• CELLMED
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• v.3 no.2
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• pp.10.1-10.8
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• 2013

Cancer is one of the major dreaded diseases causing high mortality. Lung cancer is second in position of all cancer related deaths and mainly divided into two morphologic sub-types: small-cell lung cancer and non-small cell lung cancer (NSCLC). NSCLC is an aggressive neoplasm which hardly responds to any conventional chemotherapy. Epidermal growth factor receptor (EGFR) belongs to the ErbB family of receptor tyrosine kinase that is mainly over-expressed in NSCLC. EGFR is mainly involved in the pathogenesis and progression of different carcinoma. In vivo and in vitro studies suggest that EGFR and EGF like peptides are often over-expressed in human NSCLC and these proteins are able to induce cell transformation. The conventional therapies mostly inhibit the EGFR activity and expression level in human NSCLC with the use of some EGFR-inhibitors like HKI-272, EKB569, CL-387785 etc. and some synthetic chemotherapeutic drugs like erlotinib, gefitinib, plumbagin, docetaxel, cisplatin etc., alone or in combination of two or more drugs. These therapies selectively act by competitive inhibition of the binding of adenosine triphosphate to the tyrosine kinase domain of the EGFR, resulting in inhibition of the EGFR signaling pathway. But these chemotherapeutic drugs have some cytotoxic activities to the normal cells and have some adverse side-effects. Recent studies on some traditional alternative therapies including some herbal and plant extracts, active ingredients like curcumin, different homeopathic drugs, etc. can target EGFR-signalling in NSCLC with less toxic side-effects are being currently developed.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1099-1106
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• 2013

Tuta absoluta (Povolny, 1994) is a devastating moth to the Solanaceae plants. It is a challenging pest to control, especially on tomatoes. In this work, we studied the entomopathogenic activity of the Cry-forming ${\delta}$-endotoxins produced by Bacillus thuringiensis strain KS and B. thuringiensis kurstaki reference strain HD1 against T. absoluta. These strains carried the cry2, cry1Ab, cry1Aa/cry1Ac, and cry1I genes, and KS also carried a cry1C gene. The ${\delta}$-endotoxins of KS were approximately twofold more toxic against the third instar larvae than those of HD1, as they showed lower 50% and 90% lethal concentrations (0.80 and 2.70 ${\mu}g/cm^2$ (${\delta}$-endotoxins/tomato leaf)) compared with those of HD1 (1.70 and 4.50 ${\mu}g/cm^2$) (p < 0.05). Additionally, the larvae protease extract showed at least six caseinolytic activities, which activated the KS and HD1 ${\delta}$-endotoxins, yielding the active toxins of about 65 kDa and the protease-resistant core of about 58 kDa. Moreover, the histopathological effects of KS and HD1 ${\delta}$-endotoxins on the larvae midgut consisted of an apical columnar cell vacuolization, microvillus damage, and epithelial cell disruption. These results showed that the KS strain could be a candidate for T. absoluta control.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.13 no.12
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• pp.6208-6214
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• 2012

The most essential but missing components to understand and use toxic substances from marine microalgae are developing the fast, easy and economical determining technology for detecting it. In this paper we produced the antibodies against saxitoxin (STX). Mariculture keyhole limpet hemocyanin (mcKLH) and ovalbumin (OVA) were used as carrier proteins. mcKLH-STX conjugates were injected into the peritonial cavity of BALB/c mouse for immunization. After bleeding from mouse, anti-STX antiserum was isolated. Indirect enzyme-linked immunosorbent assays (ELISA) was performed to determine antiserum titer using the microtiter plate coated with free STX and OVA-STX. A goat anti-mouse IgG-phosphatase conjugate was used as secondary antibody to enable chromogenic reaction. Reactions of anti-STX antiserum were very specific on the OVA-STX and free STX. Sensitivity of anti-STX antiserum on STX was very high and STX detection limit was to be 64.9 ng/kg for indirect ELISA.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.1989-1996
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• 2015

Ice-binding proteins (IBPs) can inhibit ice recrystallization (IR), a major cause of cell death during cryopreservation. IBPs are hypothesized to improve cell viability after cryopreservation by alleviating the cryoinjury caused by IR. In our previous studies, we showed that supplementation of the freezing medium with the recombinant IBP of the Arctic yeast Glaciozyma sp. (designated as LeIBP) could reduce post-thaw hemolysis of human red blood cells and increase the survival of cryopreserved diatoms. Here, we showed that LeIBP could improve the viability of cryopreserved mammalian cells. Human cervical cancer cells (HeLa), mouse fibroblasts (NIH/3T3), human preosteoblasts (MC3T3-E1), Chinese hamster ovary cells (CHO-K1), and human keratinocytes (HaCaT) were evaluated. These mammalian cells were frozen in dimethyl sulfoxide (DMSO)/fetal bovine serum (FBS) solution with or without 0.1 mg/ml LeIBP at a cooling rate of -1℃/min in a -80℃ freezer overnight. The minimum effective concentration (0.1 mg/ml) of LeIBP was determined, based on the viability of HeLa cells after treatment with LeIBP during cryopreservation and the IR inhibition assay results. The post-thaw viability of mammalian cells was examined. In all cases, cell viability was significantly enhanced by more than 10% by LeIBP supplementation in 5% DMSO/5% FBS: viability increased by 20% for HeLa cells, 28% for NIH/3T3 cells, 21% for MC3T3-E1, 10% for CHO-K1, and 20% for HaCaT. Furthermore, addition of LeIBP reduced the concentrations of toxic DMSO and FBS down to 5%. Therefore, we demonstrated that LeIBP can increase the viability of cryopreserved mammalian cells by inhibiting IR.

• Environmental Analysis Health and Toxicology
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• v.19 no.2
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• pp.191-200
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• 2004

Zinc (Zn) is an essential element in biological process, however inadequate Zn status in general population have been recognized. To update the knowledge for Zn-cadmium (Cd) interaction, we studied the intestinal uptake and transport, and the expression of metal transporter proteins (divalent metal transporter 1, DMT1 ; metal transporter protein 1, MTP1 ; zinc transporter 1, ZnTl ; metallothionein 1 , MT1) in duodenum after Cd exposure using Zn deficient animal model. Rats were led Zn deficient (ZnD, 0.5-1.0 mgZn/kg) or Zn supplemented (ZnS, 50mg Zn/kg) diet for 4 weeks, and followed single administration of $^{109}$ CdCl$_2$orally. The body Zn flatus and tissue Cd concentration were determined at 24 hrs after Cd administration. Total body burden of Cd and Cd absorption index (AI, %) were estimated based on the tissue Cd analyzed. DMT1, MTP1, ZnTl and MT1 mRNA were analyzed by using RT-PCR method. Feeding of Zn deficient diet for 4 weeks produced a reduced body weight gain and a depletion of body Zn. Tissue Cd concentration, body burden of Cd and Cd absorption index were higher in the ZnD diet fed rats than the ZnS diet red rats. Especially, Cd concentration in the small intestine (duodenum, jejunum and ileum) and the colon of FeD diet fed rats were higher markedly than in the FeS diet group. The expression levels of DMT1, MTP1 and ZnT1 mRNA in FeD diet fed rats were similar to the FeS diet. The level of MT1 mRNA expression was significantly lower in the FeD than the FeS diet fed rats. Taken together, theses results indicate that Zn deficiency in diet induce an increased intestinal absorption and tissue retention of Cd, and down -regulate the MT1 expression in the intestine which might be play a part of role in Cd absorption and transport in mammalian. These findings suggest that deficiency of essential metal could be enhanced the toxicity of toxic, non-esstial metals through the metal-metal interaction.

• Nutrition Research and Practice
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• v.8 no.3
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• pp.267-271
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• 2014

BACKGROUND/OBJECTIVES: Overproduction of nitric oxide (NO) by the inducible nitric oxide synthase (iNOS) enzyme can cause inflammation. Cyclooxygenase-2 (COX-2) is also involved in the inflammatory response through regulation of nuclear factor-kappa B $NF-{\kappa}B$(). Areca catechu is one of the known fruit plants of the Palmaceae family. It has been used for a long time as a source of herbal medicine in Indonesia. In this study, we explored the effect of Indonesian Areca catechu leaf ethanol extract (ACE) in lipopolysaccharide (LPS)-induced inflammation and carrageenan-induced paw edema models. Recently, this natural extract has been in the spotlight because of its efficacy and limited or no toxic side effects. However, the mechanism underlying its anti-inflammatory effect remains to be elucidated. MATERIALS/METHODS: We measured NO production by using the Griess reagent, and determined the expression levels of inflammation-related proteins, such as iNOS, COX2, and $NF-{\kappa}B$, by western blot. To confirm the effect of ACE in vivo, we used the carrageenan-induced paw edema model. RESULTS: Compared to untreated cells, LPS-stimulated RAW 264.7 cells treated with ACE showed reduced NO generation and reduced iNOS and COX-2 expression. We found that the acute inflammatory response was significantly reduced by ACE in the carrageenan-induced paw edema model. CONCLUSION: Taken together, these results suggest that ACE can inhibit inflammation and modulate NO generation via downregulation of iNOS levels and $NF-{\kappa}B$ signaling in vitro and in vivo. ACE may have a potential medical benefit as an anti-inflammation agent.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.2
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• pp.127-135
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• 2006

Objectives: The extent of bone formation that occurs at the interface of metallic implants and bone is determined by the number and activity of osteoblastic cells. Stress proteins may be contributing determinants of cell viability in altered environments. Hsp27 is a small Mr hsp which is known as a molecular chaperone. Methods: To better understand how heat shock protein 27 contributes to endosseous implant - associated metal ions affects on osteoblastic cell viability, the effect of chromium and titanium ions were compared to effects of cadmium ions in the ROS17/2.8 osteoblastic cell line. Results: ROS17/2.8 osteoblastic cell line demonstrated ion - specific reductions in growth; reductions were significantly greater for cadmium than for chromium or titanium. Chromium impaired growth of cultures without altering cell viability measured using the MTT assay. A stable transformed cell line expressing additional hsp27(clone "A7") was resistant to the toxic effects of titanium and partially protected from cadmium toxicity. Conclusions: A role for hsp27 in protection of osteoblastic cells from metal ion toxicity is supported by the chromium - induced elevations in hsp27 abundance and the behavior of the A7 cell line in response to metal ions in culture. Similar biochemical responses to altered cellular environments may contribute to the fate of tissues adjacent to select metallic implants.

• Journal of Periodontal and Implant Science
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• v.34 no.4
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• pp.771-780
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• 2004

Chronic exposure to high levels of manganese (Mn) leads a pronounced and debilitating disorder known as manganism. Research on the toxic manifestation of manganese have focused primarily on its neurological effects because exposure to high levels of the metal produces a distinct and irreversible extrapyramidal dysfunction resembling the dystonic movements associated with Parkinson's physiological and biochemical systems in the body. The purpose of this study is to determine the effects of Mn on mineralization in primary rat calvarial cells. The experimental groups were in concentration of 0, 10, 30 and 60 ${\mu}M$. The results were as follows: 1. ALP activity was decreased in concentration of 30 and 60 ${\mu}M$ (p<0.01). 2. Bone nodule formation was depressed in concentration of 30 and 60 ${\mu}M$ at day 14 and 21 (p<0.01). 3. RT-PCR results showed an altered expression of bone matrix proteins. These result suggested that manganese might decrease or alter the expression of the osteoblast phenotype.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.61-68
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• 1999

Rapid accumulation of reactive oxygen species (ROS) and their toxic reaction products with lipids and proteins significantly contributes to the damage of crop plants under biotic and abiotic stresses. We have identified several stress activated alfalfa genes, including the gene of the alfalfa ferritin and a novel NADPH-dependent aldose/aldehyde reductase enzyme. Transgenic tobacco plants that synthesize alfalfa ferritin in vegetative tissues-either in its processed form in chloroplast or in the cytoplasmic non-processed form-retained photosynthetic function upon free radical toxicity generated by paraquat treatment and exhibited tolerance to necrotic damage caused by viral and fungal infections. We propose that by sequestering intracellular iron involved in generation of the very reactive hydroxyl radicals through a Fenton reaction, ferritin protects plant cells from oxidative damage. Our preliminary results with the other stress-inducable alfalfa gene (a NADPH-dependent aldo-keto reductase) indicate, that the encoded enzyme may play role in the stress response of the plant cells. These studies reveal new pathways in plants that can contribute to the increased stress resistance with a potential use in crop improvement.

• Molecular & Cellular Toxicology
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• v.3 no.3
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• pp.208-213
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• 2007

With the help of a development and popularization of microarray technology that enable to us to simultaneously investigate the expression pattern of thousands of genes, the toxicogenomics experimenters can interpret the genome-scale interaction between genes exposed in toxicant or toxicant-related environment. The ultimate and primary goal of toxicogenomics identifies functional context among the group of genes that are differentially or similarly coexpressed under the specific toxic substance. On the other side, public reference databases with transcriptom, proteom, and biological pathway information are needed for the analysis of these complex omics data. However, due to the heterogeneous and independent nature of these databases, it is hard to individually analyze a large omics annotations and their pathway information. Fortunately, several web sites of the public database provide information linked to other. Nevertheless it involves not only approriate information but also unnecessary information to users. Therefore, the systematically integrated database that is suitable to a demand of experimenters is needed. For these reasons, we propose SOP (Search of Omics Pathway) database system which is constructed as the integrated biological database converting heterogeneous feature of public databases into combined feature. In addition, SOP offers user-friendly web interfaces which enable users to submit gene queries for biological interpretation of gene lists derived from omics experiments. Outputs of SOP web interface are supported as the omics annotation table and the visualized pathway maps of KEGG PATHWAY database. We believe that SOP will appear as a helpful tool to perform biological interpretation of genes or proteins traced to omics experiments, lead to new discoveries from their pathway analysis, and design new hypothesis for a next toxicogenomics experiments.