• Title/Summary/Keyword: Assessment Frames

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Some characters of bacterial cellulases in goats' rumen elucidated by metagenomic DNA analysis and the role of fibronectin 3 module for endoglucanase function

  • Nguyen, Khanh Hoang Viet;Dao, Trong Khoa;Nguyen, Hong Duong;Nguyen, Khanh Hai;Nguyen, Thi Quy;Nguyen, Thuy Tien;Nguyen, Thi Mai Phuong;Truong, Nam Hai;Do, Thi Huyen
    • Animal Bioscience
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    • v.34 no.5
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    • pp.867-879
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    • 2021
  • Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.

Development of a Critical Pathway for Patients with Coronary Artery Bypass Graft (관상동맥 우회술 환자를 위한 Critical Pathway개발)

  • 김기연
    • Journal of Korean Academy of Nursing
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    • v.28 no.1
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    • pp.117-131
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    • 1998
  • The purpose of this study was to develop a critical pathway for case management for patients who have received Coronary Artery Bypass Graft (CABG) because of Ischemic Heart Disease(IHD) which is a factor of rising medical expenses. For this study. a conceptual framework was developed through a review of the literature including six critical pathways which are currently being used in USA. In order to identify the overall service contents required by these patients and to draw up a preliminary critical pathway, 30 cases of medical records of patients who had CABG because of IHD between January, 1995 to June. 1996 at the Cardiovascular Center of Yonsei Medical Center in Seoul were analyzed. An expert validity test was done for the preliminary critical pathway and clinical validity test was also done using seven IHD patients with CABG between November 11 and 23, 1996. After these processes. the final critical pathway was developed. The results of this study are summarized as follows : 1. The vertical axis of the critical pathway includes the following eight items : tests, nutrition, medications, consultations, activity, assessments, treatments, education discharge planning and the horizontal axis includes the time from the start of hospitalization to discharge. 2. Analysis of the 30 medical records indicated that the average length of stay was 20.2days with the average length of stay from hospitalization day to operation day being 6.2 days, and the average length of stay from operation day to discharge day was 13. 9 days. Analysis of the service contents showed that the horizontal axis of the preliminary critical pathway was set from hospitalization to the 14th post operation day and the vertical axis was set to include eight items, the contents which ought to have occurred, according to the time frames of the horizontal axis. 3. As a result of the experts validity, it was found that among the total of 571 items. there was over 83% agreement for 482 items, less than 83% for 89 items, which were then deleted and a revision of the critical pathway was done. 4. A clinical validity test was done using seven IHD patients with CABG. During the process, three patients were deleted because they were out of the criteria the investigator set. Finally, four patients were used. The result of study indicated that only one patient was discharged on the tenth post operation day, which was one day later than the expected day. Three patients were discharged later than the expected day from three days to nine days. All the cases progressed on schedule until the operation day and the first post operation day, but from the second post operation days, there were differences between the critical pathway and the actual practice. The differences came from tests, assessments, and treatments. 5. On the basis of the results of the clinical validity test. the following revisions in the final critical pathway were made : the transfer from ICU to step down ward would be the second post operation day, and the transfer to a general ward, the fifth post operation day, for patients who complained of lack of sleep from the fifth post operation day to discharge, a sleeping pill would be prescribed, skin observations would be performed routinely from immediately after the operation until the third post operation day, and would continue if there was a sign of skin injury on the fourth post operation day, and assessment of chest pain would be done from the third post operation day, and the “stairs climbing” item, expected to be done on the ninth post operation day would be deleted. In conclusion, this critical pathway is partially applicable to the care of patients with CABG but there are some parts needed to be further investigated.

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