Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 121
Filter
2.
Rev. peru. med. exp. salud publica ; 36(2): 275-280, abr.-jun. 2019. tab, graf
Article in Spanish | LILACS | ID: biblio-1020808

ABSTRACT

RESUMEN El objetivo del estudio fue comparar diferentes métodos de concentración para recuperar la mayor cantidad de quistes de Giardia spp. a partir de muestras coprológicas. Se analizaron 100 muestras procedentes de hospitales de referencia nacional y se aplicaron cuatro métodos parasitológicos: concentración por sedimentación espontánea en tubo (TSET), Faust, gradiente de sucrosa de una fase y gradiente de sucrosa de dos fases. Se encontró que el método de gradiente de sucrosa de dos fases alcanzó resultados significativamente mejores en concentración de quistes (121 903 quistes/ml) y cantidad de detritos (6%), en comparación con los métodos de Faust (35 355 quistes/ml), concentración por sedimentación espontánea en tubo (20,145 quistes/ml) y gradiente de sucrosa de una fase (18 702 quistes/ml). Se concluye que el método más eficaz para la concentración y purificación de quistes de Giardia spp. a partir de muestras coprológicas es el método de gradiente de sucrosa de dos fases, lo que facilitaría los cultivos in vitro de Giardia spp.


ABSTRACT The aim of this study was to compare different methods of concentration to recover the largest number of Giardia spp. cysts from coprological samples. One hundred (100) samples from national reference hospitals were analyzed and four parasitological methods were applied: spontaneous tube sedimentation concentration (TSET), Faust, single-phase sucrose gradient, and two-phase sucrose gradient. The two-phase sucrose gradient method was found to achieve significantly better results in cyst concentration (121,903 cysts/ml) and amount of debris (6%), compared to Faust methods (35,355 cysts/ml), spontaneous tube sedimentation concentration (20,145 cysts/ml), and single-phase sucrose gradient (18,702 cysts/ml). It is concluded that the most effective method for the concentration and purification of Giardia spp. cysts from coprological samples is the two-phase sucrose gradient method, which would facilitate in vitro culture of Giardia spp.


Subject(s)
Humans , Centrifugation, Density Gradient/methods , Feces/parasitology , Giardia/isolation & purification , Peru
3.
Clinical and Experimental Reproductive Medicine ; : 147-151, 2019.
Article in English | WPRIM | ID: wpr-785645

ABSTRACT

OBJECTIVE: The aim of this study was to investigate DNA fragmentation status in human spermatozoa according to specific tail swelling patterns determined via hypo-osmotic swelling test (HOST).METHODS: Frozen semen samples from 21 healthy donors were thawed and prepared by the swim-up technique for use in intracytoplasmic sperm injection. The semen samples were treated for 5 minutes as part of the HOST procedure and then underwent the sperm chromatin dispersion test using a Halosperm kit. DNA fragmentation status (large halo, medium halo, small halo, no halo, or degraded) and the specific tail swelling pattern (“a”–“g”) were assessed at the level of a single spermatozoon. A total of 42,000 spermatozoa were analyzed, and the percentage of spermatozoa without DNA fragmentation (as evidenced by a large or medium halo) was assessed according to the specific tail swelling patterns observed.RESULTS: The HOST examinations showed that >93% of spermatozoa across all types displayed no DNA fragmentation. The percentage of spermatozoa without DNA fragmentation was 100% in type “d”, 98.67% in type “g”, and 98.17% in type “f” spermatozoa.CONCLUSION: We found that the type “d” spermatozoa displayed no DNA fragmentation, but the other types of spermatozoa also displayed very low rates of DNA fragmentation. This result may be associated with the processing of the spermatozoa by density gradient centrifugation and the swim-up technique.


Subject(s)
Humans , Centrifugation, Density Gradient , Chromatin , DNA Fragmentation , DNA , Infertility , Semen , Semen Preservation , Sperm Head , Sperm Injections, Intracytoplasmic , Spermatozoa , Tail , Tissue Donors
4.
Clinical and Experimental Reproductive Medicine ; : 8-13, 2019.
Article in English | WPRIM | ID: wpr-763350

ABSTRACT

OBJECTIVE: Density gradient centrifugation (DGC) is frequently used to isolate high-motility fractions of spermatozoa. We compared the efficacy of four DGC media in terms of the percentage of morphologically normal spermatozoa, DNA fragmentation level, and hyaluronic acid (HA) binding ability. METHODS: Thirty men with a total motile spermatozoa count >80 million participated. Semen samples were divided into four aliquots, which were processed using PureSperm, PureCeption, Sidney, and SpermGrad media, respectively. The DNA fragmentation level was measured using the Halosperm assay kit and HA binding ability was measured using the HBA assay kit. RESULTS: The mean percentage of morphologically normal spermatozoa was significantly enhanced after DGC using all four media (10.3%, 9.9%, 9.8%, and 10.7%, respectively; p0.05). HA binding ability did not change after DGC using any of the four media. CONCLUSION: The four media were equally effective for obtaining a sperm fraction with highly motile, morphologically normal sperm. PureSperm, PureCeption, and SpermGrad media were equally effective for acquiring a sperm fraction with less DNA fragmentation.


Subject(s)
Humans , Male , Centrifugation, Density Gradient , DNA Fragmentation , DNA , Hyaluronic Acid , Semen , Spermatozoa
5.
Int. j. morphol ; 35(4): 1495-1501, Dec. 2017. tab, graf
Article in English | LILACS | ID: biblio-893161

ABSTRACT

SUMMARY: Freeze/thawing process reduces sperm survival and fertilizing ability of cat spermatozoa, with sperm motility being the most sensitive sperm parameter altered, due to cryo-damage. In this context, swim-up and density gradient processing methods can help to recover high motile and normal spermatozoa. Maximizing the use of frozen semen sample is essential, especially in endangered felids or high value cats in which sample size, number of samples or access to semen collection is reduced. To our knowledge, there is no previous report describing an in depth analysis of sperm motility improvement, after sperm selection techniques in frozen cat semen. Accordingly, we evaluated the effect of percoll gradient (PG) and swim up (SU) sperm selection techniques on sperm motility parameters and sperm recovery rate in frozen/thawed spermatozoa of domestic cat. Next, we evaluated the individual effect of the cat over sperm motility after PG sperm selection of frozen/thawed spermatozoa. SU and PG improved significantly all sperm motility parameters of frozen/thawed cat spermatozoa compared to simple washing. However, PG allows better sperm recovery from the original frozen sample and works mostly homogeneously among individual cats. This new information could help to maximize the use of frozen semen in endangered felids or high value domestic cats for its subsequent application on in vitro fertilization and artificial insemination.


RESUMEN: El proceso de congelación/descongelación reduce la sobrevivencia espermática y la habilidad para fertilizar en los espermatozoides de gato, siendo la motilidad espermática el parámetro más sensiblemente alterado debido al daño por frío. En este contexto, los métodos de procesamiento de swim-up y gradiente de densidad pueden ayudar a recuperar los espermatozoides normales y de alta motilidad. Maximizar el uso de una muestra de semen congelado es esencial, especialmente en felinos amenazados o en gatos de alto valor en los cuales el tamaño de muestra, número de muestras o el acceso a la colecta de semen son reducidos. Para nuestro conocimiento, no hay reportes previos que describan un análisis profundo del mejoramiento de la motilidad luego de técnicas de selección espermática en semen congelado de gato. De acuerdo a esto, evaluamos el efecto de las técnicas de selección espermática gradiente de percoll (PG) y swim up (SU) sobre los parámetros de motilidad y porcentaje de recuperación de espermatozoides congelados/descongelados de gato doméstico. Luego, evaluamos el efecto individual del gato sobre la motilidad espermática luego de la selección espermática con PG en espermatozoides congelados/descongelados. SU y PG mejoraron significativamente todos los parámetros de motilidad espermática de los espermatozoides congelados/descongelados comparado con el lavado simple. Sin embargo, PG permitió una mejor recuperación de espermatozoides desde la muestra congelada original y funcionó en su mayoría de manera homogénea entre los gatos individualmente. Esta nueva información puede ayudar a maximizar el uso del semen congelado en felinos amenazados o en gatos de alto valor para su posterior aplicación en fecundación in vitro e inseminación artificial.


Subject(s)
Animals , Male , Cats , Sperm Motility , Cryopreservation , Sperm Retrieval/veterinary , Semen Analysis/veterinary , Semen Preservation , Image Processing, Computer-Assisted , Centrifugation, Density Gradient , Semen Analysis/methods
6.
IJFS-International Journal of Fertility and Sterility. 2017; 11 (1): 55-62
in English | IMEMR | ID: emr-185831

ABSTRACT

Background: Sperm selection method is usually used to collect these cells for in vitro-assisted reproduction. Few studies reported the relationship of in vivo fertility and semen parameters after sperm selection; hence, the present study attempted to assess different semen parameters after post-thaw or sperm selection, using density gradient separation BoviPure[registered], to predict in vivo fertility


Materials and Methods: In this experimental study, frozen semen quality of four Montbeliarde bulls were assessed after post-thaw [PT] or after sperm selection [SSp], using density gradient separation BoviPure[registered], to predict the fertility rate in vivo. In addition to PT or SSp, semen was examined for concentration, motility, morphology abnormalities, viability, acrosome and plasma membrane integrities. Fertility was measured as non-return rates within 56 days after the first insemination [NRR] or as corrected NRR, expressed as CNRR, to the factors influencing fertility using linear mixed model. Non-parametric Kruskal-Wallis test was performed to compare semen parameter variables. Fertility rates were compared using Chi-square test. Pearson correlation analysis was used to test the relationship between CNRR and semen parameters. Data was analysed using SPSS package program, version 21.0


Results: Most of the examined bulls exhibited a high fertility rate [3/4 bulls, 62.1-81.8% for NRR or 67.2-98.5% for CNRR]. Fertility rate, expressed as CNRR, was significantly related to semen parameters after SSp, but not after PT. Thus, CNRR was increased with decrease of total motility, progressive spermatozoa and abaxial implantation frequencies after SSp [r=-0.999, P=0.001; r=-0.990, P=0.010; r=-0.988, P= 0.012, respectively]; while, CNRR was decreased with decrease of SSp immotile spermatozoa [r=+0.995, P=0.005], underlying that maximal limit of determined immotile spermatozoa is 47%


Conclusion: High frequencies of total and progressive motility spermatozoa, and abaxial implantation in gradient selected sperm appear to be not favorable for fertility in vivo


Subject(s)
Animals , Male , Centrifugation, Density Gradient/veterinary , Fertility , Animals
7.
Clinical and Experimental Reproductive Medicine ; : 201-206, 2017.
Article in English | WPRIM | ID: wpr-226344

ABSTRACT

OBJECTIVE: The aim of this study was to compare the efficacy of swim-up and density gradient centrifugation (DGC) for reducing the amount of sperm with fragmented DNA, sex chromosome aneuploidy, and abnormal chromatin structure. METHODS: Semen samples were obtained from 18 healthy male partners who attended infertility clinics for infertility investigations and were processed with swim-up and DGC. The percentages of sperm cells with fragmented DNA measured by the sperm chromatin dispersion test, normal sex chromosomes assessed by fluorescence in situ hybridization, and abnormal chromatin structure identified by toluidine blue staining were examined. RESULTS: The percentage of sperm cells with fragmented DNA was significantly lower in the swim-up fraction (9.7%, p=0.001) than in the unprocessed fraction (27.0%), but not in the DGC fraction (27.8%, p=0.098). The percentage of sperm cells with normal X or Y chromosomes was comparable in the three fractions. The percentage of sperm cells with abnormal chromatin structure significantly decreased after DGC (from 15.7% to 10.3%, p=0.002). The swim-up method also tended to reduce the percentage of sperm cells with abnormal chromatin structure, but the difference was not significant (from 15.7% to 11.6%, p=0.316). CONCLUSION: The swim-up method is superior for enriching genetically competent sperm.


Subject(s)
Humans , Male , Aneuploidy , Centrifugation, Density Gradient , Chromatin , DNA Fragmentation , DNA , Fluorescence , In Situ Hybridization , Infertility , Methods , Semen , Sex Chromosomes , Spermatozoa , Tolonium Chloride , Y Chromosome
8.
Clinical and Experimental Reproductive Medicine ; : 199-206, 2016.
Article in English | WPRIM | ID: wpr-54502

ABSTRACT

OBJECTIVE: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. METHODS: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. RESULTS: The sperm DFI showed a significant correlation (r=–0.347, p<0.001) with sperm motility and morphology (r=–0.114, p<0.05) but not with other semen parameters. The DFI (11.5%±2.0%) of semen samples was significantly reduced by DGC (8.1%±4.1%) or MACS alone (7.4%±3.9%) (p<0.05). The DFI was significantly further reduced by a combination of DGC and MACS (4.1%±1.3%, p<0.05). Moreover, the combination of DGC and MACS (1.6%±1.1%, p<0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC (4.4%±3.2%) or MACS alone (3.4%±2.2%). CONCLUSION: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.


Subject(s)
Apoptosis , Centrifugation , Centrifugation, Density Gradient , Chromatin , DNA Fragmentation , DNA , Methods , Product Packaging , Semen , Semen Analysis , Sperm Motility , Spermatozoa
9.
National Journal of Andrology ; (12): 432-436, 2016.
Article in Chinese | WPRIM | ID: wpr-262333

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the influence of different methods of semen preservation and processing on sperm DNA integrity.</p><p><b>METHODS</b>We collected semen samples from 100 normozoospermic male volunteers and, following homogeneous mixing, preserved them by means of snap freezing, slow freezing, or at the room temperature for 4 and 24 hours. Meanwhile we processed the semen by washing, swim-up, and density gradient centrifugation (DGC). Then we obtained the sperm DNA fragmentation index (DFI) by sperm chromatin dispersion test and measured total sperm motility and DFI after cultured for 24 hours following processing.</p><p><b>RESULTS</b>The sperm DFIs after 4 hours of preservation by snap freezing, slow freezing, and at the room temperature were (27.3 ± 6.4)%, (26.9 ± 6.1)%, and (24.7 ± 6.8)%, respectively, and that after preserved at the room temperature for 24 hours was (35.6 ± 9.0)%, with statistically significant differences between the first three and the 24-hour room temperature preservation groups (P < 0.05) but not among the former three groups (P > 0.05). The sperm DFI was significantly higher in the samples processed by washing ([13.7 ± 2.0]%) than in those processed by swim-up ([9.1 ± 1.3]%) and DGC ([8.0 ± 2.5]%) (P < 0.05), and it was the lowest in the DGC group after 24-hour culture ([11.5 ± 4.2]%) as compared with the other groups (P < 0.05).</p><p><b>CONCLUSION</b>Sperm DNA integrity is influenced by different semen preservation conditions and processing methods.</p>


Subject(s)
Humans , Male , Centrifugation, Density Gradient , DNA Fragmentation , Semen , Semen Analysis , Semen Preservation , Methods , Sperm Motility , Spermatozoa , Cell Biology
10.
Braz. j. microbiol ; 46(4): 1265-1268, Oct.-Dec. 2015. graf
Article in English | LILACS | ID: lil-769661

ABSTRACT

Abstract Noroviruses (NVs) are responsible for most cases of human nonbacterial gastroenteritis worldwide. Some parameters for the purification of NV virus-like particles (VLPs) such as ease of production and yield were studied for future development of vaccines and diagnostic tools. In this study, VLPs were produced by the expression of the VP1 and VP2 gene cassette of the Brazilian NV isolate, and two purification methods were compared: cesium chloride (CsCl) gradient centrifugation and ion-exchange chromatography (IEC). IEC produced more and purer VLPs of NV compared to CsCl gradient centrifugation.


Subject(s)
Child , Humans , Centrifugation, Density Gradient/methods , Chromatography, Ion Exchange/methods , Norovirus/genetics , Viral Structural Proteins/genetics , Virosomes/isolation & purification , Brazil , Viral Structural Proteins/metabolism , Virosomes/genetics , Virosomes/metabolism
11.
Dental press j. orthod. (Impr.) ; 20(2): 119-125, Mar-Apr/2015. graf
Article in English | LILACS | ID: lil-745849

ABSTRACT

INTRODUCTION: The finite element method (FEM) is an engineering resource applied to calculate the stress and deformation of complex structures, and has been widely used in orthodontic research. With the advantage of being a non-invasive and accurate method that provides quantitative and detailed data on the physiological reactions possible to occur in tissues, applying the FEM can anticipate the visualization of these tissue responses through the observation of areas of stress created from applied orthodontic mechanics. OBJECTIVE: This article aims at reviewing and discussing the stages of the finite element method application and its applicability in Orthodontics. RESULTS: FEM is able to evaluate the stress distribution at the interface between periodontal ligament and alveolar bone, and the shifting trend in various types of tooth movement when using different types of orthodontic devices. Therefore, it is necessary to know specific software for this purpose. CONCLUSIONS: FEM is an important experimental method to answer questions about tooth movement, overcoming the disadvantages of other experimental methods. .


INTRODUÇÃO: o Método de Elementos Finitos (MEF) é um recurso da Engenharia empregado para calcular o estresse e a deformação de estruturas complexas, e tem sido amplamente utilizado nas pesquisas em Ortodontia. Apresenta a vantagem de ser um método não-invasivo e preciso, que fornece dados quantitativos e detalhados acerca das reações fisiológicas que podem ocorrer nos tecidos. OBJETIVO: esse artigo pretende realizar uma revisão da literatura sobre as etapas para realização do Método de Elementos Finitos, bem como de sua aplicabilidade na Ortodontia. RESULTADOS: o MEF é capaz de avaliar a distribuição do estresse na interface entre o ligamento periodontal e o osso alveolar, bem como a tendência de deslocamento em diversos tipos de movimentos dentários, quando utilizados diferentes tipos de aparelhos. Para tanto, é necessário conhecimento de softwares específicos para esse fim. CONCLUSÕES: o MEF é um importante método experimental que pode esclarecer questionamentos acerca da movimentação dentária, superando as desvantagens de outros métodos experimentais. .


Subject(s)
Cell Fractionation/methods , Saccharomyces cerevisiae/chemistry , Centrifugation, Density Gradient , Organelles/chemistry , Reproducibility of Results , Subcellular Fractions , Time Factors
12.
Rev. Soc. Bras. Med. Trop ; 48(1): 18-25, jan-feb/2015. tab, graf
Article in English | LILACS | ID: lil-742970

ABSTRACT

INTRODUCTION: Dengue is the most prevalent arboviral disease in tropical areas. In Mato Grosso, outbreaks are reported every year, but studies on dengue in this state are scarce. METHODS: Natural transovarial infection of Aedes aegypti by a flavivirus was investigated in the Jardim Industriário neighborhood of Cuiabá, Mato Grosso. Eggs were collected with ovitraps during the dry, intermediate, and rainy seasons of 2012. After the eggs hatched and the larvae developed to adulthood, mosquitoes (n = 758) were identified and allocated to pools of 1-10 specimens according to the collection location, sex, and climatic period. After RNA extraction, multiplex semi-nested RT-PCR was performed to detect the four dengue virus (DENV) serotypes, yellow fever virus, West Nile virus and Saint Louis encephalitis virus. RESULTS: DENV-4 was the only flavivirus detected, and it was found in 8/50 pools (16.0%). Three of the positive pools contained females, and five contained males. Their nucleotide sequences presented 96-100% similarity with DENV-4 genotype II strains from Manaus, Amazonas. The minimum infection rate was 10.5 per 1000 specimens, and the maximum likelihood estimator of the infection rate was 11.6 (95% confidence interval: 4.8; 23.3). CONCLUSIONS: This study provides the first evidence of natural transovarial infection by DENV-4 in Ae. Aegypti in Mato Grosso, suggesting that this type of infection might serve as a mechanism of virus maintenance during interepidemic periods in Cuiabá, a city where dengue epidemics are reported every year. These results emphasize the need for efficient vector population control measures to prevent arbovirus outbreaks in the state. .


Subject(s)
Animals , Humans , Mice , Kinesins/metabolism , Protein Biosynthesis , Cell Line , Centrifugation, Density Gradient , Gene Knockdown Techniques , Immunoprecipitation , Interphase , Kinesins/antagonists & inhibitors , Kinesins/genetics , Microtubules/metabolism , Peptide Chain Initiation, Translational , Protein Binding , Pyrimidines/pharmacology , RNA Interference , Ribosomes/metabolism , Thiones/pharmacology
13.
National Journal of Andrology ; (12): 532-535, 2015.
Article in Chinese | WPRIM | ID: wpr-276063

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the influence of the time interval from the end of semen processing to artificial intrauterine in semination with husband's sperm (AIH-IUI) on the rate of clinical pregnancy.</p><p><b>METHODS</b>This study involved 191 AIH-IUI cycles with the same ovulation induction protocol. After Percoll density gradient centrifugation, we divided the sperm into four groups based on the incubation time: 0-19, 20-39, 40-59, and 60-80 min, and again into another four groups according to the total progressively motile sperm count (TPMC): (0-9), (10-20), (21-30), and > 30 x 10(6). We analyzed the correlation of the clinical pregnancy rate with the time interval from the end of sperm processing to AIH-IUI and with other influencing factors, such as maternal age, infertility duration, and semen quality.</p><p><b>RESULTS</b>The rate of clinical pregnancy was significantly higher in the 20-39 min group (18.3%) than in the 0-19, 40-59, and 60-80 min groups (12.7, 11.4 and 9.1%) (all P < 0.05). The (10-20) x 10(6) group achieved a remarkably higher pregnancy rate (16.7%) than the (0-9), (21-30), and > 30 x 10(6) groups (0, 11.4, and 8.3%) (all P < 0.05). Logistic multivariate analysis showed that the rate of clinical pregnancy was decreased with the increased age of the women (OR 0.89, 95% CI 0.83-0.94) but significantly elevated in the 20-39 min group (OR 2.11, 95% CI 1.34-3.13) and of (10-20) x 10(6) group (OR 2.06, 95% CI 1.32-3.46).</p><p><b>CONCLUSION</b>The time interval from the end of sperm processing to AIH-IUI is a most significant factor influencing the rate of clinical pregnancy of AIH-IUI.</p>


Subject(s)
Female , Humans , Male , Pregnancy , Centrifugation, Density Gradient , Infertility , Therapeutics , Insemination, Artificial, Homologous , Pregnancy Rate , Semen , Semen Analysis , Sperm Count , Spermatozoa , Time Factors
14.
Acta Physiologica Sinica ; (6): 423-430, 2015.
Article in Chinese | WPRIM | ID: wpr-255930

ABSTRACT

To improve a fast and high-quality isolation method for culturing the primary cardiomyocyte and fibroblast in vitro, the neonatal Wistar rats were decapitated accordingly and left ventricles were isolated under the sterile condition. The ventricles were chopped and digested in the enzyme solution containing 0.5 mg/mL type II collagenase. During this process, the digesting time, frequency and stirring speed, centrifuging frequency and speed were strictly controlled. The cardiomyocytes were separated from the cardiac fibroblast by using the Percoll density gradient centrifugation. The cell viability was tested by staining with 0.2% trypan blue. The purity of cardiomyocytes and fibroblasts were determined by immunoflourescent staining with anti-cTnI, anti-Vimentin and anti-α-SMA antibodies. The results indicated that with this protocol, the viability and purity of cardiomyocytes were 92% and 95%. The automobile pulse of the adhered cardiomyocyte was visible. For fibroblasts, the cell viability and purity were 96% and 94%. Our results demonstrate that this advanced isolation method is reproducible, and can simultaneously produce high-quality primary cardiomyocytes and fibroblasts for the future study.


Subject(s)
Animals , Rats , Cell Separation , Methods , Cell Survival , Centrifugation, Density Gradient , Fibroblasts , Heart Ventricles , Cell Biology , Myocytes, Cardiac , Povidone , Rats, Wistar , Silicon Dioxide
15.
Clinical and Experimental Reproductive Medicine ; : 22-29, 2015.
Article in English | WPRIM | ID: wpr-64633

ABSTRACT

OBJECTIVE: Sperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF. METHODS: Patients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123). RESULTS: There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates. CONCLUSION: We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.


Subject(s)
Female , Humans , Centrifugation, Density Gradient , Embryonic Structures , Fertilization , Fertilization in Vitro , Infertility , Oocytes , Pregnancy Rate , Reproductive Techniques , Reproductive Techniques, Assisted , Retrospective Studies , Semen , Spermatozoa
16.
Assiut University Bulletin for Environmental Researches. 2015; 18 (2): 45-50
in English | IMEMR | ID: emr-177675

ABSTRACT

A method for isolation and purification of plant viruses was described. The method involved use of organic solvents, differential centrifugation and density gradient centrifugation on step and linear caesium sulphate gradients with which a sucrose cushion was used. The method was found suitable for purification of a range of plant viruses belonging to different virus groups and representing the different shapes reported for plant viruses. The plant viruses purified through this method included Potato virus Y [PVY], Tobacco etch virus [TEV], Alfalfa mosaic virus [AMV], Cucumber mosaic virus [CMV], Tobacco mosaic virus [TMV] and Tomato yellow leaf curl virus [TYLCV]. Both the step and linear gradients of caesium sulphate gave similar results in isolation and purification of these viruses and no appreciable differences were observed between them when either of which was used


Subject(s)
Centrifugation , Sucrose , Centrifugation, Density Gradient , Viruses
17.
National Journal of Andrology ; (12): 225-228, 2015.
Article in Chinese | WPRIM | ID: wpr-319515

ABSTRACT

<p><b>OBJECTIVE</b>To explore the expression patterns of the chemokine CXCL12 and its receptor CXCR4 in human sperm.</p><p><b>METHODS</b>We collected semen samples from 10 fertile men, performed density gradient centrifugation, and then determined the expressions of both CXCL12 and CXCR4 in the sperm by RT-PCR and immunofluorescence staining.</p><p><b>RESULTS</b>RT-PCR revealed the mRNA expressions of CXCL12 (0.641 +/- 0.180) and CXCR4 (0.464 +/- 0.100) in the sperm. However, only CXCR4 rather than CXCL12 was expressed at the protein level, and the positive staining for CXCR4 was observed mainly in the posterior part of the acrosome.</p><p><b>CONCLUSION</b>CXCL12 and CXCR4 are involved as important molecules in regulating the function of human sperm.</p>


Subject(s)
Humans , Male , Acrosome , Metabolism , Centrifugation, Density Gradient , Chemokine CXCL12 , Metabolism , Receptors, CXCR4 , Metabolism , Signal Transduction , Spermatozoa , Metabolism
18.
Braz. dent. j ; 25(6): 571-575, Nov-Dec/2014. graf
Article in English | LILACS | ID: lil-732261

ABSTRACT

This case report describes root canal filling performed over a large S1 ProTaper file fragment in a second mandibular molar with irreversible pulpitis. An S1 ProTaper file was fractured during the instrumentation of the mesiobuccal canal. Approximately 10 mm of file fragment remained in the apical and middle thirds of the canal. The obturation was performed over this fragment using thermomechanically compacted gutta-percha and sealer. Radiographic findings and the absence of clinical signs and symptoms at 3-year follow up indicated successful treatment. Cone-beam computed tomography images revealed absence of periapical lesion and details of intracanal file fragment related to root fillings and apex morphology. In this case, the presence of a large intracanal fractured instrument did not have a negative impact on the endodontic prognosis during the follow up evaluation period.


Este relato de caso descreve a obturação do canal radicular realizada sobre um grande fragmento da lima ProTaper S1 em um segundo molar inferior com pulpite irreversível. Uma lima ProTaper S1 fraturou durante a instrumentação do canal mésio-vestibular. Aproximadamente 10 mm de remanescente do fragmento da lima permaneceu nos terços apical e médio do canal. A obturação foi realizada sobre este fragmento usando guta-percha compactada termomecanicamente e cimento endodôntico. Achados radiográficos e ausência de sinais e sintomas clínicos após 3 anos de acompanhamento indicaram o sucesso do tratamento. Imagens de tomografia computadorizada de feixes cônicos revelaram a ausência de lesão periapical e detalhes do fragmento da lima intracanal relacionados à obturação do canal radicular e à morfologia do ápice. Neste caso, a presença de grande instrumento fraturado intracanal não teve impacto negativo no prognóstico endodôntico durante o período de acompanhamento.


Subject(s)
Bacteriological Techniques , Campylobacter/ultrastructure , Centrifugation, Density Gradient , Cell Membrane/analysis , Cell Membrane/drug effects , Electrophoresis, Polyacrylamide Gel , Edetic Acid/pharmacology , Octoxynol , Polyethylene Glycols/pharmacology , Sarcosine/analogs & derivatives , Sarcosine/pharmacology
19.
Braz. j. phys. ther. (Impr.) ; 18(6): 538-543, 09/01/2015. tab, graf
Article in English | LILACS | ID: lil-732351

ABSTRACT

BACKGROUND: The adapted arcometer has been validated for use in adults. However, its suitability for use in children can be questioned given the structural differences present in these populations. OBJECTIVE: To verify the concurrent validity, repeatability, and intra- and inter-reproducibility of the adapted arcometer for the measurement of the angles of thoracic kyphosis and lumbar lordosis in children. METHOD: Forty children were evaluated using both sagittal radiography of the spine and the adapted arcometer. The evaluations using the arcometer were carried out by two trained evaluators on two different days. In the statistical treatment, the intraclass correlation coefficient (ICC), Pearson's product moment correlation, Spearman's rho, the paired t test, and Wilcoxon's test were used (α=.05). RESULTS: A moderate and significant correlation was found between the x-ray and the adapted arcometer regarding thoracic kyphosis, but no correlation was found regarding lumbar lordosis. Repeatability and intra-evaluator reproducibility of the thoracic kyphosis and lumbar lordosis were confirmed, which was not the case of inter-evaluator reproducibility. CONCLUSION: The adapted arcometer can be used to accompany postural alterations in children made by the same evaluator, while its use for diagnostic purposes and continued evaluation by different evaluators cannot be recommended. Further studies with the aim of adapting this instrument for use in children are recommended. .


Subject(s)
Bacterial Proteins/analysis , Membrane Glycoproteins/analysis , Membrane Glycoproteins/genetics , Prodigiosin/biosynthesis , Serratia marcescens/metabolism , Centrifugation, Density Gradient , Electrophoresis, Polyacrylamide Gel , Molecular Weight , Membrane Glycoproteins/biosynthesis , Solubility , Sarcosine/analogs & derivatives , Serratia marcescens/analysis
20.
Salud pública Méx ; 56(4): 363-370, jul.-ago. 2014. ilus, tab
Article in English | LILACS | ID: lil-733301

ABSTRACT

Objective. To analyze the association between daily mortality from different causes and acute exposure to particulate matter less than 10 microns in aerodynamic diameter (PM10), in Bogota, Colombia. Materials and methods. A time-series ecological study was conducted from 1998 to 2006. The association between mortality (due to different causes) and exposure was analyzed using single and distributed lag models and adjusting for potential confounders. Results. For all ages, the cumulative effect of acute mortality from all causes and respiratory causes increased 0.71% (95%CI 0.46-0.96) and 1.43% (95%CI 0.85-2.00), respectively, per 10µg/m³ increment in daily average PM10 with a lag of three days before death. Cumulative effect of mortality from cardiovascular causes was -0.03% (95%CI -0.49-0.44%) with the same lag. Conclusions. The results suggest an association between an increase in PM10 concentrations and acute mortality from all causes and respiratory causes.


Objetivo. Analizar la asociación entre la mortalidad diaria debida a distintas causas y la exposición aguda a partículas menores de 10 micras de diámetro aerodinámico (PM10), en Bogotá, Colombia. Material y métodos. Se realizó un estudio ecológico de series de tiempo (1998-2006). La asociación entre mortalidad y exposición se analizó ajustando modelos de retraso simple y retraso distribuido para diferentes causas de mortalidad. Resultados. En todas las edades, el riesgo acumulado en la mortalidad aguda por todas las causas y causa respiratoria aumentó 0.71% (IC95% 0.46-0.96) y 1.43% (IC95% 0.85-2.00), respectivamente, por incremento de 10µg/m³ en el promedio diario de PM10, tomando un retraso de tres días anteriores al deceso, mientras el riesgo acumulado en la mortalidad por causa cardiovascular fue de -0.03% (IC95% -0.49-0.44), para el mismo retraso. Conclusiones. Los resultados sugieren asociación entre el incremento de las concentraciones de PM10 y la mortalidad aguda por todas las causas y causa respiratoria.


Subject(s)
Animals , Cattle , Bacterial Outer Membrane Proteins/isolation & purification , Mannheimia haemolytica/classification , Autoradiography/methods , Cattle Diseases , Cell Membrane/chemistry , Centrifugation, Density Gradient/methods , Detergents , Electrophoresis, Polyacrylamide Gel , Iodine Radioisotopes , Mannheimia haemolytica/chemistry , Membrane Proteins/isolation & purification , Molecular Weight , Pasteurella Infections/microbiology , Pasteurella Infections/veterinary , Sarcosine/analogs & derivatives , Solubility , Sucrose
SELECTION OF CITATIONS
SEARCH DETAIL