The genomic structure and phylogenetic relationships of HSP88 genes from P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa are described. The HSP88 genomic DNA from P. tenuipes Jocheon-1, P. tenuipes and C. militaris all contain 5 introns and 6 exons with the length of 13, 62, 32, 1438, 306, 288 bp, encoding 713 amino acid residues. C. pruinosa HSP88 genomic DNA contains 4 introns and 5 exons encoding 713 amino acids. The length of each exon of C. pruinosa HSP88 is 13, 62, 32, 1744, 288 bp and the length of exon 4 is identical to the total length of exon 4 and exon 5 of HSP88 of P. tenuipes Jochoen-1, P. tenuipes, and C. militaris. The deduced amino acid sequence of P. tenuipes Jocheon-1 HSP88 showed 99% identity with the P. tenuipes, 97% identity with the Cordyceps militaris, and 98% identity with the C. pruinosa. Phylogenetic analysis confirmed that the P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa HSP88 are placed together within the ascomycetes group of fungal clade.

In this study, a full-length heat shock protein88 complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening of P. tenuipesJocheon-1 Uni-Zap cDNA library and 5' RACE polymerase chain reaction. The Paecilomyces tenuipes Jocheon-1 heat shock protein88 cDNA contains an open reading frame of 2,139 bp encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipes Jocheon-1 HSP88 cDNA showed 77% identity to N. haematococca HSP88 and 45-76% identity to other fungi HSP88. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade and P. tenuipes Jocheon-1 HSP88 also contains the conserved ATPase domain at the N-terminal. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as a 88 kDa polypeptide in baculovirus-infected insect Sf9 cells. Under different stress conditions, mRNA expression of P. tenuipes Jocheon-1 HSP88 were quantified by real-time PCR and the result showed that heat shock stress affected the mRNA expression levels of P. tenuipes Jocheon-1 HSP88.

Presently, We have constructed an olig-d(T) primed directional cDNA library from the silkworm Dongchunghacho, an entomopathogenic fungus, of which species is belonging to Paecilomyces tenuipes Jocheon-1. To isolate and screen genes in the fungus, 626 expressed sequence tags(ESTs) were generated by a partial sequencing from the cDNA library. Paecilomyces tenuipes Jocheon-1 cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase(Pt-GAPDH) of Paecilomyces tenuipes Jocheon-1 was cloned from the above cDNA library. The complete cDNA sequence of Pt-GAPDHis comprised of 1,014bp encoding 338 amino acid residues. The deduced protein sequence of Pt-GAPDH showed higher homology with Beauberia bassiana-GAPDH(93% amino acid identity). Hydropathy analysis revealed that Pt-GAPDH protein is hydrophilic. The major three amino acids in its composition of amino acid residues were alanine(11.54%), valine(9.47%) and glycine(8.88%). The cDNA encoding Pt-GAPDH was expressed as a 37 kDa polypeptide in baculovirus-infected insect Sf9 cells. The Pt-GAPDH gene of Paecilomyces tenuipes entomopathogenic fungus consisted of three exons and two introns coding for 338 amino acid residues, and the genomic DNA length of the gene spans 1302bp. The accession number of the gene in GenBank are GU997099 for Pt-GAPDH cDNA and GU997102 for Pt-GAPDH genomic DNA.

Fungi belonging to the Paecilomyces spp. have recently been used as food and herbal medicines in Korea and are greatly popular as commercially available powdered supplement or dried fruiting body. Despite this acceptance and its use, little is known of the genes related to its reactive agents. Presently, We have constructed an olig-d(T) primed directional cDNA library from the silkworm Dongchunghacho, an entomopathogenic fungus, of which species is belonging to Paecilomyces spp. based on the previous identification of ITS1 and ITS2 at the molecular level and collected from Jocheon Miryang, Korea. To isolate and screen genes in the fungus, 626 expressed sequence tags(ESTs) were generated by a partial sequencing from the cDNA library. cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase(Pt-GAPDH) of Paecilomyces tenuipes- Jocheon was cloned from the above cDNA library. The complete cDNA sequence of Pt-GAPDH is comprised of 1,014bp encoding 338 amino acid residues. The deduced protein sequence of Pt-GAPDH showed higher homology with Beauberia bassiana-GAPDH(93% amino acid identity). Hydropathy analysis revealed that Pt-GAPDH protein is hydrophilic. The major three amino acids in its composition of amino acid residues were alanine(11.54%), valine(9.47%) and glycine(8.88%). The Pt-GAPDH gene of Paecilomyces tenuipes entomopathogenic fungus consisted of three exons and two introns coding for 338 amino acid residues, and the genomic DNA length of the gene spans 1302bp. The accession number of the gene in GenBank are GU997099 for Pt-GAPDH cDNA and GU997102 for Pt-GAPDH genomic DNA. More investigation works including gene expression, immunological analysis etc. will be carried continuously without hesitation after this presentation.

A full genomic DNA microarray technique was employed to investigate the effects of Dongchunghacho on aortal and hepatic gene expression in apolipoprotein E knockout mice fed a high-fat/high-cholesterol diet. Male 8- week - old ApoE-/- mice were randomly divided into two groups, control(high cholesterol group; HC) and supplementation of Dongchunghacho (SD). All of the mice were fed a high-fet/high cholesterol diet with or without Dongchunghacho supplemented by 1% for 6 weeks. At first, lipid profile of the Dongchunghacho was measured by biochemical analysis. No differences were observed in serum triglyceride and total cholesterol levels between the two groups. Antigenotoxic effect of the Dongchunghacho was measured by the single cell gel electrophoresis assay (Comet assay) and quantified as % fluorescence in tail. Dongchunghacho supplementation decreased significantly leukocytic DNA damage and also there was a tendency of reduction in hepatic DNA damage in Dongchunghacho group compared with the control group. In up regulated genes in liver and aorta of the mice, genes with 0 to 2- fold difference in expression level between the two group (HD and SD) was very much more in liver than in aorta, on the contrary, those with 2-fold to 16-flod difference increased greatly rather in aorta than in liver. Also, almost the same results were observed in down regulated genes in liver and aorta between the two groups. These results suggested that supplementation of Dongchunghacho might be helpful in preventing leukocytic DNA damage induced by high fat diet, and has a more crucial roles in aortal gene expression.

과학기술의 발달과 경제수준 향상 및 운동량의 부족으로 인한 비만증의 발생율이 높아지는 반면 체중 감량에 대한 관심도 증가하고 있다. 특히 이런 관심은 젊은 여성들 사이에서 발생하는 경향이 있으며, 지나친 체중감량은 건강에도 해를 미치지만 체중감량에 대한 자료가 거의 전무한 실정이다. 본 연구는 다이어트가 영양상태에 미치는 영향을 알아보고자 하여 제1보고로써 다이어트 실태에 대해 조사한 것이다. 1. 조사 대상자들은 19세(27%), 20세(25.3%)의 연령층이 가장 두터웠으며, 체중은 50.32±0.91kg, 신장은 160.90±0.47cm이었고, BMI는 18.73±4.89로 나타나 평균적으로 여윈 상태임을 보였다. 2. 조사 대상자들의 한달 용돈은 10∼15만원이 가장 많았으며, 부모님의 체형은 정상이거나 마른 분이 절반을 넘었다. 한편 아버지만 뚱뚱하신 경우보다는 어머니만 뚱뚱하신 분이 더 많은 것으로 나타났다. 3. 조사 대상자들의 식태도는 하루 한, 두끼만 섭취하며, 불규칙적으로 섭취하는 사람이 많았고, 과반수가 자신의 체형을 과대인식하는 것으로 나타나 체형의 기준을 바르게 선정할 수 있는 영양교육이 필요함을 보였다. 4. 조사 대상자들의 실제 체중은 50.32±0.91kg이었으나 이상체중은 46.94±0.39kg으로써 이상체중이 유의적으로 낮았다(p=0.0114). 5. 조사 대상자의 37.54%가 1997년 한 해 중 다이어트를 실시한 경험이 있었으며, 처음 다이어트를 실시한 시기는 고등학교, 대학생 때, 고등학교와 대학 사이의 순으로 나타나 주로 고등학교 이후에 다이어트를 하려는 이유로는 자기 신체에 대한 기대 및 느낌 등의 주관적인 이유가 주된 원인이었으며, 과반수 이상의 여대생이 다이어트를 실시해 본 경험(79.3%)을 가지고 있었고, 다이어트 실시 빈도는 가끔 실시하는 경우(38.25%)가 가장 많았다. 또한 다이어트의 실시기간은 일주일 미만인 사람이 가장 많았으며, 평균 체중감소량은 1∼2kg(37.69%), 2∼4kg(29.44%)이었으며, 과반수 이상이(76.65%) 감량된 체중을 유지하지 못하는 것으로 나타났다. 6. 가장 선호하는 다이어트 방법은 하루 한끼 혹은 그 이상을 굶는 방법, 운동, 하루 3끼를 모두 섭취하되 열량을 줄이는 방법 등의 순으로 나타나 결국 섭취열량을 제한하거나 소비열량을 늘이는 방법을 사용하는 것으로 나타났다. 7. 다이어트에 대한 정보는 주로 신문, 잡지(30.86%), 가족, 친구, 친척(28.86%) 등의 대중매체나 주위의 사람들로부터 얻는 것으로 나타났다. 8. 영양상태 지각도가 보통 이상인 경우는 아버지, 어머니 두 분 다 뚱뚱하지 않은 경우일 때는 116명(42.65%)이며, 두 분 다 뚱뚱한 경우는 8명(2.95%)로 나타나 부모가 정상체중이거나 마른 경우 그 자녀의 영양자각도가 그렇지 않은 부모의 자녀에 비해 높은 것을 알 수 있었다(p=0.004). 9. 영양상태 자각도는 자신의 체형을 적당하다, 약간 뚱뚱하다고 대답한 group이 다른 group에 비해 높았다(p=0.001).

양파를 생채로 착즙하여 분리한 액즙을 Brix 70%까지 감압농축시킨 것과 이어서 잔사에 ethyl alcohol을 가하여 추출한 것을 농축시켜 같이 합하고, 갈변방지를 위하여 1% cysteine의 첨가와 유화제로서 2% PGDR을 첨가하여 양파 oleoresin을 제조하고 저장중 품질 안정성을 검토하였다. 양파로부터 oleoresin에 추출된 당은 저장온도 5℃, 25℃ 및 40℃에서 매우 안정하여 저장 60일후까지도 함량변화는 거의 일어나지 않았다. 양파 oleoresin 추출 직후 갈변도를 나타내는 흡광도가 0.38로 담갈색을 띄었는데 5℃에서는 저장 60일 후에도 변화가 거의 없었으나, 25℃ 및 40℃에서 저장한 경우는 60일후 흡광도가 1.53 및 3.32로 증가하였고, cysteine을 1% 첨가한 경우는 대조구에 비하여 갈색 억제효과가 상당히 나타나 40℃에서 60일간 저장하였을 때의 흡광도는 대조구의 절반 수준으로 나타났다. 유화 안정성은 5℃ 저장의 경우 20일, 40일 및 60일후에 분리되지 않는 emulsion 층의 부피가 각각 96.8%, 94.1% 및 90.6%로 매우 안정하였고, 25℃ 및 40℃에서 60일 저장후는 83.2% 및 75.4%로 유화상태가 다소 불안정하였다. 생채 양파로부터 추출한 oleoresin을 저장시키지 않고 바로 대두유에 1% 첨가한 경우와 0.02% BHA를 첨가하였을 때 A.I. 는 각각 1.39 및 1.72를 나타내어 0.02% BHA를 첨가한 대두유의 유도기간 연장효과에 대하여 80.8%에 해당하는 항산화 활성을 나타내었고, 5℃ 및 25℃에서 60일 동안 저장한 양파 oleoresin을 대두유에 각각 1% 첨가하였을 때의 A.I. 는 1.37 및 1.30을 나타내어 실온 또는 그 이하의 온도에서는 양파 oleoresin이 가지는 항산화 활성은 매우 안정하였으며, 40℃에서 저장 60일후의 양파 oleoresin의 A.I. 는 1.08로 나타나 고온에서 저장기일이 길어질수록 양파 oleoresin의 항산화활성은 다소 불안정하였다. Overall odor intensity의 지표로서 total pyruvate 함량변화는 5℃, 25℃ 및 40℃에서 저장 60일후 잔존율이 각각 89.9%, 79.7% 및 65.2%로 저온에서는 상당히 안정하였다. 양파 oleoresin의 저장중 pyruvate 감소에 대한 반응속도 상수는 저장온도 범위 5∼10℃에서 1.381∼4.735mmol/ℓ·hr, Q10 값은 1.537∼1.694, 그리고 활성화 에너지는 11.649㎉/g mole이었다.

K14 환자는 유방암 수술, 방사선 치료, 피부이식 수술, 무릅 관절통의 기왕력이 있는 고령의 여성 호흡곤란증 환자로서 문진과 진찰, 혈압, 비만도 측정, 임상병리, 흉부 X-선, 심전도와 심초음파, 폐기능 검사 등 체계적인 심폐기 질환의 진단 결과 비만증과 고혈압, 좌심실 비대, 만성 폐질환의 소견이 있어 심폐기능 모두에서 문제성이 발견되었다. 검사 결과들의 정밀한 판독법 및 장기-치료 관찰 결과를 임상문헌과 함께 고찰하였다.

Cytochrome c oxidase subunit Ⅱ gene(COⅡ gene) is subunit of cytochrome oxidase, which is complex Ⅳ of mitochondria electron transport system. It has been frequently used in molecular phylogenetic studies because the speed of its DNA variation is faster than that of nucleus. It is especially useful in phylogenetic study of molecular biology in insects. In this study, we cloned and sequenced COⅡ gene of mitochondria DNA from Rhabditidae family nematode. Our results showed that this gene is comprised of 696 base pairs(bp). In the analysis of similarity of this gene with other known genes of 14 species of nematodes in Rhabditida order, we identified that this gene has high similarity with that of Caenorhabditis briggsae(86.0%) and C. elegans(85.6%) in Rhabditidae family. On the meanwhile, it has very low similarity with that of Angiostrongylus cantonensis(31.8%) in Angiostrongylidae family and Metastrongylus salmi(31.6%) in Metastrongylidae family. Based on the results of this study, we suggest that this nematode is closely related with that of Caenorhabditis genus in Rhabditidae family.

The aim of this study was to isolate chicken feather-degrading bacteria with high keratinolytic activity and to investigate cultural conditions affecting keratinolytic enzyme production by a selected isolate. A chicken feather-degrading bacterial strain CH3 was isolated from poultry wastes. Isolate CH3 degraded whole chicken feather completely within 3 days. On the basis of phenotypical and 16S rDNA studies, isolate CH3 was identified as Bacillus thuringiensis CH3. This strain is the first B. thuringiensis described as a feather degrader. The bacterium grew with an optimum at pH 8.0 and 37℃, where maximum keratinolytic activity was also observed. The composition of optimal medium for keratinolytic enzyme production was feather 0.1%, sucrose 0.7%, casein 0.3%, K2HPO4 0.03%, KH2PO4 0.04%, MgCl2 0.01% and NaCl 0.05%, respectively. The keratinolytic enzyme had a pH and temperature optima 9.0 and 45℃, respectively. The keratinolytic activity was inhibited ethylenediaminetetraacetic acid, phenylmethylsulfonyl fluoride, and metal ions like Hg2+, Cu2+ and Zn2+. The enzyme activated by Fe2+, dithiothreitol and 2-mercaptoethanol.

Feathers are produced in huge quantities as a waste product at commercial poultry processing plants. Since feathers are almost pure keratin protein, feather wastes represent an alternative to more expensive dietary ingredients for animal feedstuffs. Generally they become feather meal used as animal feed after undergoing physical and chemical treatments. These processes require significant energy and also cause environmental pollutions. Therefore, biodegradation of feather by microorganisms represents an alternative method to prevent environment contamination. The aim of this study was to investigate cultural conditions affecting keratinolytic protease production by Bacillus pumilus RS7. We also assessed the nutritive value of microbial and alkaline feather hydrolysates. The composition of optimal medium for the keratinolytic protease was fructose 0.05%, yeast extract 0.3%, NaCl 0.05%, K2HPO4 0.03%, KH2PO4 0.04% and MgCl2ㆍ6H2O 0.01%, respectively. The optimal temperature and initial pH was 30℃ and 9.0, respectively. The keratinolytic protease production under optimal condition reached a maximum after 18 h of cultivation. Total amino acid content of feather hydrolysates treated by NaOH and B. pumilius RS7 was 113.8 ㎍/ml and 504.9 ㎍/ml, respectively. Essential amino acid content of feather hydrolysates treated by NaOH and B. pumilius RS7 was 47.2 ㎍/ml and 334.0 ㎍/ml, respectively. Thus, feather hydrolysates have the potential for utilization as an ingredient in animal feed.

The aim of this study was to isolate mesophilic chicken feather-degrading bacteria and to evaluate feather hydrolysate as alternative biofertilizer. Isolate RS7 was isolated from compost and identified as Bacillus pumilus according to API analysis and 16S rDNA sequencing analysis. Chicken feathers were completely degraded after 5 days of cultivation at 30℃. Feather hydrolysate treated by B. pumilius RS7 positively influenced Helianthus sannuus L. (sunflower) growth (e.g. growth rate, number and dry weight of leave, and flowering rate). These results suggest that feather hydrolysate prepared using B. pumilius RS7 could successfully be used as alternative biofertilizer, thereby reducing the environmental impact of feather waste from the poultry industry.

To investigate the variations of physico-chemical factors and microbial population, in ten stations at water region of coastal area of Chagwi-Do, Nutritive salts, water temperature, transparency, suspended solid, salinity, COD, DO, pH, heterotrophic bacteria, coliform group and Vibrio spp. were analysed three times in September, November in 2004 and February in 2005. Heterotrophic bacteria in surface water was 3.5×101～1.16×103 cfu/ml, 1.0×102～5.2×101 cfu/ml, 2.0×101～7.6×101 and bottom water counted 7.0×102～1.0×103 cfu/ml, 1.4×101～2.5×102 cfu/ml, 2.0×102～4.2×101 cfu/ml in September, November in 2004 and February in2005, respectively. The cell number of total coliform bacteria in the surface water amounted to 0～4.3×102 cfu/ml, 0～6.0×101 cfu/ml, 0～1.0×101 cfu/ml and bottom water amounted 0～2.2×102 cfu/ml, 0～5.4×102 cfu/ml, 0～2.0×101 cfu/ml in September, November in 2004 and February in 2005, respectively. As for Vibrio spp., the cell number in the surface water was 1.0×101～2.5×102 cfu/ml, 1.0×101～2.0×101 cfu/ml, 0 cfu/㎖ and bottom water counted 1.0×101～5.2×102 cfu/ml, 0 cfu/㎖, 2.0×101 cfu/ml in September, November in 2004 and February in 2005, respectively.

Pseudomonas sp. EL-04J was previously isolated from phenol-acclimated activated sludge. This bacterium was capable of degrading phenol and cometabolizing trichloroethylene (TCE). After precultivation in the mineral salts medium containing phenol as a sole carbon source, Pseudomonas EL-04J degraded 90% of TCE 25μM within 20 hours. Thus, phenol-induced Pseudomonas sp. EL-04J cells can degrade TCE. Following a transient lag period, Pseudomonas sp. EL-04J cells degraded TCE at concentrations of at least 250μM with no apparent retardation in rate, but the transformation capacity of such cells was limited and depended on the cell concentration. The degradation rate of TCE followed the Michaelis-Menten kinetic model. The maximum degradation rate (Vmax) and saturation constant (Km) were 7nmo ℓ/min·㎎ cell protein and 11μM, respectively. Cometabolism of TCE by phenol fed experiment was evaluated in 50·mℓ serum, vial that contained 10 ㎖ of meneral sals medium supplemented with 10μM TCE. TCE degradation was inhibited in the initial period of 1 mM phenol addition, but after that time Pseudomonas sp. EL-04J cells degraded TCE and showed cell growth.

This research was performed to investigate the dynamics of microbial community by RBC (Rotating Biological Contactor) using Rhodococcus sp. EL-GT and activated sludge. Cell counts revealed by DAPI were compared with culturable bacterial counts from nutrient agar. Colony counts on nutrient agar gave values 20∼25% and 1∼15% of cell counts (DAPI). The cell counts for the dynamics of bacterial community were determined by combination of in situ hybridization with fluorescently-labelled oligonucleotide probes and epifluorescence microscopy. Around 90∼80% of total cells visualized by DAPI were also detected by the bacteria probe EUB 338. For both reactors proteobacteria belonging to the gamma subclass were dominant in the first stage (1 and 2 stage) and proteobacteria belonging to the gamma subclass were dominant in the last stage (3 and 4 stage).