, VSV-G-mediated DNA transfection was assayed by flow cytometry and luciferase measurement. Incubation of VSV-G lysate with the pGL3-control DNA complex increased the luciferase activity in Ad293 and HeLa cells by about 3-fold. Likewise, incubation of VSV-G lysate with the pCMV-DsRed DNA complex improved the transfection efficiency into Ad293 by 10% and into HeLa cells by about 1-fold. In conclusion, these results demonstrate that VSV-G could be produced from P. pastoris with biofunctionalities, demonstrating that large-scale production of the viral glycoprotein is feasible.

• Nutrition Research and Practice
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• 제14권3호
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• pp.188-202
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• 2020

BACKGROUND/OBJECTIVES: Brain aging is a major risk factor for severe neurodegenerative diseases. Conversely, L-histidine and L-carnosine are known to exhibit neuroprotective effects. The aim of this study was to examine the potential for L-histidine, L-carnosine, and their combination to mediate anti-brain aging effects in neuronal cells subjected to D-galactose-induced aging. MATERIALS/METHODS: The neuroprotective potential of L-histidine, L-carnosine, and their combination was examined in a retinoic acid-induced neuronal differentiated SH-SY5Y cell line exposed to D-galactose (200 mM) for 48 h. Neuronal cell proliferation, differentiation, and expression of anti-oxidant enzymes and apoptosis markers were subsequently evaluated. RESULTS: Treatment with L-histidine (1 mM), L-carnosine (10 mM), or both for 48 h efficiently improved the proliferation, neurogenesis, and senescence of D-galactose-treated SH-SY5Y cells. In addition, protein expression levels of both neuronal markers (β tubulin-III and neurofilament heavy protein) and anti-oxidant enzymes, glutathione peroxidase-1 and superoxide dismutase-1 were up-regulated. Conversely, protein expression levels of amyloid β (1-42) and cleaved caspase-3 were down-regulated. Levels of mRNA for the pro-inflammatory cytokines, interleukin (IL)-8, IL-1β, and tumor necrosis factor-α were also down-regulated. CONCLUSIONS: To the best of our knowledge, we provide the first evidence that L-histidine, L-carnosine, and their combination mediate anti-aging effects in a neuronal cell line subjected to D-galactose-induced aging. These results suggest the potential benefits of L-histidine and L-carnosine as anti-brain aging agents and they support further research of these amino acid molecules.

• ALGAE
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• 제35권2호
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• pp.167-176
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• 2020

Iodine exists as a trace element in seawater, with total iodine being generally constant at about 0.45-0.55 μM. Almost all of this iodine occurs in two main forms: iodate and iodide. Iodate is the thermodynamically stable form under normal seawater conditions, and thus should be the only iodine-containing species in the water column. However, iodate concentrations are found to vary considerably, being generally greater at depth and lower at the surface, while iodide concentrations follow the reverse pattern, being anomalously accumulated in the euphotic zone and decreasing with depth. The fact that iodide concentrations follow a depth dependence corresponding to the euphotic zone suggests that biological activity is the source of the reduced iodine. Nonetheless, the nature and source of iodate reduction activity remains controversial. Here, using a combination of field and laboratory studies, we examine some of the questions raised in our and other previous studies, and seek further correlations between changes in iodine speciation and the presence of marine macro- and microalgae. The present results indicate that microalgal growth per se does not seem to be responsible for the reduction of iodate to iodide. However, there is some support for the hypothesis that iodate reduction can occur due to release of cellular reducing agents that accompany cell senescence during phytoplankton bloom declines. In addition, support is given to the concept that macroalgal species such as giant kelp (Macrocystis pyrifera) can take up both iodide and iodate from seawater (albeit on a slower time scale). We propose a mechanism whereby iodate is reduced to iodide at the cell surface by cell surface reductases and is taken up directly as such without reentering the bulk solution.

• Molecules and Cells
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• 제45권9호
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• pp.660-672
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• 2022

The target of rapamycin complex (TORC) plays a key role in plant cell growth and survival by regulating the gene expression and metabolism according to environmental information. TORC activates transcription, mRNA translation, and anabolic processes under favorable conditions, thereby promoting plant growth and development. Tomato fruit ripening is a complex developmental process promoted by ethylene and specific transcription factors. TORC is known to modulate leaf senescence in tomato. In this study, we investigated the function of TORC in tomato fruit ripening using virus-induced gene silencing (VIGS) of the TORC genes, TOR, lethal with SEC13 protein 8 (LST8), and regulatory-associated protein of TOR (RAPTOR). Quantitative reverse transcription-polymerase chain reaction showed that the expression levels of tomato TORC genes were the highest in the orange stage during fruit development in Micro-Tom tomato. VIGS of these TORC genes using stage 2 tomato accelerated fruit ripening with premature orange/red coloring and decreased fruit growth, when control tobacco rattle virus 2 (TRV2)-myc fruits reached the mature green stage. TORC-deficient fruits showed early accumulation of carotenoid lycopene and reduced cellulose deposition in pericarp cell walls. The early ripening fruits had higher levels of transcripts related to fruit ripening transcription factors, ethylene biosynthesis, carotenoid synthesis, and cell wall modification. Finally, the early ripening phenotype in Micro-Tom tomato was reproduced in the commercial cultivar Moneymaker tomato by VIGS of the TORC genes. Collectively, these results demonstrate that TORC plays an important role in tomato fruit ripening by modulating the transcription of various ripening-related genes.

• Journal of Life Science
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• 제26권6호
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• pp.646-650
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• 2016

The influence of chromosome number on cell growth and cell aging was investigated in various yeast strains that have many artificial chromosomes constructed using a chromosome manipulation technique. Host strain FY833 and the YKY18, YKY18R, YKY24, and YKY30 strains harboring 16 natural chromosomes, 18 chromosomes, 18 chromosomes containing rDNA chromosome, 24 chromosomes, and 30 chromosomes, respectively, were used, and the specific growth rate of each strain was compared. The specific growth rates in the YKY18 and YKY24 strains were indistinguishable from that in the host strain, while those of the YKY18R and YKY30 strains were reduced to approximately 25% and 40% of the host strain level, respectively. Subsequently, the replicative life span was examined to investigate the relationship between the number of chromosomes and cell aging, and the life span was decreased to approximately 14% and 45% of the host strain level in the YKY24 and YKY30 strains, respectively. Moreover, telomere length, well known as a senescence factor, was shorter and more diversified in the strain, showing decreased life span. Therefore, these results suggest the possibility that an increase in the number of chromosomes containing artificial chromosomes caused cell aging, and we expected these observations would be applied to improve industrial strain harboring of versatile and special artificial chromosomes.

• Molecules and Cells
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• 제40권8호
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• pp.598-605
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• 2017

Human mesenchymal stem cells (MSCs) are currently being evaluated as a cell-based therapy for tissue injury and degenerative diseases. Recently, several methods have been suggested to further enhance the therapeutic functions of MSCs, including genetic modifications with tissue- and/or diseasespecific genes. The objective of this study was to examine the efficiency and stability of transduction using an adenoviral vector in human MSCs. Additionally, we aimed to assess the effects of transduction on the proliferation and multipotency of MSCs. The results indicate that MSCs can be transduced by adenoviruses in vitro, but high viral titers are necessary to achieve high efficiency. In addition, transduction at a higher multiplicity of infection (MOI) was associated with attenuated proliferation and senescence-like morphology. Furthermore, transduced MSCs showed a diminished capacity for adipogenic differentiation while retaining their potential to differentiate into osteocytes and chondrocytes. This work could contribute significantly to clinical trials of MSCs modified with therapeutic genes.

• KOREAN JOURNAL OF CROP SCIENCE
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• 제49권2호
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• pp.110-115
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• 2004

Rice (Oryza sativa L.) plants were cultivated to examine changes in antioxidative defence mechanism induced by elevated ozone levels. Catalase activities in tolerant Jinpumbyeo and susceptible Chucheongbyeo under ozone fumigation were reduced at 5 hrs and 3 hrs after ozone fumigation, respectively. With the increased ozone supply, peroxidase activity in Jinpumbyeo was steadily enhanced whereas in Chucheongbyeo it was not changed. Four SOD-isozymes were detected by NBT staining of native-PAGE. Two isozymes of them were obviously induced by ozone supply, particularly in Jinpumbyeo. The continuous ozone fumigation increased remarkably putrescine levels in leaves whereas it did not affect the levels of spermidine and spermine. In this study, it was implied that ozone in cell inhibits strongly diamine oxidase and thus promotes ethylene biosynthesis which will cause the senescence in rice plants.

• Journal of Animal Reproduction and Biotechnology
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• 제35권1호
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• pp.102-111
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• 2020

Nitric oxide (NO)-induced protein S-nitrosylation triggers mitochondrial dysfunction and was related to cell senescence. However, the exact mechanism of these damages is not clear. In the present study, to investigate the relationship between in vitro aging and NO-induced protein S-nitrosylation, oocytes were treated with sodium nitroprusside dihydrate (SNP), and the resultant S-nitrosylated proteins were detected through biotin-switch assay. The results showed that levels of protein S-nitroso thiols (SNO)s and expression of S-nitrosoglutathione reductase (GSNOR) increased, while activity and function of mitochondria were impaired during oocyte aging. Addition of SNP, a NO donor, to the oocyte culture led to accelerated oocyte aging, increased mitochondrial dysfunction and damage, apoptosis, ATP deficiency, and enhanced ROS production. These results suggested that the increased NO signal during oocyte aging in vitro, accelerated oocyte degradation due to increased protein S-nitrosylation, and ROS-related redox signaling.

• Radiation Oncology Journal
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• 제36권3호
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• pp.172-181
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• 2018

Successful anticancer strategies require a differential response between tumor and normal tissue (i.e., a therapeutic ratio). In fact, improving the effectiveness of a cancer therapeutic is of no clinical value in the absence of a significant increase in the differential response between tumor and normal tissue. Although radiation dose escalation with the use of intensity modulated radiation therapy has permitted the maximum tolerable dose for most locally advanced cancers, improvements in tumor control without damaging normal adjacent tissues are needed. As a means of increasing the therapeutic ratio, several new approaches are under development. Drugs targeting signal transduction pathways in cancer progression and more recently, immunotherapeutics targeting specific immune cell subsets have entered the clinic with promising early results. Radiobiological research is underway to address pressing questions as to the dose per fraction, irradiated tumor volume and time sequence of the drug administration. To exploit these exciting novel strategies, a better understanding is needed of the cellular and molecular pathways responsible for both cancer and normal tissue and organ response, including the role of radiation-induced accelerated senescence. This review will highlight the current understanding of promising biologically targeted therapies to enhance the radiation therapeutic ratio.

• Molecular & Cellular Toxicology
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• 제5권2호
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• pp.133-140
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• 2009

Caveolin may be a molecular target for modulation of aging process in cochlear hair cells and have association with oxotoxicity. First we investigated the basal expression of caveolin-1, caveolin-2, caveolin-3, nitric oxide synthase, and superoxide dismutase in UB/OC-1 cochlear hair cell line. By using a RNA interference technique, we investigated whether down-regulation of caveolin influenced telomerase activity and reactive oxygen species (ROS) production in cochlear hair cells. In addition, cisplatin and gentamycin, known ototoxic drugs, were administered to the cochlear cells to determine their impact on caveolin expression. Further attempts at elucidating cellular aging mechanism with caveolin and ototoxic drugs were carried out. The main discoveries were the presence of caveolin-1 in UB/OC-1 cells and that down-regulation of caveolin-1 reduced protein kinase A activity. Telomerase was activated by caveolin down-regulation and caveolin down-regulation inhibited oxidative stress at the mitochondrial level. When cisplatin and gentamycin were administered to the cochlear hair cells during a caveolin expression state, a decrease in telomerase activity and increase ROS activity was observed. Caveolin-1 may modulate the senescent mechanisms in cochlear cells. An increase in caveolin-1 levels can lead to ROS production in the mitochondria which may cause ototoxicity.