Isolation and characterization of bacteria residing in the oral, gut, and fecal samples of different pheasant species / Isolamento e caracterização de bactérias residentes nas amostras orais, intestinais e fecais de diferentes espécies de faisão

Abstract There is a paucity of research conducted on microbial prevalence in pheasants. The microbiota of captive birds has zoonotic significance and must be characterize. Present study is therefore planned to assess the microbiota from oral, fecal and gut content of captive avian species. It will be helpful in characterization of harmful microbes. Different samples taken from oral, gut and feces of ring-necked pheasants (Phasianus colchicus), green pheasants (Phasianus versicolor), golden pheasant (Chrysolophus pictus) and silver pheasant (Lophura nycthemera). Samples were collected, diluted, and inoculated onto different agar plates (MacConkey, SS agar, MSA and nutrient agar) for cultivation of bacterial species. Colonies of E.coli, Staphylococcus spp. Brachyspira spp. and Campylobacter spp were observed based on colony morphology. Colony forming unit showed E. coli as frequently found bacteria in fecal, oral and gut contents of all the above pheasants. The overall significance difference was found among bacterial species of golden pheasants, green pheasant, ring-necked pheasant, and silver pheasants. It was concluded that E.coli is predominant isolated from heathy pheasants followed by Campylobacter, Staphylococcus and Brachyspira.

Resumo Há uma escassez de pesquisas realizadas sobre a prevalência microbiana em faisões. A microbiota de aves em cativeiro tem significado zoonótico e deve ser caracterizada. O presente estudo está, portanto, planejado para avaliar a microbiota do conteúdo oral, fecal e intestinal de espécies aviárias em cativeiro. Será útil na caracterização de micróbios nocivos. Diferentes amostras retiradas da boca, intestino e fezes de faisões de pescoço redondo (Phasianus colchicus), faisões verdes (Phasianus versicolor), faisões dourados (Chrysolophus pictus) e faisão prateado (Lophura nycthemera). As amostras foram coletadas, diluídas e inoculadas em diferentes placas de ágar (MacConkey, ágar SS, MSA e ágar nutriente) para o cultivo de espécies bacterianas. Colônias de E. coli, Staphylococcus spp., Brachyspira spp. e Campylobacter spp foram observados com base na morfologia da colônia. A unidade formadora de colônia mostrou E. coli como bactéria frequentemente encontrada no conteúdo fecal, oral e intestinal de todos os faisões acima. A diferença de significância geral foi encontrada entre as espécies bacterianas de faisões dourados, faisões verdes, faisões de pescoço anelado e faisões prateados. Verificou-se que a E.coli é predominantemente isolada de faisões saudáveis, seguida por Campylobacter, Staphylococcus e Brachyspira.

Isolation and characterization of bacteria residing in the oral, gut, and fecal samples of different pheasant species / Isolamento e caracterização de bactérias residentes nas amostras orais, intestinais e fecais de diferentes espécies de faisão

There is a paucity of research conducted on microbial prevalence in pheasants. The microbiota of captive birds has zoonotic significance and must be characterize. Present study is therefore planned to assess the microbiota from oral, fecal and gut content of captive avian species. It will be helpful in characterization of harmful microbes. Different samples taken from oral, gut and feces of ring-necked pheasants (Phasianus colchicus), green pheasants (Phasianus versicolor), golden pheasant (Chrysolophus pictus) and silver pheasant (Lophura nycthemera). Samples were collected, diluted, and inoculated onto different agar plates (MacConkey, SS agar, MSA and nutrient agar) for cultivation of bacterial species. Colonies of E.coli, Staphylococcus spp. Brachyspira spp. and Campylobacter spp were observed based on colony morphology. Colony forming unit showed E. coli as frequently found bacteria in fecal, oral and gut contents of all the above pheasants. The overall significance difference was found among bacterial species of golden pheasants, green pheasant, ring-necked pheasant, and silver pheasants. It was concluded that E.coli is predominant isolated from heathy pheasants followed by Campylobacter, Staphylococcus and Brachyspira.

Há uma escassez de pesquisas realizadas sobre a prevalência microbiana em faisões. A microbiota de aves em cativeiro tem significado zoonótico e deve ser caracterizada. O presente estudo está, portanto, planejado para avaliar a microbiota do conteúdo oral, fecal e intestinal de espécies aviárias em cativeiro. Será útil na caracterização de micróbios nocivos. Diferentes amostras retiradas da boca, intestino e fezes de faisões de pescoço redondo (Phasianus colchicus), faisões verdes (Phasianus versicolor), faisões dourados (Chrysolophus pictus) e faisão prateado (Lophura nycthemera). As amostras foram coletadas, diluídas e inoculadas em diferentes placas de ágar (MacConkey, ágar SS, MSA e ágar nutriente) para o cultivo de espécies bacterianas. Colônias de E. coli, Staphylococcus spp., Brachyspira spp. e Campylobacter spp foram observados com base na morfologia da colônia. A unidade formadora de colônia mostrou E. coli como bactéria frequentemente encontrada no conteúdo fecal, oral e intestinal de todos os faisões acima. A diferença de significância geral foi encontrada entre as espécies bacterianas de faisões dourados, faisões verdes, faisões de pescoço anelado e faisões prateados. Verificou-se que a E.coli é predominantemente isolada de faisões saudáveis, seguida por Campylobacter, Staphylococcus e Brachyspira.

Isolation and characterization of bacteria residing in the oral, gut, and fecal samples of different pheasant species

Abstract There is a paucity of research conducted on microbial prevalence in pheasants. The microbiota of captive birds has zoonotic significance and must be characterize. Present study is therefore planned to assess the microbiota from oral, fecal and gut content of captive avian species. It will be helpful in characterization of harmful microbes. Different samples taken from oral, gut and feces of ring-necked pheasants (Phasianus colchicus), green pheasants (Phasianus versicolor), golden pheasant (Chrysolophus pictus) and silver pheasant (Lophura nycthemera). Samples were collected, diluted, and inoculated onto different agar plates (MacConkey, SS agar, MSA and nutrient agar) for cultivation of bacterial species. Colonies of E.coli, Staphylococcus spp. Brachyspira spp. and Campylobacter spp were observed based on colony morphology. Colony forming unit showed E. coli as frequently found bacteria in fecal, oral and gut contents of all the above pheasants. The overall significance difference was found among bacterial species of golden pheasants, green pheasant, ring-necked pheasant, and silver pheasants. It was concluded that E.coli is predominant isolated from heathy pheasants followed by Campylobacter, Staphylococcus and Brachyspira.

Resumo Há uma escassez de pesquisas realizadas sobre a prevalência microbiana em faisões. A microbiota de aves em cativeiro tem significado zoonótico e deve ser caracterizada. O presente estudo está, portanto, planejado para avaliar a microbiota do conteúdo oral, fecal e intestinal de espécies aviárias em cativeiro. Será útil na caracterização de micróbios nocivos. Diferentes amostras retiradas da boca, intestino e fezes de faisões de pescoço redondo (Phasianus colchicus), faisões verdes (Phasianus versicolor), faisões dourados (Chrysolophus pictus) e faisão prateado (Lophura nycthemera). As amostras foram coletadas, diluídas e inoculadas em diferentes placas de ágar (MacConkey, ágar SS, MSA e ágar nutriente) para o cultivo de espécies bacterianas. Colônias de E. coli, Staphylococcus spp., Brachyspira spp. e Campylobacter spp foram observados com base na morfologia da colônia. A unidade formadora de colônia mostrou E. coli como bactéria frequentemente encontrada no conteúdo fecal, oral e intestinal de todos os faisões acima. A diferença de significância geral foi encontrada entre as espécies bacterianas de faisões dourados, faisões verdes, faisões de pescoço anelado e faisões prateados. Verificou-se que a E.coli é predominantemente isolada de faisões saudáveis, seguida por Campylobacter, Staphylococcus e Brachyspira.

Novel compound FLZ alleviates rotenone-induced PD mouse model by suppressing TLR4/MyD88/NF-

Parkinson's disease (PD) is the second most common neurodegenerative disease, but none of the current treatments for PD can halt the progress of the disease due to the limited understanding of the pathogenesis. In PD development, the communication between the brain and the gastrointestinal system influenced by gut microbiota is known as microbiota-gut-brain axis. However, the explicit mechanisms of microbiota dysbiosis in PD development have not been well elucidated yet. FLZ, a novel squamosamide derivative, has been proved to be effective in many PD models and is undergoing the phase I clinical trial to treat PD in China. Moreover, our previous pharmacokinetic study revealed that gut microbiota could regulate the absorption of FLZ

Comparison of Sysmex UF-5000 with urine culture for the diagnosis of urinary tract infections / Эрүүл мэндийн лаборатори

Introduction@#Urinary tract infections (UTI) are at the second place in the frequency of all causes of infection after respiratory ones. The UTI requires appropriate antibiotic treatment. 85% of UTI predictive antibiotic treatment without confirmation by bacteriological analysis. This is one of the major causes of drug resistance, especially in K.coli. Urine bacteriological tests do not show bacterial culture in all cases where the number of bacteria in the urine exceeds the reference level. Therefore, there was a need to establish criteria for urine bacteriology test based on the results of urine sediment analysis.</br> In 20I7, a new fully automated Sysmex UF-5000 urine sediment analyzer was installed in the laboratory department of Medipas Hospital. The features of this analyzer include counting the number of bacteria in the urine, distinguishing between gram-positive and negative, homogeneous and mixed forms, and counting the formed elements in the urine. This feature made it possible to compare the number of bacteria and leukocytes in the urine with the results of urine bacteriology tests.@*Goal@#Determine the relationship between the number of white blood cells and bacteria in the urine measured by the Sysmex UF-5000 urine sediment analyzer and the results of the urinary bacteriological test.@*Objectives@#Compare the number of urine bacteriaand leukocyte measured by using the Sysmex UF-5000 urine sediment analyzer with the urine bacterial culture, and calculate the correlation.@*Materials and methods@#The study is analytic cross-sectional study, analyzed the results of a total of 159 people who analyzed a urinalysis and urine bacteriological test at the Medipas Hospital Laboratory in 2017-2019 years.Urine samples were collected in a 100 ml, disposable sterile container in accordance with the instructions for taking urine midstream.Urine analysis was performed within 2 hours of sampling with a fully automatic urine sediment analyzer Sysmex UF-5000 Japan. Urine bacteriological analysis was performed on a lul sterile loop of urine specimens, inoculated into 5% blood agar from Hungary's BioLab, Sabouraud agar, and Chromogen agar from Biomerieux France, and incubated for 24 hours in an incubator at 37°C. Bacterial identification and antibiotic susceptibility tests were analyzed using the "Vitek-2" analyzer from the manufacturer Biomerieux France. Bacterial and leukocyte counts data measured by the Sysmex UF-5000 analyzer and urinary bacteriological analysis data were performed using SPSS23 software.@*Results@#A total of 159 urine samples were tested for bacteriological analysis, of which 81 (50.9%) were bacteria over 105 CFU/ml or urine positive culture UTIs, 78 (49.1%) were nonsignificant bactcruria and urine negative culture.The average number of bacteria measured in the urine of 81 samples with urine positive culture above 105 CFU/ ml was 46491/ul (1168-100000 BACT/ul). </br> The average number of bacteria measured by the urine sediment analyzer of 78 samples with urine negative culture was 2645 BACT/ ul (2-57280 BACT/ul). To calculate more accurately estimate the average number of bacteria in 81 urine specimens with positive culture, the average number of bacteria in 17 (21%) samples was 4753 BACT/ul, measured in relatively low bacteria numbers of 1168-9450BACT/ul. The average leukocyte number in the urine of 81 samples with positive culture was 472.2 WBC/ul, and the average leukocyte number in the urine of 78 samples with negative culture was 87.7 WBC/ul.There is a strong correlation between the number of bacteria measured by the urine sediment analyzer and urine bacterial positive culture, which is 0.8 or statistically significant (p<0.001).The correlation coefficient of the number leukocytes measured by the urine sediment analyzer with in the urine positive cultureof bacteriological tests was 0.6 or moderately of statistically significant (p=0.005).There is a statistically significant relationship (p=0.001) between the number of bacteria in the bacterial positive culture population and the number of leukocytes.@*Discussion@#Of the 81 cases of urine bacterial positive culture, 78 (96%) were female, indicating a high prevalence of UTI among women. According to the results of the Fabio Manon's study, the number of leukocytes in the urine is 160-340 WBC/uL and the number of bacteria is 15000-30000 BACT/u,L in the case of UTI, which is approximate results compared to the our study results.Based on the results of the urine sediment analysis, indications for a urine bacteriological test should be made.</br> Based on the results of urinary bacteriological tests, the choice of antibiotic treatment is the best treatment for urinary tract infections and a way to prevent of antibiotic resistance to UTI.@*Conclusions@#The number of bacteria measured by a Sysmex UF-5000 urine sediment analyzer is directly related to the bacterial culture urine bacteriological test. If the number of bacteria in the urine is measured above 4753 BACT/ul, it can be considered as an indication for urine bacteriological analysis. Although the number of leukocytes in the urine measured by the Sysmex UF-5000 urine analyzer is moderately correlated with bacterial culture in urine bactcriologucal tests, it is a key indicator of the degree of inflammation of the urinary tract.

Probiotics modulate the microbiota-gut-brain axis and improve memory deficits in aged SAMP8 mice

ProBiotic-4 is a probiotic preparation composed of , , , and . This study aims to investigate the effects of ProBiotic-4 on the microbiota-gut-brain axis and cognitive deficits, and to explore the underlying molecular mechanism using senescence-accelerated mouse prone 8 (SAMP8) mice. ProBiotic-4 was orally administered to 9-month-old SAMP8 mice for 12 weeks. We observed that ProBiotic-4 significantly improved the memory deficits, cerebral neuronal and synaptic injuries, glial activation, and microbiota composition in the feces and brains of aged SAMP8 mice. ProBiotic-4 substantially attenuated aging-related disruption of the intestinal barrier and blood-brain barrier, decreased interleukin-6 and tumor necrosis factor- at both mRNA and protein levels, reduced plasma and cerebral lipopolysaccharide (LPS) concentration, toll-like receptor 4 (TLR4) expression, and nuclear factor-B (NF-B) nuclear translocation in the brain. In addition, not only did ProBiotic-4 significantly decreased the levels of -H2AX, 8-hydroxydesoxyguanosine, and retinoic-acid-inducible gene-I (RIG-I), it also abrogated RIG-I multimerization in the brain. These findings suggest that targeting gut microbiota with probiotics may have a therapeutic potential for the deficits of the microbiota-gut-brain axis and cognitive function in aging, and that its mechanism is associated with inhibition of both TLR4-and RIG-I-mediated NF-B signaling pathway and inflammatory responses.

Microbiological Profile of Air and Surface Contamination in Operation Theatres and Intensive Care Units of a Teaching Hospital in Rural North India

Background: Harbouring of potential pathogens in operationtheatres (OTs) and intensive care units (ICUs) of hospital is amajor cause of patient’s morbidity and mortality. Environmentalmonitoring by the microbiological testing of surfaces andequipments is useful to detect changing trends of types andcounts of microbial flora. High level of microbial contaminationindicates the needs for periodic surveillance aimed at earlydetection of bacterial contamination levels and prevention ofhospital acquired infections.Aim: The aims of the study were to count CFU (colony formingunit) rate of indoor air, to identify bacterial colonization ofsurface and equipments isolated from Operation theatres, ICUsand Labour room of a teaching hospital in district Kangra,Himachal Pradesh.Methods: This retrospective study, analyzing themicrobiological surveillance data from OTs over a period of 2years from January2017 to December2018 was conducted at atertiary care hospital. Air sampling of 8 OT’s, 4 ICU’s and 1 LRwere done by settle plate method. Swabs were taken fromdifferent sites, equipments and bacterial species were isolatedand identified from them as per standard guidelines.Result: A total of 105 air samples were collected for 2 yearfrom 8 OT’s, 4 ICU’s and 1 LR. The bacterial CFU/m3 /mincounts of air from all OTs ranged from Superspeciality OTSshowed less bacterial CFU rate of air (0-5 CFU/m3) followed byOpthalmology OT (5-8 CFU/m3) and highest in Gynae (30-46CFU/m3). CCU showed less bacterial CFU rate (10-15CFU/m3) followed by Surgery ICU (28-35 CFU/m3) and highestin PICU (38-42 CFU/m3), Labour room showed 42-51 CFU/m3.Bacterial species were isolated from 43.85 % out of total 157swab samples taken from all OTs and ICUs. The mostcommon isolate was Bacillus species 46% followed by CONS(22%). Pathogenic organisms isolated were 10% Gramnegative bacilli which included 3% Non-Fermenters, thecommon isolate was Klebsiella spp. amongst gram negatives.

Correlation between optical density and colony forming units of Mycobac-terium tuberculosis suspension / 中国感染控制杂志

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.

Effect of cryopreservation on umbilical blood cells and its mechanism / 中南大学学报(医学版)

Objective:To evaluate the effect of cryopreservation on clonogenic ability and apoptosis rate of mono-nuclear cells and CD34+cells in umbilical blood (UB), and to choose the index to present the freezing injury and optimize the cryopreservation of UB. Methods:hTe mono-nuclear cells (MNC) and CD34+cells were separated from UB and frozen.Atfer 30 days, they were thawed in warm water. Clonogenic capacity and clonogenic recovery before and atfer the cryopreservation was compared. We also used Annexin V-FITC-PI to investigate the apoptosis rate of the cells before and atfer the cryopreservation of these 2 types of cells. Results:hTe number of colony forming unit-granulocyte/monocyte (CFU-GMs) was not changed atfer freezing and thawing in both MNCs and CD34+cells, while the number of colony forming unit-granulocyte, erythrocyte, monocyte and megakaryocyte (CFU-GEMM) was obviously reduced after freezing in CD34+cells. The 2 types of cryopreserved cells had certain degree of apoptosis before the cryopreservation. MNC-type cryopreservation increased the cells apoptosis a little, while CD34+-type cryopreservation increased more. Conclusion:hTe cells have certain degree of apoptosis before the cryopreservation. hTe freezing and thawing procedure does affect the early stage progenitor cells-CFU-GEMM in the CD34+-type cryopreserved cells in UB. hTe damage may be induced by the cell apoptosis.

Cuantificación de bacterias cultivables mediante el método de Goteo en Placa por Sellado (o estampado) Masivo / Quantification of cultivable bacteria by the Massive Stamping Drop Plate method

Cuando se desea cuantificar el número de bacterias presentes en múltiples muestras, los procedimientos de rutina suelen consumir mucho tiempo. En ese periodo las muestras podrían sufrir modificaciones en su población. En el presente trabajo se evaluó una metodología alternativa para cuantificar bacterias cultivables de forma masiva, rápida y económica en la que se implica el sellado, estampado o impresión de diluciones seriadas de muestras de diversa procedencia. El tiempo requerido para preparar 22 muestras para su sellado en placa es de 15 minutos. El método se basó en realizar diluciones seriadas (base 10) de las muestras líquidas originales contenidas en una placa multipozos con la ayuda de una pipeta multicanal. Después, con un replicador se tomó un volumen (aproximadamente 1,65 µl) de muestra de cada pozo, que se inoculó por sellado en un medio de crecimiento gelificado de interés. Las placas se incubaron el tiempo necesario, se contó el número de colonias presentes en la dilución contable y se calculó el número de Unidades Formadoras de Colonia por mililitro (UFC/ml) para cada muestra. La metodología se denominó "Goteo por Sellado en Placa Masivo" (GSPM) y ha sido aplicada para cuantificar exitosamente bacterias provenientes de diferentes muestras de laboratorio, por ejemplo, de cultivos líquidos, muestras clínicas (como exudados y secreciones) y bacterias presentes en la rizósfera de plantas de maíz. Sin embargo, la metodología GSPM podría aplicarse para contabilizar masivamente a bacterias de cualquier otra procedencia.

In an attempt to quantify the number of bacteria present in a high number of samples, routine procedures are usually very time-consuming. During this period of time, bacterial population could be modified. In this work, an alternative for a massive, quick and economic method was evaluated in order to count viable bacteria, consisting in the sealing or stamping of serial dilutions performed in samples from different origins. The time required to prepare 22 samples for plate stamping is only 15 minutes. The quantification was based in performing serial dilutions (10-fold) of the original liquid samples contained in a multiwell plate using a multichannel micropipette. Afterwards, using a replicator, the same volume of each sample (approximately 1,65 µl) was recovered from each well, and then it was inoculated and sealed in a solid growth media of interest. Plates were incubated as needed, colonies were counted in the quantifiable dilution and Colony Forming Units per milliliter (CFU/ml) was calculated for each sample. We called this method "Massive Stamping Drop Plate" (MSDP) and it has been successfully applied to count bacteria from different lab samples, including liquid cultures, clinical samples (exudates and secretions) and bacteria recovered from the rhizosphere of corn plants. However, MSDP could also be applied to massively count bacteria from any other source.

Degradation of Benzo [a] Pyrene by a novel strain Bacillus subtilis BMT4i (MTCC 9447)

Benzo [a] Pyrene (BaP) is a highly recalcitrant, polycyclic aromatic hydrocarbon (PAH) with high genotoxicity and carcinogenicity. It is formed and released into the environment due to incomplete combustion of fossil fuel and various anthropogenic activities including cigarette smoke and automobile exhausts. The aim of present study is to isolate bacteria which can degrade BaP as a sole source of carbon and energy. We have isolated a novel strain BMT4i (MTCC 9447) of Bacillus subtilis from automobile contaminated soil using BaP (50 ìg /ml) as the sole source of carbon and energy in basal salt mineral (BSM) medium. The growth kinetics of BMT4i was studied using CFU method which revealed that BMT4i is able to survive in BaP-BSM medium up to 40 days attaining its peak growth (10(29) fold increase in cell number) on 7 days of incubation. The BaP degradation kinetics of BMT4i was studied using High Performance Liquid Chromatography (HPLC) analysis of BaP biodegradation products. BMT4i started degrading BaP after 24 hours and continued up to 28 days achieving maximum degradation of approximately 84.66 percent. The above findings inferred that BMT4i is a very efficient degrader of BaP. To our best of knowledge, this is the first report showing utilization of BaP as a sole source of carbon and energy by bacteria. In addition, BMT4i can degrade a wide range of PAHs including naphthalene, anthracene, and dibenzothiophene therefore, it could serve as a better candidate for bioremediation of PAHs contaminated sites.

Sangue periférico como fonte de células para terapia celular / Peripheral blood as a source of stem cells

O sangue periférico tem sido utilizado como fonte de células progenitoras hematopoéticas para o transplante de medula óssea, única aplicação clínica bem estabelecida até o momento para as células-tronco. Mais recentemente, além das células progenitoras hematopoéticas, estudos têm identificado também no sangue periférico a presença de células-tronco mesenquimais. Estas células apresentam as mesmas características e marcadores de superfície que as células-tronco mesenquimais da medula óssea e são capazes de diferenciação em células do tecido conjuntivo como osteócitos, condrócitos, adipócitos e miócitos. Embora sua origem e destino ainda sejam desconhecidos, a presença destas células no sangue periférico de indivíduos adultos representa um importante instrumento na área de medicina regenerativa e terapia celular. O conhecimento de marcadores imunofenotípicos que possam caracterizar as CTM de forma mais prática e objetiva e de possíveis estratégias capazes de aumentar o número destas células na circulação são fundamentais para o avanço de pesquisas clínicas baseadas na sua utilização.

Peripheral blood has been routinely used as a source of hematopoietic progenitor cells for allogeneic and autologous bone marrow transplantation. Recent studies have demonstrated that a low number of mesenchymal stem cells are also present in the peripheral blood. They share the same surface markers as bone marrow-derived mesenchymal stem cells and are capable of differentiating into mesenchymal lineage cells including osteocytes, adipocytes and chondrocytes. Although their origin and destination are unclear, their presence in the peripheral blood of adults seems to represent an important and powerful tool for regenerative medicine and cell therapy.

The single itraconazole and combination with interferon-? against candida albicans infection / 重庆医科大学学报

Objective:To study the efficacy of single itraconazole and combination with interferon-? against Candida albicans infection.Methods:Murine models of systemic candida albicans infection were established, and treated with different concentrations of itraconazol alone and combination with INF-?. Then the concentrations of colonyforming units(cfu) and histolo gical sections were observed to evaluate the emcacy. Results:The combination of 105U INF-? and itraconazole at the dosage of 10mg/kg showed better effect against Candida albicans than the combination of itraconazole at the dosage of 50mg/kg with 105U INF-?. Conclusion:The combination of itraconazole at a low dose with INF-? shows a good effect against Candida albicans.The single use of itraconazole at a high dose is also anti-infection.

Effect of lead acetate on growth of murine mesenchymal stem cells / 吉林大学学报(医学版)

Objective To explore the effect of lead acetate on the growth of murine mesenchymal stem cells in vitro.Methods 40.00,60.00 and 100.00 ?mol?L-1 of lead acetate were used in the culture of colony-forming unit-fibroblast(CFU-F),the effect on the rate of colony-forming and the rule of variation were observed.Results The rates of colony-forming were(3.30?0.20),(2.40?0.10) and(1.57?0.21)/105,when the doses of lead acetate were 40.00,60.00 and 100.00 ?mol?L-1 and there were significant differences compared with control group(4.20?0.20)/105,P

Distribution and Characteristics of CD133+, CD34+ Cells in Counterflow Centrifugal Elutriation Fraction of Cord Blood and Bone Marrow / 대한소아혈액종양학회지

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.

Study of Bacterial Distribution on the Skin of Psoriasis by Tape Method / 대한피부과학회지

BACKGROUND: Psoriasis is one of the relatively common chronic relapsing cutaneous disorders. The etiology and pathogenesis of the psoriatic skin lesion are still unknown. A colonization of microbacterial organisms especially Staphylococcus aureus (S. aureus) have been considered as a factor for development and exacerbation of psoriatic skin lesion. OBJECTIVES: The purpose of this study was to observe the bacteria on the skin of the patients with psoriasis and healthy normal persons, and evaluate the relations between bacterial density, S. aureus colonization, and severity of psoriatic skin lesions. MATERIALS & METHODS: Twenty two psoriasis patients and 25 healthy normal persons were involved for this study. Psoriasis patients were classified according to a severity estimated by PASI (Psoriasis Area and Severity Index) and activity of psoriatic skin lesions. Microbial sampling by tape method (3M, 5x5 cm) were performed on the psoriatic skin lesion and uninvolved skin in the patients of psoriasis, and on the inner forearm of the normal healthy person. Microbial sampling by a swab were also carried out from nasal mucosa. The tapes were gently contacted on the blood agar plate, and cultured in aerobic condition(30 degrees C) during 2-5 days and the numbers of colony forming unit (CFU) were estimated. RESULTS: The results were as follows; Total numbers of CFU in the lesion and uninvolved skin of psoriasis patients were significantly higher than those of the healthy controls (p0.05). The activity of psoriasis was relatively correlated with PASI score and total number of CFU (p0.05). There was a significant correlation between the numbers of S. aureus on the lesional and uninvolved skin and that on the nasal mucosa. (p<0.05). CONCLUSION: This study demonstrates that bacterial density is significantly higher on the psoriatic skin lesions, which suggests that bacterial colonization on the skin has a role in the development and exacerbation of the psoriatic lesion.

The Effect of Vasoactive Intestinal Peptide on Cord Blood CD34 (+) Cells / 대한소아혈액종양학회지

PURPOSE: We investigated the expression of vasoactive intestinal peptide (VIP), VIP receptor 1 (VPAC1), VIP receptor 2 (VPAC2) genes in the human umbilical cord blood CD34 cells, and the ability of VIP to stimulate human primitive as well as monopotent hematopoietic progenitors. METHODS: We isolated RNA from umbilical cord blood CD34 cells, and then performed RT-PCR, and sequencing. The umbilical cord blood CD34 cells were cultured with the various concentrations of VIP for burst-forming unit of erythrocyte (BFU-E), colony-forming unit of granulocyte/monocyte (CFU-GM), colony-forming unit of graulocyte/erythrocyte/monocyte/megakaryocyte (CFU-GEMM), and colony-forming unit of megakaryocyte (CFU-Mk). RESULTS: The RNA coding for VPAC1 was detected in human umbilical cord blood CD34 cells. VIP significantly stimulated the growth of CFU-GEMM and CFU-Mk. CONCLUSION: The present results suggest that VIP is an important neuropeptide in the early proliferation of human primitive as well as megakaryocyte progenitors.

Usefulness of the IMI Channel of Sysmex SE-9000(TM) Automated Hematology Analyzer Predicting the Optimal Timing of Peripheral Blood Stem Cell Harvest / 임상검사와정도관리

BACKGROUND: An essential prerequisite for successful procurement of sufficient peripheral blood stem cells (PBSC) for engraftment is the optimal timing of collection. The Sysmex SE-9000 automated hematology analyzer provides the immature information (IMI) channel for the identification and counting PBSC. In this study, The optimal timing of PBSC collection was studied using IMI channel. METHODS: 193 peripheral blood stem cell collections were performed from 52 patients with hematologic disorders or solid tumors and 15 donors. Pre-harvest peripheral blood WBC, mononuclear cells (MNC) and IMI were tested and compared with CD34+ cell count and CFU-GM count of harvested products. RESULTS: Peripheral blood WBC and MNC count showed a weak correlation with CD34+ cell yield (r=0.38, P1x10(6)/kg with sensitivity of 88.7%. Positive and negative predictive values of IMI >465/microliter for CD34+ cell >1x10(6)/kg were 65.5% and 87.5%, respectively. CONCLUSIONS: The automated IMI might be used as a simple and efficient indicator of PBSC mobilization and applying variable cutoff values of IMI would be a useful tool to predict the optimal timing of PBSC collection.

Measuring Intracellular Mycobacterial Killing Using a Human Whole Blood Assay / 결핵

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.

Megakaryocyte Colony Formation of Fetal Liver Cells

PURPOSE: This study was undertaken to obtain basic data about the megakaryocyte colony formation of fetal liver cells by using immunocytochemical staining and ex vivo culture with growth factors. METHODS: The mononuclear cells were isolated from fetal liver and bone marrow with idiopathic thrombocytopenic purpura(ITP) and pancytopenia. These mononuclear cells were cultured in MegaCultTM-C(Stem Cell Tech, Canada) media in the presence of growth factors and CFU-Megakaryocyte(CFU-Mk) colonies were counted on day 12. The expansion of CD34+ and CD41+ cell was analyzed by flow cytometry after 5 days incubation using flask culture. RESULTS: The numbers of CFU-Mk colonies of mononuclear cells obtained from fetal liver in the 11th week gestational age were more than those in the 19th week specimens; growth factors could not enhance the colony expansion in all cases. Total numbers of CFU-Mk colony of fetal liver cells were higher than bone marrow from ITP or pancytopenia groups. The numbers of pure or large CFU-Mk colonies of fetal liver cells were also higher than bone marrow specimens. The rate of CD34+ cell expression of fetal liver was increased after flask culture and the enhancement effect of epression was seen only in cases which added thrombopoietin. The rate of CD41+ cell expression of fetal liver was increased after incubation, but the enhancement effect of growth factors was unclear. CONCLUSION: This study revealed good results about the megakaryocyte colony assay of fetal liver mononuclear cells using MegaCultTM-C media. This study suggests that the fetal liver could be a good source of megakaryocytic progenitor cells for clinical application in hematopoietic stem cell transplantation.