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International Journal of Molecular... May 2024Cutaneous field cancerization (CFC) refers to a skin region containing mutated cells' clones, predominantly arising from chronic exposure to ultraviolet radiation (UVR),...
Cutaneous field cancerization (CFC) refers to a skin region containing mutated cells' clones, predominantly arising from chronic exposure to ultraviolet radiation (UVR), which exhibits an elevated risk of developing precancerous and neoplastic lesions. Despite extensive research, many molecular aspects of CFC still need to be better understood. In this study, we conducted ex vivo assessment of cell differentiation, oxidative stress, inflammation, and DNA damage in CFC samples. We collected perilesional skin from 41 patients with skin cancer and non-photoexposed skin from 25 healthy control individuals. These biopsies were either paraffin-embedded for indirect immunofluorescence and immunohistochemistry stain or processed for proteins and mRNA extraction from the epidermidis. Our findings indicate a downregulation of p53 expression and an upregulation of Ki67 and p16 in CFC tissues. Additionally, there were alterations in keratinocyte differentiation markers, disrupted cell differentiation, increased expression of iNOS and proinflammatory cytokines IL-6 and IL-8, along with evidence of oxidative DNA damage. Collectively, our results suggest that despite its outwardly normal appearance, CFC tissue shows early signs of DNA damage, an active inflammatory state, oxidative stress, abnormal cell proliferation and differentiation.
Topics: Humans; Oxidative Stress; DNA Damage; Cell Differentiation; Skin Neoplasms; Inflammation; Male; Female; Middle Aged; Ultraviolet Rays; Aged; Keratinocytes; Nitric Oxide Synthase Type II; Adult; Tumor Suppressor Protein p53; Skin; Ki-67 Antigen; Cyclin-Dependent Kinase Inhibitor p16; Interleukin-6
PubMed: 38891963
DOI: 10.3390/ijms25115775 -
International Journal of Molecular... May 2024The mechanism underlying podocyte dysfunction in minimal change disease (MCD) remains unknown. This study aimed to shed light on the potential pathophysiology of MCD...
The mechanism underlying podocyte dysfunction in minimal change disease (MCD) remains unknown. This study aimed to shed light on the potential pathophysiology of MCD using glomerular proteomic analysis. Shotgun proteomics using label-free quantitative mass spectrometry was performed on formalin-fixed, paraffin-embedded (FFPE) renal biopsies from two groups of samples: control (CTR) and MCD. Glomeruli were excised from FFPE renal biopsies using laser capture microdissection (LCM), and a single-pot solid-phase-enhanced sample preparation (SP3) digestion method was used to improve yield and protein identifications. Principal component analysis (PCA) revealed a distinct separation between the CTR and MCD groups. Forty-eight proteins with different abundance between the two groups (-value ≤ 0.05 and |FC| ≥ 1.5) were identified. These may represent differences in podocyte structure, as well as changes in endothelial or mesangial cells and extracellular matrix, and some were indeed found in several of these structures. However, most differentially expressed proteins were linked to the podocyte cytoskeleton and its dynamics. Some of these proteins are known to be involved in focal adhesion (NID1 and ITGA3) or slit diaphragm signaling (ANXA2, TJP1 and MYO1C), while others are structural components of the actin and microtubule cytoskeleton of podocytes (ACTR3 and NES). This study suggests the potential of mass spectrometry-based shotgun proteomic analysis with LCM glomeruli to yield valuable insights into the pathogenesis of podocytopathies like MCD. The most significantly dysregulated proteins in MCD could be attributable to cytoskeleton dysfunction or may be a compensatory response to cytoskeleton malfunction caused by various triggers.
Topics: Humans; Nephrosis, Lipoid; Proteomics; Podocytes; Kidney Glomerulus; Male; Female; Adult; Proteome; Laser Capture Microdissection; Middle Aged
PubMed: 38891801
DOI: 10.3390/ijms25115613 -
Animals : An Open Access Journal From... Jun 2024The objectives of the present study were to describe specific anatomical structures of the dromedary udder. Eighty-six dromedary udders were examined, taking...
The objectives of the present study were to describe specific anatomical structures of the dromedary udder. Eighty-six dromedary udders were examined, taking morphological measurements and producing injection casts using resin, gelatin, and paraffin. The udder of the dromedaries consists of four quarters. The shape of the udders and teats varies considerably between animals and is influenced by age, breeding, and lactation status. The most frequently found udder form was the globular udder (48.8%) and the most common teat form in this study was the funnel teat (44.2%). The most common teat tip shape was a smooth or a slightly rough ring teat (61.6%). Injection casts showed a complete separation of the teat canals. There is also no communication between tributary mammary complexes. Resin injections of the glandular tissue adjacent to the teat cistern showed an extensive branching into large, medium, and small milk ducts. Frozen sections of the udder revealed complete separation of the right and left mammary complex through the . The teat sections showed longitudinal folds from the tip of the teat to the base of the teat. A ring fold at the transition from the to the teat cistern was present. The results of this study increase the knowledge of the anatomical structures of the dromedary udder, which may be useful for breeding a selection of dairy dromedaries.
PubMed: 38891721
DOI: 10.3390/ani14111674 -
Foods (Basel, Switzerland) May 2024The application of organic fertilizers is one of the most important agricultural measures aimed at improving the flavor and productivity of , with the granulation and...
The application of organic fertilizers is one of the most important agricultural measures aimed at improving the flavor and productivity of , with the granulation and coating of organic fertilizers, which can reduce seepage losses of great significance to the ecosystem. In this study, Jingcai 8 tomato was selected as the test material. Headspace solid-phase microextraction and gas chromatography-mass spectrometry (HS-SPME-GC-MS) methods were used to investigate the effects of different pelletized organic fertilizers and various coating materials on the flavor profile of the tomatoes. The results indicated that 67 volatile organic compounds (VOCs) were identified in the tomato fruits and 62 volatile compounds were identified in the leaves under different fertilizer treatments. The volatile compound content of the fruits in the BP treatment group was 35.38 μg/g, which was higher than that in other treatment groups, and the volatile compound content of the leaves was lower. A differential compound analysis with log|fold change| ≥ 1 and variable important in projection (VIP) > 1 highlighted styrene, 3-methyl-1-butanol, and (E, E)-2,4-hexadienal as the major up-regulated compounds and methyl salicylate as the major down-regulated compound in the tomato fruit BCK (control) vs. BP. Moreover, the α-phellandrene content decreased in the tomato leaves. In addition, an analysis of the tomato fruit differential compounds and compounds with odor activity values (OAV) of ≥ 1, considering the OAV values of characteristic aroma compounds, identified key compounds affecting the flavor of the tomato fruits under the BP treatment. These included 2-nonenal, (E)-2-pentylfuran, trans-β-ionone, 1-penten-3-one, (E, E)-2,4-hexadienal, and 3-hexenol (fruity, floral, and herbaceous odors), (E, E)-2,4-heptadienal (fatty odor), and hexanal (green odor). The combined results analysis of the volatile compound content, differential compounds, and OAV values of characteristic aroma compounds aimed to clarify that the BP treatment group, which applied pelletized, large-grain organic fertilizer with polyurethane (pozzolanic + small-grain oil-coated + 2% paraffinic + 4% polyurethane) as a coating material, proved to be most effective in influencing the flavor of the tomato fruits. This finding lays the foundation for its potential commercial application in artificial orchards.
PubMed: 38890880
DOI: 10.3390/foods13111653 -
Open Forum Infectious Diseases Jun 2024Among combat injured, invasive fungal infections (IFIs) result in significant morbidity. Cultures and histopathology are the primary diagnostic methods for IFIs, but...
BACKGROUND
Among combat injured, invasive fungal infections (IFIs) result in significant morbidity. Cultures and histopathology are the primary diagnostic methods for IFIs, but they have limitations. We previously evaluated a panfungal polymerase chain reaction assay, which was 83% sensitive and 99% specific for angioinvasive IFIs. Here, we evaluated 3 less resource-intensive seminested assays targeting clinically relevant fungi in the order Mucorales and genera and .
METHODS
Formalin-fixed paraffin-embedded tissue specimens from a multicenter trauma IFI cohort (2009-2014) were used. Cases were US military personnel injured in Afghanistan with histopathologic IFI evidence. Controls were patients with similar injury patterns and no laboratory IFI evidence (negative culture and histopathology). Seminested assays specific to Mucorales (V4/V5 regions of 18S rDNA), (mitochondrial tRNA), and (internal transcribed spacer [ITS]/28A regions of DNA) were compared with a panfungal assay amplifying the internal transcribed spacer 2 region of rDNA and to histopathology.
RESULTS
Specimens from 92 injury sites (62 subjects) were compared with control specimens from 117 injuries (101 subjects). We observed substantial agreement between the seminested and panfungal assays overall, especially for the order Mucorales. Moderate agreement was observed at the genus level for and . When compared with histopathology, sensitivity and specificity of seminested assays were 67.4% and 96.6%, respectively (sensitivity increased to 91.7% when restricted to sites with angioinvasion).
CONCLUSIONS
Prior studies of seminested molecular diagnostics have focused on culture-negative samples from immunocompromised patients. Our findings underscore the utility of the seminested approach in diagnosing soft-tissue IFIs using formalin-fixed paraffin-embedded tissue samples, especially with angioinvasion.
PubMed: 38887475
DOI: 10.1093/ofid/ofae078 -
EXCLI Journal 2024Commonly used to decode the human brain's structural complexity, dissection focuses on a given structure or region but cannot depict the whole brain organization (for...
Commonly used to decode the human brain's structural complexity, dissection focuses on a given structure or region but cannot depict the whole brain organization (for example, its arterial distribution territories). Where dissection reaches its limit, the combination of tissue sectioning and 3D reconstruction may provide a volume for the assessment of structures from any view angle, following them dynamically to understand their spatial relationships. However, to produce sections, standard histological tissue processing protocols for paraffin embedding cannot be applied to a cerebral hemisphere as the latter is extensively larger than the conventional specimens. This paper presents a protocol for paraffin embedding of the whole human cerebral hemisphere and a method to reconstruct 3D volumes from serially sectioned and photographed paraffin blocks containing embedded hemispheres. Seven whole human cerebral hemispheres were included, two were serially sectioned. Main cerebral arteries were injected with colored media to label arterial territories. A detailed description of every step, from tissue processing to image acquisition of cut blockfaces and volume reconstruction, is provided. Tissue processing and section cutting were reproducible, and the former provided complete and homogeneous paraffin wax impregnation. 3D visualization of the reconstructed whole human cerebral hemisphere successfully showed the distribution territories of the main cerebral arteries. In addition, we discuss the challenges we faced and overcame while developing the presented method and highlight its originality.
PubMed: 38887398
DOI: 10.17179/excli2023-6601 -
ACS Omega Jun 2024This study aims to explore the effects of augmenting the mass proportion of a composite comprising paraffin and beeswax (PBPCM) within plaster, which influences the...
This study aims to explore the effects of augmenting the mass proportion of a composite comprising paraffin and beeswax (PBPCM) within plaster, which influences the thermal insulation of a dual wall. This work is primarily based on the thermal properties of the composite material PBPCM/plaster with varying percentages of PBPCM. Various essential parameters, such as density, thermal conductivity, specific heat capacity, thermal diffusivity, and latent heat, were assessed and juxtaposed with those of conventional plaster for the PBPCM/plaster composite material. The evaluation of this composite material was executed through an experimental device on a laboratory scale. The obtained results show that the increase in the mass fraction of PBPCM in the plaster decreased the thermal conductivity of plaster more than 3 times, whereas this increase of the PBPCM fraction in plaster enhances heat retention, specifically in specific heat capacity under constant conditions. Nevertheless, in a dynamic state, thermal effusivity has the lowest value for 50% PBPCM. The recommendation is to utilize 50% PBPCM, as it yields an optimal thermal effusivity, and significant values of specific heat capacity and latent heat have been noted for this percentage of PBPCM, measuring 1263.77 kJ/kg K and 18.9 kJ, respectively. Additionally, an increase in the PBPCM percentage narrows the temperature range suitable for effective thermal energy storage.
PubMed: 38882154
DOI: 10.1021/acsomega.4c01481 -
Translational Cancer Research May 2024The efficacy of immunotherapy for brain metastases from small cell lung cancer (SCLC) is relatively low, and the tumor microenvironment of SCLC brain metastases is still...
BACKGROUND
The efficacy of immunotherapy for brain metastases from small cell lung cancer (SCLC) is relatively low, and the tumor microenvironment of SCLC brain metastases is still unknown. Therefore, we investigated the distribution of tumor-infiltrating lymphocytes (TILs) and the expression of programmed cell death-ligand 1 (PD-L1) in patients with brain metastases from SCLC to explore the tumor microenvironment of SCLC brain metastases.
METHODS
A retrospective analysis was performed on 12 surgical specimens of brain metastases from patients with SCLC treated in the Department of Neurosurgery of The First Affiliated Hospital of Anhui Medical University from June 2017 to June 2022. The inclusion criteria for this study were the following: (I) a pathologically confirmed diagnosis of SCLC brain metastases; (II) surgical resection of brain metastases; (III) age >18 years; (IV) and complete clinical data. Patient-related data were retrieved from the inpatient medical record system, telephone follow-up of patients date of death, and overall survival (OS). The immunofluorescence-based tissue microenvironment analysis panel (MAP) was utilized for the detection of TILs, including CD3, CD8, programmed cell death 1 (PD-1), and PD-L1, in formalin-fixed and paraffin-embedded archival specimens of brain metastases. The expression levels of PD-L1 in tumor cells were detected by immunohistochemistry. The correlation between the OS and the above-mentioned markers was analyzed in the 12 patients.
RESULTS
Twelve patients were included in the study. The patients' ages ranged from 51-78 years with a median of 68 years, with 1 female and 11 males. Among 12 patients with SCLC brain metastases: positive rates of CD3 TILs in the tumor parenchyma tumor stroma were 0.60%±0.94% 1.76%±2.72% (P=0.01), respectively; positive rates of CD8 TILs in the tumor parenchyma tumor stroma were 0.80%±0.78% 2.46%±3.72% (P=0.02), respectively. There was no co-expression of CD8 and PD-1 TILs in the tumor parenchyma of 11 cases, and the infiltration density of coexpressed CD3 and PD-1 TILs was more than 10/mm in only 1 case. There was no coexpression of CD3 and PD-1 TIL in the stroma of 10 cases, and the infiltration density of CD8 and PD-1 TILs was more than 10/mm in 2 cases. Immunohistochemistry was used to detect the expression of PD-L1 in 12 cases of SCLC metastatic lesions, and 3 cases (25%) were positive. Survival analysis showed that patients with positive intraepithelial CD3 TILs had significantly longer OS [hazard ratio 3.383, 95% confidence interval (CI): 0.959-11.940; P=0.04].
CONCLUSIONS
Our study further demonstrated the immune microenvironment of SCLC brain metastases. The distribution of TILs in SCLC brain metastases is low and mainly distributed in the stroma, with the expression of PD-L1 in these tumor tissues being low. Further exploration of the immune microenvironment of SCLC brain metastases is of great significance for potential treatment.
PubMed: 38881925
DOI: 10.21037/tcr-24-552 -
Analytica Chimica Acta Aug 2024Chlorinated paraffins (CPs) are industrial chemicals categorised as persistent organic pollutants because of their toxicity, persistency and tendency to long-range...
BACKGROUND
Chlorinated paraffins (CPs) are industrial chemicals categorised as persistent organic pollutants because of their toxicity, persistency and tendency to long-range transport, bioaccumulation and biomagnification. Despite having been the subject of environmental attention for decades, analytical methods for CPs still struggle reaching a sufficient degree of accuracy. Among the issues negatively impacting the quantification of CPs, the unavailability of well-characterised standards, both as pure substances and as matrix (certified) reference materials (CRMs), has played a major role. The focus of this study was to provide a matrix CRM as quality control tool to improve the comparability of CPs measurement results.
RESULTS
We present the process of certification of ERM®-CE100, the first fish reference material assigned with certified values for the mass fraction of short-chain and medium-chain chlorinated paraffins (SCCPs and MCCPs, respectively). The certification was performed in accordance with ISO 17034:2016 and ISO Guide 35:2017, with the value assignment step carried out via an intercomparison of laboratories of demonstrated competence in CPs analysis and applying procedures based on different analytical principles. After confirmation of the homogeneity and stability of the CRM, two certified values were assigned for SCCPs, depending on the calibrants used: 31 ± 9 μg kg and 23 ± 7 μg kg. The MCCPs certified value was established as 44 ± 17 μg kg. All assigned values are relative to wet weight in the CRM that was produced as a fish paste to enhance similarity to routine biota samples.
SIGNIFICANCE AND NOVELTY
The fish tissue ERM-CE100 is the first matrix CRM commercially available for the analysis of CPs, enabling analytical laboratories to improve the accuracy and the metrological traceability of their measurements. The certified CPs values are based on results obtained by both gas and liquid chromatography coupled with various mass spectrometric techniques, offering thus a broad validity to laboratories employing different analytical methods and equipment.
Topics: Hydrocarbons, Chlorinated; Paraffin; Reference Standards; Animals; Fishes
PubMed: 38879205
DOI: 10.1016/j.aca.2024.342757 -
Genomics Jun 2024Cynanchum thesioides, a xerophytic species utilized both as a medicinal herb and a food source, plays a significant role in arid and desert ecosystem management. Its...
Cynanchum thesioides, a xerophytic species utilized both as a medicinal herb and a food source, plays a significant role in arid and desert ecosystem management. Its inflorescence is an umbellate cyme, each carrying nearly a thousand flowers; however, its fruiting rate remains remarkably low. The normal development of the anther is a necessary prerequisite for plants to produce seeds. However, our understanding of the anther development process in Cynanchum thesioides remains limited. To better understand the pollen development process in Cynanchum thesioides, the stages of pollen development were determined through paraffin sectioning, and observations were made on the distribution characteristics of polysaccharides and lipid droplets in the pollen development of Cynanchum thesioides using Periodic Acid-Schiff stain (PAS) and 0.5% Sudan Black B tissue staining. Concurrently, the gene expression patterns and metabolite profiles were delineated across various developmental stages of Cynanchum thesioides anthers (T1: microspore stage, T2: tetrad stage, T3: mononuclear stage, and T4: maturation stage). The findings revealed that Cynanchum thesioides pollen is in an aggregate form. Polysaccharides gradually accumulate during maturation and lipid droplets form a surrounding membrane, thereby preventing pollen dispersion. Furthermore, transcriptomic and metabolomic analyses across distinct developmental phases uncovered a plethora of differentially expressed genes and metabolites associated with the flavonoid biosynthesis pathway. Flavonoid levels exhibited dynamic changes concurrent with anther development, aligning with the gene regulatory patterns of the corresponding biosynthetic pathways. The study identified 63 differentially accumulated flavonoid compounds and 21 differentially expressed genes associated with flavonoid biosynthesis. Weighted gene co-expression network analysis revealed six MYB and ten bHLH transcription factors as key candidates involved in flavonoid biosynthesis, with CtbHLH (Cluster-6587.1050) and CtMYB (Cluster-6587.31743) specifically regulating structural genes within the pathway. These findings underscore the pivotal role of flavonoid biosynthesis in anther development of Cynanchum thesioides. In conclusion, this research offers a comprehensive insight into the anther development process in Cynanchum thesioides.
PubMed: 38878835
DOI: 10.1016/j.ygeno.2024.110884