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Introduction: Silver diamine fluoride (SDF) is employed as an adjunct cariostatic agent in the management of dental caries in high-risk population. Other than fluorides, chlorhexidine (CHX) is the most potent antimicrobial and efficacious agent against Streptococcus mutans. Aim: The aim of this study is to evaluate and differentiate the efficacy of 38% silver diamine fluoride, CHX varnish, and fluoride varnish on carious primary teeth. Materials and Methods: Ninety children having a count of ?1 carious lesion were recruited. Thirty-eighty percent silver diamine fluoride or fluoride varnish and CHX varnish were topically applied on the lesion. The primary outcome measured was the arrest of carious lesion (lesion rendered inactive as per the Nyvad criteria) after a follow-up of 14–21 days. Dental biofilm sample was obtained from each child and subsequently assessed for microbial composition by colony-forming unit method before and after treatment followed by protein analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis method. Results: Average proportion of arrested caries lesions in the SDF group was higher followed by CHX and fluoride varnish groups. Decreased total protein amount was found in SDF group. This proves that there is decrease in microbial load posttreatment in SDF group. Conclusion: Thirty-eight percent SDF is more effective than CHX varnish and fluoride varnish in arresting dentin carious lesions in young children.
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Background: Fixed orthodontic appliances are considered to jeopardize dental health due to the accumulation of oral microorganisms that may cause enamel demineralization. Oil pulling involves the use of edible vegetable oils as oral antibacterial agents. It is a practice of swishing oil in the mouth for oral and systemic health benefits. Aims and Objectives: To assess the effect of oil pulling therapy with virgin coconut oil on Streptococcus mutans count in patients undergoing orthodontic treatment. Methods: A total of thirty subjects were included in the study. They were divided in to 2 groups. Group A subjects were asked to swish coconut oil and Group B normal saline for a week. Streptococcus mutans colony forming units were estimated and compared. Results: A statistically significant reduction in S. mutans CFU was seen with Group A after oil pulling with coconut oil when compared to saline group (P = 0.0003. Conclusion: Edible oil-pulling therapy is natural, safe and has no side effects. Hence, it can be considered as a preventive therapy at home to maintain oral hygiene
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Background@#The intestinal microbiota of Mongolians and its composition is of great interest of researchers, a few studies have did in this fields. Maybe Mongolian encompass a uniquely wide range of environmental conditions, ethno geographical cohorts and traditional nomadic lifestyles.@*Goal@#We aimed to determine the amount of gut microbiota, including Lactobacillus and Bifidobacterium in the fecal samples of relative healthy Mongolian adults residing in various regions of Mongolia by conventional culture method and PCR. @*Material and Methods@#The study was performed population based cross sectional study in healthy volunteers. In this study, 256 relative healthy Mongolian adults with no history of gastrointestinal associated diseases were enrolled between July 2018 and April 2019. Each participants was asked to complete a questionnaire containing 164 questions about demographics, physical activity, dietary habits. Fecal samples were collected for Lactobacillus and Bifidobacterium analysis using culture method and determination of genus of Bifidobacterium sрp and Lactobacillus spp by PCR. ResultsParticipants had a mean age of 38.9±12.8 years. The mean values of Lactobacillus by culture method were 5.9±1.28 and 6.24±0.94 log10 CFU/ml (4.67х106 , 4.66х106 CFU/ml), respectively. The abundance of Lactobacillus had a positive correlation with grams for fiber and amount of bifidobacterium ((r= 0.495, р<0.001, r=0.288, p<0.05), respectively). Significant difference were observed between groups of milk frequency per day for amounts of lactobacillus. In adult intestinal tracts, B.Bifidum was the most common taxon 31 (29%) followed by B. angulatum 14 (13.1%), B. adolescentis 10 (9.3%), B. catenulatum group 10 (9.3%), B. longum 9 (8.4%). B. lactis, B. breve, B. dentium and B. gallicum were subdominant species. @*Conclusion: @#The mean amount of Bifidobacterium and Lactobacillus of all participants were 6.24±0.94 and 5.9±1.28 log10 CFU/ml (4.66*106 , 4.67*106 CFU/ml) respectively. The Lactobacillus abundance of healthy adults was higher in region of Khangai, East and West of Mongolian than other regions. The composition of lactobacillus altered with ageing. Significant correlations were found between fiber, fats, potato and amount of Lactobacillus. Keywords: Bifidobacterium, Colony forming unit, Gut microbiota, Lactobacillus
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Background and Objectives: Bacteria can cause allergic asthma and seasonal allergies, diseases which are increasingly prevalent in developing nations. Allergic asthma is currently affecting millions of people in Nepal. Therefore, the objective of this study was designed to measure the bacterial load in outdoor air. Materials and Methods: Airborne outdoor bacteria were assessed during the spring season using conventional methods to investigate the enumeration of airborne microorganisms. This was determined by sampling air using the ‘settle plate technique’. The air samples were collected during the spring season (February-March) from 10 different areas of Janakpur. Counts of airborne bacteria were measured as CFUs collected by gravity onto Nutrients Agar plates. Samples were taken periodically over a period of 2 months of February and March 2017. Results: A total of 7,404 bacterial colonies were counted on 30 Petri plates that were exposed for 1 hour. The maximum number of colonies of bacteria was 412. Similarly, the least number of bacterial colonies was 32. Higher numbers of CFUs were found in the petri plates which were exposed for 1 hour in comparison to the petri plates which were exposed for 30 minutes. According to the measurement, 36.6% of total CFUs of bacteria were collected during morning hours, 28.4% during day time and 35% during evening hours. Also, the highest numbers of colonies of bacteria were found in the petri plates that were exposed in ward number 7 and the least number of bacterial colonies were obtained in ward number 9. Conclusion: The bacteriological quality of air in janakpur was very poor. Very high microbial load was found in the outdoor air in Janakpur. The microbial count was found higher in morning than the noon and evening.
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Objective To understand effect of application of disposable shoe covers on the culture result of air-borne bacteria in intensive care unit (ICU).Methods According to Hygienic standards for disinfection in hospi-tals (GB 15982-2012)and Technical standards for disinfection in medical institutions (WS/T 367-2012),the col-ony forming units of bacteria in ICU air were detected for 15 consecutive days when disposable shoe covers were used and unused,bacterial culture results were analyzed.Results The number of patients,number of visitors, temperature,and humidity in ICU when disposable shoe covers were used and unused were not significantly differ-ent (all P >0.05).The total number of bacterial colonies in ICU air when using disposable shoe covers was (median ± interquartile range:[1.20±2.20]CFU/15 min.·Φ9 cm.plate),which was significantly higher than air when disposable shoe covers were not used(median ± interquartile range:[0.60±1.10]CFU/15 min.·Φ9 cm.plate), difference was statistically significant(P <0.05).The qualified rate of bacterial culture result of air when disposable shoe covers were not used was higher than that when disposable shoe covers were used (96.67% vs 86.67%,P <0.05).Conclusion The use of disposable shoe covers can not improve ICU air quality.
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Objective To understand effect of application of disposable shoe covers on the culture result of air-borne bacteria in intensive care unit (ICU).Methods According to Hygienic standards for disinfection in hospi-tals (GB 15982-2012)and Technical standards for disinfection in medical institutions (WS/T 367-2012),the col-ony forming units of bacteria in ICU air were detected for 15 consecutive days when disposable shoe covers were used and unused,bacterial culture results were analyzed.Results The number of patients,number of visitors, temperature,and humidity in ICU when disposable shoe covers were used and unused were not significantly differ-ent (all P >0.05).The total number of bacterial colonies in ICU air when using disposable shoe covers was (median ± interquartile range:[1.20±2.20]CFU/15 min.·Φ9 cm.plate),which was significantly higher than air when disposable shoe covers were not used(median ± interquartile range:[0.60±1.10]CFU/15 min.·Φ9 cm.plate), difference was statistically significant(P <0.05).The qualified rate of bacterial culture result of air when disposable shoe covers were not used was higher than that when disposable shoe covers were used (96.67% vs 86.67%,P <0.05).Conclusion The use of disposable shoe covers can not improve ICU air quality.
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@#BACKGROUND: In the recent years, mesenchymal stem cells have become increasingly utilized in regenerative medicine and tissue engineering applications because of their properties for self-renewal, differentiation and immunoregulation. The use of stem cells of various clinical applications is highly expected and the production of good quality stem cells is very critical for basic studies. In the bone marrow, hematopoietic and mesenchymal stem cells from an unique niche in which the oxygen tension is low. Hypoxia may have a role in maintaining stem cell fate, self renewal and multi-potency. We investigated whether low oxygen culture would be beneficial for hematopoietic stem cell and mesenchymalstemcell. MATERIAL: BMCs from 8-12 week aged, 15 mice were subjected to hypoxic conditioning by culture for 8-10 days in 20%, 3%, 1% oxygen. For culture 1x105cell/ml were seeded in colony forming assay and 2x106cell/ml were seeded in L-glutamin mediain chamber slide. We counted cell colonies under different hypoxic condiontins by Olympus IX71 fluorescence microscope. After cell culture in chamber slide, we stained cells by anti-CD90 and anti-CD105 then counted positive cells by Olympus IX71 fluorescence microscope. RESULTS: Compared to normoxic cells and hypoxic cells well morphologically differentiated and counted by Olympus IX71 microscope. More colonies were observed at 3%, 1% oxygen. Statistical significances were identified with granulocytes and macrophage colony (p<0.05) in hypoxic condition. More anti-CD90 and anti-CD105 markers were observed at 3% oxygen condition. Statistical significances were identified in 3% oxygen condition with cell markers(p<0.001). CONCLUSIONS: Our data suggests low physiological oxygen culture could improve the stemness of macrophage and granulocytes colony and improve the differentiation of mesenchymal cells. Long term culturewith additional cell markers will be necessary to confirm whether low physiological oxygen levels also improve genomic stability
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Objective To establish a reliable approach for quantification of colony forming unit(CFU)of Mycobac-terium tuberculosis (M.tb)by measuring optical density(OD).Methods M.tb suspension H37Ra was prepared using low-power ultrasonic or glass bead beating methods,and was two-fold serially diluted,OD at 600nm (OD600)of each dilution ratio was measured respectively,OD600 and dilution curve were analyzed to determine the optimum approach for preparing bacterial suspension,linear range of OD600,as well as linear relationship between OD600 and CFU.Results OD600 was 0.1 -0.6,linear regression analysis of OD600 and dilution ratio within linear range revealed that correlation coefficient (R2 )of glass bead beating and low-power ultrasonic methods were 0.98 and 1 .00 respectively,both presented a good correlation,low-power ultrasonic method was better than glass bead beat-ing method,bacterial suspension dispersed more evenly.Linear regression analysis results of OD600 and CFU val-ues showed that the regression equation of glass bead beating method and low-power ultrasonic method were CFU=2.35×107 ×OD600+4.42×105 and CFU=3.26×107 ×OD600+6.89×105 respectively.Conclusion Low-power ultrasonic method is a good method for preparation of M.tb suspension,combined the measurement of OD600 value, it can be a reliable and rapid method for quantitative analysis of M.tb.
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BACKGROUND: The number of CD34+ cells in a peripheral blood stem cell collection is the key factor in predicting successful treatment of hematologic malignancies. Korean Red Ginseng (KRG) (Panax ginseng C.A. Meyer) is the most popular medicinal herb in Korea. The objective of this study was to determine the effect of KRG on hematopoietic colony formation. METHODS: Bone marrow (BM) samples were obtained from 8 human donors after acquiring informed consent. BM mononuclear cells (MNCs) were isolated, and CD34+ cells were sorted using magnetic beads. The sorted CD34+ cells were incubated with or without total extract of KRG (50 microg/mL, 100 microg/mL) or Ginsenoside Rg1 (100 microg/mL), and the hematopoietic colony assay was performed using methylcellulose semisolid medium. The CD34+ cell counts were measured by a single platform assay using flow cytometry. RESULTS: The numbers of human BM-MNCs and CD34+ cells obtained after purification were variable among donors (5.6x10(7) and 1.3-48x10(7) and 8.9x10(4) and 1.8-80x10(4), respectively). The cells expanded 1,944 times after incubation for 12 d. Total extract of KRG added to the hematopoietic stem cell (HSC)-specific medium increased CD34+ cell counts 3.6 times compared to 2.6 times when using HSC medium alone. Total numbers of hematopoietic colonies in KRG medium were more than those observed in conventional medium, especially that of erythroid colonies such as burst forming unit-erythroid. CONCLUSION: Total extract of KRG facilitated CD34+ cell expansion and hematopoietic colony formation, especially of the erythroid lineage.
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Humans , Antigens, CD34 , Bone Marrow , Cell Count , Flow Cytometry , Hematologic Neoplasms , Hematopoietic Stem Cells , Informed Consent , Korea , Medicine, Korean Traditional , Methylcellulose , Panax , Plants, Medicinal , Stem Cells , Tissue DonorsABSTRACT
Aims: The aim of this study is to verify the disinfection of diode laser, following chemo-mechanical procedures against Enterococcus fecalis. Materials and Methods: Crowns of 30 extracted premolar teeth were sectioned at the cemento- enamel junction. The canals were shaped using step-back technique to K-file #40. The teeth were randomly assigned to three groups and placed into nutrient broth containing bacterial suspension of Enterococcus fecalis. Group A received no laser radiation. Specimens of group B and C were treated with diode laser (Sirona) with energy set at 1.5 and 3 W, respectively. After laser irradiation, the teeth were placed in vials, which contained 2 mL of the nutrient broth. The vials were incubated at 37°C for 24 h. Grown colonies were identified by standard methods. Statistical Analysis Used: Statistical analysis used was the nonparametric Kruskal-Wallis test, with comparison using the Bonferroni methods of means. Results: Higher mean CFU/mL is recorded in Group A (without laser disinfection) followed by Group B (with 1.5 W laser disinfection) and Group C (with 3 W laser disinfection), respectively. The difference in CFU/mL between the three groups is found to be statistically significant ( P < 0.001). Conclusions: The results of this research show that the 980 nm diode laser can eliminate bacteria that has immigrated into dentin, thus being able to increase the success rate in endodontic therapy.
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BACKGROUND: The application of mesenchymal stem cells derived from umbilical cord cultured in serum-containing medium has some obstacles. OBJECTIVE: To establish serum free medium system for primary culture of umbilical cord-derived mesenchymal stem cells. METHODS: Wharton’s Jel y was isolated from umbilical cord, minced, 1-3 mm3, and subsequently incubated in either serum containing medium or serum free medium. Some cells were harvested on days 11, 14 and 17 for some detection. Based on the minimal criteria established by the International Society for Cel ular Therapy in 2006, mesenchymal stem cells were assayed with colony forming unit-fibroblast. RESULTS AND CONCLUSION: The mesenchymal stem cells grew rapidly in serum free medium condition compared with serum containing medium. On day 11, the number of colonies was larger in serum free medium condition than that in serum containing medium. Thus, serum free medium could maintain properties of mesenchymal stem cells and provide possibility a credible alternative to serum containing medium for primary culture of umbilical cord-derived mesenchymal stem cells.
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A study on physico-chemical parameters and pathogenic bacterial community was carried out at the coastal waters of Pulau Tuba island, Langkawi. The physico-chemical parameters such as temperature (27.43-28.88oC), dissolved oxygen (3.79-6.49 mg l-1), pH (7.72-8.20), salinity (33.10-33.96 ppt), total dissolved solids (32.27-32.77 g l-1) and specific conductivity (49.83-51.63 mS cm-1) were observed. Station 3 and station 4 showed highest amount of nitrates (26.93 and 14.61 µg at N l-1) than station 1 (2.04 µg at N l-1) and station 2 (4.18 µg at N l-1). The highest concentration (12.4± µg l-1) of chlorophyll a was observed in station 4 in October 2005. High phosphorus content (561mg P l-1) was found in the station 2. Thirteen bacterial isolates were successfully identified using API 20E system. The highest amount of bacteria was observed at Station 4 (3400 CFU ml-1) and the lowest number was at Station 2 (890 CFU ml-1). Out of identified 13 Gram-negative bacterial isolates dominant species were Aeromonas hydrophila, Klebsiella oxytoca, Pseudomonas baumannii, Vibrio vulnificus, Proteus mirabilis, Providencia alcalifaciens and Serratia liquefaciens. Apart from this, oil biodegrading Pseudomonas putida were also identified. The study reveals the existing status of water quality is still conducive and the reasonably diverse with Gram-negative bacteria along the Pulau Tuba Langkawi.
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PURPOSE: Endothelial progenitor cells (EPCs), which mediates neovascularization of uterine endometrium may be involved in the neovascularization in the utero-placental circulation. Low numbers of endothelial progenitor colony-forming unit (CFU) in culture are predictive biomarker of vascular disease. The aim of the present study was to evaluate whether the number of CFU in preeclampsia differed from that in normal pregnancy. MATERIALS AND METHODS: Women with singleton normal (n=26) or preeclamptic (n=20) pregnancies were studied during the third trimester. The number of EPCs was quantified by CFU methodology. Plasma levels of angiogenic factors, vascular endothelial growth factor (VEGF), soluble fms-like tyrosine kinase-1 (sFlt-1), and placental growth factor (PlGF) were determined by enzyme-linked immunoassay. RESULTS: CFU numbers were significantly decreased in the preeclamptic patients compared with the controls (median, 3; range 1-12 vs. 31; 3-81 CFU/well, P<0.001). A majority of the cells comprising individual colonies were positive for endothelial characteristics (Ulex europaeus lectin staining and acetylated low-density lipoprotein uptake). Plasma levels of the sFlt-1 were highly elevated (P<0.001) in patient with preeclampsia compared to controls, whereas PlGF were highly reduced (P=0.004), but these factors did not associate with CFU numbers. CONCLUSION: Our results suggest that reduced numbers of CFU obtained from maternal peripheral blood may contribute to the development of preeclampsia.
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Female , Humans , Pregnancy , Angiogenesis Inducing Agents , Endometrium , Lipoproteins , Plasma , Pre-Eclampsia , Pregnancy Trimester, Third , Stem Cells , Tyrosine , Vascular Diseases , Vascular Endothelial Growth Factor AABSTRACT
Human cytomegalovirus (HCMV) late mRNA expression in megakaryoblast and in turn the pathogenesis of idiopathic thrombocytopenic purpura (ITP) patients with HCMV infection, and effectiveness of ganciclovir were investigated. Colony forming unit-megakaryocytes (CFU-MK) of 46 ITP patients with HCMV infection were incubated from patients' bone marrow mononuclear cells (MNC). Reverse transcriptase-polymerase chain reaction (RT-PCR) was subsequently used for CFU-MK for HCMV-late mRNA detection. Ganciclovir therapy was given to both HCMV-late mRNA positive and negative groups for comparison of therapeutic effectiveness. The results in 19 of 46 CFU-MK culture cells specimens with positive HCMV-DNA by PCR or positive CMV-IgM by enzyme linked immunosorbent assay (ELISA) in the correspondent serum of peripheral blood were positive for HCMV-late mRNA. Sixteen out of 19 patients with positive HCMV-late mRNACFU-MK had a positive response to ganciclovir. Amongst 27 patients with negative HCMV-late mRNA CFU-MK, only 4 positive responders to ganciclovir therapy were observed. Curative effectiveness of ganciclovir in HCMV-late mRNA positive group was significantly higher than that in HCMV-late mRNA negative group (P＜0.01). It was suggested that HCMV could directly infect CFU-MK, which might be one of the mechanisms responsible for HCMV related ITP. The ganciclovir is an effective therapy in resulting in the increases in thrombocyte in the ITP patients whose HCMV- late mRNA was positive in their CFU-MK.
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flow cytometry.A strong linear relationship was observed in the standard curve of real-time PCR of each bacteria. Conclusion These three non-culture methods can be used in the quantitative analysis of oral microorganisms.Real-time PCR and laser scanning confocal microscopy are better than the traditional culture-based CFU count,and real-time PCR is the most sensitive method.
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PURPOSE: Because of the unavailability of marrow transplantation, umbilical cord blood (CB) is increasingly being used. We evaluated the potential of ex vivo expansion and clonality in CD34+ cells separated from cord blood source and mobilized peripheral blood (PB) in a serum-free media. METHODS: The CD34+ cells, selected from CB and mobilized PB, were expanded with hematopoietic growth factors. They were then cultured for burst-forming units of erythrocytes (BFU-E), colony-forming units of granulocytes and monocytes (CFU-GM) and colony-forming units of megakaryocytes (CFU-Mk) at culture days 0, day 4, day 7, and day 14 with various growth factors. RESULTS: The CB-selected CD34+ cells showed significantly higher total cell expansion than those from the PB at day 7 (2 fold increase than PB). The CB-selected CD34+ cells produced more BFU-E colonies than did the PB on culture at days 7 and at day 14. Also, the CB-selected CD34+ cells produced more CFU-Mk colonies than did the PB on culture at day 4 and at day 7. CONCLUSION: The ex vivo expansion of the CB cells may be promising in producing total cellular expansion, CFU-Mk and BFU-E compared with PB for 7 to 14 days. The growth factors combination including megakaryocyte growth and development, flt3-ligand and interleukin-3 showed more expansion in the view of total cells and clonal maintenance compared with less combination.
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Bone Marrow , Culture Media, Serum-Free , Erythrocytes , Erythroid Precursor Cells , Fetal Blood , Granulocytes , Growth and Development , Intercellular Signaling Peptides and Proteins , Interleukin-3 , Megakaryocytes , Monocytes , Stem CellsABSTRACT
PURPOSE: Many studies for hematopoietic stem cell have investigated CD133, instead of CD34, as a new surrogate stem cell marker. Counterflow centrifugal elutriation (CCE) is a physical separation of a homogeneous cell population through cell sedimentation characteristics. We evaluated the stem cell distribution and hematopoietic function from cord blood (CB) and bone marrow (BM) through CCE. METHODS: We obtained total nucleated cells from CB and BM, and separated the cell fractions according to media infusion flow rates (17 mL/min (FR 17), 24 mL/min (FR 24), 29 mL/min (FR 29), and rotor off (R/O) ) by CCE. We analyzed the proportion of CD34+ and CD133+ cells in each fraction, and performed methylcellulose-based colony assay. RESULTS: In CB, the cell recovery rates after CCE were 5.9+/-4.3% in FR 17, 4.2+/-2.1% in FR 24, 19.4+/-11.9% in FR 29, and 61.9+/-11.7% in R/O. In BM, they were 14.9+/-8.2% in FR 17, 17.4+/-13.4% in FR 24, 23.6+/-6.11% in FR 29, and 27.1+/-8.9% in R/O. The distributions of CD133+ and CD34+ cells in CB were more abundant in R/O (2.91%, 1.85%) than in other fractions. In BM, CD133+ and CD34+ cell rates in R/O (5.40%, 2.75%) were similar with those in unmanipulated BM (5.48%, 2.78%). In both CB and BM, there was more CFU-GM and BFU-E in R/O than in other fractions. CONCLUSION: We suggested that the distribution of CD34+ and CD133+ cells might be different between CB and BM. However, the R/O containing relatively large cells could have an effective clonogenicity compared with the unmanipulated sample in both CB and BM.
Subject(s)
Bone Marrow , Erythroid Precursor Cells , Fetal Blood , Granulocyte-Macrophage Progenitor Cells , Hematopoietic Stem Cells , Stem CellsABSTRACT
A limpeza é, inegavelmente, o núcleo central de todas as atividades relacionadas ao reprocessamento de artigos médico-hospitalares. A lavadora desinfectadora é uma nova tecnologia, que trouxe grandes vantagens, como padronização dos procedimentos de limpeza, documentação do processo e diminuição do risco ocupacional de ordem biológica. Atualmente, existem equipamentos que disponibilizam programas com diferentes tempo e temperatura. Para subsidiar a escolha de programas, propôs-se nesta pesquisa investigar o desempenho das lavadoras desinfectadoras nos distintos programas (Norma Alemã, BGA 94ºC/10 minutos; Norma da Grã-Bretanha, DHSS/HTM 90ºC/1segundo; Norma da Grã-Bretanha, DHSS/HTM 82ºC/2minutos; Norma da Holanda, RIVM 90ºC/5 minutos; Norma da Suécia, SPRI/SIS 85ºC/1minuto; Norma da Suécia, SPRI/SIS 85ºC/3 minutos; ciclo com temperatura 70ºC e tempo 30 minutos para Pasteurização), avaliando-se o desempenho dos testes de limpeza e termodesinfecção em equipamento validado. Conforme as recomendações das Normas ISO 15883-1/1999 e HTM2030 (NHS States,1997), para avaliação do desempenho da limpeza foram utilizados três testes: Soil Test, Biotrace Pro-tect e Test Kit Proteína. Como resultado dos testes do desempenho da limpeza, constatou-se resíduo de sujidade após avaliação com Soil-Test em 1,3% dos instrumentais, do total de 313 avaliados (cinco instrumentais de complexidade crítica, dois deles não desmontáveis-Kerrison e Goiva). Na avaliação de resíduo de proteína com teste Biotrace Pro-tect constatou-se que, dos 65 instrumentais avaliados, 60 (92%) apresentaram resultado satisfatório. Os cinco instrumentais (8%) que apresentaram resíduo de sujidade com Soil-Test, também apresentaram, após avaliação com teste Biotrace Pro-tect, resíduo de proteína. Na avaliação realizada com o Test Kit Proteína, 141(100%) instrumentais apresentaram ausência de proteína. Para avaliação do desempenho da termodesinfecção, ) os instrumentais escolhidos para experimento foram intencionalmente contaminados com sangue humano e em seguida submetidos aos processos de termodesinfecção em diferentes programas. A contagem de UFC dos microrganismos viáveis foi feita antes e após a termodesinfecção, partindo-se da contaminação inicial de 107 e 108 UFC. Quanto aos resultados destes testes, de modo uniforme, todos os ciclos apresentaram desempenho satisfatório de <102 UFC, resultado esse entendido como ausência de crescimento microbiano, considerando-se a diluição empregada. Nos cálculos dos valores da Letalidade Mínima e DAL - Nível de Segurança de Desinfecção, os protocolos aprovados foram: Norma Alemã, BGA 94ºC/10 minutos; Norma da Grã-Bretanha, DHSS/HTM 90ºC/1 segundo; Norma da Holanda, RIVM 90ºC / 5 minutos; Norma da Suécia, SPRI/SIS 85ºC / 1 minuto; Norma da Suécia, SPRI/SIS 85ºC/ 3 minutos. Os protocolos que não alcançaram os valores preconizados da Letalidade Mínima de 10 minutos e DAL =10-2 após validação foram: Norma da Grã-Bretanha, DHSS/HTM 82ºC / 2 minutos; Temperatura 70ºC e Tempo de 30 minutos para Pasteurização. Como conclusão, a presente pesquisa evidenciou desempenhos satisfatórios das Máquinas Lavadoras Desinfectadoras tanto na limpeza mecânica quanto na desinfecção, em todos os programas testados, com diferentes tempos e temperaturas apesar do DAL e A0 de alguns programas terem sido reprovados. Evidenciou que em instrumentais de conformação complexa, especialmente quando não desmontáveis, a remoção completa dos resíduos não ocorre nas máquinas, sugerindo que a limpeza destes seja manual, utilizando-se artefatos adequados.
Cleaning is undeniably the main focus of all activities related to the reprocessing of medical and hospital supplies. The washer-disinfector machine represents a new technology that has brought great advantages as the standardization of the cleaning procedures, the process registering and the reduction of occupational hazards coming from biological sources. Nowadays, there are equipments with programs presenting different time and temperature. In order to subside the choice of programs, this essay aims to investigate the performance of washer-disinfectors in the following different programs (German Federal Health Authority BGA, 94ºC/10 minutes; British Standard DHSS/HTM, 90º C/1 second; British Standard DHSS/HTM, 82º C/2 minutes; The Netherlands RIVM, 90º C/5 minutes; Swedish Standards Institute SPRI/SIS, 85ºC/1 minute; Swedish Standard Institute SPRI/SIS, 85ºC/3minutes; cycle temperature 70ºC and pasteurization time 30 minutes), evaluating the performance of cleaning and thermo disinfection tests in validated equipment. According to recommendations in ISO 15883-1/1999 and HTM 2030 (NHS States, 1997) for evaluating the cleaning and disinfection processes, three tests were performed: Soil Test, Biotrace Pro-tect and Test Kit Protein. As a result of the cleaning tests performance, soiling was found after evaluation with Soil Test performed in 1,3% of the instruments (5 of them presenting critical complexity, 2 of them not dismountable - Kerrison and Rongeur). In 65 instruments evaluated by Biotrace Pro-tect, 60 instruments (92%) presented good results. Five instruments (8%) presented soiling after evaluation with Soil Test, and after Biotrace Pro-tect evaluation presented protein contamination. After the evaluation performed with Test Kit Protein no instrument presented protein contamination (141 instruments, 100%). To evaluate the thermo disinfection performance the instruments chosen for the test were intentionally contaminated with human blood and after that were under thermo disinfection processes in different programs. The counting of possible present microorganisms was done before and after the thermo disinfection process, starting from an initial contamination of 107 and 108 UFC. Concerning the results of these tests, in general all cycles presented a good performance (<102 UFC), which was understood as the absence of microbiological growth, considering the dilution given. To calculate the Minimal Lethality and DAL - Disinfection Assurance Level, the approved protocols were: German Federal Health Authority BGA, 94ºC/10 minutes; British Standard DHSS/HTM, 90ºC/1 second; The Netherlands RIVM, 90ºC/5 minutes; Swedish Standards Institute SPRI/SIS, 85ºC/1 minute and Swedish Standards Institute SPRI/SIS, 85ºC/3 minutes. The protocols that did not reach the professed values for Minimal Lethality of 10 minutes and DAL = 10 -2 after validation were: British Standard DHSS/HTM, 82ºC/2 minutes; temperature 70ºC and pasteurization time 30 minutes. Concluding, this study evidenced the good performance of the washer-disinfector machines for mechanic cleaning as for disinfection in all tested programs with different time and temperature. It also demonstrated that for instruments with complex shape, specially the not dismountable ones, the complete soiling remove do not occur on the machines and suggests that this cleaning requires handicraft employing adequate material.
Subject(s)
Operating Room Nursing , Housekeeping, Hospital , Surgical Equipment , Employee Performance AppraisalABSTRACT
BACKGROUND: The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. METHOD: The PPD positive subjects were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis H37Ra for 4 days by rotating the culture in a 37degrees C, 5% CO2 incubator. In some experiments, methylprednisolone- or pentoxifylline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The delta log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. RESULTS: 1. A trend was noted toward the improved killing of mycobacteria in PPD+subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifylline adversely affected the killing in the PPD+subjects, umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis H37Ra by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis H37Ra. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the delta logKR continuously decreased in a 3 and 4 days of whole blood culture. CONCLUSION: The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.
Subject(s)
Humans , Lung NeoplasmsABSTRACT
BACKGROUND: Rotor off (R/O) fraction obtained by counterflow centrifugal elutriation (CCE) contains small number of T cells and many hematopoietic stem cells. Since megakaryocytes and its progenitors are larger than other cells in bone marrow, it may be easier to be separated by CCE and newly applied to hematopoietic stem cell transplantation (HSCT) for early megakaryocytes reconstitution. METHODS: The marrow cells of BALB/c mice in each group (17, 25, 28mL/min, and R/O fraction) were cultured for quantifying CFU-MK and measured after 10 days. BALB/c mice were lethally irradiated and transplanted with R/O cells. The dosages of transplanted cells were 5x104 in Group A, 5x105 in Group B, and 5x106 in Group C. The platelet counts in peripheral blood were measured up to post-transplant day 14. RESULTS: After CCE, recovery rate of the loaded cells was 82.2% and the R/O fraction was 35.9%. Most CFU-MK were formed in R/O fraction, and Group C showed the fastest recovery. Group A couldn't reach the level of 100x109/L until post-transplant day 14, and Group B showed slower recovery compared to Group C. All 5 mice survived in Group C, but 2 out of 5 mice survived in Group A and B. CONCLUSIONS: R/O fraction contains higher number of megakaryocyte progenitors, and CCE could be an effective method for separating megakaryocyte progenitors essential for reconstituting platelet after HSCT. The cell dose of 5x106 was required for the effective recovery of platelet and the survival of BALB/c mice in syngeneic bone marrow transplantation with R/O cells.