Recommandations techniques et règles de bonne pratique pour la coloration de May-Grünwald-Giemsa : revue de la littérature et apport de l’assurance qualité

Piaton, Éric; Fabrè, Monique; Goubin-Versini, Isabelle; Bretz-Grenier, M.-F.; Courtade-Saïdi, Monique; Vincent, S.; Belleannée, Geneviève; Thivolet, Françoise; Boutonnât, Jean; Debaque, Hervé; Fleury‐Feith, Jocelyne; Vielh, Philippe; Cochand‐Priollet, Béatrix; clinique, pour la Société française de cytologie; Egele, C.; Bellocq, Jean‐Pierre; Michiels, Jean‐François; pathologiques, pour l’Association française d’assurance qualité en anatomie et cytologie

doi:10.1016/j.annpat.2015.05.019

Cited by 28 publications

(11 citation statements)

References 16 publications

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“…Thus, live cells will be fluorescent green and dead cells, fluorescent red [20]. May-Grunewald-Giemsa staining was used to visualize cytopathic effects in neutrophils [21].…”

Section: Leukotoxin Detection By Stainingmentioning

confidence: 99%

Biological and molecular characterization of a sheep pathogen isolate of Mannheimia haemolytica and leukotoxin production kinetics

et al. 2021

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Background and Aim: Mannheimia haemolytica (Mha) is a common agent of pneumonia in ruminants globally, causing economic losses by morbidity, mortality, and treatment costs. Infection by Mha is often associated with or promoted by respiratory viral pathogens and environmental conditions. Infections due to Mha have rarely been described in small ruminants. This study reports the biological and molecular characteristics of a new Moroccan Mha isolate from small ruminants presenting typical respiratory symptoms. We also studied the cultural parameters, growth kinetics, and Lkt excretion of the isolate and its pathogenicity on laboratory animals and small ruminants. Materials and Methods: Suspected pasteurellosis cases in sheep and goat flocks in Morocco were investigated. A local strain of Mha was isolated and identified using biochemical and molecular methods. Polymerase chain reaction-targeting specific genes were used for serotyping and phylogenetic analyses; further, leukotoxin production, cytotoxicity, and pathogenicity of the isolate in mice, goats, and sheep were investigated. Results: Phylogeny analysis revealed 98.76% sequence identity with the USA isolate of 2013; the strain growth with a cycle of 9-10 h with leukotoxin secretion was detected by NETosis and quantified by cytotoxicity and mortality of mice. Goat and sheep infections cause hyperthermia, with characteristic postmortem lesions in the trachea and lung. Conclusion: A local isolate of Mha from sheep that died of pneumonia was characterized for the 1st time in North Africa using biological and molecular methods. Although growth on appropriate culture media is accompanied by intense leukotoxin secretion, experimental infections of sheep and goats cause hyperthermia and typical lesions of pneumonia.

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“…Thus, live cells will be fluorescent green and dead cells, fluorescent red [20]. May-Grunewald-Giemsa staining was used to visualize cytopathic effects in neutrophils [21].…”

Section: Leukotoxin Detection By Stainingmentioning

confidence: 99%

Biological and molecular characterization of a sheep pathogen isolate of Mannheimia haemolytica and leukotoxin production kinetics

et al. 2021

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“…For differential counts, blood films were done from K3-EDTA blood samples. After fixation in methanol, the blood films were stained using the May-Grünwald-Giemsa technique ( 32 ). White blood cells were counted at 100× magnification to determine the differential counts.…”

Section: Methodsmentioning

confidence: 99%

Development of an innovative in vivo model of PJI treated with DAIR

Poilvache

Bambeke²

2022

Front. Med.

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IntroductionProsthetic Joint Infection (PJI) are catastrophic complications of joint replacement. Debridement, implant retention, and antibiotic therapy (DAIR) is the usual strategy in acute infections but fails in 45% of MRSA infections. We describe the development of a model of infected arthroplasty in rabbits, treated with debridement and a course of vancomycin with clinically relevant dosage.Materials and methodsA total of 15 rabbits were assigned to three groups: vancomycin pharmacokinetics (A), infection (B), and DAIR (C). All groups received a tibial arthroplasty using a Ti-6Al-4V implant. Groups B and C were infected per-operatively with a 5.5 log10 MRSA inoculum. After 1 week, groups C infected knees were surgically debrided. Groups A and C received 1 week of vancomycin. Pharmacokinetic profiles were obtained in group A following 1st and 5th injections. Animals were euthanized 2 weeks after the arthroplasty. Implants and tissue samples were processed for bacterial counts and histology.ResultsAverage vancomycin AUC0–12 h were 213.0 mg*h/L (1st injection) and 207.8 mg*h/L (5th injection), reaching clinical targets. All inoculated animals were infected. CFUs were reproducible in groups B. A sharp decrease in CFU was observed in groups C. Serum markers and leukocytes counts increased significantly in infected groups.ConclusionWe developed a reproducible rabbit model of PJI treated with DAIR, using vancomycin at clinically relevant concentrations.

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“…Passage 4 cells were stained with Giemsa staining. [ 19 ] Cells with and without staining were observed under a microscope and the images were captured using Nikon camera. Cell transfections were performed using Lipofectamine 2000 (Life Technologies) according to the manufacturers instructions.…”

Section: Methodsmentioning

confidence: 99%

Experimental study of expression profile and specific role of human microRNAs in regulating atrophic bone nonunion

Wei

Chen

et al. 2020

Medicine

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The expression profile and specific roles of microRNAs (miRNAs) in regulation of atrophic bone nonunion are not fully understood. Here, we present evidence that miRNAs are involved in regulation of several osteogenic genes and may contribute to the development of atrophic bone nonunion. The miRNA expression profile of repairing tissues in atrophic bone nonunion patients (group A) and in callus tissues from patients with healed fractures (group B) were quantitatively measured. microRNA microarrays were used to identify differentially expressed miRNAs, and the bioinformatics methods were used to predict the potential target genes. Quantitative real-time polymerase chain reaction (qRT-PCR), western blot, and dual-luciferase reporter assay were performed in human bone marrow stromal cells (hBMSCs) to validate the microarray results. Nine miRNAs in group A were up-regulated 1.5 times compared to group B, while the other 9 miRNAs in group A were down-regulated 1.5 times. Several target regions of these miRNAs were identified in the osteogenic genes, as well as in the other genes in their families or related regulatory factors. Four miRNAs (hsa-miR-149 ∗ , hsa-miR-221, hsa-miR-628-3p, and hsa-miR-654-5p) could play important roles in regulating bone nonunion development. hBMSCs transfected with these miRNAs significantly decreased mRNA levels of alkaline phosphatase, liver/bone/kidney ( ALPL ), platelet derived growth factor subunit A ( PDGFA ), and bone morphogenetic protein 2 ( BMP2 ). Lower protein expression levels were observed using western blotting, confirming that ALPL , PDGFA , and BMP2 were directly targeted by hsa-miR-149 ∗ , hsa-miR-221, and hsa-miR-654-5p, respectively. In summary, hsa-miR-149 ∗ , hsa-miR-221, and hsa-miR-654-5p may play important biological roles by repressing osteogenic target genes ALPL , PDGFA , and BMP2 , and, therefore, contributing to progression of atrophic bone nonunion.

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Recommandations techniques et règles de bonne pratique pour la coloration de May-Grünwald-Giemsa : revue de la littérature et apport de l’assurance qualité

Cited by 28 publications

References 16 publications

Biological and molecular characterization of a sheep pathogen isolate of Mannheimia haemolytica and leukotoxin production kinetics

Biological and molecular characterization of a sheep pathogen isolate of Mannheimia haemolytica and leukotoxin production kinetics

Development of an innovative in vivo model of PJI treated with DAIR

Experimental study of expression profile and specific role of human microRNAs in regulating atrophic bone nonunion

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