Formalin, an aqueous solution of formaldehyde, has been the gold standard for fixation of histological samples for over a century. Despite its considerable advantages, growing evidence points to objective toxicity, particularly highlighting its carcinogenicity and mutagenic effects. In 2016, the European Union proposed a ban, but a temporary permission was granted in consideration of its fundamental role in the medical-diagnostic field. In the present study, we tested an innovative fixative, glyoxal acid-free (GAF) (a glyoxal solution deprived of acids), which allows optimal tissue fixation at structural and molecular level combined with the absence of toxicity and carcinogenic activity. An open-label, non-inferiority, multicentric trial was performed comparing fixation of histological specimens with GAF fixative vs standard phosphate-buffered formalin (PBF), evaluating the morphological preservation and the diagnostic value with four binary score questions answered by both the central pathology reviewer and local center reviewers. The mean of total score in the GAF vs PBF fixative groups was 3.7 ± 0.5 vs 3.9 ± 0.3 for the central reviewer and 3.8 ± 0.5 vs 4.0 ± 0.1 for the local pathologist reviewers, respectively. In terms of median value, similar results were observed between the two fixative groups, with a median value of 4.0. Data collected indicate the non-inferiority of GAF as compared to PBF for all organs tested. The present clinical performance study, performed following the international standard for performance evaluation of in vitro diagnostic medical devices, highlights the capability of GAF to ensure both structural preservation and diagnostic value of the preparations.
- Klíčová slova
- Fixation, Formalin, Glyoxal acid-free (GAF), Trial,
- MeSH
- fixace tkání * metody MeSH
- fixativa * chemie MeSH
- formaldehyd * chemie MeSH
- glyoxal * MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- hodnocení ekvivalence MeSH
- multicentrická studie MeSH
- srovnávací studie MeSH
- Názvy látek
- fixativa * MeSH
- formaldehyd * MeSH
- glyoxal * MeSH
With the onset of the COVID-19 pandemic, a problem arose with classic body donation programmes for obtaining cadavers for anatomical dissections, science and research. The question has emerged whether bodies of individuals who died of COVID-19 or were infected by SARS-CoV-2 could be admitted to Departments of Anatomy. To determine the risk of SARS-CoV-2 transmission to employees or students, the presence and stability of SARS-CoV-2 RNA in cadavers after fixation agents' application and subsequent post-fixation baths over time were examined. The presence of viral RNA in swabs from selected tissues was assessed by the standardized routine RNA isolation protocol and subsequent real-time PCR analysis. To support the results obtained from the tissue swabs, samples of RNA were exposed in vitro to short and long-term exposure to the components of the injection and fixation solutions used for the bodies' conservation. Substantial removal of SARS-CoV-2 RNA was observed in post-mortem tissue following perfusion with 3.5% phenol, 2.2% formaldehyde, 11.8% glycerol and 55% ethanol, and subsequent post-fixation in an ethanol bath. In vitro experiments showed significant effects of formaldehyde on SARS-CoV-2 RNA, while phenol and ethanol showed only negligible effects. We conclude that cadavers subjected to fixation protocols as described here should not pose a considerable risk of SARS-CoV-2 infection while being handled by students and staff and are, therefore, suitable for routine anatomical dissections and teaching.
- Klíčová slova
- Coronavirus, Dissection, Embalming, Infections, Polymerase chain reaction,
- MeSH
- COVID-19 * MeSH
- ethanol MeSH
- fenoly MeSH
- formaldehyd MeSH
- lidé MeSH
- mrtvola MeSH
- pandemie MeSH
- RNA virová MeSH
- SARS-CoV-2 * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- ethanol MeSH
- fenoly MeSH
- formaldehyd MeSH
- RNA virová MeSH
The reactions of 2-naphthylamine and methyl 6-amino-2-naphthoate with formalin and paraformaldehyde were studied experimentally, spectrally, and by quantum chemical calculations. It was found that neither the corresponding aminals nor imines were formed under the described conditions but could be prepared and spectrally characterized at least in situ under modified conditions. Several of the previously undescribed intermediates and by-products were isolated or at least spectrally characterized. First principle density functional theory (DFT) calculations were performed to shed light on the key aspects of the thermochemistry of decomposition and further condensation of the corresponding aminals and imines. The calculations also revealed that the electrophilicity of methanal was significantly greater than that of ordinary oxo-compounds, except for perfluorinated ones. In summary, methanal was not behaving as the simplest aldehyde but as a very electron-deficient oxo-compound.
- Klíčová slova
- DFT, Tröger’s base, mechanisms, methanal, naphthylamine, quinazoline, spiro-Tröger’s base,
- MeSH
- 2-naftylamin * MeSH
- formaldehyd * MeSH
- iminy MeSH
- spektrální analýza MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 2-naftylamin * MeSH
- formaldehyd * MeSH
- iminy MeSH
The utilisation of waste wood from furniture production brings new problems connected with an incomplete thermochemical decomposition of additives (chemicals for improving properties of plastics) in small heating with the addition of sources. Unique organic compounds produced by the combustion of waste wood allow the identification of the type of fuel. The organic compounds contained in the char deposits were analysed by pyrolysis gas chromatography with mass spectrometry. The deposits from the combustion of briquettes from furniture production contain organic compounds originating by decomposition of phenolic resins, aminoplasts (urea-formaldehyde, resorcinol-formaldehyde and melamine), polyurethanes and wood glue. Additives contained in the deposits include plasticisers such as phthalates (DEHP, dibutyl phthalate and diisobutyl phthalate), flame retardants (2-propanol, 1-chlorophosphate (3:1) and p-terphenyl). Deposits from the combustion of briquettes from virgin wood do not contain these compounds. The total amount of compounds identified in the deposits from the boiler, which do not come from virgin wood combustion, varies in the range between 4.25 and 6.25 g/kg. Phthalates (55.5%) and PVAc adhesives (18.6%) are the main anthropogenic compounds in the deposits from domestic boilers.
- Klíčová slova
- Air emissions, Domestic heating, Furniture waste wood, Polymer wood composites, Py-GC/MS,
- MeSH
- dřevo chemie MeSH
- formaldehyd analýza MeSH
- látky znečišťující vzduch * analýza MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- polyurethany analýza MeSH
- vytápění MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- formaldehyd MeSH
- látky znečišťující vzduch * MeSH
- polyurethany MeSH
The determination of secondary volatile degradation products in drying oil extracts is substantial to prevent formation of undesirable metal formates in paintings and/or other artefacts. This study develops a simple, cost-effective, and reliable, high-performance liquid chromatography with diode array detector (HPLC-DAD) method to determine three secondary volatile degradation products (methanol, formaldehyde, and formic acid) in drying oils, including linseed, poppy-seed, and walnut oil. Extraction of analytes was performed using QuEChERS-based procedure followed by metal oxide-based dispersive solid-phase extraction (d-SPE) clean-up and presented a good performance for all of the volatile analytes of interest with recoveries in the range of 90-120% after application of the nanostructured cerium oxide-based (CeO2) and zirconia-based (ZrO2) sorbents prepared by favorable and ecological-friendly methods. With a new clean-up solution for samples with high-fat content, it was possible to achieve higher recoveries than with commercial Z-Sep/C18 sorbent. In all cases, relative standard deviations (RSD) of less than 10% were achieved. No significant matrix interference was observed due to the application of effective sorbents in nanostructured form. The developed method was applied to samples of drying oils, and it was found that after storage for three months, all methanol was most likely oxidized to formaldehyde and formic acid. The concentrations of formaldehyde were in the range of 260 - 304 μg∙g-1, while formic acid concentrations ranged between 72 - 386 μg∙g-1. The highest concentration of formaldehyde (304 μg∙g-1) and formic acid (386 μg∙g-1) was found in linseed oil.
- Klíčová slova
- Drying oil, HPLC-DAD, Methanol-derivates, Nanostructured sorbents,
- MeSH
- extrakce na pevné fázi metody MeSH
- formaldehyd MeSH
- formiáty MeSH
- lněný olej * MeSH
- methanol * MeSH
- oleje MeSH
- oxidy MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- formaldehyd MeSH
- formiáty MeSH
- formic acid MeSH Prohlížeč
- lněný olej * MeSH
- methanol * MeSH
- oleje MeSH
- oxidy MeSH
Mitochondrial oxidative phosphorylation (OXPHOS) generates ATP, but OXPHOS also supports biosynthesis during proliferation. In contrast, the role of OXPHOS during quiescence, beyond ATP production, is not well understood. Using mouse models of inducible OXPHOS deficiency in all cell types or specifically in the vascular endothelium that negligibly relies on OXPHOS-derived ATP, we show that selectively during quiescence OXPHOS provides oxidative stress resistance by supporting macroautophagy/autophagy. Mechanistically, OXPHOS constitutively generates low levels of endogenous ROS that induce autophagy via attenuation of ATG4B activity, which provides protection from ROS insult. Physiologically, the OXPHOS-autophagy system (i) protects healthy tissue from toxicity of ROS-based anticancer therapy, and (ii) provides ROS resistance in the endothelium, ameliorating systemic LPS-induced inflammation as well as inflammatory bowel disease. Hence, cells acquired mitochondria during evolution to profit from oxidative metabolism, but also built in an autophagy-based ROS-induced protective mechanism to guard against oxidative stress associated with OXPHOS function during quiescence.Abbreviations: AMPK: AMP-activated protein kinase; AOX: alternative oxidase; Baf A: bafilomycin A1; CI, respiratory complexes I; DCF-DA: 2',7'-dichlordihydrofluorescein diacetate; DHE: dihydroethidium; DSS: dextran sodium sulfate; ΔΨmi: mitochondrial inner membrane potential; EdU: 5-ethynyl-2'-deoxyuridine; ETC: electron transport chain; FA: formaldehyde; HUVEC; human umbilical cord endothelial cells; IBD: inflammatory bowel disease; LC3B: microtubule associated protein 1 light chain 3 beta; LPS: lipopolysaccharide; MEFs: mouse embryonic fibroblasts; MTORC1: mechanistic target of rapamycin kinase complex 1; mtDNA: mitochondrial DNA; NAC: N-acetyl cysteine; OXPHOS: oxidative phosphorylation; PCs: proliferating cells; PE: phosphatidylethanolamine; PEITC: phenethyl isothiocyanate; QCs: quiescent cells; ROS: reactive oxygen species; PLA2: phospholipase A2, WB: western blot.
- Klíčová slova
- ATG4B, biosynthesis, cell death, electron transport chain, endothelial cells, mitochondria, oxidative phosphorylation, oxidative stress, reactive oxygen species,
- MeSH
- adenosintrifosfát metabolismus MeSH
- autofagie * MeSH
- cystein metabolismus MeSH
- dextrany metabolismus MeSH
- dýchání MeSH
- endoteliální buňky metabolismus MeSH
- fibroblasty metabolismus MeSH
- formaldehyd metabolismus MeSH
- fosfatidylethanolaminy metabolismus MeSH
- idiopatické střevní záněty * metabolismus MeSH
- isothiokyanatany MeSH
- lidé MeSH
- lipopolysacharidy metabolismus MeSH
- mitochondriální DNA metabolismus MeSH
- mitochondrie metabolismus MeSH
- mTORC1 metabolismus MeSH
- myši MeSH
- proteinkinasy aktivované AMP metabolismus MeSH
- proteiny asociované s mikrotubuly metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- sirolimus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- adenosintrifosfát MeSH
- cystein MeSH
- dextrany MeSH
- formaldehyd MeSH
- fosfatidylethanolaminy MeSH
- isothiokyanatany MeSH
- lipopolysacharidy MeSH
- mitochondriální DNA MeSH
- mTORC1 MeSH
- phenethyl isothiocyanate MeSH Prohlížeč
- proteinkinasy aktivované AMP MeSH
- proteiny asociované s mikrotubuly MeSH
- reaktivní formy kyslíku MeSH
- sirolimus MeSH
AIM: Activating mutations in the epidermal growth factor receptor (EGFR) are predominantly detected in pulmonary adenocarcinoma and have been reported in small cell lung cancer (SCLC) for decades. This retrospective single-center study aimed to determine the frequency and types of EGFR mutations in SCLC in Taiwan. METHODS: This study comprises a consecutive cohort of 161 patients histologically diagnosed with SCLC between January 1992 and August 2014 at the Department of Pathology in Keelung Chang Gung Memorial Hospital, Taiwan. Archived formalin-fixed paraffin-embedded sections from 71 patients were eligible for molecular analysis. EGFR mutation analysis was performed using a fully-automated IdyllaTM EGFR Mutation Test and confirmed a comparable result through Qiagen Therascreen® EGFR RGQ PCR. In addition, EGFR gene copy number was assessed in EGFR-mutated tumors by fluorescence in situ hybridization (FISH). RESULTS: Mutational status of the EGFR gene was successfully analyzed in 63 specimens by both IdyllaTM and Qiagen platforms. Both methods detected L858R point mutation in exon 21 in an 81-year-old female and a 47-year-old male non-smoker. Both tumors show no concurrent EGFR gene amplification. The overall agreement between results obtained with the Idylla™ EGFR Mutation Test and Qiagen Therascreen® EGFR RGQ PCR was 100% Conclusions. Our results showed that EGFR mutation is a rare mutation type in a consecutive series of de novo SCLC. Furthermore, the performance of Idylla™ EGFR Mutation Test and Qiagen Therascreen® EGFR RGQ PCR on archived paraffin sections of limited quantities is available with the high agreement of results.
- Klíčová slova
- IdyllaTM, epidermal growth factor receptor, gene amplification, gene mutation, small cell lung cancer,
- MeSH
- erbB receptory * genetika MeSH
- formaldehyd MeSH
- hybridizace in situ fluorescenční MeSH
- lidé MeSH
- malobuněčný karcinom plic * diagnóza genetika MeSH
- mutace MeSH
- mutační analýza DNA metody MeSH
- nádory plic * diagnóza MeSH
- nemalobuněčný karcinom plic diagnóza MeSH
- parafín MeSH
- retrospektivní studie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- Názvy látek
- EGFR protein, human MeSH Prohlížeč
- erbB receptory * MeSH
- formaldehyd MeSH
- parafín MeSH
Biomarker testing is crucial for treatment selection in advanced non-small cell lung cancer (NSCLC). However, the quantity of available tissue often presents a key constraint for patients with advanced disease, where minimally invasive tissue biopsy typically returns small samples. In Part 1 of this two-part series, we summarise evidence-based recommendations relating to small sample processing for patients with NSCLC. Generally, tissue biopsy techniques that deliver the greatest quantity and quality of tissue with the least risk to the patient should be selected. Rapid on-site evaluation can help to ensure sufficient sample quality and quantity. Sample processing should be managed according to biomarker testing requirements, because tissue fixation methodology influences downstream nucleic acid, protein and morphological analyses. Accordingly, 10% neutral buffered formalin is recommended as an appropriate fixative, and the duration of fixation is recommended not to exceed 24-48 h. Tissue sparing techniques, including the 'one biopsy per block' approach and small sample cutting protocols, can help preserve tissue. Cytological material (formalin-fixed paraffin-embedded [FFPE] cytology blocks and non-FFPE samples such as smears and touch preparations) can be an excellent source of nucleic acid, providing either primary or supplementary patient material to complete morphological and molecular diagnoses. Considerations on biomarker testing, reporting and quality assessment are discussed in Part 2.
- Klíčová slova
- Best practice, Biopsy, Cytological techniques, Histology, Molecular diagnostics, Non-small cell lung carcinoma,
- MeSH
- biologické markery MeSH
- fixace tkání metody MeSH
- fixativa MeSH
- formaldehyd MeSH
- lidé MeSH
- nádory plic * diagnóza patologie MeSH
- nemalobuněčný karcinom plic * diagnóza patologie MeSH
- nukleové kyseliny * MeSH
- zalévání tkání do parafínu MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- biologické markery MeSH
- fixativa MeSH
- formaldehyd MeSH
- nukleové kyseliny * MeSH
BACKGROUND: Clinical laboratories routinely use formalin-fixed paraffin-embedded (FFPE) tissue or cell block cytology samples in oncology panel sequencing to identify mutations that can predict patient response to targeted therapy. To understand the technical error due to FFPE processing, a robustly characterized diploid cell line was used to create FFPE samples with four different pre-tissue processing formalin fixation times. A total of 96 FFPE sections were then distributed to different laboratories for targeted sequencing analysis by four oncopanels, and variants resulting from technical error were identified. RESULTS: Tissue sections that fail more frequently show low cellularity, lower than recommended library preparation DNA input, or target sequencing depth. Importantly, sections from block surfaces are more likely to show FFPE-specific errors, akin to "edge effects" seen in histology, while the inner samples display no quality degradation related to fixation time. CONCLUSIONS: To assure reliable results, we recommend avoiding the block surface portion and restricting mutation detection to genomic regions of high confidence.
- Klíčová slova
- Cancer genomics, FFPE, Next-generation sequencing, Oncopanel sequencing, Preanalytics, Precision medicine,
- MeSH
- fixace tkání MeSH
- formaldehyd * MeSH
- lidé MeSH
- sekvenční analýza DNA MeSH
- vysoce účinné nukleotidové sekvenování * MeSH
- zalévání tkání do parafínu MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- formaldehyd * MeSH
Nutrient disorder and presence of disease-causing agents in soilless media negatively influence the growth of muskmelon. To combat these issues, use of environmentally-friendly sanitation techniques is crucial for increased crop productivity. The study was conducted under greenhouse and field conditions to investigate the effect of two different sanitation techniques: steaming and formalin fumigation on various media's characteristics and their impact on muskmelon yield. Media: jantar, guar, wheat straw and rice hull and peat moss of 10% air-filled porosity and sanitized with formalin and steaming. Steaming of guar, jantar, and wheat straw increased the phosphorus (P) and potassium (K) concentrations by 13.80-14.86% and 6.22-8.45% over formalin fumigation. Likewise, P and K concentrations in muskmelon were higher under steaming. Steaming significantly inhibited the survival of Fusarium wilt sp. melonis, root knot nematode sp. meloidogyne and nitrifying bacteria in media than formalin fumigation. In conclusion, steaming decreased the prevalence of nitrifying bacteria and pathogens which thus improved the NO3--N:NH4+-N ratios, P and K nutritional balance both in the media and muskmelon transplants. Hence, steaming as an environment-friendly approach is recommended for soilless media. Further, optimization of steaming for various composts with different crops needs to be investigated with steaming teachnique.
