Cultivo de Acanthamoeba spp. en agua apirógena / Acanthamoeba spp. culture in pyrogen-free water / Cultura de Acanthamoeba spp. em água apirogênica

Resumen El objetivo de este estudio fue evaluar el cultivo de cepas de Acanthamoeba spp. en agua destilada estéril apirógena de uso farmacéutico. Se utilizaron dos cepas de genotipo T4 [un aislamiento de encefalitis granulomatosa amebiana (EGA) y una ambiental] y cepas correspondientes a los genotipos T5 y T15. Los quistes de cada una de las cepas se sembraron en placas de Petri con agar no nutritivo con diferentes soluciones (agua destilada estéril apirógena uso médico para preparaciones inyectables, agua destilada filtrada, medio Page) y combinados con suspensiones de Escherichia coli. Las placas se incubaron a 37 °C y se monitorearon diariamente durante 15 días para la detección de trofozoítos. El crecimiento se evaluó mediante examen microscópico directo. Cada cultivo contó con cuatro repeticiones para cada una de las cepas (n=96). En conclusión, se hallaron diferencias estadísticamente significativas en el crecimiento de las cepas por día. Las cepas T5 y T4 (encefalitis amebiana granulomatosa) desarrollaron mayor cantidad de trofozoítos en el primer día respecto de la cepa T15 (H=16,42; p=0,001). En el agua apirógena con E. coli se obtuvo un crecimiento igual a la solución de Page con E. coli (H=24,64; p=0,0001). No se hallaron diferencias estadísticamente significativas en la cantidad de trofozoítos obtenidos en agua apirógena con E. coli y solución de Page con E. coli en la cepa T4 (EGA) (U=4; p<0,05) pero sí en la cepa T4 ambiental (U=0; p<0,05).

Abstract The objective of this study was to evaluate the culture of strains of Acanthamoeba spp. in sterile apyrogenic distilled water for pharmaceutical use. Two T4 genotype strains [one isolate of granulomatous amebic encephalitis (GAE) and one environmental], a T5 and T15 genotype strains were used. The cysts of each of the strains were seeded in Petri dishes with non-nutritive agar with different solutions (pyrogenic sterile distilled water for medical use for injectable preparations, filtered distilled water, Page medium) and combined with Escherichia coli suspensions. Plates were incubated at 37 °C and monitored daily for 15 days for the detection of trophozoites. Growth was assessed by direct microscopic examination. Each medium culture counted four replicates for each of the strains (n=96). Concluding, statistically significant differences were found in the growth of the strains per day. Strains T5 and T4 (granulomatous amebic encephalitis) developed a greater number of trophozoites on the first day compared to strain T15 (H=16.42; p=0.001). In apyrogenic water with E. coli, a growth equal to Page's solution with E. coli was obtained (H=24.64; p=0.0001). No statistically significant differences were found in the amount of trophozoites obtained in apyrogenic water with E. coli and Page's solution with E. coli in strain T4 (GAE) (U=4; p<0.05), but significant differences were found in the environmental T4 strain (U=0; p<0.05).

Resumo O objetivo deste trabalho foi avaliar o cultivo de cepas Acanthamoeba spp. em água destilada apirogênica estéril para uso farmacêutico. Foram utilizadas duas cepas de genótipo T4 [um isolamento de encefalite granulomatosa amebiana (EGA) e uma ambiental], e cepas correspondentes aos genótipos T5 e T15. Os cistos de cada uma das cepas foram semeados em placas de Petri com ágar não nutritivo com diferentes soluções (água destilada estéril apirogênica para uso médico para preparações injetáveis, água destilada filtrada, meio Page) e combinados com suspensões de Escherichia coli. As placas foram incubadas a 37 °C e monitoradas diariamente durante 15 dias para detecção de trofozoítos. O crescimento foi avaliado através de exame microscópico direto. Cada cultura contou com quatro réplicas para cada uma das cepas (n=96). Em conclusão, diferenças estatisticamente significativas foram encontradas no crescimento das cepas por dia. As cepas T5 e T4 (encefalite amebiana granulomatosa) desenvolveram maior número de trofozoítos no primeiro dia em relação à cepa T15 (H=16,42; p=0,001). Em água apirogênica com E. coli, foi obtido crescimento igual ao da solução de Page com E. coli (H=24,64, p=0,0001). Não foram encontradas diferenças estatisticamente significativas na quantidade de trofozoítos obtidos em água apirogênica com E. coli e solução de Page com E. coli na cepa T4 (EGA) (U=4; p%<0,05), mas sim na cepa T4 ambiental (U=0; p<0,05).

Inhibitory effect of distilled water on viability of human lens epithelial cells in vitro / 中华实验眼科杂志

Objective:To investigate the effect of distilled water on the viability of human lens epithelial cells (LECs) cultured in vitro. Methods:A total of 156 anterior capsule specimens were collected from 156 patients (156 eyes) who were diagnosed with age-related cataract during phacoemulsification combined with intraocular lens implantation from May to December 2020 in Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School.The 156 specimens were divided into 312 small pieces.Of the 312 pieces, 157 pieces were divided into normal control group (23 pieces), positive control group (10 pieces), balanced salt solution (BSS) immersion group (61 pieces) and distilled water immersion group (63 pieces) using computer-generated random numbers.Normal control group received no treatment.Positive control group was directly fixed with a mass fraction of 4% histiocytes fixative solution.For the 61 pieces in BSS immersion group, 20 pieces were soaked for 1 minute, 21 pieces for 2 minutes, and 20 pieces for 3 minutes.For the 63 pieces in distilled water immersion group, 20 pieces were soaked for 1 minute, 23 pieces for 2 minutes, and 20 pieces for 3 minutes.Another 125 pieces were selected to simulate the cataract aspiration-irrigation according to the treatment in BSS immersion group and distilled water immersion group respectively, plus rinse in a bottle containing BSS at a height of 70 cm for 1 minute.Cell viability was detected by trypan blue-eosin staining.LECs density, dead cell count, cell death rate and percentage of shedding (%) were calculated.Of the remaining 30 pieces, every 15 pieces were divided into normal control group, BSS immersion group, and distilled water immersion for 1, 2 and 3 minutes groups, with 3 pieces in each group.BSS immersion group was immersed for 3 minutes, and the other four groups were treated as mentioned above, and the LECs structure of the four groups was observed by light microscopy and transmission electron microscopy.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School (No.2019-248-01). Written informed consent was obtained from each subject.Results:The boundaries of LECs in BSS treatment groups were clear, and there was no significant difference in morphology compared with normal control group.With time increasing, LECs in distilled water treatment groups gradually swelled, and the boundaries of dead cells were not clear.There were significant differences in LECs density, dead LECs count and LECs mortality ( F=13.459, 98.918, 130.600; all at P<0.001). The LECs density was lower in 2-minute and 3-minute distilled water treatment groups than in normal control group, showing statistically significant differences (both at P<0.05). The dead LECs count and LECs mortality were higher in 1-minute, 2-minute and 3-minute distilled water treatment groups than in normal control group and BSS treatment groups for the same time, and the differences were statistically significant (all at P<0.05). Only a few shed LECs were seen in normal control group, 1-minute, 2-minute and 3-minute BSS treatment groups, and BSS immersion combined rinse group.After different time of soaking, there were more shed LECs in distilled water immersion combined rinse group, and the range of LECs shedding increased with the extension of distilled water immersion.There was a significant difference in the shedding percentage of LECs among different groups ( F=123.670, P<0.001). The shedding percentages of LECs at different time points were higher in distilled water immersion groups and distilled water immersion combined rinse groups than in normal control group, and the difference was statistically significant (all at P<0.05). The shedding percentage of LECs increased significantly in distilled water immersion groups with the extension of immersion.Light microscopy showed that the cells were destroyed in 1-minute, 2-minute and 3-minute distilled water treatment groups, and some LECs shed in the 2-minute and 3-minute treatment groups.Transmission electron microscopy showed cell lysis and destruction, suborganelles swelling, disruption of intercellular junctions in 1-minute, 2-minute and 3-minute distilled water treatment groups, loose attachment between cells and capsule in the 2-minute and 3-minute treatment groups, and cell detachment from capsule in the 3-minute treatment group. Conclusions:Distilled water immersion leads to LECs death in a time-dependent manner, and distilled water immersion combined with rinse can remove LECs on the lens capsule.

Antibacterial efficacy of Carica papaya leaf extract, probiotics, kidodent, and placebo mouthwashes in reduction of salivary Streptococcus mutans A double-blinded parallel designed randomized controlled trial

Aim: To compare the antibacterial efficacy of Kidodent, Probiotics, and Carica papaya Leaf extract mouthwashes in reducing Streptococcus mutans count in 8–12 years' old school children. Methodology: Sixty children of age group of 8–12 years were nominated and grouped as Group A (Kidodent mouthwash), Group B (probiotics mouthwash) Group C (C. papaya leaf extract mouthwash), and Group D (distilled water placebo). Probiotics sachets (Prebact) of about 1 g were diluted in 10 ml of water and given as mouthwash. C. papaya leaf extract was obtained by Soxhlet extraction using ethanol as a solvent. Participants were asked to rinse with mouthwashes for 30 s once daily for up to 15 days. Saliva samples were collected and inoculated using Salivarius Mitis and Agar Agar Type I at 38°C for 24 h and incubated, later colony-forming units per milliliter were determined by serial dilution and calculated using colony counter manually. Statistical Analysis: Data were statistically analyzed using the one-way ANOVA and t-test using the SPSS V.20 software. Results: Probiotics and C. papaya leaf extract mouthwashes were equally effective as Kidodent in reducing S. mutans count in saliva. Conclusion: Probiotics and C. papaya leaf extract mouthwashes manifested potential efficacy in reduction of S. mutans.

A comparative study on the effect of nano-pearl powder ground with different dispersion media / 中国组织工程研究

BACKGROUND: To retain more biological activity of organic matter and materials, it is necessary to grind and refine the pearl powder by physical method. The ball grinding method can retain the organic matter in the pearl powder and its activity to the greatest extent. The nanomaterials prepared by ball milling in different dispersion media exhibit different effects. OBJECTIVE: To compare nano-pearl powder milled with distilled water and anhydrous ethanol. METHODS: Nano-pearl powder was prepared by grinding with anhydrous ethanol and water as dispersion medium respectively. The prepared nano-pearl powder was compared by scanning electron microscope, transmission electron microscope, X-ray diffraction, Kjeldahl method and by determining amino acid content in foods. RESULTS AND CONCLUSION: (1) The nano-pearl powder prepared with anhydrous ethanol as dispersion medium was mainly round particles of different sizes (range, 30-50 nm), with the average grain size of 20 nm. The relative percentage of calcite calcium carbonate increased to 7%. The contents of protein and amino acid did not change obviously. (2) The nano-pearl powder prepared with distilled water as dispersion medium was mainly round particles of different sizes with the average grain size of 30 nm. There were irregular grain-like or block-like particles. The relative percentage of calcite calcium carbonate increased to 10%. The contents of protein and amino acid decreased. (3) These results showed that there was a significant difference in the particle size of the pearl powder ground with distilled water and anhydrous ethanol. The pearl powder prepared with anhydrous ethanol as the dispersion medium had a finer more uniform particle size.

The efficacy of 3 irrigating solution in surgical removal of lower wisdom tooth: a pilot study

Surgical removal of impacted lower wisdom tooth has become increasingly costly to patient while still remains as the most common dental surgical procedure that is performed on outpatient basis. In the present study, a total of 23 patients with impacted lower wisdom tooth were surgically removed under local anaesthesia by using different irrigating solution namely, normal saline, distilled water and chlorhexidine. The samples underwent standard operating procedures and medication. Post operative complications in terms of pain, swelling, infection and delayed wound healing were assessed and compared on Day 1 and Day 7 after surgery. The result of this study showed that there is no significant difference between the three irrigating solution used in surgical removal of impacted lower wisdom tooth in terms of postoperative complication. A bigger scale of research with more samples is recommended to evaluate the most efficacy irrigating solution during surgical removal of impacted lower wisdom tooth.

Water and Physiological Saline to Prevent the Formation of P-Chloroaniline / Agua y Suero Fisiológico para Prevenir la Formación de Paracloroanilina

This study determined if p-chloroaniline (PCA) can be minimized by using distilled water and physiological saline solution following sodium hypochlorite but before chlorhexidine. Hypochlorite 5%, 0.5%, 0.05%, 0.005% and 0.0005% dissolved in 0.9% NaCl and in distilled water were mixed with 2% chlorhexidine for the formation of PCA. The PCA was determined using UV-VISIBLE spectrometry, with spectral curve was determined the wavelength of maximum absorption of PCA. Formed PCA absorbance was measured between 0.025%, 0.02%, 0.015%, 0.01%, 0.005% and 0.0025% hypochlorite and 2% chlorhexidine. 2% chlorhexidine and hypochlorite with physiological saline form a white precipitate which prevents the measurement of PCA. Colored PCA is formed with 0.05%, 0.005% hypochlorite aqueous dilutions and 2% chlorhexidine. The lwavelength of maximum absorption obtained was 470 nm and absorbance of PCA showed a linear decrease. 0.005% NaClO produces the least amount of PCA. The best solvent to prevent the formation of PCA during the interaction of sodium hypochlorite with chlorhexidine is distilled water.

Este estudio determinó si la p-cloroanilina (PCA) puede ser minimizada mediante el uso de agua destilada y solución salina fisiológica seguido de la aplicación de hipoclorito de sodio, previo a la aplicación de clorhexidina. Hipoclorito al 5%, 0,5%, 0,05%, 0,005% y 0,0005% fue disuelto en 0,9% de NaCl y en agua destilada se mezcló con 2% de clorhexidina para la formación de PCA. El PCA se determinó mediante espectrometría UV-Visible, y con curva espectral se determinó la longitud de onda máxima del PCA. La absorbancia del PCA formado se midió con 0,025%, 0,02%, 0,015%, 0,01%, 0,005% y 0,0025% de hipoclorito y 2% de clorhexidina. La combinación de 2% de clorhexidina e hipoclorito en solución salina fisiológica forman un precipitado blanco que impide la medición del PCA. El PCA coloreado es formado con 0,05%, 0,005% diluciones acuosas de hipoclorito y 2% de clorhexidina. La longitud de onda máxima obtenida fue de 470 nm y la absorbancia del PCA mostró una disminución lineal. NaClO al 0,005% produce menor cantidad de PCA. El mejor disolvente para evitar la formación de PCA durante la interacción de hipoclorito de sodio con clorhexidina es agua destilada.

Conservación de cultivos de hongos de importancia médica en agua destilada / Preservation of fungal cultures of medical importance in distilled water

Introducción: en la colección de cultivos de hongos patógenos del Instituto de Medicina Tropical Pedro Kourí, como centro de referencia nacional, se conservan los agentes causales de las principales micosis humanas, con fines de diagnóstico, investigaciones y enseñanza de la microbiología médica. La conservación de cultivos fúngicos en agua destilada estéril o método de Castellani ha sido avalada como un método que garantiza porcentajes elevados de viabilidad, pureza y estabilidad de las cepas; esto unido a su bajo costo y sencillez, la convierte en una alternativa ventajosa para el mantenimiento de los microorganismos en muchos laboratorios. Objetivo: mostrar los resultados en la preservación en agua destilada estéril de las principales especies fúngicas que integran esta colección. Métodos: se evaluó la viabilidad, pureza y estabilidad de las principales características morfológicas y fisiológicas de 240 cepas de diferentes especies fúngicas pertenecientes a la colección, conservadas en agua destilada estéril. Resultados: de los cultivos, 80 por ciento se conservó en estado viable y sin contaminaciones. Los mejores resultados se obtuvieron en la preservación de los hongos productores de abundantes conidios (97 por ciento de recuperación) y las levaduras (83 por ciento), mientras que con los dermatofitos y hongos dimórficos fue de 69 y 68 por ciento, respectivamente. En 17 géneros, la recuperación de cepas viables fue superior a 60 por ciento, mientras que en 8 resultó de 100 por ciento. El tiempo de conservación fue de 15 a 20 años. Se implementó una base de datos en formato digital de la colección sobre plataforma web. Conclusiones: el método de Castellani es un método de elección para la conservación de cultivos fúngicos en laboratorios de recursos limitados

Introduction: causal agents of the main human myicoses have been preserved in the culture collection of pathogenic fungi at Pedro Kourí Tropical Medicine Institute, a national reference center, with the purpose of using them for medical microbiology diagnoses, research and teaching. Preservation of fungal cultures in sterile distilled water, or Castellani's method, has been endorsed as a method ensuring high rates of strain viability, purity and stability, low costs and great simplicity. It is therefore an advantageous alternative for the maintenance of microorganisms in many laboratories. Objective: present the results obtained from the preservation in sterile distilled water of the main fungal species included in the collection. Methods: an evaluation was conducted of the viability, purity and stability of the main morphological and physiological characteristics of 240 strains of different fungal species from the collection, which had been preserved in sterile distilled water. Results: 80 percent of the cultures had retained their viable status without any contamination. The best results corresponded to the preservation of fungi producing abundant conidia (97 percent recovery) and yeasts (83 percent), followed by dermatophytes (69 percent) and dimorphic fungi (68 percent). In 8 genera, recovery of viable strains was 100 percent, and in 17 it was above 60 percent. Preservation time was 15-20 years. A web-based digital database was created for the collection. Conclusions: Castellani's is the method of choice for the preservation of fungal cultures in resource-limited laboratories

A study on the rapid eradication of tumor cells on the surgical tools used in hepatocellular carcinoma resection / 中华肝胆外科杂志

Objective This study investigated a method to rapidly inactivate tumor cells on surgical instruments intraoperatively.Methods Tumor cells were collected by immersing and washing the surgical instruments in 37 ℃ saline.The precipitation was collected by low speed centrifugation and then was cultured to harvest the tumor cells.The tumor cells were immersed in saline and distilled water of different temperatures for different duration of time.Inverted microscopy was used to investigate the changes in morphology.Results After immersion in 55 ℃ distilled water for 60 seconds,the tumor cells were swollen,the cell membranes disappeared,the sizes of the nuclei were reduced,the chromatin was condensed,and some cells lysed and separated from each other.Additionally,these tumor cells floated in the culture medium and lacked any living cells adhering to the walls of the bottle.In the group of tumor cells treated with 55 ℃ saline for 60 seconds,there were no obvious morphological changes of the tumor cell or nucleus.Conclusion The intraoperative immersion of surgical instruments in 55 ℃ distilled water for 60 seconds could completely inactivate tumor cells.

An Evaluation of The Bio Compatibility of Grade 1 Commercial Pure Titanium For Orthodontic Implants.

The future of orthodontic and prosthodontic research will involve replacement materials and methods that evaluate compatibility of the materials to living tissues. The biocompatibility of grade1 comercially pure titanium is examined by using cytotoxic study. The extraction liquid of test implant material resulted from incubation in the medium at 37degree Celsius for 120 hours. Cells used are rhesus monkey fibroblasts and the experimental alloy did not cause any cytopathic changes in VERO cells.

Interference of the assessment method in pH values of an epoxy-based cement

Introduction :Alkalinization potential is a fundamental property of endodontic epoxy-based cements containing calcium hydroxide. Studies have shown discrepant pH results for same materials at different evaluation periods. A possible reason accounting for these differences may be the assessment procedures. Objective: To evaluate the pH value of an epoxy-based cement (Sealer 26) in different periods of analysis, using two assessment methods. Material and methods:Sealer 26 was manipulated and immediately placed into polyethylene tubes (n=10, each group) and immersed in distilled water. In G1, the tubes were kept in the same water during all experiment; and in G2, the tubes were removed and placed into another flask with an equal amount of water after the pH evaluation. The pH of these solutions was measured at 24 hours, 7, 14 and 28 days. Analysis were made within the same group according to the experimental periods and between groups in each experimental period. Data were submitted to ANOVA (a = 5%) and t test, respectively.Results:For G1 and G2, all periods showed different pH values (p < 0.05), except between 14 and 28 days (p > 0.05) and between 7 and 14 days (p > 0.05), respectively. In each period, no significant differences were observed between the groups.Conclusion: The method to obtain the pH values in different experimental periods no interfered in the final results. However, difference was observed when the results were analyzed at same group.

Conservación de cultivos fúngicos de alto riesgo de Histoplasma y Cryptococcus / Preservation of high risk fungal cultures of Histoplasma and Cryptococcus

Introducción: las colecciones de cultivos microbianos son las encargadas de garantizar el material biológico requerido para el desarrollo de las ciencias biológicas. Entre los métodos de conservación de cultivos fúngicos, la inmersión en agua destilada, por su bajo costo y sencillez, constituye una ventajosa alternativa. Objetivo: evaluar la utilidad de este método de conservación en los cultivos fúngicos de Histoplasma y Cryptococcus. Métodos: se realizó una evaluación del estado de conservación de las especies de mayor riesgo biológico, pertenecientes a los géneros Histoplasma y Cryptococcus de la colección de cultivos de hongos del Instituto de Medicina Tropical "Pedro Kourí". Se analizaron 102 cepas conservadas en agua destilada, de las cuales 92 por ciento estaba preservado por más de 10 años. Resultados: los porcentajes de recuperación para H. capsulatum, C. neoformans y C. gattii fueron 64,3; 79,1 y 100 por ciento, respectivamente. Se demostró que este método de conservación resulta satisfactorio para cultivos fúngicos en laboratorios de recursos limitados. Se implementó sobre plataforma web una base de datos digital con la información de interés de la colección. Se hizo una valoración de la importancia del estricto cumplimiento de las medidas de bioseguridad para el trabajo de las colecciones, especialmente frente a patógenos de alto riesgo. Conclusiones: la conservación de cultivos de hongos en agua destilada es un método de gran utilidad en laboratorios de recursos limitados. El trabajo de las colecciones de cultivos debe considerarse una actividad imprescindible para enfrentar los nuevos retos del desarrollo de las ciencias biomédicas.

Introduction: culture collections are responsible for providing the microbial resources for development of biological sciences. Storage in distilled water is one of the easiest and least expensive method for long-term fungal preservation. Objective: to evaluate the usefulness of this preservation method in fungal culture of Histoplasma and Cryptococcus. Methods: the preservation condition of the highest biological risk species from Histoplasma y Cryptococcus genera, included in the fungal culture collection of "Pedro Kourí" Institute of Tropical Medicine in Havana, was evaluated in this study. One hundred and two strains stored in distilled water, 92 percent of which had been preserved for more than 10 years, were analyzed. Results: the percentages of recovered strains from H. capsulatum, C. neoformans and C. gattii were 64.3 percent; 79.1 percent and 100 percent respectively. This method of preservation proved to be satisfactory for fungal culture in labs with limited financial resources. A web-based database with interesting information about the collection was made. The importance of strict compliance with the biosafety measures in these collections, particularly with high risk pathogens. Conclusions: preservation of fungal cultures in distilled water is a very useful method for laboratories with limited resources. Culture collections should be assumed as an essential activity in order to solve increasing challenges in the development of biomedical sciences.

Dynamic study on fluoride release from glass ionomer cement in distilled water and artificial saliva / 中国基层医药

Objective In this study,the fluorine release behavior of six glass ionomer cements (GIC) samples was investigated in distilled water and artificial saliva.Methods The fluorine release behavior in GIC was measured by gas chromatography.Results Various GIC materials had the maxium fluorine release rate in the beginning,and the rate decreased rapidly.After 56 days,although the fluorine release quantity was in a trend of decreasing,the release process can be retained during a long period.For No3,No5,and No6 sampler,the fluorine release quantity in artificial saliva was lower than distilled water,whereas no significant difference was observed for Nol and No2 materials.The No4 sample produced a larger fluorine release quantity in artificial saliva.It can also be conducted that the immersible time of GIC and the fluorine release quantity met the equation of log ( Y ) =a + b x log ( t ).Conclusion The fluorine release behavior of six GIC matched the logarithmic regulation.

Establecimiento de una línea celular primaria a partir de huevos con embrión de Toxocara canis / Establishing a primary cell line from Toxocara canis embryonated eggs

Introducción. Toxocara canis es el segundo nematelminto más prevalente en perros a nivel regional y entre los tres más frecuentes en algunos países de la región. Debido a que la fuente de contaminación es el perro, éste se convierte en un nematodo con gran potencial zoonótico. Por esta razón, consideramos importante disponer de una línea celular de este helminto para el estudio de los aspectos básicos, así como para el desarrollo de técnicas diagnósticas. Objetivo. Obtener una línea celular primaria a partir de huevos con embrión de T. canis. Métodos. Los parásitos se extrajeron del intestino delgado de perros menores de un año. Las células embrionarias se obtuvieron mediante la embriogénesis de los huevos de los nematodos adultos, en cuatro diferentes medios; dos ricos en sustancias nutritivas, el tercero con formol al 1 % y el cuarto con agua destilada. Las células se obtuvieron mediante disociación mecánica de los huevos con embrión mediante la utilización de jeringas 30G. Resultados. El tiempo estimado de obtención de la línea celular fue de 15 días, en los que siete eran utilizados en la embriogénesis de los huevos. Las células respondieron positivamente a los métodos de crioconservación luego de dos días, e inclusive dos meses después, permitiendo fases de replicación de cuatro pases. Conclusiones. Se logró obtener una línea celular de T. canis a partir de huevos con embrión de este helminto. Esta línea celular ayudará al entendimiento de las relaciones patógenas, posibles blancos terapéuticos y para el desarrollo de métodos diagnósticos.

Introduction: Toxocara canis is the second most prevalent nemathelminthes in dogs at regional level and among the three most frequent in some countries in the region. Due to the fact that the dog is the contamination source, it becomes a nematode with a high zoonotic potential, so we consider it important to be able to use the cell line of this helminth to study the basic aspects, as well as the development of diagnostic techniques. Objective: To obtain a primary cell line from embryonated eggs of T.canis. Methods: The parasites were extracted from the small intestines of dogs under one year old. Embryonic cells were obtained by embryogenesis of the eggs secreted by adult worms in four different media; two were rich in nutrients, one was 1% formaldehyde, and the other was distilled water. The cells were obtained by mechanical dissociation of embryonated eggs using 30G needles. Results: The estimated time for obtaining the cell line was fifteen days, from which seven were used for egg embryogenesis. The cells responded positively to the cryopreservation methods after two days or even two months, allowing a replication phase with four passes. Conclusions: We managed to obtain a cell line from T. canis embryonated eggs. This cell line will help the understanding of pathogenic relationships, potential therapeutic targets and for developing diagnostic methods.

Pseudomonas spp. y Staphylococcus spp. de origen alimentario y su conservación en agua destilada / Pseudomonas spp. and Staphylococcus spp. of food origin and its conservation in distilled water

INTRODUCCIÓN: El agua destilada ha sido utilizada como medio de soporte para preservar cepas fúngicas, fundamentalmente por existir poca información sobre sus beneficios para mantener otros microorganismos. Con esta premisa, se decidió evaluar su utilidad para conservar bacterias, de origen alimentario, en la colección de cultivos microbianos del Instituto Nacional de Higiene, Epidemiología y Microbiología. MÉTODOS: Un total de 12 cepas (seis pertenecientes a Pseudomonas spp. y seis a Staphylococcus spp.) fueron ensayadas. Los datos de viabilidad obtenidos durante el año de conservación fueron procesados con el paquete estadístico SPSS versión 11.5. El análisis estadístico incluyó el análisis de la varianza para la comparación de las medias del recobrado de viables para las variables tiempo de conservación y dilución, y el test de Scheffé de comparaciones múltiples post hoc para la discriminación de las medias. Fueron controladas las características fisiológicas y la respuesta a la tinción de Gram de las cepas. RESULTADOS: No se encontraron diferencias significativas entre los grupos microbianos conservados en agua destilada. Los factores tiempo de conservación y dilución ejercieron su influencia sobre el recobrado de los cultivos. Se obtuvo estabilidad en la recuperación de viables a partir de los siete días de estudio. Se manifestaron diferencias significativas respecto a las diluciones de mayor valor. Se obtuvo 100 por ciento de estabilidad en las características fisiológicas y en la respuesta a la tinción de Gram para todas las cepas. CONCLUSIONES: La conservación en agua destilada resulta adecuada para preservar las cepas de Pseudomonas spp. y Staphylococcus spp. aisladas de muestras de alimentos durante un año con una buena viabilidad.

INTRODUCTION: The distilled water has been used as a support means to preserve fungal strains, mainly due to lack of information on its benefits to maintain other microorganisms. Thus, authors assessed its usefulness to preserve bacteria of food origin in the collection of microbial cultures in the National Institute of Hygiene, Epidemiology and Microbiology. METHODS: A total of 12 strains (six from Pseudomonas spp and six from Staphylococcus spp) were assayed. Data on viability obtained during the year of conservation were processed using the SPSS statistical package version 11.5. The statistical analysis included that of variance to compare the means of recovery of viable for the following variables: time of conservation and dilution, the Scheffé's test of post hoc multiples comparisons for discrimination of means. The physiological characteristics and the response to Gram's tincture of strains were controlled. RESULTS: There were not significant differences among microbial groups conserved in distilled water. The factors time of conservation and dilution had influence on the recovery of the cultures. There was a good stability in vials recovery from the 7 study days. There were significant differences regarding the dilutions of a greater value. It was possible to obtain a 100 percent of stability in the physiological characteristics and in the response to Gram's tincture for all the strains. CONCLUSIONS: The conservation in distilled water is appropriate to preserve the strains of Pseudomonas spp and of Staphylococcus spp isolated from samples of foods during a year achieving a good viability.

Two Cases of Corneal Edema Caused by Distilled Water during Cataract Surgery

PURPOSE: To report two cases of toxic corneal reaction induced by infusion of distilled water into anterior chamber during cataract surgery. CASE SUMMARY: The first case was a 60-year-old female who was inadvertently infused with distilled water for 20 minutes during phacoemulsification in place of balanced salt solution (BSS). The second case was a 70-year-old male who received anterior chamber irrigation with distilled water for approximately 1 minute then and then immediately irrigated with BSS as soon as the mistake was identified. In both cases, topical 1% prednisolone acetate and 5% NaCl solution was immediately administered every hour as well as oral prednisolone at 1 mg/kg for one week after which the dose was slowly tapered. The first case completely returned to normal after 3 months, whereas the second case only requied 1 month to return to pre-surgery conditions. CONCLUSIONS: Patients who were exposed to distilled water within the anterior chamber resulted in corneal endothelial damage and corneal edema proportionate to the amount irrigated. However, The corneal edema gradually healed with treatment and eventually regained translucency without complications, completely.

Research about prevent contamination of water system in department of stomatology by replace holding tank of water and distilled water for each person / 中国实用护理杂志

Objective To know a kind of method to prevent contamination of water system in department of stomatology. Methods Divided 120 patients with dental diseases into the control group and the observation group randomly, there were 60 cases in each group. Replace the holding tank of wa-ter and distilled water per 24 hours was used in the control group, while the method of replace the hold-ing tank of water and distilled water for each person was used in the observation group, and then com-pared the contamination condition of water system between the two groups. Results The percent of pass of water system in the observaiton group was 100%, which significant higher than the 68.33% in the control group. Conclusions Replace the holding tank of water and distilled water for each person is a kind of method to avoid contamination of water system in department of stomatology.

A case of HME obstruction by distilled water from incidentally mounted heated wire circuit kit: A case report / 대한마취과학회지

Heat-moisture exchanger (HME) is an inexpensive and effective device used to prevent respiratory complications that can be caused by endotracheal tube insertion during general anesthesia. But, HME can increase airway resistance and be occluded by the patient's secretions. Whether a HME could be occluded by clear fluids such as condensate in the airway circuit is not certain yet. In vitro, a case of HME occlusion by normal saline was reported. We report a case of HME obstruction by distilled water came from the heated wire circuit which was unintentionally connected to the HME.

Effect of rewetting agent on dentinal microtensile bond strength / 대한치과보존학회지

This study investigated that the effect of rewetting agent on dentinal microtensile bond strength (microTBS). Human molars were sectioned to expose the superficial dentin surfaces. Samples were divided into two groups according to type of adhesives-Single Bond (S) and One-Step (O)], and again subdivided into five groups by different dentin surface treatment-dry for 15s (D), blot dry (BD) or dry for 15s, and rewet with different rewetting agents [distilled water (DW), Gluma Desensitizer (GD) and Aqua-Prep (AP)] for 30s. After application of adhesive, composite resin was built up on the bonding surface. Each tooth was sectioned to obtain stick with 1 mm2 cross sectional area and the microTBS was determined by EZ test. In the S group, the mean microTBS of GD, AP and BD group was significantly higher than that of DW and D group (p < 0.05). In the O group, the mean microTBS of AP, GD, BD and DW group was significantly higher than that of D group (p < 0.05). The data suggested that Gluma Desensitizer and Aqua-Prep could be successfully used as rewetting agents, and Distilled water could be acceptable in aceton based adhesive system only.

A Case of Anterior Chamber Irrigation with Distilled Water During Cataract Operation

PURPOSE: To report a case of anterior chamber irrigation with distilled water during cataract operation. METHODS: During the cataract operation of 56 year-old male patient, corneal edema and anterior chamber hazziness were noted after anterior chamber irrigation with distilled water for a minute. Distilled water was replaced rapidly balanced salt solution (BSS) as irrigation solution and operation was completed. At postoperative one day, corneal edema and anterior chamber exudative membrane were formed. After topical 5% NaCl, 1% prednisolone treatment, corneal edema and exudative membrane disappeared at postoperative sixth week. BCVA was 0.6. At postoperative ninth week, the patient complained of decreased visual acuity. On fundus exmination and flourescein angiography, cystoid macular edema (CME) was detected. RESULTS: After prednisolone oral administration and diclofenac eyedrop instillation, CME improved. At postoperative 24th month, BCVA was 0.8 and CME disappeared. CONCLUSIONS: From our experience of a case of anterior chamber irrigation with distilled water during cataract operation, if balanced salt solution replaces hypotonic solution rapidly as irrigation solution, corneal and other complications are managed properly, long term visual acuity appears good.

Toxic Corneal Reaction Induced by Distilled Water Infused during Cataract Operation

PURPOSE: To report a case of toxic corneal reaction induced by accidentally infused distilled water into the anterior chamber during cataract operation. METHODS: 67-year-old female patient was admitted due to corneal edema and opacity which had been developed instantly by distilled water infused into anterior chamber during cataract operation at a private eye clinic. Cataract surgery was performed after 1 week, and the patient was treated with 5% NaCl solution and artificial eyedrop. RESULTS: On the sixth month after operation, the corneal edema and opacity were decreased remarkably. CONCLUSIONS: When acute corneal edema is developed during cataract operation, the surgeon should stop the procedure and examine all intraocular solutions and irrigation fluids. The prognosis must be estimated after use of hypertonic saline solution at least 6 months.