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1.
Pathogens ; 12(7)2023 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-37513765

RESUMO

Pseudomonas aeruginosa, an opportunistic pathogen causing infections in immunocompromised patients, usually shows pronounced antimicrobial resistance. In recent years, the frequency of carbapenemases in P. aeruginosa has decreased, which allows use of new beta-lactams/combinations in antimicrobial therapy. Therefore, the in vitro evaluation of these drugs in contemporary isolates is warranted. We evaluated the antimicrobial susceptibility and genomic aspects of 119 clinical P. aeruginosa isolates from 24 different hospitals in Brazil in 2021-2022. Identification was performed via MALDI-TOF-MS, and antimicrobial susceptibility was identified through broth microdilution, gradient tests, or disk diffusion. Whole-genome sequencing was carried out using NextSeq equipment. The most active drug was cefiderocol (100%), followed by ceftazidime-avibactam (94.1%), ceftolozane-tazobactam (92.4%), and imipenem-relebactam (81.5%). Imipenem susceptibility was detected in 59 isolates (49.6%), and the most active aminoglycoside was tobramycin, to which 99 (83.2%) isolates were susceptible. Seventy-one different sequence types (STs) were detected, including twelve new STs described herein. The acquired resistance genes blaCTX-M-2 and blaKPC-2 were identified in ten (8.4%) and two (1.7%) isolates, respectively. Several virulence genes (exoSTUY, toxA, aprA, lasA/B, plcH) were also identified. We found that new antimicrobials are effective against the diverse P. aeruginosa population that has been circulating in Brazilian hospitals in recent years.

2.
Neuroimage Clin ; 18: 932-942, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29876278

RESUMO

Background: White matter (WM) structural changes, particularly affecting the corpus callosum (CC), seem to be critically implicated in psychosis. Whether such abnormalities are progressive or static is still a matter of debate in schizophrenia research. Aberrant maturation processes might also influence the longitudinal trajectory of age-related CC changes in schizophrenia patients. We investigated whether patients with first-episode schizophrenia-related psychoses (FESZ) would present longitudinal CC and whole WM volume changes over the 5 years after disease onset. Method: Thirty-two FESZ patients and 34 controls recruited using a population-based design completed a 5-year assessment protocol, including structural MRI scanning at baseline and follow-up. The linear effects of disease duration, clinical outcome and antipsychotic (AP) use over time on WM and CC volumes were studied using both voxelwise and volume-based morphometry analyses. We also examined maturation/aging abnormalities through cross-sectional analyses of age-related trajectories of total WM and CC volume changes. Results: No interaction between diagnosis and time was observed, and clinical outcome did not influence CC volumes in patients. On the other hand, FESZ patients continuously exposed to AP medication showed volume increase over time in posterior CC. Curve-estimation analyses revealed a different aging pattern in FESZ patients versus controls: while patients displayed a linear decline of total WM and anterior CC volumes with age, a non-linear trajectory of total WM and relative preservation of CC volumes were observed in controls. Conclusions: Continuous AP exposure can influence CC morphology during the first years after schizophrenia onset. Schizophrenia is associated with an abnormal pattern of total WM and anterior CC aging during non-elderly adulthood, and this adds complexity to the discussion on the static or progressive nature of structural abnormalities in psychosis.


Assuntos
Antipsicóticos/uso terapêutico , Corpo Caloso/efeitos dos fármacos , Corpo Caloso/patologia , Esquizofrenia/tratamento farmacológico , Esquizofrenia/patologia , Adulto , Idoso , Estudos Transversais , Feminino , Humanos , Estudos Longitudinais , Imageamento por Ressonância Magnética/métodos , Masculino , Transtornos Psicóticos/diagnóstico , Transtornos Psicóticos/patologia , Esquizofrenia/diagnóstico
3.
Biochem Biophys Res Commun ; 487(1): 28-33, 2017 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-28365155

RESUMO

Human Leucocyte Antigen-G (HLA-G) is a non classical major histocompatibility complex (MHC) molecule that through RNA splicing can encode seven isoforms which are membrane bound (-G1, -G2, -G3 and -G4) and soluble (-G5, -G6 and -G7). HLA-G is described as important immune suppressor endogenous molecule to favor maternal-fetal tolerance, transplant survival and tumor immune scape. HLA-G shows low protein variability and a unique structural complexity that is related with the expression of different isoforms followed by biochemical processes, such as, proteolytic cleavage, molecular interactions, and protein ubiquitination. Studies with HLA-G have shown difficult to assess the role of the individual isoforms. Thus, the aim of this work was to obtain a HLA-G6 recombinant form. The results indicated the production of high homogeneous preparations of soluble recombinant HLA-G6 (srHLA-G6) with molecular mass 23,603.76 Da, determined by MALD-TOF/TOF. In addition, native and denatured srHLA-G6 were detected by ELISA, using commercial monoclonal antibodies. Finally, we developed a suitable methodology to express srHLA-G6 that could contribute in structural and functional studies involving specific isoforms.


Assuntos
Antígenos HLA-G/química , Antígenos HLA-G/imunologia , Proteínas Recombinantes/química , Sítios de Ligação , Humanos , Peso Molecular , Ligação Proteica , Solubilidade
4.
Biopreserv Biobank ; 15(4): 321-331, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28394173

RESUMO

The vitrification of preantral follicles followed by in vitro growth (IVG) could be valuable to produce fertilizable oocytes. However, the meiotic resumption rates of oocytes cultured from vitrified secondary follicles (SF) have been reported as suboptimal. This study aimed to verify two base media (alpha modification of minimum essential medium, α-MEM, and tissue culture medium 199, TCM199) on vitrified SF regarding different requirements during IVG. Sheep ovarian fragments were divided in six groups: (1) Fresh groups (Control α-MEM and TCM199): SF without vitrification; (2) Follicle-Vitrified (Follicle-Vit α-MEM and TCM199): SF vitrified after isolation; and (3) Tissue-Vitrified (Tissue-Vit α-MEM and TCM199): SF vitrified enclosed in ovarian fragments and, subsequently, isolated. The isolated SF were submitted to IVG for 18 days. Thereafter, the recovered cumulus-oocyte complexes (COCs) underwent in vitro maturation (IVM) and evaluation of chromatin configuration. Follicular granulosa cells were analyzed for their gene expression of Bax, Bcl2, and Connexins (CX) 37 and 43. COCs from in vivo antral follicles were used as in vivo control. Data were analyzed by analysis of variance, Tukey, and chi-square tests. Differences were considered significant if p-value is <0.05. Follicle-Vit groups had higher (p < 0.05) percentage of antrum formation compared with Tissue-Vit groups. Vitrification did not affect (p > 0.05) oocyte diameter postmaturation. Oocytes from Follicle-Vit in α-MEM reached metaphase II stage after IVM. Gene expression for CX37, CX43, and Bax was lower in Tissue-Vit groups. For Bcl2, the gene expression was the opposite. In conclusion, during IVG for 18 days, maximal oocyte meiotic resumption was not negatively impacted by vitrification and was greatest for isolated SF using α-MEM as a medium.


Assuntos
Oócitos/citologia , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Vitrificação , Animais , Células Cultivadas , Feminino , Perfilação da Expressão Gênica , Metáfase , Compostos Orgânicos/metabolismo , Ovinos , Fatores de Tempo
5.
Acta sci. vet. (Impr.) ; 45: 1-8, 2017. tab, ilus
Artigo em Português | VETINDEX | ID: biblio-1457638

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at 7°C and evaluated up until four h after DMSO addition. Individual progressive motility (MIP), sperm vigor (V), percentage of spermatozoa reactive to hypoosmotic test (HO) and morphologically normal (NOR) were evaluated. IPM, vigor and NOR remained within normal standards for the caprine species in all treatments test. Percentage results of spermatozoa reactive to hypoosmotic was higher in Tris yolk with values ranging between 34.66% to 46.33%. Sperm vigor was positively correlated (r = 0.85) with IPM in the MEM diluted pool of sperm.[...]


Assuntos
Animais , Antivirais , Dimetil Sulfóxido/uso terapêutico , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Ruminantes , Solventes/toxicidade , Extratos Vegetais , Vírus da Artrite-Encefalite Caprina
6.
Artigo em Inglês | LILACS-Express | VETINDEX | ID: biblio-1457678

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at

7.
Acta sci. vet. (Online) ; 45: 1-8, 2017. tab, ilus
Artigo em Português | VETINDEX | ID: vti-20225

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at 7°C and evaluated up until four h after DMSO addition. Individual progressive motility (MIP), sperm vigor (V), percentage of spermatozoa reactive to hypoosmotic test (HO) and morphologically normal (NOR) were evaluated. IPM, vigor and NOR remained within normal standards for the caprine species in all treatments test. Percentage results of spermatozoa reactive to hypoosmotic was higher in Tris yolk with values ranging between 34.66% to 46.33%. Sperm vigor was positively correlated (r = 0.85) with IPM in the MEM diluted pool of sperm.[...](AU)


Assuntos
Animais , Ruminantes , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Solventes/toxicidade , Antivirais , Dimetil Sulfóxido/uso terapêutico , Extratos Vegetais , Vírus da Artrite-Encefalite Caprina
8.
Artigo em Inglês | VETINDEX | ID: vti-733516

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at

9.
Artigo em Inglês | VETINDEX | ID: vti-732067

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at

10.
Artigo em Inglês | VETINDEX | ID: vti-731411

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at

11.
Artigo em Inglês | VETINDEX | ID: vti-730889

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at

12.
Artigo em Inglês | VETINDEX | ID: vti-730238

RESUMO

Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at

13.
Rev. salud pública Parag ; 6(1): 24-28, ene-jun. 2016.
Artigo em Espanhol | LILACS, BDNPAR | ID: biblio-908530

RESUMO

Introducción: muchas veces los hombres jóvenes se definen a sí mismos y a su identidad masculina a través de relaciones románticas y sexuales; sus decisiones sexuales pueden afectar su transición hacia la madurez, así como la adquisición de infecciones de transmisión sexual, VIH y tasas de embarazos. Objetivo: Este artículo analiza cómo influyen en el modo en que actúan en las relaciones sexuales y románticas y la formación de su identidad masculina, los grupos de amigos de hombres jóvenes paraguayos, sus familias. Metodología: En el 2010 llevamos a cabo cinco charlasen grupo en Asunción, Paraguay, en las que se examinaron las normas comunitarias con grupos de adolescentes entre 14 y 19 años de edad. Luego entrevistamos a la mitad de los miembros de cada grupo para examinar sus relaciones con amigos, la familia, mujeres jóvenes y sus creencias sobre las normas de género existentes. Resultados: Los jóvenes describieron dos tipos de normas masculinas (“pareja/proveedor” y “macho”) y dos tipos de relaciones románticas (“casual” y “formal”). Muchas veces se encontró una concordancia en el lenguaje utilizado para describir cada abanico de conductas, lo que pone de relieve la relación existente entre las normas masculinas y las relaciones románticas.


Introduction: Many times young men define themselves and their male identity through romantic and sexual relationships; their sexual decisions can affect their transition to adulthood as well as the acquisition of sexually transmitted infections, HIV and pregnancy rates. Objective:This article discusses how to influence the waythey act in sexual and romantic relationshipsand the formation of their male identity, groupsof young men Paraguay an friends, their families. Methodology: In 2010 we conducted five talks group in Asuncion, Paraguay, in which Community rules were examined with groups of teenagers between 14 and 19 years old. Then we interviewed half of the members of each group to examine their relationships with friends, family, young women and their beliefs about existing gender norms. Results: Young described two types of male norms(“partner / supplier” and “male”) and two types of romantic relationships (“casual” and “formal”).Many times a match was found in the languageused to describe each range of behaviors, which highlights the relationship between male normsand romantic relationships. The rules received bythe neighborhood as models that were more machobehavior by young people themselves informedcharacteristics. The rules can not be changed unlessyoung people talk about their behavior clearly notnormative. This demonstrates that further studieson the formation, meaning and transformation of male standards are needed.


Assuntos
Humanos , Masculino , Adolescente , Comportamento do Adolescente , Masculinidade , Psicologia do Adolescente , Marginalização Social , Paraguai
14.
Cytotechnology ; 68(4): 1623-32, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25634598

RESUMO

To design an estrogen and phenol red free medium for cell culture and check its effectiveness and safety on osteoblast growth it is necessary to maintain the estrogen receptors free for tests. For this purpose, we tested some modifications of the traditional culture media: estrogen depleted fetal bovine serum; estrogen charcoal stripped fetal bovine serum and phenol red free α-MEM. The aim of this work is to examine the effects of its depletion in the proliferation, differentiation, and toxicity of mesenchymal stromal cells differentiated into osteoblasts to obtain an effective interference free culture medium for in vitro studies, focused on non-previously studied estrogen receptors. We performed viability tests using the following techniques: MTT, alkaline phosphatase specific activity, formation of mineralized matrix by Alizarin technique and analysis of SEM/EDX of mineralized nodules. The results showed that the culture media with estrogen free α-MEM + phenol red free α-MEM did not impact viability, alkaline phosphatase activity and mineralization of the osteoblasts culture compared to control. In addition, its nodules possess Ca/P ratio similar to hydroxyapatite nodules on the 14th and 21st day. In conclusion, the modified culture medium with phenol red free α-MEM with estrogen depleted fetal bovine serum can be safely used in experiments where the estrogen receptors need to be free.

15.
Anim. Reprod. (Online) ; 11(4): 567-572, Oct.-Dec.2014. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: biblio-1461137

RESUMO

The aim of this study was to evaluate the effect of culture media (Alpha Minimum Essential Medium, α-MEM; and Tissue Culture Medium-199, TCM-199) in the absence or presence of Epidermal Growth Factor (EGF) on an in vitro culture of goat and sheep preantral follicles enclosed in ovarian tissue. The fragments of ovarian cortex from both species were immediately analyzed after collection (non-cultured control group) or cultured for 1 or 7 days in α-MEM+ or TCM-199+in the absence or presence of EGF (10 ng/ml). Before and after the culture, the fragments of ovarian cortex were analyzed by classical histology and fluorescence microscopy. After 1 day of culture, all treatments decreased the percentage of morphologically normal follicles when compared to non-cultured control in both species (P 0.05). Regarding follicle activation, an increase in the percentage of growing follicles was observed in all treatments after 7 days of culture when compared to the control group in both species. However, in sheep, after 7 days, only the treatments α-MEM+/EGF and TCM-199+showed larger(P < 0.05) oocytes than the control group. In conclusion, the TCM-199+ Preserved goat preantral follicle viability after in vitro culture. Furthermore, the media α-MEM+/EGF and TCM-199+ increased the oocyte diameter after 7 days of culture in sheep. Therefore, it isrecommended to use TCM-199+ In the culture of preantral follicles in both species.


Assuntos
Feminino , Animais , Cabras , Células Epidérmicas , Hormônio Foliculoestimulante , Meios de Cultura/análise , Ovinos
16.
Anim. Reprod. ; 11(4): 567-572, Oct.-Dec.2014. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: vti-29041

RESUMO

The aim of this study was to evaluate the effect of culture media (Alpha Minimum Essential Medium, α-MEM; and Tissue Culture Medium-199, TCM-199) in the absence or presence of Epidermal Growth Factor (EGF) on an in vitro culture of goat and sheep preantral follicles enclosed in ovarian tissue. The fragments of ovarian cortex from both species were immediately analyzed after collection (non-cultured control group) or cultured for 1 or 7 days in α-MEM+ or TCM-199+in the absence or presence of EGF (10 ng/ml). Before and after the culture, the fragments of ovarian cortex were analyzed by classical histology and fluorescence microscopy. After 1 day of culture, all treatments decreased the percentage of morphologically normal follicles when compared to non-cultured control in both species (P < 0.05). In fluorescence microscopy, viable sheep follicles were observed to decrease in all treatments after 7 days of culture when compared to non-cultured controls. However, in goats, the culture with TCM-199+maintained follicle viability after 7 days of culture, similar to fresh tissue (P > 0.05). Regarding follicle activation, an increase in the percentage of growing follicles was observed in all treatments after 7 days of culture when compared to the control group in both species. However, in sheep, after 7 days, only the treatments α-MEM+/EGF and TCM-199+showed larger(P < 0.05) oocytes than the control group. In conclusion, the TCM-199+ Preserved goat preantral follicle viability after in vitro culture. Furthermore, the media α-MEM+/EGF and TCM-199+ increased the oocyte diameter after 7 days of culture in sheep. Therefore, it isrecommended to use TCM-199+ In the culture of preantral follicles in both species.(AU)


Assuntos
Animais , Feminino , Meios de Cultura/análise , Hormônio Foliculoestimulante , Cabras , Ovinos , Células Epidérmicas
17.
Gene ; 532(2): 186-91, 2013 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-24076352

RESUMO

Gaucher disease (GD) is caused by mutations in the GBA gene that confer a deficient level of activity of glucocerebrosidase (GCase). This deficiency leads to accumulation of the glycolipid glucocerebroside in the lysosomes of cells of monocyte/macrophage system. Type I GD is the mildest form and is characterized by the absence of neuronopathic affection. Bone compromise in Gaucher disease patients is the most disabling aspect of the disease. However, pathophysiological aspects of skeletal alterations are still poorly understood. The homeostasis of bone tissue is maintained by the balanced processes of bone resorption by osteoclasts and formation by osteoblasts. We decided to test whether bone resorption and/or bone formation could be altered by the use of a chemical in vitro murine model of Gaucher disease. We used two sources of cells from monocyte/macrophages lineage isolated from normal mice, splenocytes (S) and peritoneal macrophages (PM), and were exposed to CBE, the inhibitor of GCase (S-CBE and PM-CBE, respectively). Addition of both conditioned media (CM) from S-CBE and PM-CBE induced the differentiation of osteoclasts precursors from bone marrow to mature and functional osteoclasts. TNF-α could be one of the factors responsible for this effect. On the other hand, addition of CM to an osteoblast cell culture resulted in a reduction in expression of alkaline phosphatase and mineralization process. In conclusion, these results suggest implication of changes in both bone formation and bone resorption and are consistent with the idea that both sides of the homeostatic balance are affected in GD.


Assuntos
Doença de Gaucher/patologia , Osteoblastos/metabolismo , Osteoclastos/fisiologia , Animais , Antígenos de Diferenciação/metabolismo , Células da Medula Óssea/fisiologia , Calcificação Fisiológica , Diferenciação Celular , Células Cultivadas , Meios de Cultivo Condicionados , Doença de Gaucher/induzido quimicamente , Doença de Gaucher/metabolismo , Inositol/análogos & derivados , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Osteólise/metabolismo , Fator de Necrose Tumoral alfa/fisiologia
18.
Biochim Biophys Acta ; 1828(11): 2646-53, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23899501

RESUMO

Large procyanidins (more than three subunits) are not absorbed at the gastrointestinal tract but could exert local effects through their interactions with membranes. We previously showed that hexameric procyanidins (Hex), although not entering cells, interact with membranes modulating cell signaling and fate. This paper investigated if Hex, as an example of large procyanidins, can selectively interact with lipid rafts which could in part explain its biological actions. This mechanism was studied in both synthetic membranes (liposomes) and Caco-2 cells. Hex promoted Caco-2 cell membrane rigidification and dehydration, effects that were abolished upon cholesterol depletion with methyl-ß-cyclodextrin (MCD). Hex prevented lipid raft structure disruption induced by cholesterol depletion/redistribution by MCD or sodium deoxycholate. Supporting the involvement of cholesterol-Hex bonding in Hex interaction with lipid rafts, the absence of cholesterol markedly decreased the capacity of Hex to prevent deoxycholate- and Triton X-100-mediated disruption of lipid raft-like liposomes. Stressing the functional relevance of this interaction, Hex mitigated lipid raft-associated activation of the extracellular signal-regulated kinases (ERK) 1/2. Results support the capacity of a large procyanidin (Hex) to interact with membrane lipid rafts mainly through Hex-cholesterol bondings. Procyanidin-lipid raft interactions can in part explain the capacity of large procyanidins to modulate cell physiology.


Assuntos
Colesterol/metabolismo , Microdomínios da Membrana/metabolismo , Proantocianidinas/metabolismo , Western Blotting , Células CACO-2 , Detergentes , Ativação Enzimática , Humanos , Lipossomos , Sistema de Sinalização das MAP Quinases , Transdução de Sinais
19.
West Indian med. j ; West Indian med. j;61(8): 778-783, Nov. 2012. ilus, tab
Artigo em Inglês | LILACS | ID: lil-694340

RESUMO

AIMS: The aim of this study was to detect and characterize the presence of metallo-β-lactamase (MBL) production in multidrug resistant (MDR) P aeruginosa collected from clinical samples in a tertiary care hospital. METHODS AND MATERIALS: A total of 67 non-repetitive isolates of MDR P aeruginosa recovered from various clinical specimens were screened for MBL production by IPM/MEM-EDTA combined disc test. Polymerase chain reaction was performed on all isolates using blaIMP and blaVIM consensus primers to characterize them genotypically. RESULTS: Among 67 P aeruginosa isolates, 62.7% (42/67) and 70.1% (47/67) were resistant to imipenem and meropenem respectively and 47 (70.1%) were found to be MBL producers. Among this 47 MBL-producing isolates, 41 (61.1%) strains carried the blaVIM gene and 2 (3%) strains carried the blaIMP gene. Three strains were phenotypically negative but positive genotypically for blaVIM gene. One strain was resistant to both imipenem and meropenem but did not show phenotypic positivity. CONCLUSION: This study confirms the dissemination of blaVIM genes among MDR Pseudomonas aeruginosa and hence it is indispensible to identify and aptly control the threat of horizontal and vertical transfer.


OBJETIVO: El objetivo de este estudio es descubrir y caracterizar la presencia de producción de metallo-betalactamasa (MBL) en P aeruginosa resistente a los multifármacos (RMF), recogida de muestras clínicas de un hospital de atención terciaria. MÉTODO: Un total de 67 aislados no repetitivos de P aeruginosa RMF obtenidos de varios specímenes clínicos, fueron tamizados en busca de producción de MBL, mediante una prueba de disco combinado IPM/MEM-EDTA. Se efectuó una reacción en cadena de la polimerasa sobre todos los aislados, usando iniciadores de consenso blaIMP y blaVIM para la caracterización genotípica. RESULTADOS: Entre los aislados de P aeruginosa, 62.7% (42/67) y 70.1% (47/67) fueron resistentes al Imipenem y al Meropenem respectivamente, mientras que se halló que 47 (70.1%) eran productores de MBL. De los 47 aislados productores de MBL, 41 (61.1%) cepas eran portadoras del gen blaVIM en tanto que 2 (3%) cepas eran portadoras del gen blaIMP. Tres cepas fueron fenotípicamente negativas, pero genotípicamente positivas con respecto al gen blaVIM. Una cepa fue resistente tanto al Imipenem como al Meropenem, pero no mostró positividad fenotípicamente. CONCLUSIÓN: El presente estudio confirma la diseminación de los genes blaVIM entre las Pseudomonas aeruginosa RMF. Es importante identificar así como controlar adecuadamente la amenaza de la transferencia horizontal y vertical.


Assuntos
Farmacorresistência Bacteriana Múltipla , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/genética , beta-Lactamases/genética , beta-Lactamases/metabolismo , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Genótipo , Imipenem/farmacologia , Fenótipo , Pseudomonas aeruginosa/efeitos dos fármacos , Atenção Terciária à Saúde , Tienamicinas/farmacologia
20.
Colomb. med ; 40(4): 399-407, nov.-dic. 2009. tab, graf
Artigo em Espanhol | LILACS | ID: lil-573465

RESUMO

Objetivo: Evaluar la reproducibilidad de la retinoscopia dinámica monocular y su nivel de acuerdo con la retinoscopia estática binocular y monocular, retinoscopia de Nott y Método Estimado Monocular (MEM). Métodos: Se determinó la reproducibilidad entre los evaluadores y entre los métodos por medio del coeficiente de correlación intraclase (CCI) y se establecieron los límites de acuerdo de Bland y Altman.Resultados: Se evaluaron 126 personas entre 5 y 39 años y se encontró una baja reproducibilidad interexaminador de la retinoscopia dinámica monocular en ambos ojos CCI ojo derecho: 0.49 (IC95% 0.36; 0.51); ojo izquierdo 0.51 (IC95% 0.38; 0.59). El límite de acuerdo entre evaluadores fue ±1.25 D. Al evaluar la reproducibilidad entre la retinoscopia dinámica monocular y la estática se observó que la mayor reproducibilidad se obtuvo con la estática binocular y monocular y, en visión próxima, entre el método estimado monocular y la retinoscopia de Nott. Conclusiones: La retinoscopia dinámica monocular no es una prueba reproducible y presenta diferencias clínicas significativas para determinar el estado refractivo, en cuanto a poder dióptrico y tipo de ametropía, por tanto, no se puede considerar dentro de la batería de exámenes aplicados para determinar diagnósticos y correcciones refractivas tanto en la visión lejana como en la visión próxima.


Objective: To assess the reproducibility of monocular dynamic retinoscopy and its level of agreement with binocular static retinoscopy, the Nott retinoscopy, and the Monocular Estimated Method (MEM). Materials and methods: The reproducibility was determined among evaluators and among the methods through the intraclass coefficient of correlation (ICC) and set the limits of agreement of Bland and Altman. Results: A total of 126 subjects between 5 and 39 years of age were evaluated and low inter-rater reproducibility of the monocular dynamic retinoscopy was found in both eyes: ICC right eye: 0.49 (IC 95% 0.36; 0.61) left eye 0.51 (IC 95% 0.38; 0.59). The limit of agreement between evaluators was ±1.25D. In assessing reproducibility between the monocular dynamic retinoscopy and the static retinoscopy, it was observed that the greatest reproducibility was obtained with the binocular and monocular static retinoscopy and in near vision between the estimated method monocular and Nott retinoscopy. Conclusions: Monocular dynamic retinoscopy is not a reproducible test and presents clinical differences meaningful to determine the refractive state, regarding power dial and type of ametropy; therefore, it could not be considered within the battery of tests used to determine diagnoses and refractive corrections both in distant vision and in near vision.


Assuntos
Reprodutibilidade dos Testes , Retinoscopia/métodos , Retinoscopia
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