7 research outputs found
SARS-CoV-2-fehérjék kimutatása immunhisztokémiai módszerrel emberi szövetekben : Patológiai körvizsgálat
SARS-CoV-2-fehérjék kimutatása immunhisztokémiai módszerrel emberi szövetekben : patológiai körvizsgálat
BevezetĂ©s: A SARS-CoV-2 (sĂşlyos akut lĂ©gzĹ‘szervi szindrĂłmát elĹ‘idĂ©zĹ‘ koronavĂrus) okozta COVID–19 világszerte
sajnálatosan nagy halálozással jár. A fertőzés kimutatása elsősorban polimeráz-láncreakcióval (PCR) történik élőben
vagy a halál után, amely azonban nem ad informáciĂłt arrĂłl, hogy a vĂrus mely sejtekben, szövetekben van jelen.
A SARS-CoV-2 tĂĽske- Ă©s nukleokapszid-proteinjeinek, valamint a vĂrus-ribonukleinsavnak (RNS) az in situ kimutatása
igazolhatja a vĂrus jelenlĂ©tĂ©t, valamint adatot szolgáltathat annak direkt vagy indirekt sejtpusztulást okozĂł mechanizmusárĂłl. Jelenleg számos SARS-CoV-2-tĂĽske- Ă©s -nukleokapszid fehĂ©rjeellenes antitest van kereskedelmi forgalomban, melyek eltĂ©rĹ‘ eredmĂ©nnyel kĂ©pesek a megfelelĹ‘ antigĂ©nek kimutatására.
CĂ©lkitűzĂ©s: A jelen munka cĂ©lja a megfelelĹ‘, megbĂzhatĂłan működĹ‘ antitest kiválasztása volt.
MĂłdszer: COVID–19-ben elhunyt 3 egyĂ©n formalinfixált, paraffinba ágyazott, SARS-CoV-2-PCR-pozitĂv tĂĽdejĂ©nek
anyagai, valamint fertĹ‘zött placenta anonim mĂłdon jelölt mintái kerĂĽltek vizsgálatra, megfelelĹ‘ negatĂv kontrollal.
Az immunhisztokĂ©miai reakciĂłk intenzitását Ă©s specificitását hasonlĂtották össze nĂ©gy hazai orvostudományi egyetemi
patolĂłgiai intĂ©zet rĂ©szvĂ©telĂ©vel, kĂĽlönbözĹ‘ antitesteket Ă©s hĂgĂtásokat alkalmazva. Az elvĂ©gzett immunhisztokĂ©miai
reakciĂłk szkennelt, kĂłdolt metszeteken kerĂĽltek Ă©rtĂ©kelĂ©sre, majd az eredmĂ©nyek összesĂtĂ©se után statisztikai elem-
zésre.
EredmĂ©nyek: A vizsgálatok alapján meghatározhatĂłk voltak azon antitestek, amelyek a jelölt hĂgĂtásban Ă©s mĂłdszerrel
megfelelĹ‘ intenzitásĂş, megbĂzhatĂł eredmĂ©nyt adtak.
Következtetés: A vizsgálat alapot ad arra, hogy a SARS-CoV-2 egyes komponensei biopsziás/sebészi anyagban és az
elhunytak szöveteiben nagy pontossággal és reprodukálható módon kimutathatók legyenek a COVID–19-ben megbe-
tegedett, elhunyt egyĂ©nek Ă©lĹ‘ben vagy halál után eltávolĂtott szöveteiben, sejtjeiben
A COVID–19 patológiája
A SARS-CoV-2-pandĂ©mia Ăłta a Semmelweis Egyetemen Ă©s egyĂ©b intĂ©zmĂ©nyekben rendszeresen vĂ©geznek boncolá-sokat, melyek feltárták a COVID–19 jellegzetessĂ©geit. A legsĂşlyosabb kĂ©p a tĂĽdĹ‘ben mutatkozik, melynek lĂ©gtelen-sĂ©ge változĂł kiterjedĂ©sű, oka összetett, Ăgy tĂĽdĹ‘vizenyĹ‘, fehĂ©rjĂ©ben gazdag izzadmány, az erek vĂ©rrög okozta elzárĂł-dása Ă©s gyulladás. A szĂv, a vese, az agy Ă©s a máj változĂł mĂ©rtĂ©kben Ă©rintett, Ă©rrögösödĂ©s, elhalás, degeneratĂv elváltozások mutatkoznak. A SARS-CoV-2-vĂrus fehĂ©rjĂ©i (tĂĽske, nukleokapszid) Ă©s a vĂrus genetikai anyaga (RNS) kimutathatĂł az egyes szervekben, leginkább a tĂĽdĹ‘ben. KlinikopatolĂłgiai elemzĂ©ssel megállapĂthatĂł, hogy a halál a SARS-CoV-2-fertĹ‘zĂ©s mint közvetlen kĂłrok következmĂ©nye, vagy egyĂ©b krĂłnikus megbetegedĂ©s, melyet sĂşlyosbĂ-tott a SARS-CoV-2-fertĹ‘zĂ©s, vagy a halál a vĂrusfertĹ‘zĂ©stĹ‘l fĂĽggetlenĂĽl következett be
Dynamic Interplay in Tumor Ecosystems: Communication between Hepatoma Cells and Fibroblasts
Tumors are intricate ecosystems where cancer cells and non-malignant stromal cells, including cancer-associated fibroblasts (CAFs), engage in complex communication. In this study, we investigated the interaction between poorly (HLE) and well-differentiated (HuH7) hepatoma cells and LX2 fibroblasts. We explored various communication channels, including soluble factors, metabolites, extracellular vesicles (EVs), and miRNAs. Co-culture with HLE cells induced LX2 to produce higher levels of laminin β1, type IV collagen, and CD44, with pronounced syndecan-1 shedding. Conversely, in HuH7/LX2 co-culture, fibronectin, thrombospondin-1, type IV collagen, and cell surface syndecan-1 were dominant matrix components. Integrins α6β4 and α6β1 were upregulated in HLE, while α5β1 and αVβ1 were increased in HuH7. HLE-stimulated LX2 produced excess MMP-2 and 9, whereas HuH7-stimulated LX2 produced excess MMP-1. LX2 activated MAPK and Wnt signaling in hepatoma cells, and conversely, hepatoma-derived EVs upregulated MAPK and Wnt in LX2 cells. LX2-derived EVs induced over tenfold upregulation of SPOCK1/testican-1 in hepatoma EV cargo. We also identified liver cancer-specific miRNAs in hepatoma EVs, with potential implications for early diagnosis. In summary, our study reveals tumor type-dependent communication between hepatoma cells and fibroblasts, shedding light on potential implications for tumor progression. However, the clinical relevance of liver cancer-specific miRNAs requires further investigation
Are Artificial Intelligence-Assisted Three-Dimensional Histological Reconstructions Reliable for the Assessment of Trabecular Microarchitecture?
Objectives: This study aimed to create a three-dimensional histological reconstruction through the AI-assisted classification of tissues and the alignment of serial sections. The secondary aim was to evaluate if the novel technique for histological reconstruction accurately replicated the trabecular microarchitecture of bone. This was performed by conducting micromorphometric measurements on the reconstruction and comparing the results obtained with those of microCT reconstructions. Methods: A bone biopsy sample was harvested upon re-entry following sinus floor augmentation. Following microCT scanning and histological processing, a modified version of the U-Net architecture was trained to categorize tissues on the sections. Detector-free local feature matching with transformers was used to create the histological reconstruction. The micromorphometric parameters were calculated using Bruker’s CTAn software (version 1.18.8.0, Bruker, Kontich, Belgium) for both histological and microCT datasets. Results: Correlation coefficients calculated between the micromorphometric parameters measured on the microCT and histological reconstruction suggest a strong linear relationship between the two with p-values of 0.777, 0.717, 0.705, 0.666, and 0.687 for BV/TV, BS/TV, Tb.Pf Tb.Th, and Tb.Sp, respectively. Bland–Altman and mountain plots suggest good agreement between BV/TV measurements on the two reconstruction methods. Conclusions: This novel method for three-dimensional histological reconstruction provides researchers with a tool that enables the assessment of accurate trabecular microarchitecture and histological information simultaneously
Investigating the Prognostic Relevance of Tumor Immune Microenvironment and Immune Gene Assembly in Breast Carcinoma Subtypes
We hypothesized that different BC subtypes are characterized by spatially distinct tumor immune microenvironment (TIME) and that immune gene assembly of metastatic (Met) and non-metastatic (Ctrl) BCs vary across subtypes. Peritumoral, stromal and intratumoral TIL was assessed on 309 BC cases. Hot, cold and immune-excluded groups were defined, and the prognostic role of this classification was assessed. CD4+/CD8+ positivity was analyzed in 75 cases in four systematically predefined tumor regions. Immune gene expression of Met and Ctrl HER2-negative BCs was compared by using NanoString nCounter technology. The amount of TIL infiltration varied greatly within all BC subtypes. Two-third of the cases were cold tumors with no significant survival difference compared to hot tumors. A lower CD4+/CD8+ ratio at the stromal internal tumor region was significantly associated with longer distant metastasis-free survival. The differentially expressed immune genes between Met and Ctrl varied across the studied BC subtypes with TNBC showing distinct features from the luminal subtypes. The TIME is characterized by a considerable heterogeneity; however, low level of TILs does not equate to disease progression. The differences in immune gene expression observed between Met and Ctrl breast carcinomas call attention to the important role of altered immune function in BC progression