The purpose of this study was to evaluate importance and performance of dietitian's task at long term care hospitals foodservices in the Busan.Kyongnam area. The research was performed through using questionnaires and conducted from June 11 to July 16, 2010 for 186 dietitians at 141 long-term care hospitals. Seventy-two percent of hospitals had two dietitians and 69% of them had a dietitian's office. Fifty-two percent of dietitians has worked for less than 2 years at long term care hospital, and 37.1% of them worked additional tasks. Seventy-three percent of hospitals conducted a therapeutic diet program and the therapeutic diets frequently provided were diabetic diet > tube feeding diet > dysphasia diet > sodium controlled diet. Mean score for the importance (4.36/5.00) and performance (3.91/5.00) of dietitian's tasks were significantly different (p < 0.001). The importance and performance grid showed that the purchase-inspection management and sanitation-safety management were high scores to the importance and performance (doing great area), menu-foodservice management and cooking-working management were low scores to the importance and high scores to the importance (overdone area), and nutrition management was low scores to the importance and performance (low priority). Forty-three percent of dietitians agreed with the needs for role separation between foodservice dietitian and clinical dietitian.
Aphasia ; Diet, Diabetic ; Diet ; Enteral Nutrition ; Long-Term Care ; Surveys and Questionnaires ; Sodium

BACKGROUND: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(Polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. METHODS: We used the InstaGene(TM) DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1, 245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2, 536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. RESULTS: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value.. CONCLUSION: Even though both methods had lower possibility of cross contamination, shorter time requrirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usfulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.
DNA* ; Electrophoresis, Agar Gel ; Endopeptidase K ; Ion Exchange* ; Microwaves ; Polymerase Chain Reaction* ; Sensitivity and Specificity ; Sputum ; Tuberculosis*

In this study, we investigated profiles of the cytokines IFN-g, IL-12, and IL-10 in active pulmonary tuberculosis (EAPTB) patients, HIV-negative patients with multidrug-resistant tuberculosis (MDR-TB) and in healthy tuberculin reactors (HTR). We studied the responses of peripheral blood mononuclear cells (PBMC) from 12 EAPTB patients and 15 MDR-TB patients to stimulation with a purified protein derivatives (PPD) antigen (Ag), and compared them with those from 14 HTR. Using ELISA, IFN-g production was found to be significantly depressed, while IL-10 was significantly elevated in both MDR-TB and EAPTB after in vitro stimulation with PPD, compared with those in HTR. Although there was no significant difference in IL-12 production among the three groups, mean IL-12 production was highest in patients with MDR-TB. In these patients, IL-12 production was significantly correlated with IL-10 expression, but not IFN-g production. In addition, neutralization of endogenous IL-10 led to enhanced IFN-g and IL-12Rb2 mRNA expression in TB patients. Our findings suggest that both groups of TB patients may have a similar disregulated pattern of IL-12, IL-10, and IFN-g production during M. tuberculosis infection. Furthermore, the results suggest a potentially pathogenic role for IL-10 in impaired Th1 immune responses in TB patients.
Cytokines ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interferon-gamma ; Interleukin-10* ; Interleukin-12* ; Mycobacterium tuberculosis ; RNA, Messenger ; Tuberculin ; Tuberculosis ; Tuberculosis, Multidrug-Resistant* ; Tuberculosis, Pulmonary

PURPOSE: This study examined a rapid isolation method decreasing the time and cost of the clinical application of adipose tissue-derived stem cells (ASCs). MATERIALS AND METHODS: Aliquots (10 g) of the lipoaspirates were stored at 4degrees C without supplying oxygen or nutrients. At the indicated time points, the yield of mononuclear cells was evaluated and the stem cell population was counted by colony forming unit-fibroblast assays. Cell surface markers, stem cell-related transcription factors, and differentiation potentials of ASCs were analyzed. RESULTS: When the lipoaspirates were stored at 4degrees C, the total yield of mononuclear cells decreased, but the stem cell population was enriched. These ASCs expressed CD44, CD73, CD90, CD105, and HLA-ABC but not CD14, CD31, CD34, CD45, CD117, CD133, and HLA-DR. The number of ASCs increased 1x1014 fold for 120 days. ASCs differentiated into osteoblasts, adipocytes, muscle cells, or neuronal cells. CONCLUSION: ASCs isolated from lipoaspirates and stored for 24 hours at 4degrees C have similar properties to ASCs isolated from fresh lipoaspirates. Our results suggest that ASCs can be isolated with high frequency by optimal storage at 4degrees C for 24 hours, and those ASCs are highly proliferative and multipotent, similar to ASCs isolated from fresh lipoaspirates. These ASCs can be useful for clinical application because they are time- and cost-efficient, and these cells maintain their stemness for a long time, like ASCs isolated from fresh lipoaspirates.
5'-Nucleotidase/metabolism ; Adipose Tissue/*cytology ; Adult ; Antigens, CD/metabolism ; Antigens, CD44/metabolism ; Antigens, Thy-1/metabolism ; Cell Differentiation/physiology ; Cells, Cultured ; Female ; Humans ; Immunoblotting ; Immunohistochemistry ; Immunophenotyping ; Mesenchymal Stem Cells/metabolism ; Muscle Development/genetics/physiology ; Osteogenesis/genetics/physiology ; Receptors, Cell Surface/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells/*cytology/metabolism ; Young Adult

Understanding human immune responses in chronic refractory tuberculosis (CRTB) is important for developing immunotherapy against the disease. The aim of this study was to examine cytokine responses [interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-10] by peripheral blood mononuclear cells (PBMCs) in CRTB patients after in vitro stimulation with the 30-kDa or purified protein derivative (PPD) antigen (Ag). Most of the CRTB cases were multidrug-resistant (MDR) TB. The results were compared with those from early TB (E-TB) patients and healthy tuberculin reactors (HTR). IFN-gamma production was significantly depressed in both CRTB and E-TB groups compared with HTR. In response to the 30-kDa Ag, TNF-alpha levels were significantly depressed only in CRTB patients, while greatly increased in E-TB patients. In addition, IL-10 production was significantly increased in E-TB patients, and PBMC from both E-TB and CRTB patients secreted more IL-6 than HTR. IL-10 neutralization significantly increased TNF-alpha levels, whereas anti-TNF-alpha did not alter IL-10 induction significantly in PBMC from HTR and CRTB patients. Our findings suggest that CRTB patients have depression in both IFN-gamma and TNF-alpha reponses, which might play important roles during chronic M. tuberculosis infection.
Depression* ; Humans ; Immunotherapy ; Interferon-gamma* ; Interleukin-10 ; Interleukin-12 ; Interleukin-6 ; Interleukins ; Mycobacterium tuberculosis ; Tuberculin ; Tuberculosis* ; Tuberculosis, Multidrug-Resistant ; Tumor Necrosis Factor-alpha*

BACKGROUND: On performing umbilical cord blood (UCB) transplantation, faster engraftment may lead better clinical outcome. Because transplanted viable cell count in UCB is related to the engraftment, accurate evaluation of viability of CD34+cells in cryopreserved UCB has clinical implication. We examined the difference in viability of cells in cryopreserved UCB according to the duration of cryopreservation and different methods. METHODS: A total of 60 UCB samples which were cryopreserved for 1 to 4 years were used in this study. Viability of cryopreserved cells were examined with trypan blue exclusion assay, DNA contents analysis, caspase-3 activation test, intracellular esterase activity and Annexin-V/PI staining. RESULTS: After thawing the cryopreserved UCB, 89% of the total MNCs and 84% of CD34+cells were viable as identified by trypan blue exclusion assay. In the CD34+cell population, the cell death rate was found to be 47% by Annexin-V/PI staining and less than 5% by DNA contents analysis. However, cspase-3 activity failed to document apoptosis. The intracellular esterase activity test also showed a cell death rate of about 10~20% at 2, 4, and 6 hours after thawing. CONCLUSION: Viable cells in UCB should be measured by several compensatory techniques rather than a single method. Discordance among Annexin-V/PI staining versus trypan blue exclusion, DNA contents analysis, and the caspase-3 activation test or intracellular esterase activity should be clarified in order to apply these techniques for actual cord blood transplantation.
Apoptosis ; Caspase 3 ; Cell Count ; Cell Death ; Cryopreservation ; Diminazene ; DNA ; Fetal Blood ; Transplants ; Trypan Blue