scholarly journals Conteúdo de peptídeos e avaliação morfofisiológica dos espermatozóides do epidídimo e ejaculado de bovinos

2003 ◽  
Vol 32 (6 suppl 2) ◽  
pp. 1890-1900 ◽  
Author(s):  
Antonio Emidio D. Feliciano Silva ◽  
André Lima Dias ◽  
Maria Marina Unanian ◽  
Alfredo Ribeiro de Freitas ◽  
Carlos Bloch Junior

Objetivou-se com este estudo a identificação de alguns fatores protéicos envolvidos na qualidade funcional dos espermatozóides epididimais (SPZEP) e ejaculados (SPZEJ) de bovinos. Foram avaliadas as características morfofisiológicas e analisado o conteúdo peptídico destas estruturas de 11 animais mestiços Nelore, de 24 a 30 meses de idade. As avaliações morfofisiológicas foram motilidade progressiva (MOT, %), vigor, patologias espermáticas, integridade acrossômica e da cromatina. Foi observado que, os SPZEJ, na média, apresentaram MOT maior do que os SPZEP, 72,3 e 46,4%, respectivamente. Considerando as patologias espermáticas, taxas de defeitos maiores (DEFMAI), menores (DEFMEN) e totais (DEFTOT), houve diferença significativa entre as taxas dos DEFMEN e DEFTOT dos SPZEP e SPZEJ, sendo, em média, 91,1 e 8,5% e 95,4 e 11,8%, respectivamente. As taxas dos DEFMEN e DEFTOT dos SPZEP foram maiores em função da presença de espermatozóides com gotas citoplasmáticas distais. A análise das protéinas dos SPZEP e SPZEJ foi realizada por espectrometria de massa, método MALDI-TOF (matrix -assisted laser desorption/ionization - time of flight), e revelou presença de peptídeos de massa molecular variando de 1,1 a 26,3 kDa nos SPZEJ e de 1,1 a 11,6 kDa nos SPZEP. Foram identificados peptídeos de 10,6 e 13,4 kDa somente nos SPZEJ e de 6,8 kDa somente nos SPZEP. Foi observada relação do peptídeo de massa molecular de 7,4 kDa dos SPZEP e de 4,7 kDa dos SPZEJ, com a MOT Ê 80%, destas estruturas. Os resultados sugerem o envolvimento destes peptídeos nos processos funcionais das células espermáticas do epidídimo e ejaculado. O estudo utilizou o método MALDI/TOF para espectrômetro de massa, para identificar peptídeos em espermatozóides do epidídimo de bovinos, pela primeira vez no País.

2016 ◽  
Vol 55 (1) ◽  
pp. 90-96 ◽  
Author(s):  
Julie Denis ◽  
Marie Machouart ◽  
Florent Morio ◽  
Marcela Sabou ◽  
Catherine Kauffmann-LaCroix ◽  
...  

ABSTRACT The genus Malassezia comprises commensal yeasts on human skin. These yeasts are involved in superficial infections but are also isolated in deeper infections, such as fungemia, particularly in certain at-risk patients, such as neonates or patients with parenteral nutrition catheters. Very little is known about Malassezia epidemiology and virulence. This is due mainly to the difficulty of distinguishing species. Currently, species identification is based on morphological and biochemical characteristics. Only molecular biology techniques identify species with certainty, but they are time-consuming and expensive. The aim of this study was to develop and evaluate a matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) database for identifying Malassezia species by mass spectrometry. Eighty-five Malassezia isolates from patients in three French university hospitals were investigated. Each strain was identified by internal transcribed spacer sequencing. Forty-five strains of the six species Malassezia furfur , M. sympodialis , M. slooffiae , M. globosa , M. restricta , and M. pachydermatis allowed the creation of a MALDI-TOF database. Forty other strains were used to test this database. All strains were identified by our Malassezia database with log scores of >2.0, according to the manufacturer's criteria. Repeatability and reproducibility tests showed a coefficient of variation of the log score values of <10%. In conclusion, our new Malassezia database allows easy, fast, and reliable identification of Malassezia species. Implementation of this database will contribute to a better, more rapid identification of Malassezia species and will be helpful in gaining a better understanding of their epidemiology.


2021 ◽  
Vol 30 (3) ◽  
pp. 65-69
Author(s):  
Marwa N. Mohamed ◽  
Ahmed F. Azmy ◽  
Ehab M. Fahmy ◽  
Mervat G. Elanany ◽  
Nesreen M. Kamel

Background: Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) is a novel technique for identification of microbes. This new method led to a new era in microbial identification because of its rapid, accurate, valid, simple and relatively decreased cost. Objectives: The aim of this study was identification of predominant pathogens by MALDI-TOF technique. Methodology: Pathogens were identified by both conventional methods and MALDI-TOF. Results: From July till December 2018, predominant pathogens were Klebsiella pneumoniae (21%), Pseudomonas aeroguinosa and Candida each constitutes (17%), E-coli (10%), Staph. aureus (9%), Acinetobacter (9%). Identification of isolates (from September to December 2018) by MALDI-TOF revealed a total agreement of (94.1%) with conventional method at genus level, (88.2%) at level of species. Kappa agreement revealed almost perfect correlation between both techniques. Conclusion: The MALDI-TOF results might suggest that its’ usage may be dependable for microbiological identification.


2013 ◽  
Vol 34 (9) ◽  
pp. 990-995 ◽  
Author(s):  
Pranita D. Tamma ◽  
Kennard Tan ◽  
Veronique R. Nussenblatt ◽  
Alison E. Turnbull ◽  
Karen C. Carroll ◽  
...  

We evaluated 222 hospitalized patients whose clinical isolates were tested using standard methods and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). MALDI-TOF could have reduced time to appropriate therapy for 28.8% and 44.6% patients based on the treating physician's choices and stewardship team recommendations, respectively. Clinicians should be aware of scenarios in which MALDI-TOF can optimize antibiotic therapy.


2015 ◽  
Vol 54 (1) ◽  
pp. 5-6 ◽  
Author(s):  
Neil W. Anderson

An article in this issue of theJournal of Clinical Microbiology(J.-H. Youn, S. K. Drake, R. A. Weingarten, K. M. Frank, J. P. Dekker, and A. F. Lau, J Clin Microbiol 53:35–42, 2015,http://dx.doi.org/10.1128/JCM.01643-15) describes the use of matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry for the detection of organisms carrying ablaKPC-containing plasmid. This powerful and promising application highlights the challenges of using MALDI-TOF mass spectrometry for purposes other than organism identification.


2003 ◽  
Vol 19 (1) ◽  
pp. 49-53 ◽  
Author(s):  
Masakazu SHIMIZU ◽  
Noriyuki OJIMA ◽  
Hitoshi OHNISHI ◽  
Tomoteru SHINGAKI ◽  
Yasuyuki HIRAKAWA ◽  
...  

2015 ◽  
Vol 53 (7) ◽  
pp. 2163-2171 ◽  
Author(s):  
Camille Lasserre ◽  
Luc De Saint Martin ◽  
Gaelle Cuzon ◽  
Pierre Bogaerts ◽  
Estelle Lamar ◽  
...  

The recognition of carbapenemase-producingEnterobacteriaceae(CPE) isolates is a major laboratory challenge, and their inappropriate or delayed detection may have negative impacts on patient management and on the implementation of infection control measures. We describe here a matrix-assisted laser desorption ionization−time of flight (MALDI-TOF)-based method to detect carbapenemase activity inEnterobacteriaceae. After a 20-min incubation of the isolate with 0.5 mg/ml imipenem at 37°C, supernatants were analyzed by MALDI-TOF in order to identify peaks corresponding to imipenem (300 Da) and an imipenem metabolite (254 Da). A total of 223 strains, 77 CPE (OXA-48 variants, KPC, NDM, VIM, IMI, IMP, and NMC-A) and 146 non-CPE (cephalosporinases, extended-spectrum β-lactamases [ESBLs], and porin defects), were tested and used to calculate a ratio of imipenem hydrolysis: mass spectrometry [MS] ratio = metabolite/(imipenem + metabolite). An MS ratio cutoff was statistically determined to classify strains as carbapenemase producers (MS ratio of ≥0.82). We validated this method first by testing 30 of our 223 isolates (15 CPE and 15 non-CPE) 10 times to calculate an intraclass correlation coefficient (ICC of 0.98), showing the excellent repeatability of the method. Second, 43 strains (25 CPE and 18 non-CPE) different from the 223 strains used to calculate the ratio cutoff were used as external controls and blind tested. They yielded sensitivity and specificity of 100%. The total cost per test is <0.10 U.S. dollars (USD). This easy-to-perform assay is time-saving, cost-efficient, and highly reliable and might be used in any routine laboratory, given the availability of mass spectrometry, to detect CPE.


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