Comparative evaluation of antimicrobial efficacy of silver diamine fluoride, chlorhexidine varnish with conventional fluoride varnish as a caries arresting agent. An in vivo sodium dodecyl sulfate-polyacrylamide gel electrophoresis study

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.

Biocompatibility with the Ocular Surface and Antimicrobial Activity of a New Multi-Purpose Contact Lens Solution / Biocompatibilidad con la superficie ocular y actividad antimicrobiana de una nueva solución multipropósito para lentes de contacto / Biocompatibilidade com a superfície ocular e atividade antimicrobiana de uma nova solução de lentes de contato multipropósito

Abstract Introduction: Multipurpose solutions (MPS) for soft contact lenses (SCL) play an essential role in inhibiting potentially pathogenic agents. Their antimicrobial effectiveness is assessed in vitro and their safety in vivo, with clinical trials that include a combination of different solutions and lens materials. The objective is to assess the biocompatibility of a new SCL MPS produced in Colombia that contains polyhexamethylene biguanide (PHMB) and to determine its antimicrobial activity. Materials and Methods: This was a crossover study with 25 subjects who did not wear lens and who were fitted with different combinations of five SCL materials with either MPS or control physiological saline solution (CS). Corneal thickness, conjunctival hyperemia, corneal staining, and comfort were assessed after two hours of wearing SCL. Antimicrobial effectiveness was measured using ISO 14729 standard assays. Results: When considering SCL material, there was a statistically significant difference between the new MPS and the CS for Comfilcon A (p < 0.05). There was no statistical or clinically significant difference for corneal thickness or corneal staining between the combination of lens material and new MPS with the CS (p > 0.05). After two hours of lens insertion, comfort scores were higher than 7.8. The MPS reduced bacteria colony forming units (CFU) in over 3 log, and fungal CFU in over 1.0 log. Conclusions: The new MPS met the antimicrobial standards of ISO 14729, is considered safe and biocompatible with the ocular surface and retains high comfort levels.

Resumen Introducción: las soluciones multipropósito (SMP) para lentes de contacto blandos (LCB) desempeñan un papel esencial en la inhibición de agentes potencialmente patógenos. Su efectividad antimicrobiana se evalúa in vitro, y su seguridad, in vivo, con ensayos clínicos que incluyen una combinación de diferentes soluciones y materiales para lentes. El objetivo es evaluar la biocompatibilidad de una nueva SMP producida en Colombia que contiene polihexametileno biguanida (PHMB) y determinar su actividad antimicrobiana. Materiales y métodos: estudio cruzado con 25 sujetos no usuarios de lentes, que fueron adaptados con cinco combinaciones diferentes de materiales de LCB con una nueva SMP o solución salina fisiológica de control (CS). El grosor corneal, la hiperemia conjuntival, la tinción corneal y la comodidad se evaluaron después de dos horas de uso del LC. La efectividad antimicrobiana se midió utilizando ensayos estándar ISO 14729. Resultados: considerando el material del LCB, solo hubo una diferencia estadísticamente significativa entre la nueva SMP y el CS para el Comfilcon A (p < 0.05). Tampoco hubo diferencias estadísticamente o clínicamente significativas para el grosor corneal o la tinción corneal, entre la combinación del material del lente y la nueva SMP con el CS (p > 0.05). Después de dos horas de uso del lente, las puntuaciones de confort fueron superiores a 7.8. La SMP redujo las unidades formadoras de colonias (UFC) de bacterias en más de 3 log, y las UFC fúngicas en más de 1.0 log. Conclusiones: la nueva SMP cumplió con los estándares antimicrobianos de ISO 14729, y se considera segura y biocompatible con la superficie ocular, con altos niveles de confort.

Resumo Introdução: as soluções multipropósito (SMP) para lentes de contato macias (LCM) apresentam um papel essencial na inibição de agentes potencialmente patógenos. Sua eficácia como agente antimicrobiano se valia in vitro, e sua segurança, in vivo, como ensaios clínicos que incluem uma combinação de diferentes soluções e materiais para lentes. O objetivo é avaliar a biocompatibilidade de uma nova SMP produzida na Colômbia a base de polihexametileno biguanida (PHMB) e determinar seu potencial antimicrobiano. Materiais e métodos: estudo cruzado com 25 indivíduos não usuários de lentes, que foram adaptados com cinco combinações diferentes de LCM como uma nova SMP ou solução salina fisiológica como controle (CS). A espessura da córnea, a hiperemia conjuntival, a coloração da córnea e a comodidade, foram avaliadas após duas horas de uso da LCB. A eficácia antimicrobiana foi medida com ensaios padrão ISO 14729. Resultados: considerando o material da LCB, houve apenas uma diferença estatisticamente significativa entre a nova SMP e o CS, paro o Comfilcon A (p <0.05). Não houve diferença estatisticamente ou clinicamente significativa para a espessura da córnea ou a coloração da córnea, entre a combinação do material da lente e a nova SMP com o controle CS (p > 0.05). Após duas horas de uso, as pontuações de conforto foram superiores a 7,8. A SMP reduziu as unidades formadoras de colônias (UFC) de bactérias em mais de 3 log, e as UFC fúngicas em mais de 1.0 log. Conclusões: a nova SMP cumpriu com os padrões antimicrobianos ISO 14729, é considerada segura e biocompatível com a superfície ocular, com altos níveis de conforto.

Optimización de unidades formadoras de colonias y detección del virus de papiloma humano en sangre de cordón umbilical / Colony-forming units optimization and human papillomavirus detection in umbilical cord blood

Resumen Introducción: Se requiere analizar diversos parámetros para el control de calidad adecuado de las unidades de sangre de cordón umbilical (USCU) cuando se utilizan con fines terapéuticos. Objetivo: Optimizar las unidades formadoras de colonias (UFC) de cultivos clonogénicos y detectar el genoma del virus del papiloma humano (VPH) en USCU. Métodos: Se incluyeron 141 muestras de sangre de cordón umbilical (SCU), de segmento y de UFC de cultivos clonogénicos de USCU. Se realizó extracción de ADN, cuantificación y amplificación por PCR del gen endógeno GAPDH. Se detectó el gen L1 del VPH con los oligonucleótidos MY09/MY11 y GP5/GP6+; los productos de PCR se migraron en electroforesis de agarosa. El ADN purificado de las UFC se analizó mediante electroforesis de agarosa y algunos ADN, con la técnica sequence specific priming. Resultados: La concentración de ADN extraído de UFC fue superior comparada con la de SCU (p = 0.0041) y la de segmento (p < 0.0001); así como la de SCU comparada con la de segmento (p < 0.0001). Todas las muestras fueron positivas para la amplificación de GAPDH y negativas para MY09/MY11 y GP5/GP6+. Conclusiones: Las USCU criopreservadas fueron VPH netativas; además, es factible obtener ADN en altas concentraciones y con alta pureza a partir de UFC de los cultivos clonogénicos.

Abstract Introduction: Analysis of several markers is required for adequate quality control in umbilical cord blood units (UCBU) when are used for therapeutic purposes. Objective: To optimize colony-forming units (CFU) from clonogenic cultures and to detect the human papillomavirus (HPV) genome in UCBU. Methods: One hundred and forty-one umbilical cord blood (UCB), segment or CFU samples from UCBU clonogenic cultures were included. DNA extraction, quantification and endogenous GAPDH gene PCR amplification were carried out. Subsequently, HPV L1 gene was detected using the MY09/MY11 and GP5/GP6+ oligonucleotides. PCR products were analyzed with electrophoresis in agarose gel. CFU-extracted purified DNA was analyzed by electrophoresis in agarose gel, as well as some DNAs, using the SSP technique. Results: CFU-extracted DNA concentration was higher in comparison with that of UCB (p = 0.0041) and that of the segment (p < 0.0001), as well as that of UCB in comparison with that of the segment (p < 0.0001). All samples were positive for GAPDH amplification and negative for MY09/MY11 and GP5/GP6+. Conclusions: Cryopreserved UCBUs were HPV-negative. Obtaining CFU DNA from clonogenic cultures with high concentrations and purity is feasible.

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

Developmental relationship between mesenchymal stem/progenitor cells and hematopoietic cells dur-ing human embryogenesis / 中华微生物学和免疫学杂志

Objective To analyze the developmental relationship between mesenchymal stem/pro-genitor cells (MSPCs) and hematopoietic cells during human embryogenesis. Methods Aborted embryos at different developmental stages were used in this study after medical abortion. Embryonic blood tissues were isolated and digested into single cells. These single cells were plated in semisolid medium in favor of the dif-ferentiation of colony-forming cell with high proliferative potential ( HPP-CFC) and incubated for 10 to 14 d. Individual colonies with diameter more than 0. 5 mm were picked and replated in liquid medium. Fibroblastic adherent cells appeared in the replated colonies were cultured for cell proliferation and cytokins expressed on cell surface were identified to analyze whether they had the characteristics of MSPCs. Results This study summarized the dynamic development of HPP-CFCs and other hematopoietic progenitor cells in different tis-sues including aorta-gonad-mesonephros ( AGM) region, yolk sac and embryonic liver. From the 28-somite stage, a proportion of HPP-CFCs in AGM region could give rise to adherent fibroblastic cells in addition to hematopoietic cells. The adherent cells harbored the differentiation potential of MSPCs and could inhibit the proliferation of T cells in lymphocyte transformation test. Conclusions This study suggests some prehemato-poietic precursors in AGM region can give rise to both hematopoietic progenitors and MSPCs during human embryogenesis.

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.

Developmental relationship between mesenchymal stem/progenitor cells and hematopoietic cells during human embryogenesis / 中华微生物学和免疫学杂志

Objective@#To analyze the developmental relationship between mesenchymal stem/progenitor cells (MSPCs) and hematopoietic cells during human embryogenesis.@*Methods@#Aborted embryos at different developmental stages were used in this study after medical abortion. Embryonic blood tissues were isolated and digested into single cells. These single cells were plated in semisolid medium in favor of the differentiation of colony-forming cell with high proliferative potential (HPP-CFC) and incubated for 10 to 14 d. Individual colonies with diameter more than 0.5 mm were picked and replated in liquid medium. Fibroblastic adherent cells appeared in the replated colonies were cultured for cell proliferation and cytokins expressed on cell surface were identified to analyze whether they had the characteristics of MSPCs.@*Results@#This study summarized the dynamic development of HPP-CFCs and other hematopoietic progenitor cells in different tissues including aorta-gonad-mesonephros (AGM) region, yolk sac and embryonic liver. From the 28-somite stage, a proportion of HPP-CFCs in AGM region could give rise to adherent fibroblastic cells in addition to hematopoietic cells. The adherent cells harbored the differentiation potential of MSPCs and could inhibit the proliferation of T cells in lymphocyte transformation test.@*Conclusions@#This study suggests some prehematopoietic precursors in AGM region can give rise to both hematopoietic progenitors and MSPCs during human embryogenesis.

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.

Evaluation of Efficacy of Virgin Coconut Oil in Maintaining Hygiene in Orthodontics Patients

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

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.

The effect of chlorhexidine-thymol and fluoride varnishes on the levels of Streptococcus mutans in saliva in children aged 6–8 years

Background: Streptococcus mutans is considered as the main pathogenic factor for initiation and progression of dental caries. Fluoride is one of the most effective agents used to control caries. Chlorhexidine (CHX) is the most antimicrobial agent against S. mutans and dental caries. Aims: The study aimed to compare the effectiveness of antimicrobial activity of CHX-thymol (CHX/T) and fluoride varnishes on S. mutans levels in children's saliva aged from 6 to 8 years old. Materials and Methods: The total number of children involved in this study is sixty, ages 6 and 8 years old. The participants were divided into three groups by block randomization: Group 1 CHX/T varnish, Group 2 fluoride varnish (f varnish, and Group 3 control group. Varnish was applied onto all tooth surfaces of the participants. At the baseline conditions, saliva samples were collected from the participants for bacterial examination test. This procedure was repeated in week 1, 4, and 12. Bacterial quantitative test was performed, and the number of S. mutans was estimated. Results: The results revealed the significant efficacy of the two groups (fluoride and CHX/T varnishes) in reducing salivary S. mutans numbers when compared to the control group (P < 0.05). In terms of salivary colony-forming unit counts reduction of S. mutans, no significant difference was observed between the fluoride and CHX/T varnish groups (P > 0.05). Conclusion: The outcomes showed that there was a significant reduction in S. mutans counts in children's saliva following the application of fluoride and CHX/T varnishes.

Effects of liraglutide on the biological behavior of human endothelial colony forming cell induced by high glucose via regulating sirtuin expression / 解放军医学杂志

Objective To explore whether liraglutide could affect high glucose induced-biological behavior of human endothelial colony forming cell (ECFC) by regulating sirtuin. Methods Mononuclear cells in human peripheral blood were isolated by density gradient centrifugation. ECFCs were induced and cultured with EBM-2 complete medium. The cells were randomly divided into three groups: normal glucose group (NG, 5.5 mmol/L D-glucose for 6 days), high glucose group (HG, 30 mmol/L D-glucose for 6 days), and osmotic pressure group (OSM, 5.5 mmol/L D-glucose+24.5 mmol/L mannitol for 6 days). The proliferation, migration, tube formation and senescence of ECFCs in the three groups were tested by EdU, Transwell, tube formation and SA-β-gal experiments respectively. The protein expression levels of Sirtuins (Sirtuin1-7), vascular endothelial growth factor (VEGF) and angiogenin were measured by Western blotting. Except NG and HG groups mentioned above, NG + liraglutide intervention group (5.5 mmol/L D-glucose+100 nmol/L liraglutide) and HG+liraglutide intervention group (30 mmol/L D-glucose+100 nmol/L liraglutide) were added in order to observe the effects of liraglutide on ECFC biological behavior and the expression of VEGF and angiogenin. The intervention time in all groups is 6 days. Results EdU, Transwell and tube formation experiments showed that the proliferation rate, migration ability and tube formation ability of ECFCs were lower in HG group [(30.16±12.36)%, 25.11±6.05, 27.50±3.90, respectively] than in NG group [(88.00±13.77)%, 64.89±10.73, 69.61±5.48, respectively], while the SA-β-gal experiment showed the senescence rate of ECFCs was higher in HG group than in NG group [(87.63±9.63)% vs. (71.35±7.58)%], all with statistical significance (P<0.05). Under high glucose conditions, the expression of Sirtuins family was generally reduced, and the expressions of VEGF and angiogenin significantly decreased (P<0.05). The expression of Sirtuin1 showed an upward trend with the increase of liraglutide concentration. After intervention with appropriate concentration of liraglutide, the expressions of VEGF and angiogenin significantly increased (P<0.05). EdU experiment showed the proliferation rate of ECFCs was higher in HG+liraglutide intervention group than in NG group [(54.09±27.29)% vs. (29.01±7.56)%], the Transwell and tube formation experiments showed that the migration ability and tube formation ability of ECFCs were higher in HG+liraglutide intervention group (32.25±4.99, 69.61±8.11) than in HG group (21.75±3.10, 39.85±5.78), and the SA-β-gal experiment showed the senescence rate of ECFCs was lower in HG+liraglutide intervention group than in HG group [(52.06±7.94)% vs. (69.03±1.57)%], all with statistical significance (P<0.05). Conclusions High glucose may accelerate ECFCs senescence while decrease the ability of proliferation, migration and tube formation of ECFCs, accompanied by down-regulation of Sirtuins, VEGF and angiogenin. Liraglutide may reverse the changes of ECFCs biological behavior induced by high glucose and up-regulate the expression of Sirtuin1, VEGF and angiogenin.

The study of the intestinal microbiota among Mongolian Adults, it related some factors / Монголын Анагаах Ухаан

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

Microbial air contamination with air borne bacteria in the outdoor environment at Janakpurdham, Nepal

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.

Assessment of Candida SPP. proliferation in occlusal and palatal splints / Avaliação da proliferação Candida SPP. em placas oclusais e palatais

Candida species inhabit the oral cavity of all individuals who wear complete denture and whose material is the same as that used in splints. Assess the growth of C. albicans in occlusal and palatal splints used for treatment of TMD so that the potential risks of oral microbiota can be assessed. The growth of Candida spp. was assessed in the saliva of 27 individuals wearing splints for treatment of TMD. They were divided into two groups: G1 (n = 14), individuals wearing occlusal splint; and G2 (n = 13), individuals wearing palatal splint. Saliva samples were collected during placement of the splints (T1) and after 4 months (T2), being stored in PBS (10 mL) after 60-second rinses. It was observed that patients wearing occlusal splints (G1) had an increase of 0.648 CFU/mL (Log 10), with statistically significant differences (P = 0.043) for C. albicans (42.33%), C. glabrata (5.52%), C. krusei (41.72%) and C. tropicalis (10.43%). In the group of patients wearing palatal splints (G2), there was a decrease of 0.101 CFU/mL (Log 10), was observed with (P = 0.964) only the presence of C. albicans. The results suggest that growth of Candida species was greater in patients wearing occlusal splints compared to those wearing palatal ones as the presence of different yeast species was found in the former.

Espécies de Candida habitam a cavidade oral de 60-100% de indivíduos usuários de prótese total, cujo material é o mesmo utilizado em placas miorrelaxante. Avaliar o crescimento de C. albicans. em placas relaxantes musculares oclusais e palatais, usadas para o tratamento de DTM, na intenção de verificar riscos em potencial à microbiota bucal. Avaliou-se o crescimento de Candida spp. na saliva de 27 indivíduos, usuários de placa miorrelaxante, em tratamento para DTM no ICT-UNESP. Os indivíduos foram divididos em dois grupos: G1(n=14) ­ placa com recobrimento oclusal; e G2 (n=13) ­ sem recobrimento. As coletas foram com PBS (10mL), em bochechos por 60seg, na instalação das placas (T1) e após 4 meses (T2). Observou-se que pacientes usuários da placa miorrelaxante com recobrimento oclusal (grupo G1) apresentaram aumento de 0,648 UFC/mL (Log10) com diferença estatisticamente significante (p=0,043) analisando-se 42,33% C. albicans, 5,52% C. glabrata, 41,72% C. krusei e 10,43% C. tropicalis. No grupo de pacientes que utilizaram a placa sem recobrimento (grupo G2), observou-se diminuição de 0,101 UFC/mL (Log10) com (p=0,954) apresentando apenas C. albicans. Os resultados sugerem que os pacientes que fizeram uso de placa miorrelaxante com recobrimento oclusal apresentaram maior crescimento de Candida spp. em relação aos usuários de placa sem recobrimento, verificando-se a presença de diferentes espécies da levedura.

Effect of dichloroacetate on colony-forming, invasion and migration of the T24 bladder cancer cells line and its mechanism / 实用医学杂志

Objective To study the effects of dichloroacetate (DCA) on cell colony-forming,cell invasion and cell migration of the bladder cancer cells and to study the underlying mechanism.Methods The bldder cancer cells T24 were randomly divided into two groups:the observation group and the control group.Cells in the observation groups were treated with 5 mmol/L,10 mmol/L and 20 mmol/L dichloroacetate,and the control group was treated with the same amount of dimethyl sulfoxide.Colony formation assays were detected with Giemsa staining.Cell wound scratch assay and Transwell assay were applied to evaluate the ability of the T24 cell invasion and migration.Realtime PCR and Western blot were applied to detect the expression of the epithelial-mesenchymal transition (EMT)-related marker,including E-cadherin,N-cadherin,vimentin,Snail and Slug.Results Compared with the control group,the colony formation assays of T24 cells constantly decreased along with the increased doses in the observation group(P ＜ 0.01).The cell wound scratch assay showed that the scratch width of the observation groups were significantly higher along with the increased doses and prolonged time than that in the control group (P ＜ 0.01).The transwell assay showed that the invasion ability of the observation groups were significantly discreased along with the increased doses than that in the the control group (P ＜ 0.01).The expression levels of E-cadherin mRNA and protein in combination the control group were higher than those in the the observation groups (P ＜ 0.05).However,the expression levels of N-cadherin,vimentin,Snail and Slug mRNAs and proteins in combination the control group were lower than those in the the observation groups (P ＜ 0.05).Conclusion Dichloroacetate can inhibit the colony-forming,invasion and migration of bladder cancer T24 cells,and its mechanism may inhibit the expression of epithelial mesenchymal transition in T24 cells by down-regulating the expression of nuclear transcription factor Snail and Slug.

Two-Cell Spheroid Angiogenesis Assay System Using Both Endothelial Colony Forming Cells and Mesenchymal Stem Cells

Most angiogenesis assays are performed using endothelial cells. However, blood vessels are composed of two cell types: endothelial cells and pericytes. Thus, co-culture of two vascular cells should be employed to evaluate angiogenic properties. Here, we developed an in vitro 3-dimensional angiogenesis assay system using spheroids formed by two human vascular precursors: endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs). ECFCs, MSCs, or ECFCs+MSCs were cultured to form spheroids. Sprout formation from each spheroid was observed for 24 h by real-time cell recorder. Sprout number and length were higher in ECFC+MSC spheroids than ECFC-only spheroids. No sprouts were observed in MSC-only spheroids. Sprout formation by ECFC spheroids was increased by treatment with vascular endothelial growth factor (VEGF) or combination of VEGF and fibroblast growth factor-2 (FGF-2). Interestingly, there was no further increase in sprout formation by ECFC+MSC spheroids in response to VEGF or VEGF+FGF-2, suggesting that MSCs stimulate sprout formation by ECFCs. Immuno-fluorescent labeling technique revealed that MSCs surrounded ECFC-mediated sprout structures. We tested vatalanib, VEGF inhibitor, using ECFC and ECFC+MSC spheroids. Vatalanib significantly inhibited sprout formation in both spheroids. Of note, the IC₅₀ of vatalanib in ECFC+MSC spheroids at 24 h was 4.0 ± 0.40 μM, which are more correlated with the data of previous animal studies when compared with ECFC spheroids (0.2 ± 0.03 μM). These results suggest that ECFC+MSC spheroids generate physiologically relevant sprout structures composed of two types of vascular cells, and will be an effective pre-clinical in vitro assay model to evaluate pro- or anti-angiogenic property.

Advanced tube formation assay using human endothelial colony forming cells for in vitro evaluation of angiogenesis

The tube formation assay is a widely used in vitro experiment model to evaluate angiogenic properties by measuring the formation of tubular structures from vascular endothelial cells (ECs). in vitro experimental results are crucial when considered the advisability of moving forward to in vivo studies. Thus, the additional attentions to the in vitro assay is necessary to improve the quality of the pre-clinical data, leading to better decision-making for successful drug discovery. In this study, we improved the tube formation assay system in three aspects. First, we used human endothelial colony forming cells (ECFCs), which are endothelial precursors that have a robust proliferative capacity and more defined angiogenic characteristics compared to mature ECs. Second, we utilized a real-time cell recorder to track the progression of tube formation for 48 hours. Third, to minimize analysis error due to the limited observation area, we used image-stitching software to increase the microscope field of view to a 2×2 stitched area from the 4× object lens. Our advanced tube formation assay system successfully demonstrated the time-dependent dynamic progression of tube formation in the presence and absence of VEGF and FGF-2. Vatalanib, VEGF inhibitor, was tested by our assay system. Of note, IC₅₀ values of vatalanib was different at each observation time point. Collectively, these results indicate that our advanced tube formation assay system replicates the dynamic progression of tube formation in response to angiogenic modulators. Therefore, this new system provides a sensitive and versatile assay model for evaluating pro- or anti-angiogenic drugs.

Dysfunction of endothelial progentior cells in the angiographically defined coronary atherosclerosis / Монголын Анагаах Ухаан

Introduction@#Endothelial progenitor cells (EPC) have a role in the maintenance and promotion of vascular repair and are negatively correlated with coronary atherosclerosis. @*Goal@#To culture of EPC-CFUs during coronary atherosclerosis, evaluate endothelial nitric oxide synthetase (eNOS) enzyme levels in the culture.@*Materials and Methods@#The 10 ml blood was drawn from the peripheral vein of 12 man patients that stable angina 4, acute myocardial infarction (AMI) 4 and healthy people 4. Peripheral blood mononuclear cells were isolated by Ficoll density-gradient centrifugation and EPC-CFUs was assayed after two platings and a 6 day culture on fibronectin coated, 72 well plates, as described. eNOS enzyme titers were determined by ELISA according to the protocol in the cells culture.@*Results@#The people were 52±2.12 years. The number of EPC-CFUs increases with accordance of patients with stable angina, AMI, healthy people with the statistical significance (F=17.3, p<0.001): stable angina (2.6±0.47 colony/well), AMI (6.7±0.81 colony/well), healthy people (10.5±1.34 colony/well). Furthermore, ANOVA test of eNOS enzyme levels in patients with stable angina (5.2±0.61 pg/ml), AMI (8.7±1.49 pg/ml) and healthy people (13.7±2.48 pg/ml). The significant difference (F=6.2, p=0.003) was observed among the three groups. The number of EPC-CFUs had direct significantly correlation (r=0.621, p<0.001) with the eNOS enzyme levels of this culture.@*Conclusion@#Number of EPC-CFUs and eNOS enzyme levels decrease at patient with stable angina, indicate more than endothelial dysfunction.@*Ethical approval@#The ethics committee of Mongolian National University of Medical Sciences (ID: 6/3/201506, approved on Jan 01, 2015)

RESULTS OF CULTURING STEM CELLS DERIVED FROM MOUSE BONE MARROW UNDER VARYING OXYGEN CONDITIONS / Шинэ санаа Шинэ нээлт

@#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