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

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.

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.

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.

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)

Expression of CD47 in de novo acute myelogenous leukemia with normal karyotype and its clinical significance / 白血病·淋巴瘤

Objective To investigate the CD47 expression in de novo acute myelogenous leukemia (AML) patients with normal karyotype and its clinical significance. Methods One hundred thirty-seven cases of de novo AML with normal karyotype and 3 healthy volunteers were selected. Relative CD47 expressions in normal bone marrow hematopoietic stem cells (HSC) and multipotent progenitor (MPP) from healthy volunteers, as well as bone marrow mononuclear cells (MNC) and leukemia stem cells (LSC, Lin-CD34+CD38-CD90-) from AML patients were determined by flow cytometry. CD47 expression on the Lin-CD34+CD38-LSC-enriched fraction of specimen was determined by flow cytometry. The FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD) was detected by using the Genome Analyzer platform. CD34+CD38-CD47hi and CD34+CD38-CD47lo expressing cells were identified and purified using FACS. Two groups of cells were inoculated with MethoCult H4445 medium on agarose-containing methylcellulose plates. After 12 days, MPP colony forming units (CFU) were counted, and 1×105 CD34+ CD38- CD47lo and CD34+ CD38-CD47hi cells were transplanted into NSG (NOD-SCID IL-2R γ null) mice irradiated by 280 cGy, and mice were sacrificed after 8 weeks. The ratio of human CD45+cells was detected by flow cytometry. Results The expression of CD47 in AML patients was higher than that in the healthy control. CD47 was expressed in all FAB (French-American-British) subtypes of AML. No significant difference in CD47 expression among different FAB subtypes was found (F=0.545, P>0.05). Among the 37 patients with CD34+CD38-CD47hi, 17 (46 %) were FLT3-ITD negative, and 20 (54 %) were FLT3-ITD positive. Among the 100 patients with CD34+CD38-CD47hi, 63 (63%) cases were FLT3-ITD negative, 37 (37%) cases were FLT3-ITD positive. The rate of FLT3-ITD positive in patients with CD34+ CD38- CD47lo had no statistical difference compared with patients with CD34+CD38-CD47hi (χ2= 3.79, P> 3.79). The CD34+CD38-CD47lo or CD34+CD38-CD47hi which was selected by FACS, was inoculated with the methylcellulose plate containing agarose for 12 days, and CD34+CD38-CD47lo cells could form CFU. The NSG mouse transplantation experiment showed that CD34+CD38-CD47lo cells could be reconstructed hematopoiesis, and CD34+CD38-CD47hi implantation failed. Conclusion CD34+CD38-CD47hi could enrich LSC, which may be a potential marker to detect minimal residual disease.

Eficacia antimicrobiana del primer ortodóncico adicionado con nanopartículas de plata: Estudio transversal in vitro / Antimicrobial efficacy of orthodontic primer added with silver nanoparticles: Cross-sectional in vitro study

Se evaluó la eficacia antimicrobiana de las nanopartículas de plata (NPsAg) incorporadas al adhesivo (primer) colocado en el esmalte dental adyacente a la aparatología ortodóncica fija (brackets). Se realizó un estudio experimental in vitro en 40 premolares, los cuales se dividieron en dos grupos con brackets, uno cementado con primer convencional y otro adicionado con NPsAg; se colocaron en medios de cultivo, previamente inoculados con Streptococcus mutans, y se tomaron muestras para hacer cultivos y conteo de unidades formadoras de colonias (UFC) al día 1, 15 y 30. Se observó una disminución de la presencia de Streptococcus mutans en las muestras a los 15 días de colocado el primer con la agregación de nanopartículas, aunque tal efecto se redujo a los 30 días. Esta reducción del efecto de las nanopartículas puede deberse a la inexistencia de limpieza mecánica, lo cual favoreció la agregación bacteriana sobre el biofilm, afectando su efecto antimicrobiano. Esto sugiere la necesidad de realizar estudios in vivo que permitan observar el comportamiento de los biomateriales en el medio bucal. Las NPsAg agregadas al primer resultan una herramienta eficaz para prevenir la desmineralización del esmalte alrededor de la aparatología ortodóncica fija.

The antimicrobial efficacy of the silver nanoparticles (NPsAg), incorporated into the adhesive (primer) placed in the enamel adjacent to fixed orthodontic appliances (brackets), was evaluated. An experimental study was performed on 40 premolars in vitro, which were divided into two groups with brackets, one cemented with conventional primer and another added with NPsAg, placed in culture media previously inoculated with Streptococcus mutans, and sampled for culturing and counting colony forming units (UFC) on days 1, 15 and 30. A decrease in the presence of Streptococcus mutans in the samples after 15 days with nanoparticle aggregation was observed, and a reduction in the effect of said nanoparticles after 30 days. This reduction of the nanoparticles effects can be due to the absence of mechanical cleaning, which favored the bacterial aggregation on the biofilm, affecting its antimicrobial effect. This suggest the need for realizing studies in “vivo” which will allow the observation of the behavior of the biometals on the buccal medium. The NPsAg added to the primer are an effective tool to prevent the demineralization of the enamel around the fixed orthodontic appliances.

Carga microbiana de las lámparas de fotocurado en el uso y desuso de las barreras adhesivas de protección

Se realizó un estudio acerca del análisis de la carga microbiana en las lámparas de fotocurado en cuanto al uso y desuso de barreras adhesivas de protección, con el fin de que los resultados sirvan para hacer conciencia sobre el uso de las barreras adhesivas de protección, para resguardar tanto al profesional como a su ambiente de trabajo, y a los pacientes, evitando además la contaminación cruzada entre ellos, como con el medio oral y los demás instrumentos. Para la investigación se tomó una muestra de 47 lámparas, 24 con barrera y 23 sin barrera, para así determinar la cantidad de unidades formadoras de colonias por superficie, y además la presencia o ausencia de Escherichia Coli como indicador de microorganismo patógeno.

It has been made a study of the analysis of the microbial load in dental curing lights in the use and disuse of adhesive protective barriers, with the goal that the results serve to raise awareness on use of adhesive protective barriers to protect the professional, their work environment and patients, while avoiding cross-contamination between them, as with the oral environment and other instruments. For this research the sample were 47 lamps, 24 with barrier and 23 without barrier, to determine the number of colony forming units per surface, and also the presence or absence of Escherichia coli as pathogen indicator.

Caries dental y microorganismos asociados a la caries en la saliva de los alumnos del primer año de la Facultad de Odontología, UNAM / Dental caries and caries-associated mechanisms found in the saliva of first year students at the School of Dentistry, UNAM (National Autonomous University of Mexico)

Se estudiaron 380 alumnos del primer año en la Facultad de Odontología (n = 380) (periodo 2012-2013) a fin de determinar el índice CPOD y relacionar si la caries está asociada con los microorganismos Streptococcus y Lactobacillus. El índice CPOD (cariado, perdido y obturado) se registró usando los parámetros de la Organización Mundial de la Salud. Se tomaron muestras de saliva de cada alumno y se determinaron las unidades formadoras de colonias de Streptococcus y Lactobacillus. La media de los índices CPOD fue de 7.25 ± 4.59. Las mujeres (n = 278) y hombres (n = 102) presentaron una media de índices CPOD de 7.11 ± 4.66 y 7.29 ± 4.57, respectivamente. Encontramos que los alumnos de 19 años presentaron menos caries que los estudiantes de otras edades. Tanto Streptococcus y Lactobacillus se correlacionaron significativamente entre sí, así como en la incidencia de caries. Un incremento en el número de estos microorganismos, especialmente de Streptococcus mutans, se asociaron con el incremento en CPOD.

Three hundred and eighty first year students of the National School of Dentistry (UNAM) (n = 380) (academic year 2012-2013), were assessed targeting determination of DMFT (decayed, missing, lost teeth) index as well as to establish a relationship of whether caries is associated to Lactobacillus and Streptococcus microorganisms. DMFT index was recorded using World Health Organization (WHO) parameters. Samples of all students were taken and colony-forming units of Streptococcus and Lactobacillus were determined. DMFT indexes mean was established at 7.25 ± 4.59. Females (n = 278) and males (n = 102) exhibited mean DMFT indexes of 7.11 ± 4.66 and 7.29 ± 4.57 respectively. Results revealed that 19 year old students exhibited lesser amounts of caries than students of other ages. Both Streptococcus and Lactobacillus were significantly correlated to each other as well as to caries incidence. Increase in the number of the aforementioned micro-organisms, especially Streptococcus mutans, were associated to DMFT increase.

Celularidade do leite e unidades formadoras de colônias nas mastites causadas por Staphylococcus coagulase positiva e coagulase negativa / Milk cellularity and colony forming units in mastitis caused by coagulase-positive staphylococci and coagulase negative

A mastite é a principal afecção do gado destinado à produção leiteira, que impacta significativamente a cadeia produtiva do leite, com reflexos ainda para a saúde pública. Estudou-se aspectos relacionados à etiologia, celularidade e de contagem bacteriana em 10 propriedades leiteiras, localizadas no Estado de São Paulo. Foram examinadas 1148 vacas em lactação, totalizando 4584 glândulas mamárias. Foram considerados os casos, em que houve isolamento de estafilococos coagulase positiva (SCP) e estafilococos coagulase negativa (SCN). Os resultados revelaram microbiota com vários patógenos e diferentes espécies de SCN (128 casos) e SCP (45), Staphylococcus aureus(90), Streptococcus agalactiae(70), Streptococcus dysgalactiae (69), Streptococcus uberis(29), Corynebacteriumspp. (230), Klebsiella pneumoniae(28), Klebsiella oxytoca(2), Escherichia coli(15), Enterobactersp. (3). Os resultados de contagem de células somáticas (CCS) relacionados aos SCP e SCN não mostraram diferenças entre as propriedades avaliadas, entretanto com diferenças significantes ao se avaliar a CCS entre os dois grupos de estafilococos, como pode ser evidenciado ao comparar SCN Discreto e SCP exuberante (P<0,01), SCP Discreto e SCP exuberante (P<0,001) e SCN moderado e SCP exuberante (P<0,01). A avaliação da CCS relacionada à intensidade da infecção, considerando-se como crescimento discreto o isolamento de até nove colônias, moderado de dez a 29 colônias e exuberante, com 30 ou mais colônias, revelou para ambos os grupos de estafilococos que quanto maior o número de unidades formadoras de colônias (UFC), a CCS é mais elevada, sendo sempre maior nos casos de SCP. Conclui-se que quando há maior número de UFC, há concomitantemente maior CCS/mL de leite, no caso dos SCP e SCN...

Mastitis is the main affection of cattle intended for dairy production, which significantly impacts the milk production chain, with consequences yet to public health. It was studied aspects related to etiology, cellularity and bacterial count in 10 dairy farms, in the State of São Paulo. There were examined 1148 milking cows, totaling 4584 mammary glands. Cases in which there was isolation coagulase positive staphylococci (CPS) and coagulase-negative staphylococci (CNS) were considered. The results showed microbiota with various different pathogens and CNS (128 cases) and CPS (45), Staphylococcus aureus (90), Streptococcus agalactiae (70), Streptococcus dysgalactiae (69), Streptococcus uberis (29), Corynebacterium spp . (230), Klebsiella pneumoniae (28), Klebsiella oxytoca (2), Escherichia coli (15), Enterobacter spp. (3). The somatic cell count (SCC) related to CPS and CNS showed no differences among the evaluated properties, although with significant differences when evaluating the SCC between both groups of staphylococci, as evidenced by comparing discreet CNS and exuberant CPS (P <0.01), discreet CPS and exuberant CPS (P <0.001) and moderate SCN and exuberant SCP (P <0.01). The evaluation of SCC was related to the intensity of infection, considering how slight growth isolation of up to nine colonies, moderate ten to 29 colonies and lush, with 30 or more colonies, revealed for both groups of staphylococci that the higher the number of colony forming units (CFU), SCC is higher being larger in cases of CPS. The results indicate that the importance of both CPS and CNS when considering the high occurrence of the evaluated properties and the fact that it raises the SCC, which compromises the quality of milk. We conclude that when there is a larger number of CFU, is concomitantly higher SCC/ml of milk, in the case of CPS and CNS...

Inhibiting effect of alcohol extract from Dioscore bulbifera on gastric cancer cells / 天津医药

Objective To investigate the inhibiting effects of alcohol extract from Dioscore bulbifera on proliferation, colony formation and migration of cancer cell lines. Methods Alcohol extract from Dioscore bulbifera was prepared using Soxhlet extraction. Human gastric cancer cell line MGC803 was treated with different concentrations(0, 60, 120 mg/L)of al?cohol extract from Dioscore bulbifera. In vitro, proliferation, colony formation and migration of gastric cancer cells were detect?ed by MTT, colony formation experiments and Transwell assay respectively. Results The proliferation(day2-day 6, F=29.130, 21.864, 67.826, 36.015, 43.656, P<0.01)and colony formation(F=11.918,P<0.01)of gastric cancer cells were significantly inhibited by administration of alcohol extract from Dioscore bulbifera at both 60 mg/L and 120 mg/L . The migra?tion(F=4.258,P<0.05)of gastric cancer cells were significantly suppressed after cells were treated with120 mg/L alcohol ex?tract from Dioscore bulbifera. Conclusion Alcohol extract from Dioscore bulbifera significantly inhibit proliferation, colony formation and migration of gastric cancer cells.

Mechanism of embryonic stem cells microenvironment enhancing stemness and inhibiting apoptosis of human limbal stem cells / 中华实验眼科杂志

Background The fate of adult stem cells is associated with its surrounding microenviroment.Our previous work found that embryonic stem cells (ESCs) micro-environment enhance the stemness of human limbal stem cells (LSCs),but its mechanism has not been elucidated.Objective This study was to explore the molecular mechanism of ESC micro-environment enhancing the stemness and inhibiting the apoptosis of LSCs.Methods Human LSCs were cultured by explant culture method with CnT-20 medium and CnT-20+20％ ES culture supernatant (ESC-CM),respectively.Colony formation assay was used to analyze the proliferation ability of cells.Telomerase reverse transcriptase (TERT) siRNA (19-25nt siRNA) or siRNA (sc-37007) was transfected into the cells of ESCCM group.Apoptosis and mitochondrial membrane potential were assayed by flow cytometry,and the expressions of telomerase and reactive oxygen species (ROS) in TERT siRNA-or siRNA-F-transfected cells by immunofluorescence and flow cytomery.RT-PCR,immunofluorescence staining and Western blot were employed to determine the expressions of p63,ATP-binding cassette transporer G2 (ABCG2),integrin β1 mRNA and proteins and cytokeratin 3 (C K3) in the cells.The levels of focal adhesion kinase (FAK),Akt,glycogen synthase kinase 3β (GSK3β) and p21 protein and phosphorylation proteins in the cells were detected by Western blot.Results The LSCs presented an increased proliferative capacity and passaged to the eighth generation with the colony-forming efficiency (CFE) of (7.6±0.6) ％ in ESC-CM group,but the cells to the sixth generation with the CFE of (5.6±0.6)％,showing a significant difference between them (t =4.454,P =0.011).The apoptotic rates of the cells from 2 through 6 generations were lower in the ESC-CM group than those in the CnT-20 group (all at P＜0.05).The apoptotic rate of the cells was (7.67± 1.31)％ in the siRNA-F transfected group,which was significantly lower than (32.33 ±3.13)％in the siRNA-TERT transfected group (t =-12.588,P =0.000).No significant differences were seen in the expression levels of p63,ABCG2,integrin β1 mRNA and proteins and TERT protein in the primary cells between the ESC-CM group and the CnT-20 group (all at P＞0.05),but significantly declined expressions of CK3 mRNA and protein were found in the ESC-CM group compared with the CnT-20 group (all at P＜0.01).However,the expressions of p63,ABCG2,integrin β1 mRNA and proteins and TERT protein in the second generation of the cells were significantly higher in the ESC-CM group compared with the CnT-20 group (all at P＜0.01).The telomerase activity was (4.83±0.67) ％ in the siRNA-TERT transfected group,which was significantly lower than (46.71±1.22) ％ of the siRNA-F transfected group (t =52.116,P =0.000).The expression of pFAK,pAkt,pGSK3β proteins were weakened,but the expression of p21 was increased in the ESC-CM group after addition of FAK inhibitor,GSK3β inhibitor and TERT-siRNA transfected group.Mitochondrial membrane potential in the second generation of cells was elevated in the ESC-CM group in comparison with the CnT-20 group and the siRNA-TERT transfected group (all at P＜0.01),and the rates of ROS positively reaction was lower in the ESC-CM group and the siRNA-F transfected group than those of the CnT-20 group and siRNA-TERT transfected group (all at P＜0.01).Conclusions ESC-CM culture system can effectively keep the stemness of LSCs and inhibit apoptosis.ESC-CM culture system plays functions probably via telomerase-p21-mitochondrial axis and the activation of the FAK/Wnt signaling pathways.

The feasibility of IFN-γ and IL-10 as surrogate markers to evaluating the activity of drugs against mycobacterium tuberculosis / 中国药学杂志

OBJECTIVE: To study the probability of IFN-γ and IL-10 being surrogate markers for evaluating the anti-TB activity of drugs. METHODS: 75 BALB/c mice infected with H37Rv using a Glas-col inhalation exposure system, were randomized into 13 groups. After 2 months of therapy, the numbers of CFU in the lungs and the levels of IFN-γ and IL-10 in blood were determined. The relevance of IFN-γ, IL-10 concentrations, the ratio of IFN-γ/IL-10 and CFU were evaluated respectively. RESULTS: (1) CFU enumeration: the CFU of treatment groups except Lfx and PAS were significantly different from that of negative control group; the CFU of every treatment groups were significantly different from that of positive group. (2) Levels of IFN-γ: the levels of IFN-γ in peripheral blood of treatment groups except Lfx and PAS were significantly different from that of negative control group; the IFN-γ of every treatment groups were significantly different from that of positive group. (3) IL-10: There were differences exiting neither between treatment groups and negative group nor between any two treatment groups. (4) HFNγ/IL-10: significantly differences were displayed between CLF, LZD+Pto, LZD+CLF, LZD+Am, HRZ and negative group respectively, while no significantly differences showed between any pair of treatment groups, (5) positive corrections were displayed between IFN-γ, IL-10, IFN-γ/IL-10 and CFU, certainty factor(r2) were 0.688, 0.237, 0.582. CONCLUSION: The levels of IFN-γ may be used to evaluate the anti-TB activity of drugs and may not be the surrogate markers to predict the CFU in vivo.

Construction of CIK cell vehicles carrying oncolytic virus and preliminary study on its antihepatoma ability / 肿瘤

Objective: To construct cytokine-induced killer (CIK) cell vehicles carrying recombinant adenovirus carrying tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene, and preliminarily observe its anti-hepatoma ability. Methods: Lymphocytes were isolated from peripheral blood to culture CIK cells. The phenotypic identification of CIK cells was performed by flow cytometry (FCM). Then, the lentiviral pLenti-hCD40L-E1AB containing CD40L promoter and the recombined adenovirus vector pAd5/35-TRAIL were constructed, respectively. The two viruses were infected into CIK cells by two-step method. After that, the secretory function and proliferative capacity of CIK cells as well as the effect on angiogenesis and the ablility of colony-formation of hepatoma cells were assessed by ELISA, MTT method, Tubule formation assay and soft agarose assay, respectively. Results: The CIK lymphocytes grew vigorously, in which the expressions of CD3, CD56, CD11a and CD226 were positive, while the expressions of CD8 and CD305 were negative. The lentiviral pLenti-hCD40L-E1AB containing active hCD40L promoter and adenovirus E1 gene was successfully constructed, and the recombined adenovirus vector pAd5/35-TRAIL containing human TRAIL gene was also constructed. In CIK cells infected with Ad5/F35-TRAIL and pLenti-hCD40L-E1AB, the expression level of interferon-? was significanly increased (P < 0.05), and the angiogenesis and the colony-formation rate of hepatoma cells were inhibited, but the proliferative capacity of CIK cells was less affected. Conclusion: CIK cell vehicles carrying adenovirus and expressing TRAIL gene are successfully constructed. The growth inhibition of hepatoma cells may be induced by CIK cells. Copyright © 2013 by TUMOR.

Measuring of Mycobacterium tuberculosis growth: a correlation of the optical measurements with colony forming units

The quantification of colony forming units (cfu), turbidity, and optical density at 600 nm (OD600) measurements were used to evaluate Mycobacterium tuberculosis growth. Turbidity and OD600 measurements displayed similar growth curves, while cfu quantification showed a continuous growth curve. We determined the cfu equivalents to McFarland and OD600 units.

Aeromycological study at the intensive care unit of the "Dr. Manuel Gea Gonzalez" General Hospital

INTRODUCTION: An aeromycological study verifies the presence and quantifies the concentration of fungal propagules in the air. It is very important in the hospital setting because of the increasing numbers of immunosuppressed and severely ill patients. The objective of this study was to determine the concentration of fungi in the air of the intensive care unit (ICU) of "Dr. Manuel Gea González" General Hospital. METHODS: This is a descriptive, observational cross-sectional study. Air samples were obtained with a single stage Thermo-Andersen Viable Particle Sampler (Thermo Electron Corporation -Massachusetts, U.S.A.) in a Petri dish with potato dextrose agar for 15 minutes at two different times (morning and afternoon) and heights (1 and 1.5 meters). The Petri dishes were incubated for five to seven days at 27ºC, the number of colonies was counted, and the total CFU/m³ was determined. The isolated fungal genera were identified by morphological features. Epi Info v. 3.4.3 © was used for statistical analysis. RESULTS: The mean concentration of fungi in the air of the ICU was 85.08 ± 29.19 CFU/m³; while in the outside air it was 84.3 ± 17.23 CFU/m³ (p = 0.96). The fungi isolated were: Cladosporium spp., Penicillium spp., Aspergillus spp. (non-fumigatus), Fusarium spp., Exophiala spp., Syncephalastrum spp., and Acremonium spp. DISCUSSION: Fungal spores were found in the air of the ICU and Cladosporium spp. was the most frequently isolated fungi. There was no difference according to sampling time or height.

Increased Circulating Endothelial Microparticles and Carotid Atherosclerosis in Obstructive Sleep Apnea

BACKGROUND AND PURPOSE: Endothelial impairment is a linking mechanism between obstructive sleep apnea (OSA) and cardiovascular diseases. Profiles of endothelial microparticles (EMPs) and endothelial progenitor cells (EPCs) reflect the degree of endothelial impairment. The aims of this study were to measure the levels of EMPs and progenitor cells in OSA, determine the correlations between these factors and OSA severity and the degree of atherosclerosis, and document any changes in these factors after therapy. METHODS: Subjects with (n=82) and without (n=22) OSA were recruited prospectively. We measured the number of colony-forming units (CFU) in cell culture as the endothelial progenitor cell index, and the number of EMPs using flow cytometry with CD31 [platelet endothelial cell adhesion molecule (PECAM)], CD42 (platelet glycoprotein), annexin V, and CD62E (E-selectin) antibodies at baseline and after 4-6 weeks of continuous positive airway pressure (CPAP) therapy. Carotid intima-media thickness (IMT) was regarded as a marker of atherosclerosis. RESULTS: The levels of PECAM+CD42- (p<0.001), PECAM+annexin V+ (p<0.001), and E-selectin+ microparticles (p=0.001) were higher in OSA subjects than in non-OSA subjects. The number of CFU did not differ between the two groups. OSA severity independently predicted the levels of PECAM+CD42- (p=0.02) and PECAM+annexin V+ (p=0.004). Carotid IMT was correlated with OSA severity (p<0.001), PECAM+CD42- (p=0.03), and PECAM+annexin V+ (p=0.01). Neither OSA severity nor carotid IMT was correlated with either the number of CFU or E-selectin+. CPAP therapy decreased the occurrence of E-selectin+ (p<0.001) in 21 of the OSA subjects, but had no effect on the other microparticles of the number of CFU. CONCLUSIONS: OSA led to the overproduction of EMPs, which moderately correlated with OSA severity and the degree of atherosclerosis, and partly responded to therapy. The endothelial impairment might contribute to future cardiovascular events.