• Toxicological Research
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• 제21권3호
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• pp.235-240
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• 2005

Pollen has been used for prevention or treatment of certain diseases such as diabetes arthritis or cancer in traditional medicine. Among various pollens, pine tree pollen is known to relieve hypertension, suppress fatty liver progression, and facilitate the digestion, but its immunological activities are less known. To evaluate immunological reactivities and immunotoxicities of pine tree pollen, BALB/c mice were administered to the poller through oral route. Pine tree pollen suspended in distilled water or extracted with methanol has been administered at the concentration of 0, 10, or 100 mg/kg five days per week for four weeks. Polyclonal activation of splenic T cells with phytohemagglutinins did not induce a significant difference in IL-4 and $IFN_{\gamma}$ production between the pollen-administered mice groups and the control mice. Furthermore, polyclonal activation of splenic B cells with lipopolysaccharides did not result a significant difference in IgG1 and IgG2a production among the groups. These findings imply that the intake of pine tree pollen does not bring any humoral and cellular immune-dysrequlation. Whereas, viability of Listeria monocytogenes was suppressed in the mice administered with 100 mg/kg bw methanol extract, indicating the potential ability of pine tree pollen to enhance cell-mediated immunity mediated by type-1 helper T cells. In addition, aberrant upregulation of plasma IgG1 level was observed in the pollen-administered mice, which suggests a possibility of allergic response induction through the pine tree pollen uptake. Overall, pine tree pollen-mediated modulation of humoral or cellular immunity is worthy of further systematic investigation.

• Journal of Microbiology and Biotechnology
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• 제29권10호
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• pp.1522-1542
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• 2019

To adapt to environmental changes and to maintain cellular homeostasis, microorganisms adjust the intracellular concentrations of biochemical compounds, including metal ions; these are essential for the catalytic function of many enzymes in cells, but excessive amounts of essential metals and heavy metals cause cellular damage. Metal-responsive transcriptional regulators play pivotal roles in metal uptake, pumping out, sequestration, and oxidation or reduction to a less toxic status via regulating the expression of the detoxification-related genes. The sensory and regulatory functions of the metalloregulators have made them as attractive biological parts for synthetic biology, and the exceptional sensitivity and selectivity of metalloregulators toward metal ions have been used in heavy metal biosensors to cope with prevalent heavy metal contamination. Due to their importance, substantial efforts have been made to characterize heavy metal-responsive transcriptional regulators and to develop heavy metal-sensing biosensors. In this review, we summarize the biochemical data for the two major metalloregulator families, SmtB/ArsR and MerR, to describe their metal-binding sites, specific chelating chemistry, and conformational changes. Based on our understanding of the regulatory mechanisms, previously developed metal biosensors are examined to point out their limitations, such as high background noise and a lack of well-characterized biological parts. We discuss several strategies to improve the functionality of the metal biosensors, such as reducing the background noise and amplifying the output signal. From the perspective of making heavy metal biosensors, we suggest that the characterization of novel metalloregulators and the fabrication of exquisitely designed genetic circuits will be required.

• Applied Biological Chemistry
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• 제36권3호
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• pp.184-190
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• 1993

Rhodobacter sphaeroides 균주가 생산하는 ${\delta}-aminolevulinic\;acid(ALA)$의 생산성에 관하여 in vivo, in vitro 상에서 기질 및 관련 화합물을 이용하여 검토하였다. $C_5\;ALA$ 생합성계에 의한 ALA yield 대비의 $C_4$ 생합성계에 의한 비율은 in vive 상에서 0.78인 반면, in vitro 상에서의 비율은 1.37이었다.$C_4\;C_5$ 각 계의 기질 첨가 배양에 의한 cell-free system의 $C_4,\;C_5$ 계의 발현도는 미첨가 배양에 의한 system과 비교하여 각각 1.35, 1.52로 증가하였으나, 증가한 계에 대한 상대계의 발현도가 억제되어, $C_4$ 와 $C_5$ 계가 각각 0.91, 0.83으로 나타났다. ${\gamma}-Glutamyl\;derivatives$의 세포내 uptake rates는 L-glutamic acid를 기준으로 비교해서 D-glutamic acid, 0.55: D-glutamine, 0.5: L-glutamine, 0.4: ${\gamma}-L-glutamyl\;ethylester$, 0.3: GSH 및 Glu-pNA, 0의 순서를 보였다. Uptake rate와 관계없이 in vivo 상에서 L-과 D-glutamine이 L-, D-glutamic acid보다 균체 외 ALA의 생산에 있어서 각각 높은 yield의 효과를 보였다.

• 동의생리병리학회지
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• 제37권1호
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• pp.1-8
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• 2023

The aim of this study is to evaluate the antidiabetic effect of the water extract of Aurantii fructus immaturus (WAF), in diabetic models using enzyme, cells and mice, and to suggest a putative mechanism explaining its antidiabetic effect. In an enzyme model using the enzyme α-glucosidase, WAF had no significant effect on α-glucosidase, as compared with acarbose, an antidiabetic drug. Nonetheless, WAF was capable of reducing the blood glucose levels during oral sucrose tolerance test and oral glucose tolerance test, implying that there would be other antidiabetic pathways in no relation to inhibition of α-glucosidase. In cell models using RIN-m5f β-cells and L6 myotubes, WAF, at its non-cytotoxic doses, augmented the secretion of insulin in RIN-m5f β-cells stimulated with 5 mM glucose. In addition, it enhanced the cellular uptake of glucose in L6 myotubes stimulated with deprivation of glucose for 12 h. Therefore, it is most likely that WAF may exert its antidiabetic effects, at least in part, by enhancing insulin secretion and glucose uptake. Meanwhile, in diabetic mice induced with peritoneal injection of streptozotocin (STZ), WAF significantly improved fast blood glucose levels, glycosylated hemoglobin levels, body weight loose, blood pressure, and diabetic adverse effects on functions of the kidney and the liver. Taken together, the water extract of Aurantii fructus immaturus may ameliorate diabetes in mice injected with STZ, at least in part, by enhancing insulin secretion and glucose uptake.

• BMB Reports
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• 제48권3호
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• pp.147-152
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• 2015

Small interfering RNA (siRNA) functions through pairing with specific mRNA sequences and results in the mRNA's degradation. It is a potential therapeutic approach for many diseases caused by altered gene expression. The delivery of siRNA is still a major problem due to its rapid degradation in the circulation. Various strategies have been proposed to help with the cellular uptake of siRNA and short or small hairpin RNA (shRNA). Here, we reviewed recently published data regarding local applications of siRNA. Compared with systemic delivery methods, local delivery of siRNA/shRNA has many advantages, such as targeting the specific tissues or organs, mimicking a gene knockout effect, or developing certain diseases models. The eye, brain, and tumor tissues are 'hot' target tissues/organs for local siRNA delivery. The siRNA can be delivered locally, in naked form, with chemical modifications, or in formulations with viral or non-viral vectors, such as liposomes and nanoparticles. This review provides a comprehensive overview of RNAi local administration and potential future applications in clinical treatment.

• 한국대기환경학회지
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• 제13권3호
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• pp.207-214
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• 1997

We collected airborne complex mixtures in a industrial area of Yeochun, and examined whether these complex mixtures could affect gap junctional intercellular communication (GJIC) in a cultured WBF-344 rat liver epithelial cells (LEC). Since the reduction of GJIC plays an important role in chemical carcinogenesis, measurement of changes of GJIC is a meaningful method to screen carcinogenicity of these mixtures. High and low volume samples were dissolved in dimethyl sulfoxide (DMSO) and tested. Blank filter extractions were also examined for exclud-ing possible toxicity of filter itself, and TPA (12-O-tetradecanoylphorbol-13-acetate) and DMSO were used as positive and negative control, respectively. When the cells were exposed to samples at concentration below that required to maintain rather than 85% cell viability based on the result of neutral red uptake assay, maximal inhibition of GJIC was observed at 1hr after treatment with both high and low volume samples by scrape-loading dye transfer assay. In fluorescence recovery after photobleaching assay, recovery rates via gap junctions were 33%/min in high volume sample and 62%/min in low volume sample. In together, airborne samples collected in Yeochun inhibited GJIC in a cultured WBF-344 rat LEC. These results suggest airborne samples tested in this experiment may attribute to cause a certain type and degree of cancers in in vivo when exposured for some periods.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.821-827
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• 2010

Gene delivery that provides targeted delivery of therapeutic genes to the cells of a lesion enhances therapeutic efficacy and reduces toxic side effects. This process is especially important in cancer therapy when it is advantageous to avoid unwanted damage to healthy normal cells. Incorporating cancer-specific ligands that recognize receptors overexpressed on cancer cells can increase selective binding and uptake and, as a result, increase targeted transgene expression. In this study, we investigated whether a peptide capable of homing to hepatocellular carcinoma (HCC) could facilitate targeted gene delivery by cationic liposomes. This homing peptide (HBP) exhibited selective binding to a human hepatocarcinoma cell line, HepG2, at a concentration ranging from 5 to 5,000 nM. When conjugated to a cationic liposome, HBP substantially increased cellular internalization of plasmid DNA to increase the transgene expression in HepG2 cells. In addition, there was no significant enhancement in gene transfer detected for other human cell lines tested, including THLE-3, AD293, and MCF-7 cells. Therefore, we demonstrate that HBP provides targeted gene delivery to HCC by cationic liposomes.

• Journal of Life Science
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• 제9권1호
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• pp.9-13
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• 1999

Reduced thiols were important compounds for the maintenance of leukemia and lymphoma cell survival (and growth). In the course of examining the microenvirn-mental effects on lymphoma and leukemia cell growth, we found that cysteine suppressed apoptosis in these cells. In a present study, in order to investigate the role of cystein on the suppression of apoptotic cell death, we used CS21, P388, and L1210 cell lines. The addition of BSO, an inhibitor of glutathione synthase, induced apoptosis of these cells by blocking the cellular uptake of cysteine in CS21 cells. Although L1210 cells underwent apoptosis without thiol compounds, the addition of these compounds suppressed the apoptosis and promoted the growth or L1210 cells. When specific inhibitors of caspase3-like proteases, but not caspase1-like proteases, were activated during the L1210 cell apoptosis but the addition of thiol compounds suppressed the activation of caspase3-like proteases. These results suggest that reduced thiols including cysteine play an important role in the suppression of cell apoptosis by inhibiting the activation of caspase3-like proteases.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1115-1119
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• 2005

The photoheterotrophic $H_2$ production of Rhodobacter sphaeroides was significantly increased through disruption of the genes coding for uptake hydrogenase and poly-${\beta}$-hydroxybutyrate (PHB) synthase (Lee et al., Appl. Microbiol. Biotechnol. 60: 147-153, 2002). In this work, we further removed the B800-850 light-harvesting (LH) complex from the strain and found an increase in $H_2$ production at the light-saturating cell growth (${\ge}10$ Watts $[W]/m^2$). Neither the mutant nor the wild-type produced more $H_2$ at the brighter light. Accordingly, light does not appear to be limited for the $H_2$ production by the presence of B800-850. However, increase in the level of the spectral complexes resulted in decrease of $H_2$ production. Thus, although the B875 is essential for light harvesting, the consumption of cellular energy for the synthesis of B800-850 and the surplus LH complexes may reduce the energy flow into the $H_2$ production of R. sphaeroides.

• 한국미생물·생명공학회지
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• 제34권4호
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• pp.329-334
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• 2006

콜레스테롤 유래의 양이온성 리피드 2-aminoethylcarbamate-cholesterol(Chol-E)를 합성하여 이의 리포좀을 제조하였다. 리포좀은 다양한 비율로 중성지방인 DOPE와 섞어서 만든 후 $100{\sim}200nm$의 membrane으로 extrusion시켜 균일한 리포좀을 제작하여 크기 및 전위를 측정하였다. 형광단백질 및 luciferase plasmid의 발현을 여러가지 세포에서 확인한 결과 우수한 발현양상을 보였으며 혈청이 있는 조건에서도 발현이 증가임을 볼수 있었으며, 합성 ODNs의 전달도 adipocyte cell 에서도 잘 이루어지는 것을 확인할 수 있었다 임상실험에 쓰이는 저독성의 DC-chol에 비교하여도 독성이 적은 리포좀임을 알 수 있으며 혈청하에서도 안정하게 유전자를 전달할 수 있는 응용성이 기대되는 새로운 리포좀을 제조하였음을 알 수 있다.