Objective
Since the guidelines of the International Committee for Standardisation in Haematology (ICSH) in 1984 and those of the European Committee for External Quality Assessment Programmes in ...Laboratory Medicine (EQALM) in 2004, no leading organisation has published technical recommendations for the preparation of air‐dried cytological specimens using May‐Grünwald–Giemsa (MGG) staining.
Data sources
Literature data were retrieved using reference books, baseline‐published studies, articles extracted from PubMed/Medline and Google Scholar, and online‐available industry datasheets.
Rationale
The present review addresses all pre‐analytical issues concerning the use of Romanowsky's stains (including MGG) in haematology and non‐gynaecological cytopathology. It aims at serving as actualised, best practice recommendations for the proper handling of air‐dried cytological specimens. It, therefore, appears complementary to the staining criteria of the non‐gynaecological diagnostic cytology handbook edited by the United Kingdom National External Quality Assessment Service (UK‐NEQAS) in February 2015.
These valuable guidelines for MGG staining will complement the NEQAS handbook and provide helpful information for anyone trouble shooting or improving their MGG staining.
Abstract The ubiquitous presence of nanoparticles (NPs) together with increasing evidence linking them to negative health effects points towards the need to develop the understanding of mechanisms by ...which they exert toxic effects. This study was designed to investigate the role of surface area and oxidative stress in the cellular effects of two chemically distinct NPs, carbon black (CB) and titanium dioxide (TiO2 ), on the bronchial epithelial cell line (16HBE14o-). CB and TiO2 NPs were taken up by 16HBE cells in a dose-dependent manner and were localized within the endosomes or free in the cytoplasm. Oxidative stress produced inside the cell by NPs was well correlated to the BET surface area and endocytosis of NPs. Contrary to intracellular conditions only CB NPs produced reactive oxygen species (ROS) under abiotic conditions. Exposure of cells to NPs resulted in an increased granulocyte macrophage colony stimulating factor (GM-CSF) mRNA expression and secretion. Inflammatory effects of NPs were dependent on the surface area and were mediated through oxidative stress as they were inhibited by catalase. It can be concluded that NP induced oxidative stress and pro-inflammatory responses are well correlated not only with the BET (Brunauer, Emmett and Teller) surface of the individual NPs but also with the internalized amount of NPs. Differences of even few nanometers in primary particle size lead to significant changes in inflammatory and oxidative stress responses.
Pleural fluid accumulation is a frequent clinical observation in diffuse malignant pleural mesothelioma (MPM). The cytological analysis of pleural fluid often reveals the presence of free spheroid ...aggregates of malignant cells, giving rise to the question of the ability of non-adherent tumor cells to resist the loss of anchorage-induced apoptosis (termed as anoikis), and to develop new tumor foci in the pleural cavity. Here, we show that MPM cells cultured under non-adherent conditions form well-organized aggregates composed of viable cells, which progressively enter in G(0). Although the PI3K/Akt, ERK and SAPK/JNK signaling pathways are activated in adherent MPM cells, loss of anchorage results in the inactivation of these pathways. By comparison, we show that the non-tumoral mesothelial cells MeT-5A enter anoikis in an SAPK/JNK-, Bim- and caspase-9-dependent pathway. The survival of MPM cells can be reversed by activating SAPK/JNK with anisomycin, according to a Bim-dependent mitochondrial pathway. Finally, our findings show that impairment of cell aggregation activates SAPK/JNK and Bim and induces anoikis. Our results underline the importance of intercellular contacts in the anoikis resistance of MPM cells.
To evaluate the incidence, clinical features, and prognosis of pulmonary complications associated with toxic epidermal necrolysis
Prospective study.
Dermatology intensive care unit in Mondor ...Hospital, France.
41 consecutive patients.
On admission, then daily, respiratory evaluation was based on clinical examination, chest X-ray, and arterial blood gas analysis. When clinical symptoms, X-ray abnormalities, or hypoxemia partial pressure of oxygen (PO2) < 80 mm Hg were present, fiberoptic bronchoscopy was performed.
10 patients presented early manifestations: dyspnea (n = 10), bronchial hypersecretion (n = 7), marked hypoxemia (n = 10) (PO2 = 59 +/- 8 mm Hg). Chest X-ray was normal (n = 8) or showed interstitial infiltrates (n = 2). In these 10 patients, fiberoptic bronchoscopy demonstrated sloughing of bronchial epithelium in proximal airways. Delayed pulmonary complications occurred in 6 of these 10 patients from day 7 to day 15: pulmonary edema (n = 2), atelectasis (n = 1), bacterial pneumonitis (n = 4). Mechanical ventilation was required in 9 patients. A fatal outcome occurred in 7 patients. Seven patients did not develop early pulmonary manifestations (PO2 on admission 87 +/- 6 mm Hg) but only delayed pulmonary symptoms related to atelectasis (n = 1), pulmonary edema (n = 4), and bacterial pneumonitis (n = 3); bronchial epithelial detachment was not observed. None of them required mechanical ventilation and all recovered with appropriate therapy.
"Specific" involvement of bronchial epithelium was noted in 27% of cases and must be suspected when dyspnea, bronchial hypersecretion, normal chest X-ray, and marked hypoxemia are present during the early stages of toxic epidermal necrosis. Bronchial injury seems to indicate a poor prognosis, as mechanical ventilation was required for most of these patients and was associated with a high mortality.
Les pathologies pleurales liées à l’exposition aux fibres sont maintenant bien connues pour les fibres d’amiante, bien que le mécanisme d’action de ces fibres ne soit pas encore totalement élucidé. ...Le développement des fibres minérales artificielles et des nanotubes de carbone, dont les caractéristiques morphologiques sont proches des fibres d’amiante, remet en première ligne les hypothèses physiopathologiques associant exposition aux fibres et pathologies pleurales. Les plaques pleurales se développent uniquement dans la plèvre pariétale, et dès les années 1990, des observations cliniques ont montré que le développement précoce du mésothéliome se produisait également au niveau de la plèvre pariétale. La particularité de la plèvre pariétale par rapport à la plèvre viscérale est la présence de « stomatas ou stomas » qui sont des orifices de communication entre la cavité pleurale et les vaisseaux lymphatiques de la plèvre pariétale. Des observations morphologiques en thoracoscopie et des études expérimentales ont permis de montrer que les fibres étaient présentes dans la plèvre pariétale au niveau de zones anthracosiques particulières (blackspots) et qu’elles s’accumulaient au niveau des stomas, allant jusqu’à les obstruer et provoquant localement une réaction inflammatoire, avec libération de cytokines, pouvant faire le lit du mésothéliome. Cependant, malgré les données expérimentales et les observations en pathologie humaine, les mécanismes responsables de la translocation des fibres dans la plèvre pariétale ne sont pas encore clairement établis.
It is now well established that some pleural diseases, pleural plaques and malignant mesothelioma are related to asbestos fibre exposure although the mechanism of action of asbestos fibres is not fully understood. The development of artificial mineral fibres and carbon nanotubes, which share some morphological characteristics similar to asbestos fibres, is a present concern in the context of pleural diseases. Pleural plaques develop only in the parietal pleura, and in the 1990s, clinical observations have shown that the early development of mesothelioma also occurred on the parietal pleura. The peculiarity of the parietal pleura in contrast to the visceral pleura is the presence of “stomas” which are communication holes between the pleural cavity and the parietal pleura lymphatics. Morphological observations by thoracoscopy and experimental studies have shown that inhaled fibres translocate to the pleural space and, in human, are present in the parietal pleura at specific anthracotic areas (blackspots). Fibres accumulate on the stomas, up to block and locally induce an inflammatory reaction with cytokines release, that can be the bed of mesothelioma. However, despite the experimental data and observations in human pathology, the mechanisms of fibre translocation into the pleura is not yet clearly established.
Objectif de l'étude. –
Étudier la signification pronostique du contenu en ADN et de la fraction de cellules en phase S de cancers du sein de stades I et II.
Patientes et méthodes. –
Une étude a été ...réalisée chez 271 patientes traitées par chirurgie suivie ou non de chimiothérapie, d'hormonothérapie et de radiothérapie. La durée médiane de suivi était de 64 mois. Une préparation cellulaire standardisée de cytométrie en flux a été utilisée à partir d'échantillons congelés avec des règles consensuelles pour l'interprétation des données. Trois classes de fraction de cellules en phase S ont été définies sur la base de terciles après ajustement au contenu en ADN. En combinant le contenu d'ADN et la fraction de cellules en phase S, quatre groupes pronostiques ont été définis : tumeurs diploïdes à fraction de cellules en phase S basse (
n
=
37), tumeurs diploïdes à fraction de cellules en phase S intermédiaire et/ou haute (
n
=
76), tumeurs aneuploïdes à fraction de cellules en phase S basse (
n
=
24), tumeurs aneuploïdes à fraction de cellules en phase S intermédiaire et/ou haute (
n
=
68). Une corrélation a été recherchée entre les taux de contrôle local, de survie sans rechute, de survie sans métastase et de survie globale d'une part, le contenu en ADN, la fraction de cellules en phase S, ces deux critères combinés, les stades T et N, le grade SBR, l'âge et à la présence de récepteurs hormonaux, d'autre part, avec une analyse uni- et multifactorielle (modèle de Cox).
Résultats. –
Après analyse unifactorielle, les taux de contrôle local et de survie sans rechute étaient significativement meilleurs en cas de tumeur diploïde. Le taux de survie sans rechute était significativement plus bas en cas de fraction de cellules en phase S élevée. Les taux de contrôle local, de survie sans rechute, de survie sans métastase et de survie globale étaient moins bons lorsque la tumeur était diploïde et la fraction de cellules en phase S intermédiaire et/ou haute que lorsqu'elle était diploïde et la fraction de cellules en phase S basse. D'après analyse multifactorielle, le groupe défini à partir de la combinaison du contenu d'ADN et la fraction de cellules en phase S était un facteur indépendant de survie sans métastase, de même que l'atteinte ganglionnaire (N+) et le stade T. Chez les patientes dont le cancer n'atteignait pas l'aisselle (N–), le groupe défini en combinant le contenu d'ADN et la fraction de cellules en phase restait le seul facteur pronostique indépendant de survie sans métastase et de survie sans rechute, alors qu'il n'avait aucune influence en cas d'atteinte axillaire. Lorsque la tumeur était de grade SBR III, il y avait une relation significative entre le groupe défini en combinant le contenu d'ADN et la fraction de cellules en phase S et la survie sans métastase et la survie globale.
Conclusion. –
Ces résultats soulignent l'intérêt de l'utilisation de la combinaison du contenu en ADN et de la fraction de cellules en phase S chez les patientes atteintes d'un cancer du sein de stades I et II, sans atteinte axillaire.
Purpose. –
To assess the significance of S-phase fraction (SPF) and DNA ploidy evaluated by DNA flow cytometry as prognostic markers in stage I or II breast cancer.
Patients and methods. –
A series of 271 patients, treated by surgery, radiotherapy
±
systemic therapy was analysed (median follow up: 64 months). Standardized flow cytometry cell preparation from frozen samples and consensus rules for data interpretation were followed. Three SPF classes were defined on the basis of tertiles after adjustment for ploidy. Four groups were defined based on combinations of DNA ploidy (DIP: diploid; ANEUP: aneuploid) and SPF: DIP and low SPF (DL,
N
=
37), DIP and medium or high SPF (DMH,
N
=
76), ANEUP and low SPF (AL,
N
=
24), ANEUP and medium or high SPF (AMH,
N
=
68). Local control rate (LCR), disease-free survival (DFS), metastasis-free survival (MFS), and overall survival (OS) were correlated with DNA ploidy, SPF, DL to AMH groups, T and N stages, SBR grading, age, and hormonal status on univariate and multivariate analysis (Cox model).
Results. –
On univariate analysis, DFS and LCR were higher for DIP tumours. High SPF values were associated with shorter DFS. LCR, MFS, DFS, and OS rates were significantly different with an increasingly poorer prognosis from DL to AMH. On multivariate analysis, groups DL to AMH, histological node involvement and T stage were independently associated with MFS, and DFS. In N– patients, DL to AMH remained independent for MFS and DFS. For SBR III tumours, MFS and OS were significantly different in DL to AMH groups. These results strongly support the use of combined evaluation of DNA ploidy and SPF as independent parameters in clinical trials for N– stage I and II breast cancer.
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