Yousef, Amany I;El-Masry, Omar S;Abdel Mohsen, Mohamed A
Asian Pacific Journal of Cancer Prevention
/
v.17
no.2
/
pp.743-748
/
2016
Background: $K^-Ras$ activation is an early event in colorectal carcinogenesis and associated mutations have been reported in about 40% of colorectal cancer patients. These mutations have always been responsible for enhancing malignancy and silencing them is associated with attenuation of tumorigenicity. Among downstream effectors are the RAF/MEK/ERK and the PI3K/Akt signaling pathways. PI3K/Akt signaling leads to reduction of apoptosis, stimulated cell growth and enhanced proliferation. Ellagic acid (EA), a naturally occurring antioxidant, has recently emerged as a promising anti-cancer agent. Purpose: To evaluate the impact of cellular genetic makeup of two colon cancer cell lines with different genetic backgrounds, HCT-116 ($K^-Ras^-/p53^+$) and Caco-2 ($K^-Ras^+/p53^-$), on response to potential anti-tumour effects of EA. In addition, the influence of $K^-Ras$ silencing in HCT-116 cells was investigated. Materials and Methods: Cellular proliferation, morphology and cell cycle analysis were carried out in addition to Western blotting for detecting total Akt and p-Akt (at Thr308 and Ser473) in the presence and absence of different concentrations of EA. Cell proliferation was also assessed in cells transfected with different concentrations of $K^-Ras$ siRNA or incubated with ellagic acid following transfection. Results: The results of the present study revealed that EA exerts anti-proliferative and dose-dependent pro-apoptotic effects. Cytostatic and cytotoxic effects were also observed. p-Akt (at Thr308 and Ser473) was downregulated. Moreover, EA treatment was found to (i) reduce $K^-Ras$ protein expression; (ii) in cells transfected with siRNA and co-treated with EA, pronounced anti-proliferative effects as well as depletion of p-Akt (at Thr308) were detected. Conclusions: Cellular genetic makeup ($K^-Ras^-/p53^-$) was not likely to impose limitations on targeting EA in treatment of colon cancer. EA had a multi-disciplinary pro-apoptotic anti-proliferative approach, having inhibited Akt phosphorylation, induced cell cycle arrest and showed an anti-proliferative potential in HCT-116 cells (expressing mutant $K^-Ras$).
Finasteride $[N-(1, 1-dimethylethyl)-3-oxo-4-aza-5{\alpha}-androst-1-ene-17{\beta}-carboxamide]$ is a 4-aza-3-oxosteroidal inhibitor of human $5{\alpha}-reductase$. The purpose of the present study was to evaluate the bioequivalence of two finasteride tablets, $Proscar^{\circledR}$ (MSD Korea Ltd.) and Procare (Hana Pharm. Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The release of finasteride from the two finasteride formulations in vitro was tested using KP VIII Apparatus II method with various dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty six healthy male subjects, $23.7\;{\pm}\;2.24$ years in age and $67.2\;{\pm}\;8.55\;kg$ in body weight, were divided into two groups and a randomized $2\;{\time}\;2$ cross-over study was employed. After two tablets containing 5 mg as finasteride was orally administered, blood samples were taken at predetermined time intervals and the concentrations of finasteride in serum were determined using HPLC with UV detector. The dissolution profiles of two formulations were similar in all tested dissolution media. The pharmacokinetic parameters such as $AUC_t,\;C_{max},\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t,\;C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, $Proscar^{\circledR}$, were 6.39, 4.65 and -13.9% for $AUC_t,\;C_{max},\;and\;T_{max},$ respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.800 to log 1.25 $(e.g.,\;log\;0.990{\sim}log\;1.14\;and\;log\;0.977{\sim}log\;1.13 for\;AUC_t\;and\;C_{max},\;respectively)$. Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Procare tablet was bioequivalent to $Proscar^{\circledR}$ tablet.
Sertraline HCl, (1S-cis)-4-(3, 4-dichloro-phenyl)-1, 2, 3, 4-tetrahydro-N-methyl-l-naphthalenamine hydrochloride, is a potent and selective serotonin reuptake inhibitor which is used in the treatment of depression and obsessivecompulsive disorders. The purpose of the present study was to evaluate the bioequivalence of two sertraline HCl tablets, Traline tablet (Myungin Pharm. Co. Ltd.) and Zoloft$^{(R)}$ tablet (Pfizer Inc.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The in vitro release of sertraline from the two sertraline HCl formulations was tested using KP VIII Apparatus II method with various dissolution media. Twenty four healthy Korean male volunteers, $23.50{\pm}1.74$ years in age and $64.09{\pm}7.10\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ crossover study was employed. After a single tablet containing 50 mg as sertraline HCl was orally administered, blood samples were taken at predetermined time intervals and the concentrations of sertraline in serum were determined using an online columnswitching HPLC method with UV/Vis detection. The dissolution profiles of two formulations were similar in all tested dissolution media. The pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated, and computer programs (Equiv Test and K-BE Test) were utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and un-transformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, Zoloft$^{(R)}$ tablet, were 0.04, 3.26 and -1.29% for $AUC_t$, $C_{max}$, and $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log0.8 to log1.25. Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Traline tablet was bioequivalent to Zoloft$^{(R)}$ tablet.
An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.
Purpose - As economic activities between different countries have rapidly spread in a world of free trade, it is inevitable that a large volume of cargo will be carried between countries. It is natural, then, that CO2 emissions and other environmental pollutions have followed, which exposes people and society to serious environmental problems and social costs, and so on. Therefore, the need to understand logistics is not only a matter of transportation but also an environmentally oriented matter. The purpose of this study is to look at some lessons and implications from the European case in terms of green logistics matters. Research design, data, and methodology - In order to look into this matter, first, it has to be established that some cargo transport volumes using different transportation modes have clearly declined because of previous economic recessions. Some transport policies produced by the European Union (EU) are based in a long history of struggling to cope with transport matters in European countries. In its recent transport policies, the EU has provided greener transportation alternatives, realizing that pollution matters affect the European transport market. This study tries to determine what policies the EU has implemented to deal with green logistics matters. This study concentrates in particular on the Marco Polo program in the EU. Results - This study found that the EU seems to consider these kinds of matters, that is, transport and the environment in the context of green logistics. The EU launched some policy instruments to solve this matter relatively earlier than other countries and reviewed them as necessary. In order to make these policy tools work, the EU provided PACT for combined transport, and then the Marco Polo I and II European transport white paper packages. These European policies deal with green logistics matters in two ways. First, some restrictions have been imposed, especially taxation, and so on. Transport subsidies are also powerful means of handling green logistics matters in Europe. Along with these two means of dealing with transport and the environment, the EU eventually targeted integration of different transport modes. Instead of employing only a single transport instrument mode to deliver the cargo to be carried, such as trucking, rail, ocean-going carrier, flight, or inland waterway transport, the EU has proposed that combining transport modes is the best alternative for transport and the environment. That is, the EU is pursuing the adoption of multimodalism as an answer to the green logistics challenge as it provides a more cost efficient and more productive means of transport. Conclusions - In conclusion, multimodal transport should be considered when applying green logistics, as it can provide an alternative way to achieve transport and environmental solutions together at the present time. Two methods can be used to encourage multimodal transport: restrictions and subsidies. These are the lessons and implications from European green logistics policies.
Processing of collagen from yellowfin tuna (Thunnus albacares) abdominal skins was optimized by response surface methodology and central composite design. The values of independent variables at optimal conditions were NaOH concentration: 0.5 N, NaOH treatment time: 36.2 hr, pepsin concentration: 1:4.9 ratio (0.245%, w/v), and digestion time: 48.1 hr, respectively. The collagen content estimated under optimal conditions was 33.1%, and the actual experimental collagen content was 32.3%. Physicochemical properties of collagen from yellowfin tuna abdominal skin were investigated by amino acids analysis, SDS-PAGE, FT-IR, viscosity and denaturation temperature. Amino acids content of the collagen was 21.0%. SDS-PAGE pattern of the collagen showed two different $\alpha$-chain (${\alpha}_1$- and ${\alpha}_2$- chain), $\beta$-component and $\gamma$-component. The spectrum of FT-IR of the collagen showed wavenumber at 3,434, 1,650, 1,542 and $1,235\;cm^{-1}$ representing the regions of amide A, I, II and III, respectively. Relative viscosity of the collagen decreased continuously on heating up to $32^{\circ}C$, and the rate of decrease was retarded in the temperature range of $35-50^{\circ}C$. Denaturation temperature (Td) of the collagen solution (0.06%, w/v) was $31^{\circ}C$ and was lower than calf skin collagen ($35^{\circ}C$).
Purpose : To propose various types of clinical research which is feasible for botanical new drug (IND) development processes, and suggest essential steps to development of study protocol for IND. Methods : Literature-based discussions and one research group's experience is given regarding domestic act, regulation, and system. Results : In order to get an approval of IND for botanical drug in Korea there are several types of clinical research to conduct. In quality control steps for standardized medicinal herbs, case reports or case series can be conducted, and for good manufacturing practice(GMP) steps, we can conduct case reports, case series, and retrospective cohort studies. In addition, as long as we gathered good laboratory practice(GLP) data we can conduct up to quasi-experimental studies and clinical trials including investigator initiated trials. In order to conduct these studies development of study protocol is essential. First, we obtain historical evidence including target disease and indication, efficacy, safety, and endpoints by reviewing medical classics. Second, we obtain clinically and statistically important data by conducting non-clinical studies, observation studies, and quasi-experimental studies. Third, we generate research hypotheses and purposes and explore methodologies, endpoints, clinical practice guidelines, cost-effectiveness, and commercial potential. Finally, we develop study protocol with aid of biostatistician or expert in contract research organization. Discussions and conclusions : This study have obvious limitations in that most thoughts, suggestions, and proposes are from one research group's experience. Therefore, we hope to see various types of research in this topic and process from other research group as well.
We fabricated sponge and poly(lactide-co-glycolide)(PLGA) scaffolds impregnated demineralized bone particle(DBP)(DBP/PLGA) and investigated proper condition to proliferation and phenotype maintenance of intervertebral disc(IVD) cells by comparison between DBP/PLGA scaffold and DBP sponge. DBP/PLGA scaffolds were prepared by solvent casting/salt leaching. Human IVD cells were seeded in scaffolds of two types. Cell viability and proliferation according to scaffolds were analyzed by WST assay and SEM. RT-PCR was assessed to measure mRNA expression of aggrecan and type II collagen of human IVD cells. In WST assay results, cell viability in scaffolds impregnated DBP/PLGA scaffold were higher than DBP sponge. We could observe that disc cell mRNA expressed better in DBP/PLGA scaffold than DBP sponge. We concluded that the using of DBP/PLGA in terms of scaffold fabrication for bio-disc with human IVD cells is helpful growth of disc cells maintenance of phenotypes.
Chromosomes of gonadal tissues from Fasciola spp, Eurytrema pancreaticum and Calicophoron calicophorum occurred Korean cattle were egamined using modified air-drying method. To compare their phenotype with three different genotypes among Fasciola spp, the adult and egg si2e were measured since they have been known as important taxonomical characters. The results obtained were as followed; Cattle liver fluke, Fasciola spp were classified into three types based on their chromosomal complements such as individual with 2o chromosome(diploid), 30 chromosome(triploid) and 20/30 mosaic constitution(mixoploid). The propotions of appearance of three types were 40.00%, 54.29% and 5.71%, respectively. The frequency of three types in type I which was regarded as F gigantica were 58.82% for diploid, 35.29% for triploid and 5.88% for mixoploid, but in type II which was regarded as F hepatica were 72.2% for triploid, 22.22% for diploid and 5.56% for mixoploid. Egg length of triploid forms was significantly larger than that of diploid forms and egg size of mixoploid forms was similiar to that of triploid forms. Worm size of triploid forms was larger than that of diploid forms and was more similar to that of mixoploid forms, but the statistical data were not significant. Diploid chromosome consisted of one pair of metacentric chromosome(m), four pairs of submetacentric chromosomes(sm), five pairs of subtelocentric chromosomes(st), while triploid chromosome consisted of one pair of metacentric chromosome, seven pairs of submetacen.tric chromosomes, one pair of subtelocentric chromosome and telocentric chromosome(t), respectively. In mixoploid chromosome, constitution of the chromosomes of diploid or triploid cell was consistent with that of diploid or triploid. Chromosomes of gonadal tissues from pancreatic fluke, Eurytrema pancreaticum consisted of 13 pairs of homologs(2n=26, n=13). The mitotic and meiotic divisions were observed frequently. In the mitotic metaphase, Karyotype consisted of five pairs of metacentric chromosomes, four pairs of submetacentric chromosomes, three pairs of subtelocentric chromosomes and one pair of telocentric chromosome. Chromosomes of gonadal tissues from stomach fluke, Calicvphoron calicophorum consisted of 9 pairs of homologs(2n=18, n=9). The meiotic divisions was frequently observed, but mitotic divisions was rare. In the mitotic metaphase, Karyotype consisted of two pairs of metacentric chromosomes, three pairs of submetacentric chromosomes and four pairs of subtelocentric chromosomes. Karyotype of Calicophoron calicophorum differed from that of Japanese C calicophorum which was similar to that of Paramphistomum cervi of Korean cattle. Though that of Calicophoron calicophorum of Korean cattle was similar to that of Paramphistomum explanatum of Korean cattle, that have been recognized to be a different species of fluke.
Recently river water pollution in Korea is given rise to serious problem in aspect of crop production, drinking well, water contamination and etc. Under these urgent situations, it is prime importance to protect water resources from pollutants. An environmental isotope survey of the groundwater form the shallow alluvial and the underlying crystalline rock aquifer of the Han River Basin has been undertaken, Analysis of the data has I) confirmed the hypothesis that the groundwater from the metropolitan area is recharged from the river whereas that form the non-urbanized region of the Basin is replenished by the infiltrating precipitation; ii) shown that crystalline rock aquifers are recharged by the ground water form the overlying alluvium. Old groundwater is a group of wells with tritium values in the range of 0 to 2 TU. These low values indicate that the water sampled was recharged much ealier, at least a few decades, than the other groundwater samples of higher tritium content. The low values in this region may, in fact, reflect the effect of the impermeable clay layers which impede infilteration from the surface. Stable isotope evidence confirmed that a recharge in the karst area occurs at a significantly greater elevation than that to the alluvial aquifer. An analysis of the tritium level collected over an annual cycle suggests that the residence time of groundwater is probably not more than a few months. There does not appear to be any correlation between the trace level of Zn, Mn and Pb in the groundwater and the mechanism of the recharge.
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