Gap junction is an ion channel forming between adjacent cells. It also acts as a membrane channel like sodium or potassium channels in a single cell. The amino acid residues up to the $10^{th}$ position in the amino (N)-terminus of gap junction hemichannel affect gating polarity as well as current-voltage (I-V) relation. While wild-type Cx32 channel shows negative gating polarity and inwardly rectifying I-V relation, T8D channel in which threonine residue at $8^{th}$ position is replaced with negatively charged aspartate residue shows reverse gating polarity and linear I-V relation. It is still unclear whether these changes are resulted from the charge effect or the conformational change of the N-terminus. To clarify this issue, we made a mutant channel harboring cysteine residue at the $8^{th}$ position (T8C) and characterized its biophysical properties using substituted-cysteine accessibility method (SCAM). T8C channel shows negative gating polarity and inwardly rectifying I-V relation as wild-type channel does. This result indicates that the substitution of cysteine residue dose not perturb the original conformation of wild-type channel. To elucidate the charge effect two types of methaenthiosulfonate (MTS) reagents (negatively charged $MTSES^-$ and positively charged $MTSET^+$) were used. When $MTSES^-$ was applied, T8C channel behaved as T8D channel, showing positive gating polarity and linear I-V relation. This result indicates that the addition of a negative charge changes the biophysical properties of T8C channel. However, positively charged $MTSET^+$ maintained the main features of T8C channel as expected. It is likely that the addition of a charge by small MTS reagents does not distort the conformation of the N-terminus. Therefore, the opposite effects of $MTSES^-$ and $MTSETT^+$ on T8C channel suggest that the addition of a charge itself rather than the conformational change of the N-terminus changes gating polarity and I-V relation. Furthermore, the accessibility of MTS reagents to amino acid residues at the $8^{th}$ position supports the idea that the N-terminus of gap junction channel forms or lies in the aqueous pore.
Ahn Sun Hee;Lee Eun Mi;Kim Dong Gyun;Hong Gyoung Eun;Park Eun Mi;Kong In Soo
Journal of Life Science
/
v.16
no.1
/
pp.131-135
/
2006
Oligopeptide is known to be an essential nitrogen nutrient for bacterial growth. Oligopeptide can be transported into cytoplasm by a specific transport system, Opp system. Opp system is composed of five proteins, which are transcribed by an operon. These are responsible for oligopeptide binding protein (OppA), permease (OppB and OppC) and energy generation system (OppD and OppF), respectively. Previously, we isolated the opp operon from Vibrio fluvialis and constructed the oppA mutant by allelic exchange method. In this study, we investigated the growth pattern and biofilm production under the different growth condition. When the cells were cultivated using brain heart infusion(BHI) medium, the wild type was faster than the mutant in growth during the exponential phase. However, it showed that the growth pattern of two strains in M9 medium is very similar. The growth of wild type showed better than that of the mutant grown at pH 8. At pH 7, there was no an obvious difference in growth. After 5 mM $H_2O_2$ was treated to the cells $(OD_{600}=1.2)$, the cell survival was examined. The oppA mutation did not affect in survivability. In the presence of $10{\mu}g/ml$ polymyxin B, the biofilm production of the oppA mutant was higher than that of the wild type.
An Internet shopping mall for clothing operates a warehouse for packing and shipping products to fulfill its orders. All the products in the warehouse are put into the boxes of same brands and the boxes are stored in a row on shelves equiped in the warehouse. To make picking and managing easy, boxes of the same brands are located side by side on the shelves. When new products arrive to the warehouse for storage, the products of a brand are put into boxes and those boxes are located adjacent to the boxes of the same brand. If there is not enough space for the new coming boxes, however, some boxes of other brands should be moved away and then the new coming boxes are located adjacent in the resultant vacant spaces. We want to minimize the movement of the existing boxes of other brands to another places on the shelves during the warehousing of new coming boxes, while all the boxes of the same brand are kept side by side on the shelves. Firstly, we define the adjacency of boxes by looking the shelves as an one dimensional series of spaces to store boxes, i.e. cells, tagging the series of cells by a series of numbers starting from one, and considering any two boxes stored in the cells to be adjacent to each other if their cell numbers are continuous from one number to the other number. After that, we tried to formulate the problem into an integer programming model to obtain an optimal solution. An integer programming formulation and Branch-and-Bound technique for this problem may not be tractable because it would take too long time to solve the problem considering the number of the cells or boxes in the warehouse and the computing power of the Internet shopping mall. As an alternative approach, we designed a fast heuristic method for this reallocation problem by focusing on just the unused spaces-empty cells-on the shelves, which results in an assignment problem model. In this approach, the new coming boxes are assigned to each empty cells and then those boxes are reorganized so that the boxes of a brand are adjacent to each other. The objective of this new approach is to minimize the movement of the boxes during the reorganization process while keeping the boxes of a brand adjacent to each other. The approach, however, does not ensure the optimality of the solution in terms of the original problem, that is, the problem to minimize the movement of existing boxes while keeping boxes of the same brands adjacent to each other. Even though this heuristic method may produce a suboptimal solution, we could obtain a satisfactory solution within a satisfactory time, which are acceptable by real world experts. In order to justify the quality of the solution by the heuristic approach, we generate 100 problems randomly, in which the number of cells spans from 2,000 to 4,000, solve the problems by both of our heuristic approach and the original integer programming approach using a commercial optimization software package, and then compare the heuristic solutions with their corresponding optimal solutions in terms of solution time and the number of movement of boxes. We also implement our heuristic approach into a storage location assignment system for the Internet shopping mall.
In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at $30^{\circ}C$ for $60{\sim}90$ min after the pretreatment of $1{\sim}2%$ 2-mercaptoethanol at $30^{\circ}C$ for $15{\sim}20$ min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/50 pV, 615 $V/256\;{\mu}sec$) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.
KIM Ji Hoe;YOON Ho Dong;PARK Hee Yun;LEE Hee Jung;CHANG Dong Suck
Korean Journal of Fisheries and Aquatic Sciences
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v.36
no.1
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pp.24-29
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2003
In the previous reports, the authors isolated two strains of marine bacteria, Shewanella sp. SR-14, which has Chaetonros sp. growth inhibition activity, and Vibrio alginolyticus, that did not affect growth of the alga. In the present study, fatty acid compositions of diatoms, Chaetoceros calcitrans and Skeletonema costatum, and marine bacteria, Shewanella sp. SR-14 and V. alginolyticus, were analyzed. Changes of fatty acid composition in the diatoms grown with the marine bacteria were also determined. Major fatty acids of Sbewanella sp. SR-14 were 16:1n-7 $(29.4\%)$ and 16:0 $(19.2\%)$ during incubation in peptone broth at $20^{\circ}C$ for 3 days. The compositions of V. alginolyticus detected were 16:0 $(23.7\%),$ 16:1n-7 $(27.7\%)$ and 18:1n-7 $(21.0\%).$ C. calcitrans consisted of 16:1n-7 $(33.3\%),$ 16:0 $(27.1\%)$ and 14:0 $(12.1\%).$ S. costatum mainly contained 16:1n-7 $(28.9\%),$ 16:0 $(21.6\%)$ and 20:5 $(19.8\%).$ When halves of cell numbers of C. calcitrans were moribund cells by Shewanella sp. SR-14, the C. calcitrans and S. costatum simultaneously cultured with the bacteria were harvested by filtration with GE/D glass microfibre filter. In the fatty acid composition of both diatoms, saturated fatty acid contents in both diatoms grown with Shewanella sp. SR-14 were decreased, but unsaturated fatty acid contents were increased. The differences were greater in C. calcitrans than those in S. costatum. During the growth of diatoms with V. alginolyticus, C. calcitrans showed increase of saturated fatty acid contents and decrease of unsaturated fatty acid contents; however, S. costatum did not show sharp difference in fatty acid content. In this study, Shewanella sp. SR-14, which showed growth inhibition activity against C. calcitrans, influenced on the changes of fatty acid contents in the diatom. It was suggested that increased unsaturated fatty acid was synergistically activated algal growth inhibition activity of Shewanella sp. SR-14.
Ten patients with deep muscle-invading bladder carcinoma (clinical stages T3a to T4b) who were not candidates for cystectomy were treated with combined modality treatment with transurethral resection, cisplatin chemotherapy and pelvic irradiation from 1989 through 1990, and were analyzed retrospectively. All patients were not candidates for cystectomy because the tumors were judged unresectable or they were not fit for a radical cystectomy. Of the patients 5 had clinical stage T3a, 3 stage T3b and 2 stage T4b disease. The minimum follow-up was 16 months. The complete response rate is 60$\%$ for all patients. The complete responses were achieved in 4 of 5(80$\%$) with stage cT3a, in 2 of 3(67$\%$)with stage cT3b and in none of 2(0$\%$) with stage cT4b. The partial responses were achieved in 2, so an overall response rate was 80$\%$. All six patients with grade I or II transitional cell carcinoma showed complete responses. Four patients with higher grade tumors showed partial responses in 2 and no response in 2, and all died of their bladder cancer. Six patients who showed complete responses after treatment are alive and only one of them showed a local recurrence 10 months after treatment. Distant metastases developed in 3 patients: lungs in 2(cT4b) of those who were never locally free of disease and spine in 1 patient (cT3b) among those with a partial response. Two patients died of metastases to lungs. During the follow-up diarrhea occurred in one which was improved after conservative treatment. On the basis of this analysis it is suggested that combined modality treatment seems to be a tolerable regimen and can be offered with a relatively high probability of success and conservation of bladder function in those with less advanced tumors by clinical stage and low grade.
Purpose: To evaluate the effectiveness of postoperative radiation therapy in cervical cancer patients and define the prognostic factors to affect survival rates. Materials and Methods: Eighty one patients with cervical cancer who were treated with postoperative radiation therapy following surgery at our institution between May 1992 and April 2000 were retrospectivelv analyzed. Forty two patients had stage IB disease, 17 had stage IIA disease, and remaining 22 had stage IIB disease, respectively. Histological examination revealed 76 squamous cell carcinoma and 5 adenocarclnoma. Sixty one patients were noted to have stromal invasion greater than 8 mm and 20 patients were noted to have stromal invasion 7 mm or less. Sixteen patients had parametrial invasion and 65 patients did not. Positive vaginal resection margin was documented in only eight patients and positive lymphovascular invasion was in twelve patients. All of the patients were treated with external beam radiation therapy alone. Majority of the patients were treated with 4 field brick technique to encompass whole pelvis. Total of 5,500 cGy was delivered to the primary surgical tumor bed. Minimum follow up period was four years. Results: Actuarial disease free survival rates for entire group of the patients were 95% and 89% at 2 and 5 years, respectively Five year disease free survival rates for patients with stage IB, IIA, and IIB disease were 97%, 87% and 70%, respectivelv. Local recurrences were documented in 5 patients. Cumulative local failure rate at 3 years was 6% Five year disease free survival rates for patients with stromal invasion greater than 8 mm and 7 mm or less were 88% and 92%, respectively (p>0.05). Five year disease free survival rate for patients with parametrial invasion was significantly lower than those with no invasion (72% vs 92%, p<0.05). Also there was significantly lower survival in patients with positive vaginal resection margin, compared with patients with negative resection margin (64% vs 94%, p<0.05). However, lymphovascular invasion was not a statistically significant prognostic factor Parametrial invasion and positive surgical resection margins were noted to be significant prognostic factors. Conclusions: Postoperative radiation therapy appears to be beneficial in controlling local disease in cervical cancer patients with high pathologic risk factors. Parametrial invasion and positive resection margins were noted to be significant prognostic factors to affect survival and more effective treatment should be investigated in these patients.
Purpose: Rapid diagnosis and prompt treatment remain the cornerstone of management of patients with meningitis. Examination of cerebrospinal fluid(CSF) for glucose, protein, cells, and organisms is necessary for accurate diagnosis, but in many parts of the world facilities do not exist to do so. We tested CSF samples from 69 children with suspected meningitis with urinary reagent strips and analyzed the results to know the usability of urinary reagent strips in diagnosis of meningitis. Methods: 69 CSF samples obtained from children with suspected meningitis were analysed. Each sample was divided into two; one was sent to the laboratory for routine diagnostic evaluation, and the other was tested with the reagent strip(Combur10 Test M). Laboratory values of CSF glucose, protein, and leucocytes for each CSF sample were plotted against the corresponding reagent-strip category. Results: There was good agreement between the dipstick and laboratory values for CSF protein and cell count but not for glucose. Conclusion: Rapid diagnosis of meningitis could be made with three component of the Combur10 test strip. It is easy to do and would be of particular value to those working in parts of the world where no laboratory facilities exist.
Purpose: We performed this study to find out short period humoral immunogenicity and safety of 47 passaged Oka strain live attenuated varicella vaccine(1,400PFU) in 12 months to 15 years aged healthy children. Methods: Ninety nine healthy chidren, who have no histories of varicella vaccination, recent chicken pox illness and contact, allergy to other vaccines and underlying severe diseases, were involved in this study from April 1997 to August 1997. 5ml blood were collected before vaccination and after vaccination from all vaccinees to measure varicella membrane antibody by FAMA, and varicella IgG antibody by EIA. And immediate reactions within 30 minutes after vaccination, local and systemic reactions within 3 days after vaccination and vaccine induced systemic illness during 6 weeks postvaccination period were observed in all vaccinees to identify side effects of study vaccine. Results: 1) 49 seronegative and 50 seropositive vaccinees were identified in both prevaccination serologic tests. 2) Serologic responses after vaccination measured by the FAMA in seronegative group showed that the mean GMT level revealed 64.0, and seroconversion rate was 97.9%. And serologic responses after vaccination measured by the FAMA in seropositive group showed that the mean GMT level(242.2) was markedly elevated comparing with the mean GMT level(9.2) of pre vaccination. 3) The results of EIA in seronegative group revealed that postvaccination mean GMT was 435.2(prevaccination GMT; 78.7), and 100% seroconversion rate. Also, the results of EIA in seropositve group showed that the mean GMT level(769.9) of postvaccination was almostly two fold hihger than the mean GMT level(419.7) of prevaccination. 4) Observed local reactions like injection sites redness, pain, hardness and itching sense were mild and disappeared within 3 days, also shorterm systemic reactions like irritability, lethargy, poor appetites and rash were not remarkable. And there were no remarkable side effects due to vaccine during study period in all vaccinees. Conclusion: We confirmed that 47 paasaged Oka strain live attenuated varicella vaccine has high shorterm humoral immunogenicity and safety. However, we need more detail and longterm humoral and cell mediated immunogenicity studies of this vaccine including clinical field trials.
Purpose : The purpose of this study was to identify useful predictors for diagnosing bacterial meningitis and performing CSF studies in febrile infants three months or younger. Methods : Six hundred and fifty two febrile infants with a rectal temperature ${\geq}38.0^{\circ}C$ presented from January 2003 to April 2008 and were retrospectively studied. The total white blood cell count (WBC), band count, absolute neutrophil count (ANC), quantitative C-reactive protein (CRP) and blood cultures were performed on admission. The clinical variables associated with bacterial meningitis were analyzed. Results : In patients with bacterial meningitis, the clinical variables including CRP (P=0.036), band count (P=0.037), ANC (P=0.036) and age (P=0.001) were significantly different. The area under the receiver-operating characteristic curve was 0.969 for CRP, 0.946 for the band count, 0.765 for the ANC and 0.235 for age. A CRP cutoff point of 8 mg/dL was determined to maximize both the sensitivity and specificity (sensitivity 83%, specificity 95%, likelihood ratio 16.6). A CRP concentration of <7 mg/dL "ruled-out" bacterial meningitis, with a likelihood ratio of 0.17, a posttest probability of <0.1% and negative predictive value 91%. A CRP concentration greater than 9 mg/dL had a much higher likelihood ratio (20.1) than the band count (16.6) and ANC (2.2). Conclusion : The CRP concentration was a useful laboratory test for the differential diagnosis of bacterial meningitis among febrile infants three months of age or younger. A CRP concentration of <7 mg/dL effectively ruled out bacterial meningitis; a value ${\geq}9mg/dL$ increased the clinical suspicion of bacterial meningitis and the need for CSF evaluation.
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