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1.
Environ Sci Technol ; 2024 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-39145972

RESUMO

This study delves into the unexplored distribution and accumulation of chlorinated paraffins (CPs), pervasive industrial contaminants used as flame retardants and plasticizers, within the hadal trenches, some of Earth's most isolated marine ecosystems. Analysis of sediments from the Mussau (MS) and Mariana trench (MT) reveals notably high total CP concentrations (∑SCCPs + ∑MCCPs) of 10,963 and 14,554 ng g-1 dw, respectively, surpassing those in a reference site in the western Pacific abyssal plain (8533 ng g-1 dw). In contrast, the New Britain Trench (NBT) exhibits the lowest concentrations (2213-5880 ng g-1 dw), where CP distribution correlates with clay content, δ13C and δ15N values, but little with total organic carbon and depth. Additionally, amphipods from these trenches display varying CP levels, with MS amphipods reaching concerning concentrations (8681-16,138 ng g-1 lw), while amphipods in the MT-1 site show the lowest (4414-5010 ng g-1 lw). These bioaccumulation trends appear to be primarily influenced by feeding behaviors (δ13C) and trophic levels (δ15N). Utilizing biota-sediment accumulation factor values and principal component analysis, we discern that CPs in sediment may come from surface-derived particulate organic matters, while those in amphipods may come from the above carrion. Our findings elucidate the profound impacts of the emerging pollutants on the Earth's least explored marine ecosystems.

2.
Case Rep Vet Med ; 2024: 1402828, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39161575

RESUMO

Canine protothecosis is a rare disease caused by saprophytic unicellular achlorophyllous aerobic algae that are ubiquitous in the environment. We report a novel case of neurological and cardiological manifestations associated with disseminated protothecosis. An adult spayed female Boxer dog was presented with a 1-week history of anorexia, progressive central vestibular signs, and a Grade III/VI systolic heart murmur. Magnetic resonance (MR) imaging revealed obstructive hydrocephalus at the level of the mesencephalic aqueduct, while echocardiography and elevated troponin levels suggested an infiltrative cardiomyopathy. No obvious cause was identified. Cerebrospinal fluid (CSF) collection was not performed due to associated procedural risks. Despite receiving symptomatic treatment and maintaining stability for 3 weeks, the dog eventually suffered cardiorespiratory arrest. Postmortem examination revealed disseminated protothecosis, predominantly affecting the heart and brain. We recommend that in cases where the cause of obstructive hydrocephalus is unclear, especially when CSF collection is not feasible, a comprehensive diagnostic method should be implemented. This includes meticulous investigations to identify infected tissues, followed by sampling and performing cytology/histology and culture tests to confirm the presence of the algal organism. Early diagnosis may allow early treatment, although long-term prognosis remains largely unfavorable due to the absence of effective treatments.

3.
Sci Rep ; 14(1): 18275, 2024 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-39107471

RESUMO

Formalin-fixed paraffin-embedded (FFPE) tissue represents a valuable source for translational cancer research. However, the widespread application of various downstream methods remains challenging. Here, we aimed to assess the feasibility of a genomic and gene expression analysis workflow using FFPE breast cancer (BC) tissue. We conducted a systematic literature review for the assessment of concordance between FFPE and fresh-frozen matched tissue samples derived from patients with BC for DNA and RNA downstream applications. The analytical performance of three different nucleic acid extraction kits on FFPE BC clinical samples was compared. We also applied a newly developed targeted DNA Next-Generation Sequencing (NGS) 370-gene panel and the nCounter BC360® platform on simultaneously extracted DNA and RNA, respectively, using FFPE tissue from a phase II clinical trial. Of the 3701 initial search results, 40 articles were included in the systematic review. High degree of concordance was observed in various downstream application platforms. Moreover, the performance of simultaneous DNA/RNA extraction kit was demonstrated with targeted DNA NGS and gene expression profiling. Exclusion of variants below 5% variant allele frequency was essential to overcome FFPE-induced artefacts. Targeted genomic analyses were feasible in simultaneously extracted DNA/RNA from FFPE material, providing insights for their implementation in clinical trials/cohorts.


Assuntos
Neoplasias da Mama , Estudos de Viabilidade , Formaldeído , Genômica , Inclusão em Parafina , Fixação de Tecidos , Humanos , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Inclusão em Parafina/métodos , Feminino , Formaldeído/química , Fixação de Tecidos/métodos , Genômica/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Perfilação da Expressão Gênica/métodos
4.
J Med Virol ; 96(8): e29837, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39105417

RESUMO

Human papillomavirus (HPV) infections are an increasing cause of oropharyngeal squamous cell carcinomas (OPSCC). Integration of the viral genome into the host genome is suggested to affect carcinogenesis, however, the correlation with OPSCC patient prognosis is still unclear. Research on HPV integration is hampered by current integration detection technologies and their unsuitability for formalin-fixed paraffin-embedded (FFPE) tissues. This study aims to develop and validate a novel targeted proximity-ligation based sequencing method (targeted locus amplification/capture [TLA/TLC]) for HPV integration detection in cell lines and FFPE OPSCCs. For the identification of HPV integrations, TLA/TLC was applied to 7 cell lines and 27 FFPE OPSCCs. Following preprocessing steps, a polymerase chain reaction (PCR)-based HPV enrichment was performed on the cell lines and a capture-based HPV enrichment was performed on the FFPE tissues before paired-end sequencing. TLA was able to sequence up to hundreds of kb around the target, detecting exact HPV integration loci, structural variants, and chromosomal rearrangements. In all cell lines, one or more integration sites were identified, in accordance with detection of integrated papillomavirus sequences PCR data and the literature. TLC detected integrated HPV in 15/27 FFPE OPSCCs and identified simple and complex integration patterns. In general, TLA/TLC confirmed PCR data and detected additional integration sites. In conclusion TLA/TLC reliably and robustly detects HPV integration in cell lines and FFPE OPSCCs, enabling large, population-based studies on the clinical relevance of HPV integration. Furthermore, this approach might be valuable for clonality assessment of HPV-related tumors in clinical diagnostics.


Assuntos
Carcinoma de Células Escamosas , Papillomavirus Humano , Neoplasias Orofaríngeas , Infecções por Papillomavirus , Integração Viral , Feminino , Humanos , Masculino , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/virologia , Linhagem Celular Tumoral , DNA Viral/genética , Formaldeído , Papillomavirus Humano/classificação , Papillomavirus Humano/genética , Papillomavirus Humano/isolamento & purificação , Neoplasias Orofaríngeas/virologia , Neoplasias Orofaríngeas/genética , Infecções por Papillomavirus/virologia , Infecções por Papillomavirus/diagnóstico , Inclusão em Parafina , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA , Fixação de Tecidos , Integração Viral/genética
5.
Tissue Cell ; 90: 102511, 2024 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-39141933

RESUMO

Precision-cut liver slices (PCLS) are tissue explants extensively used as an ex vivo model for metabolism and toxicity studies. When in vitro assays are conducted, it is imperative to perform a histomorphological evaluation as part of the viability analyses throughout the assay time. It is considered that good quality PCLS histological sections are difficult to obtain because they are hard to manipulate, and may shrink or fold during processing. Moreover, bibliography is not detailed on the embedding processes used. In this article, we propose an adjusted and rapid method for paraffin embedding of PCLS from crossbreed steers. Each PCLS was covered with a piece of gauze and placed into a histological cassette. These cassettes were submitted to a series of baths: 80 % ethanol for 10 min; 3 baths of 96 % ethanol for 10 min; 3 baths of butanol for 10 min; 1 bath of butanol-paraffin (1:1) for 20 min in a 60 °C laboratory oven; and 3 baths of paraffin for 20 min in a 60 °C laboratory oven. Folded paper boxes were used to produce paraffin blocks. It was possible to obtain complete sections with preserved cell morphology and no artifacts, and tissue appearance was similar to previous PCLS processed through the routine protocol, demonstrating the adequacy of the method implemented.

6.
J Wound Care ; 33(8): 570-576, 2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39137253

RESUMO

OBJECTIVE: Acute laceration wound (ALW) is one of the most common injuries in Indonesia with potential significant morbidities. In rural areas, povidone-iodine and honey are commonly used as wound dressings. This study aimed to identify the effectiveness of honey compared to paraffin gauze and the commonly used povidone-iodine in improving ALW healing time. METHOD: This study was a single-blind, pilot randomised controlled trial (RCT) with three intervention groups (honey, povidone-iodine, and paraffin). The outcomes were wound healing time, slow healing, secondary healing, signs of infection, wound dehiscence, oedema, maceration, necrosis, exudate and cost. RESULTS: A total of 35 patients (male to female ratio: 4:1), with a mean age of 22.5 (range: 6-47) years, were included and randomised to treatment groups using predetermined randomisation according to wound location and wound dressing selection: honey group, n=12; povidone-iodine group, n=11; paraffin group, n=12 with one patient lost to follow-up. All groups achieved timely healing, with a mean healing time of 9.45±5.31 days and 11.09±5.14 days for the povidone-iodine and paraffin groups, respectively, and a median healing time of 10 (3-19) days for the honey group (p>0.05). More wounds in the honey group achieved healing in ≤10 days compared with the other groups. Both povidone-iodine and honey groups had fewer adverse events, with the latter having the lowest cost. CONCLUSION: In this study, honey was clinically effective in accelerating healing time with a lower cost compared to paraffin, and was comparable to povidone-iodine. Future RCTs with a larger sample size should be pursued to determine honey's role in ALW treatment.


Assuntos
Anti-Infecciosos Locais , Mel , Lacerações , Povidona-Iodo , Cicatrização , Humanos , Povidona-Iodo/uso terapêutico , Cicatrização/efeitos dos fármacos , Masculino , Feminino , Adulto , Projetos Piloto , Pessoa de Meia-Idade , Adolescente , Método Simples-Cego , Anti-Infecciosos Locais/uso terapêutico , Lacerações/terapia , Adulto Jovem , Criança , Indonésia , Bandagens , Parafina/uso terapêutico , Resultado do Tratamento
7.
Med Mol Morphol ; 2024 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-39141108

RESUMO

Invasive fungal infections including invasive pulmonary aspergillosis (IPA) generally have a poor prognosis, because the fungi spread throughout various organs. Therefore, it is important to accurately identify the fungal species for treatment. In this article, we present the results of pathological and molecular morphological analyses that were performed to elucidate the cause of respiratory failure in a patient who died despite suspicion of IPA and treatment with micafungin (MCFG). Pathological analysis revealed the existence of cystic and linear fungi in lung tissue. The fungi were identified as Aspergillus fumigatus (A. fumigatus) by partial sequencing of genomic DNA. Correlative light microscopy and electron microscopy (CLEM) analysis confirmed that fungi observed with light microscopy can also be observed with scanning electron microscopy (SEM) using formalin-fixed paraffin-embedded tissue sections. SEM revealed an atypical ultrastructure of the fungi including inhomogeneous widths, rough surfaces, and numerous cyst-like structures of various sizes. The fungi showed several morphological changes of cultured A. fumigatus treated with MCFG that were previously reported. Our results indicate that integrated analysis of ultrastructural observation by SEM and DNA sequencing may be an effective tool for analyzing fungi that are difficult to identify by conventional pathological analysis.

8.
BMC Endocr Disord ; 24(1): 138, 2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39090709

RESUMO

BACKGROUND: Mitochondrial DNA (mtDNA) copy number is associated with tumor activity and carcinogenesis. This study was undertaken to investigate mtDNA copy number in papillary thyroid cancer (PTC) tissues and to evaluate the risk of PTC development. The clinicopathological features of patients and mtDNA copy number were correlated. The value of mtDNA copy number was evaluated as a biomarker for PTC. METHOD: DNA was extracted from 105 PTC tissues and 67 control thyroid tissues, and mtDNA copy number mtDNA oxidative damage were determined using qPCR techniques. RESULTS: Overall, the relative mtDNA copy number was significantly higher in PTC patients (p < 0.001). The risk of developing PTC increased significantly across the tertiles of mtDNA copy number (p trend < 0.001). The higher the mtDNA copy number tertile, the greater the risk of developing PTC. Patients with follicular variants had an odds ratio of 2.09 (95% CI: 1.78-2.44) compared to those with classical variants (p < 0.001). The level of mtDNA oxidative damage in PTC was significantly elevated compared to controls (p < 0.001). The ROC analysis of mtDNA copy number indicated an area under the curve (AUC) of 77.7% (95% CI: 0.71 to 0.85, p < 0.001) for the ability of mtDNA copy number z-scores in differentiate between PTC and controls. CONCLUSION: Our results indicated that the augmentation of mtDNA content plays a significant role during the initiation of thyroid cancer, and it might represent a potential biomarker for predicting the risk of PTC.


Assuntos
Variações do Número de Cópias de DNA , DNA Mitocondrial , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide , Humanos , DNA Mitocondrial/genética , Masculino , Feminino , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/epidemiologia , Câncer Papilífero da Tireoide/genética , Câncer Papilífero da Tireoide/patologia , Pessoa de Meia-Idade , Adulto , Estudos de Casos e Controles , Fatores de Risco , Biomarcadores Tumorais/genética , Prognóstico , Seguimentos
9.
Infect Drug Resist ; 17: 2833-2851, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39005849

RESUMO

Objective: The increasing incidence of chronic skin infections caused by Mycobacterium marinum, coupled with the time-consuming and low detection rates nature of traditional culture and histological-based diagnostic methods, underscores the need for an expedited approach. The study aims to develop a rapid and efficient method for detecting M. marinum with PCR technology. Methods: We designed four pairs of primers based on DNA sequences from GeneBank and prior studies, we utilized both PCR and Real-time PCR to identify M. marinum. Specificity and sensitivity assessments were conducted in vitro by DNAs extracted from M. marinum and other bacterial or fungal cultures. Further validation was performed through the implementation of a mouse skin infection model to optimize and confirm the efficacy of the detection method in both fresh and paraffin-embedded skin tissues. The same PCR testing system was further confirmed with paraffin-embedded skin tissues samples from patients as well. Results: The results of the study indicate promising outcomes for the four-pair primers system. It demonstrated 100% sensitivity in detecting M. marinum from purified cultures, including typical strains and nine clinical isolates, while achieving a specificity of 100%. This specificity was evidenced by the absence of PCR products from 12 bacterial species, 12 fungi species, and six other non-tuberculous mycobacterium (NTM) species. In the animal model, the PCR assay exhibited high detection efficacy for both infected fresh tissues and paraffin-embedded tissues, with a slight superiority observed in fresh tissues. However, the PCR assay exhibited high detection efficacy for clinical paraffin-embedded tissues. These findings collectively underscore the robust detection capabilities of our four-pair primers in both in vitro and in vivo settings. Conclusion: A sensitive and highly specific rapid detection system has been successfully developed that can be used to detect M. marinum in both infected fresh tissues and paraffin-embedded tissues.

10.
Heliyon ; 10(12): e32896, 2024 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-38988576

RESUMO

Whole transcriptome analysis (WTA) using RNA extracted from Formalin Fixed Paraffin Embedded (FFPE) tissue is an invaluable tool to understand the molecular pathology of disease. RNA extracted from FFPE tissue is either degraded and/or in very low quantities hampering gene expression analysis. Earlier studies described protocols applied for cellular RNA using poly-A primer-based linear amplification. The current study describes a method, LINCATRA (LINear amplifiCAtion of RNA for whole TRAnscriptome analysis). It employs random nonamer primer based method which can amplify short, fragmented RNA with high fidelity from as low as 5 ng to obtain enough material for WTA. The two-cycle method significantly amplified RNA at ∼1000 folds (p < 0.0001) improving the mean read lengths (p < 0.05) in WTA. Overall, increased mean read length positively correlated with on-target reads (Pearson's r = 0.71, p < 0.0001) in both amplified and unamplified RNA-seq analysis. Gene expression analysis compared between unamplified and amplified group displayed substantial overlap of the differentially expressed genes (DEGs) (log2 fold change cut-off < -2 and >2, p < 0.05) identified between lung cancer and asthma cohorts validating the method developed. This method is applicable in clinical molecular pathology field for both diagnostics and elucidation of disease mechanisms.

11.
Int J Mol Sci ; 25(14)2024 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-39062962

RESUMO

Postharvest fibrosis and greening of Toona sinensis buds significantly affect their quality during storage. This study aimed to clarify the effects of low-temperature storage on postharvest red TSB quality harvested in different seasons. Red TSB samples were collected from Guizhou province, China, 21 days after the beginning of spring (Lichun), summer (Lixia), and autumn (Liqiu), and stored at 4 °C in dark conditions. We compared and analyzed the appearance, microstructure, chlorophyll and cellulose content, and expression levels of related genes across different seasons. The results indicated that TSB harvested in spring had a bright, purple-red color, whereas those harvested in summer and autumn were green. All samples lost water and darkened after 1 day of storage. Severe greening occurred in spring-harvested TSB within 3 days, a phenomenon not observed in summer and autumn samples. Microstructural analysis revealed that the cells in the palisade and spongy tissues of spring and autumn TSB settled closely during storage, while summer TSB cells remained loosely aligned. Xylem cells were smallest in spring-harvested TSB and largest in autumn. Prolonged storage led to thickening of the secondary cell walls and pith cell autolysis in the petioles, enlarging the cavity area. Chlorophyll content was higher in leaves than in petioles, while cellulose content was lower in petioles across all seasons. Both chlorophyll and cellulose content increased with storage time. Gene expression analysis showed season-dependent variations and significant increases in the expression of over half of the chlorophyll-related and cellulose-related genes during refrigeration, correlating with the observed changes in chlorophyll and cellulose content. This research provides valuable insights for improving postharvest storage and freshness preservation strategies for red TSB across different seasons.


Assuntos
Celulose , Clorofila , Temperatura Baixa , Estações do Ano , Clorofila/metabolismo , Celulose/metabolismo , Regulação da Expressão Gênica de Plantas , China
12.
J Histotechnol ; : 1-7, 2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-38957981

RESUMO

H vessels are an essential link in angiogenic-osteogenic coupling and orchestrate the process of bone healing. H vessels are critically deficient in the setting of radiation-induced fractures, which have been reported to occur in up to 25% of patients undergoing radiotherapy. By increasing H-vessel proliferation, Deferoxamine (DFO) revitalizes the physiologic response to skeletal injury and accelerates irradiated fracture repair. H-vessel quantification is therefore an important outcome measure in histologic analysis of bone healing. However, an optimized protocol for staining H vessels in formalin-fixed paraffin-embedded (FFPE) tissue sections has not been reported. With this protocol, we describe a method of staining FFPE bone samples with minimal background fluorescence and high signal-to-noise ratio. We examined mandibular specimens in a rat model of bone healing from a range of fracture conditions, including healthy bone (Fx), irradiated bone (XFx), and irradiated bone with DFO treatment (XFx-DFO). Quantitative analysis revealed a significant increase of H vessels in the XFxDFO group compared to both the Fx and XFx groups. By optimizing immunofluorescent staining of H vessels in FFPE samples across a range of fracture conditions, we offer investigators an efficacious means of producing reliable imaging for quantitative analysis of H vessels in an irradiated fracture callus.

13.
Cancers (Basel) ; 16(14)2024 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-39061169

RESUMO

Pancreatic neuroendocrine neoplasms pose a growing clinical challenge due to their rising incidence and variable prognosis. The current study aims to investigate microRNAs (miRNA; miR) as potential biomarkers for distinguishing between grade 1 (G1) and grade 2 (G2) pancreatic neuroendocrine tumors (PanNET). A total of 33 formalin-fixed, paraffin-embedded samples were analyzed, comprising 17 G1 and 16 G2 tumors. Initially, literature-based miRNAs were validated via real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), confirming significant downregulation of miR-130b-3p and miR-106b in G2 samples. Through next-generation sequencing, we have identified and selected the top six miRNAs showing the highest difference between G1 and G2 tumors, which were further validated. RT-qPCR validation confirmed the downregulation of miR-30d-5p in G2 tumors. miRNA combinations were created to distinguish between the two PanNET grades. The highest diagnostic performance in distinguishing between G1 and G2 PanNETs by a machine learning algorithm was achieved when using the combination miR-106b + miR-130b-3p + miR-127-3p + miR-129-5p + miR-30d-5p. The ROC analysis resulted in a sensitivity of 83.33% and a specificity of 87.5%. The findings underscore the potential use of miRNAs as biomarkers for stratifying PanNET grades, though further research is warranted to enhance diagnostic accuracy and clinical utility.

14.
Sci Rep ; 14(1): 17077, 2024 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-39048620

RESUMO

To study the source and content change of oridonin in the ice ribbons, the contents of oridonin in the ice ribbons and bleeding sap of Isodon rubescens at different times were determined with RP-HPLC. The paraffin sectioning and electron microscopy imaging were performed to study the transport channel of oridonin in the stem. The results showed that there were abundant xylem rays and perfect pit pairs in the secondary xylem of I. rubescens stems. The oridonin content in the ice ribbons of I. rubescens stems was lower than that in the stem of I. rubescens and even decreased over time. The contents of oridonin in the bleeding sap of I. rubescens stems was equal to that in second-day ice ribbons and was lower than that in first-day ice ribbons. The water in the ice ribbons of I. rubescens stems originated from water absorbed by the roots from soil. This water was transported from the roots of I. rubescens to the stem and then transferred through efficient lateral conducting tissues to the surface of the stem. The oridonin in the phloem and cortex of I. rubescens stems dissolves in water originating from the soil and freezes in the form of ice ribbons below 0 °C.


Assuntos
Diterpenos do Tipo Caurano , Gelo , Isodon , Água , Xilema , Diterpenos do Tipo Caurano/química , Isodon/química , Água/química , Gelo/análise , Xilema/química , Xilema/metabolismo , Caules de Planta/química , Raízes de Plantas/química , Floema/química , Floema/metabolismo
15.
Indian Dermatol Online J ; 15(4): 620-623, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39050059

RESUMO

Background: When immunofluorescence on the frozen section is insufficient or unavailable, salvage immunofluorescence techniques can be used on formalin-fixed, paraffin-embedded tissue. The goal of the current investigation was to evaluate the diagnostic value of paraffin immunofluorescence following proteinase K digestion on skin biopsy samples in comparison to fresh frozen immunofluorescence. Materials and Methods: It was standardized and compared to the immunofluorescence on fresh frozen tissue (IF-Frozen) for paraffin immunofluorescence by proteinase K digestion of formalin-fixed paraffin-embedded skin biopsies (IF-FFPE). The study included 50 native skin biopsy cases, and fluorescein isothiocyanate-labeled IgA, IgG, IgM, and C3 intensity levels were evaluated in each case. Results: A total of 50 cases of native skin biopsy were included in the study, and their intensities for IgA, IgG, IgM, and C3 antibodies were compared. The average staining intensities in each disease group for the antibodies had equal intensity or had a minor difference (1+)/significant difference (2+). Paraffin immunofluorescence, proteinase K digestion had the best correlation, that is, had either equal or minor difference (1+) with fresh frozen immunofluorescence. The difference of 2+ intensity of antibodies between IF-FFPE and IF-Frozen was noted mainly in C3 antibody on paraneoplastic pemphigus. IF-FFPE showed a sensitivity of 100%, 97.6%, 100%, and 81.6% for IgA, IgG, IgM, and C3, respectively, whereas the specificity was 100% for IgA, IgG, IgM, and C3. Limitations: Small sample size and and the employment of one method of antigen retrieval in IF-FFPE. Conclusion: Immunofluorescence techniques done on formalin-fixed paraffin-embedded tissue can serve as salvage techniques in cases where immunofluorescence on the frozen section may not be adequate or may not be available.

16.
Methods Mol Biol ; 2823: 193-223, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39052222

RESUMO

Archived tumor specimens are routinely preserved by formalin fixation and paraffin embedding. Despite the conventional wisdom that proteomics might be ineffective due to the cross-linking and pre-analytical variables, these samples have utility for both discovery and targeted proteomics. Building on this capability, proteomics approaches can be used to maximize our understanding of cancer biology and clinical relevance by studying preserved tumor tissues annotated with the patients' medical histories. Proteomics of formalin-fixed paraffin-embedded (FFPE) tissues also integrates with histological evaluation and molecular pathology strategies, so that additional collection of research biopsies or resected tumor aliquots is not needed. The acquisition of data from the same tumor sample also overcomes concerns about biological variation between samples due to intratumoral heterogeneity. However, the protein extraction and proteomics sample preparation from FFPE samples can be onerous, particularly for small (i.e., limited or precious) samples. Therefore, we provide a protocol for a recently introduced kit-based EasyPep method with benchmarking against a modified version of the well-established filter-aided sample preparation strategy using laser-capture microdissected lung adenocarcinoma tissues from a genetically engineered mouse model. This model system allows control over the tumor preparation and pre-analytical variables while also supporting the development of methods for spatial proteomics to examine intratumoral heterogeneity. Data are posted in ProteomeXchange (PXD045879).


Assuntos
Formaldeído , Inclusão em Parafina , Proteômica , Fixação de Tecidos , Proteômica/métodos , Inclusão em Parafina/métodos , Fixação de Tecidos/métodos , Formaldeído/química , Animais , Camundongos , Humanos , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/genética , Microdissecção e Captura a Laser/métodos , Neoplasias/patologia , Neoplasias/metabolismo , Neoplasias/genética , Adenocarcinoma de Pulmão/patologia , Adenocarcinoma de Pulmão/metabolismo
17.
EFSA J ; 22(7): e8913, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-39045513

RESUMO

The conclusions of the European Food Safety Authority (EFSA) following the peer review of the initial risk assessments carried out by the competent authorities of the rapporteur Member State, Greece, and co-rapporteur Member State, France, for the pesticide active substance paraffin oil are reported. The context of the peer review was that required by Commission Implementing Regulation (EU) No 844/2012, as amended by Commission Implementing Regulation (EU) No 2018/1659. The conclusions were reached on the basis of the evaluation of the representative uses of paraffin oil as an acaricide and insecticide on potatoes, ornamentals (flower bulbs) and orchards (pear/apple), on pome fruit and stone fruit, on field and permanent protected fruiting vegetables and on field and permanent protected roses and on citrus. The reliable end points appropriate for use in regulatory risk assessment are presented. Missing information identified as being required by the regulatory framework is listed. Concerns are reported where identified.

18.
Methods Mol Biol ; 2825: 173-184, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38913309

RESUMO

Multitarget fluorescence in situ hybridization (mFISH) is a technique that allows the detection of multiple target sequences on the same sample using spectrally distinct fluorophore labels. The mFISH approach is currently a useful assay in the oncologic field for the detection of predictive, prognostic, and diagnostic biomarkers. In this chapter, we summarize the application of mFISH in the identification of target genetic aberrations in formalin-fixed, paraffin-embedded (FFPE) tissue samples of several tumor types. We discuss the mFISH protocols in FFPE samples, the innovative multitarget probes used, and the critical issues related to their interpretation.


Assuntos
Hibridização in Situ Fluorescente , Neoplasias , Inclusão em Parafina , Hibridização in Situ Fluorescente/métodos , Humanos , Neoplasias/genética , Neoplasias/diagnóstico , Inclusão em Parafina/métodos , Fixação de Tecidos/métodos , Biomarcadores Tumorais/genética , Formaldeído/química
19.
BMC Public Health ; 24(1): 1648, 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38902608

RESUMO

BACKGROUND: Injury due to ingestion of harmful chemicals has become an area of concern globally. In South Africa, paraffin has been widely implicated in multiple health outcomes, including severe ingestion injuries. A specific category of such injuries is those that are self-inflicted. A significant proportion of self-inflicted ingestion is reported to be intentional, although intentionality for self-infliction may be difficult to determine. Nonetheless, the identification of key explanatory risks and demographic factors of self-inflicted ingestion may contribute towards a better understanding of self-inflicted and harmful chemical ingestion injuries. METHODS: This study used secondary data that had been collected on burn injuries of all causes, including those due to the ingestion of harmful chemicals, from a sample of South Africans from low-income communities close to major metropolitan centres. The current analysis focused on the risks for self-inflicted ingestion injuries and used logistic regression to determine risks for self-inflicted ingestion as differentiated from ingestion due to the actions of another person (other-inflicted ingestion) by sex and age cohort of the victim, and the presence of alcohol, by examining paraffin ingestion versus that of other chemicals. RESULTS: The overwhelming majority of ingestion injuries (92.1%) were self-inflicted. The current findings indicate that sex (with females almost twice as likely to present with self-inflicted ingestion), age cohort (with those aged 18-29 and 30-44 years old four times more likely affected than older adults), presence of alcohol (twice as likely present than amongst individuals reporting ingestion injuries inflicted by others), and chemicals other than paraffin (three times more likely) are key explanatory factors for an increased risk for self-inflicted ingestion of harmful chemicals. CONCLUSIONS: The study empirically confirms the role of several key risk factors in what remains a relatively unreported and understudied phenomenon, but which appears to align with the demographic and risk profile reported for suicidal injuries through chemical ingestion, i.e. intentional self-inflicted ingestion. The findings may contribute towards improved safety policies on the availability and sale of chemical products and more focussed community interventions for at-risk individuals such as females and young people. It also flags the importance of assessing for alcohol use and alcohol use disorders at hospital admission of self-ingestion injuries.


Assuntos
Comportamento Autodestrutivo , Humanos , Feminino , Masculino , Adolescente , África do Sul/epidemiologia , Adulto , Comportamento Autodestrutivo/epidemiologia , Fatores de Risco , Adulto Jovem , Parafina , Pessoa de Meia-Idade , Queimaduras/epidemiologia
20.
J Hazard Mater ; 474: 134759, 2024 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-38823100

RESUMO

Short- and medium-chain chlorinated paraffins (SCCPs and MCCPs) are hazardous industrial chemicals that tend to bioaccumulate in animal-derived foodstuffs through the food supply chain. However, the lack of reliable noninvasive bioindicators hinders the monitoring of farm animal exposure to CPs. In this study, 169 cattle hair samples were collected from beef cattle farms in six Chinese provinces, with further beef, feed, and soil samples being collected in Hebei province. Geographical differences in CP concentrations were observed in the hair samples, and CP concentrations in samples collected from Hebei province decreased in the following order: hair > feed > beef > soil. C10-11Cl6-7 and C14Cl7-8 were the predominant SCCPs and MCCPs, respectively, in all the hair, beef, feed, and soil samples. CP concentrations in hair samples significantly correlated with those in beef, feed, and soil samples, indicating that hair can be used as a bioindicator of cattle exposure to CPs. The possible health risks associated with exposure to CPs through beef consumption, especially for children and high-volume beef consumers, should be further investigated.


Assuntos
Cabelo , Parafina , Animais , Bovinos , Cabelo/química , Parafina/análise , China , Hidrocarbonetos Clorados/análise , Ração Animal/análise , Monitoramento Ambiental/métodos , Fazendas , Poluentes do Solo/análise , Poluentes Ambientais/análise , Contaminação de Alimentos/análise
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