25,592 research outputs found

    Efectos de la Estimulaci贸n Magn茅tica Transcraneana repetitiva de baja frecuencia sobre la producci贸n del lenguaje en pacientes

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    La afasia se una de las complicaciones m谩s discapacitantes en la producci贸n del lenguaje, posterior a un ACV del hemisferio izquierdo. Cerca del 19% de los pacientes con ACV experimentan afasia en la fase aguda y aunque la recuperaci贸n puede ocurrir de manera espont谩nea, frecuentemente ocurre despu茅s de varias semana o meses. Algunos estudios han reportado que la (EMT-r) de baja frecuencia, en pacientes con ACV isqu茅mico izquierdo genera modulaci贸n de la excitabilidad cortical facilitando la reorganizaci贸n funcional y favoreciendo la producci贸n de lenguaje. Se realizar谩 un ensayo cl铆nico, controlado con placebo, doblemente cegado, radomizado, para evaluar los efectos de la EMT-r de baja frecuencia, en la recuperaci贸n del lenguaje en 100 pacientes con primer evento de ACV isqu茅mico entre los primeros 4-8 meses postinfarto y afasia no fuente.Departamento Administrativo de Ciencia, Tecnolog铆a e Innovaci贸n [CO] Colciencias6566-569-33492s

    Characterization and validation of SHH knock-in melanoma cell lines

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    U okviru ovog diplomskog rada identificirana je stani膷na linija dobivena ugradnjom sintetske sekvence koja kodira za protein sonic hedgehog. Aktivni oblik proteina sonic hedgehog, SHH-N, odgovoran je za autokrinu i parakrinu aktivaciju signalnog puta vezanjem za transmembranski receptor Patched 1. Provo膽enjem signala u stanicu, aktivacijski oblici transkripcijskih faktora glioma associated oncogenes ulaze u jezgru i aktiviraju transkripciju gena koji sudjeluju u ovom signalnom putu. Glavna uloga ovog signalnog puta je u organizaciji polarnosti organizma tijekom embriogeneze, dok je u odraslom organizmu ve膰inom inaktivan, osim tijekom proliferacije i diferencijacije stanica. Zbog 膷injenice da regulira rast i razvoj, promjene u ovom signalnom putu mogu dovesti do nastanka tumora, poticanja njihovog rasta i invazije. Iz pojedina膷nih transfeciranih stanica uspostavljene su klonske stani膷ne linije kojima je pomo膰u PCR-a testirana prisutnost inserta SHH u genomu, ekspresija proteina SHH i SHH-N. Na temelju tih rezultata izabrane su 膷etiri linije kojima su westernskom metodom otiska i imunofluorescencijom analizirane koli膷ine proteina HH/GLI signalnog puta (PTCH1, GLI1, GLI2, GLI3) i markera EMT (vimentin, E-kadherin). Identifikacijom prikladne stani膷ne linije koja stabilno eksprimira SHH-N bit 膰e mogu膰e daljnje istra啪ivanje ovog signalnog puta, osobito u stanicama tumora.This master thesis contains the identification of one knock-in cell line for the protein SHH. Its active form, SHH-N is responsible for autocrine and paracrine activation of the HH/GLI signaling pathway by binding to its transmembrane receptor PTCH1. Transduction of the HH/GLI signal into the cell leads to the localization of activator forms of transcription factors GLI in the nucleus where they induce transcription of GLI target genes. This signaling pathway is mostly active during embryonic development, during which it regulates the polarity of the organism. In adults, under normal conditions, the pathway is inactivated or very weakly activated, except during cell proliferation and differentiation. Because of its role in regulating growth and development, changes in this signaling pathway can lead to tumor development, stimulating their growth and invasion. Cell lines developed from individual transfected cells were tested by PCR to detect the SHH insert, and by Western blot to identify the lines with increased SHH and SHH-N protein expression. Four candidate cell lines were selected based on PCR and Western blot results and additionally tested by Western blot and immunofluorescence to analyze the levels of HH/GLI components (PTCH1, GLI1, GLI2, GLI3) and EMT markers (vimentin, E-cadherin). By finding the appropriate cell line with a stable expression of SHH-N, it becomes possible to further study this signalling pathway, especially in tumor cells

    DataSheet_1_Investigating the role of FADS family members in breast cancer based on bioinformatic analysis and experimental validation.docx

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    Breast cancer (BC) is the most common malignant tumor in women worldwide. Emerging evidence indicates the significance of fatty acid metabolism in BC. Fatty acid desaturase (FADS) is closely associated with cancer occurrence and development. Here, bioinformatic analysis and experimental validation were applied to investigate the potential functions of FADS in BC. Several public databases, including TCGA, GEO, HPA, Kaplan鈥揗eier plotter, STRING, DAVID, cBioPortal, TIMER, TRRUST, and LinkedOmics were used to determine mRNA/protein expression levels, prognostic significance, functional enrichment, genetic alterations, association with tumor-infiltrating immune cells, and related transcription factors and kinases. BC tissues showed higher and lower mRNA expression of FADS2/6/8 and FADS3/4/5, respectively. FADS1/2/6 and FADS3/4/5 showed higher and lower protein expression levels, respectively, in BC tissues. Moreover, FADS1/7 up- and FADS3/8 down-regulation predicted poor overall and recurrence-free survival, while FADS2/5 up- and FADS4 down-regulation were associated with poor recurrence-free survival. Receiver operating characteristic curves revealed that FADS2/3/4/8 were indicative diagnostic markers. FADS family members showing differential expression levels were associated with various clinical subtypes, clinical stages, lymph node metastasis status, copy number variants, DNA methylation, and miRNA regulation in BC. The mRNA expression level of FADS1/2/3/4/5/7/8 was observed to be significantly negatively correlated with DNA methylation. FADS1/2 upregulation was significantly correlated with clinical stages. FADS1/4 expression was obviously lower in BC patients with higher lymph node metastasis than lower lymph node metastasis, while FADS7/8 expression was obviously higher in BC patients with higher lymph node metastasis than lower lymph node metastasis. FADS family members showed varying degrees of genetic alterations, and Gene Ontology and KEGG pathway enrichment analyses suggested their involvement in lipid metabolism. Their expression level was correlated with immune cell infiltration levels. FADS2 was chosen for further validation analyses. We found FADS2 to be significantly over-expressed in clinical BC tissue samples. The proliferation, migration, and invasion abilities of MDA-MB-231 and BT474 cells were significantly reduced after FADS2 knockdown. Furthermore, FADS2 may promote the occurrence and development of BC cells via regulating the epithelial鈥搈esenchymal transition (EMT) pathway. Altogether, our results suggest that FADS1/2/3/4 can serve as potential therapeutic targets, prognostic indicators, and diagnostic markers in patients with BC.</p

    Annual report of the officers of the town of Jackson, New Hampshire for the fiscal year ending December 31, 2022.

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    This is an annual report containing vital statistics for a town/city in the state of New Hampshire

    Immunohistochemical expression of E-cadherin and N-cadherin in endometrioid endometrial carcinoma and its precursor lesions

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    Objectives: Endometrial carcinoma (EC), majority being endometroid endometrial carcinoma (EEC), is the most common invasive gynecological cancer world-wide. Epithelial mesenchymal transition (EMT) is the process that cells undergo to switch from a polarized epithelial phenotype to a motile mesenchymal phenotype. Loss of E-cadherin and expression of N-cadherin are a critical step for development and progression of malignant tumors. In present study we evaluated the immunohistochemical (IHC) expression of E-cadherin and N-cadherin with clinicopathological parameters of EEC and its precursors lesions. Methods: It was a prospective nested case-control study involving women aged 35鈥70 years, whose endometrial biopsy were obtained for histological diagnosis. Signed informed consent was obtained before enrolling the women in the study, and the approval to conduct the study was obtained from the institutional ethics committee. The IHC was done to measure the E-cadherin and N-cadherin. Results: Reduced expression of E-cadherin was observed in 69% of cancer and 65% of precancer, however only 32% of normal proliferative endometrium showed reduced E-cadherin expression. N-cadherin expression was observed in 82% of EEC and 81% of precancer cases, while 11% of normal proliferative endometrium showed N-cadherin expression. Statistically significant downregulation of E-cadherin and upregulation of N-cadherin were observed in EEC and precancerous lesions as compared to normal control. Conclusion: Since downregulation of E-cadherin was also associated with upregulation of N-cadherin, the integration of immuno-expression of both E-cadherin and N-cadherin might offer a potential therapeutic target for EEC

    Pathogenesis and treatment of chronic rhinosinusitis from the perspective of sinonasal epithelial dysfunction

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    BackgroundChronic rhinosinusitis (CRS) is a clinical syndrome primarily characterized by long-term mucosal inflammation of the nasal cavity and sinuses. The pathogenesis of CRS is still unclear due to its high heterogeneity. A number of studies have recently focused on the sinonasal epithelium. Thus, there has been a quantum leap in awareness of the role of the sinonasal epithelium, which is now understood as an active functional organ rather than simply an inert mechanical barrier. Undoubtedly, epithelial dysfunction plays a vital role in the onset and development of CRS.ObjectiveIn this article, we discuss the potential contribution of sinonasal epithelium dysfunction to CRS pathogenesis and explore a few current and developing therapeutic options targeting the sinonasal epithelium.ResultsImpaired mucociliary clearance (MCC) and an abnormal sinonasal epithelial barrier are usually considered to be the main causative factors in CRS. Epithelial-derived bioactive substances, such as cytokines, exosomes, and complements, play a vital role in the regulation of innate and adaptive immunity and contribute to the pathophysiological alterations of CRS. The phenomena of epithelial鈥搈esenchymal transition (EMT), mucosal remodeling, and autophagy observed in CRS offer some novel insights into the pathogenesis of this disease. In addition, existing treatment options targeting disorder of sinonasal epithelium can help to relieve the main symptoms associated with CRS to some extent.ConclusionThe presence of a normal epithelium is fundamental for maintaining homeostasis in the nasal and paranasal sinuses. Here, we describe various aspects of the sinonasal epithelium and highlight the contributions of epithelial dysfunction to CRS pathogenesis. Our review provides sound evidence of the need for in-depth study of the pathophysiological alterations of this disease and for the development of novel epithelium-targeting alternative treatments

    Demystifying inertial specifications : supporting the inclusion of grid-followers

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    Inertia provision from converters is often separated into two categories according to the control approach. Inertia from grid-followers (GFLs) is deemed to be "synthetic" due to a slow response. In contrast, grid-forming (GFM) inertia is deemed to be "true", and more useful for frequency stability, due its faster provision. This paper analyses the distinctions between GFM and GFL inertia by carrying out parametric sweeps of each approach at different operating conditions. The analysis aims to assist the ongoing efforts to quantify grid stabilising phenomena, particularly the recent adaptation of the GB grid code to incorporate GFM converters. The optimal tuning configurations are identified, showing that the GFL can achieve fast inertial provision that can contain the grid frequency as effectively as GFM inertia on strong grids, despite the opposing consensus in the literature. The simulations also highlight the importance of voltage-source behaviours in determining the initial evolution of grid frequency, that these features should be considered more explicitly by system operators, and that GFLs should not be excluded so readily. Neglecting GFL control could limit the assets available to support the grid and inhibit the rate that the net zero transition can occur

    Antimetastatic Activity of Apoptolidin A by Upregulation of N-Myc Downstream-Regulated Gene 1 Expression in Human Colorectal Cancer Cells

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    Colorectal cancer (CRC) is one of the most prevalent tumors with high metastatic potential; consequently, finding new drug candidates that suppress tumor metastasis is essential. Apoptolidin A is a macrocyclic lactone produced by Amycolatopsis sp. DW02G. It exhibits significant cytotoxicity against several cancer cell lines, but its effects on CRC cells remain unknown. Therefore, the present study investigated the antiproliferative and antimetastatic activities of apoptolidin A and its underlying molecular mechanisms in CRC cells. Apoptolidin A effectively inhibited CRC cell growth and colony formation. The induction of G0/G1 phase cell cycle arrest was associated with the downregulation of cyclin D1 and CDK4/6 expression. Long-term exposure to apoptolidin A also induced apoptosis as confirmed by the downregulation and upregulation of Bcl-2 and Bax expression, respectively. Moreover, apoptolidin A effectively upregulated the suppressed expression of N-Myc downstream-regulated gene 1 (NDRG1), a tumor suppressor gene, in a concentration-dependent manner in CRC cells. The antimetastatic potential of apoptolidin A was also correlated with the expression of epithelial鈥搈esenchymal transition (EMT) biomarkers, including the upregulation of E-cadherin and downregulation of N-cadherin, vimentin, snail, and MMP9 in CRC cells. These findings suggest that apoptolidin A exerts antiproliferative and antimetastatic activities by regulating the NDRG1-activated EMT pathway in CRC cells

    Identifizierung pr盲diktiver und prognostischer Biomarker in unterschiedlichen Tumorkompartimenten des 枚sophagealen Adenokarzinoms

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    Das 枚sophageale Adenokarzinom zeigt eine global steigende Inzidenz und hat mit einer 5-Jahres-脺berlebensrate von weniger als 25% eine schlechte Prognose. Personalisierte Therapieans盲tze sind selten und prognostische/pr盲diktive Biomarker des Tumormikromilieus sind unzureichend charakterisiert. Die kumulative Promotion n盲hert sich dieser Problematik in drei unterschiedlichen Schwerpunkten. 1. Zur Identifizierung Kompartiment-spezifischer Biomarker wurde eine Methode entwickelt, welche als kosteng眉nstige Alternative zum sc-Seq Expressionsprofile individueller Zelltypen generiert. Dabei erfolgt die Extraktion der RNA nicht aus Einzelzellen, sondern aus flowzytometrisch-getrennten Zellkompartimenten. Die Separation der Proben in Epithelzellen, Immunzellen und Fibroblasten wurde durch verschiedene Verfahren validiert und eine suffiziente Ausbeute an RNA auch f眉r kleine Gewebemengen gezeigt. 2. Biomarker des Immunzellkompartiments als therapeutische Angriffspunkte wurden in einem Patientenkollektiv von bis zu 551 Patienten auf ihre Bedeutung beim EAC 眉berpr眉ft. Es zeigte sich eine Expression der Immuncheckpoints LAG3, VISTA und IDO auf TILs durch IHC und RNA-Sonden basierte Verfahren in einem relevanten Anteil (LAG3: 11,4%, VISTA: 29%, IDO: 52,6%). Es konnte eine prognostisch g眉nstige Bedeutung der VISTA, LAG3 und IDO Expression gezeigt werden. Durch den Vergleich von Genexpressionsprofilen aus therapienaiven und vorbehandelten Tumoren konnte zudem ein immunsuppressiver Effekt von neoadjuvanten Therapiekonzepten auf das Tumormikromilieu des EACs gezeigt werden. Dabei kam es zur verminderten Expression von Checkpoints und Anzahl TILs nach (Radio-) Chemotherapie. 3. Im Tumorzellkompartiment wurde die Rolle von Amplifikationen in ErbB-Rezeptor abh盲ngigen Signalwegen durch FISH-Technik und Immunhistochemie evaluiert. Es fanden sich KRAS Amplifikationen in 17,1%, PIK3CA Amplifikationen in 5% sowie eine HER2/neu-脺berexpression in 14,9% der untersuchten Tumore

    Establishment and characterization of a HER2-enriched canine mammary cancerous myoepithelial cell line

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    Abstract Background Canine mammary tumors (CMTs) have a poor prognosis, along with tumor recurrence and metastasis. Cell lines are vital in vitro models for CMT research. Many CMT epithelial cell lines were reported. However, canine mammary myoepithelial cells, the contractile component of the canine mammary tissue were overlooked. This study aimed at establishing such a cell line. CMT-1 cell line was obtained from a canine mammary tumor CMT-1 and characterized molecularly through qPCR, western blotting, immunochemistry and immunofluorescence. Its doubling time, cytogenetic analysis and migration rate were evaluated using growth study, karyotype analysis and wound healing assay respectively. To determine its tumorigenesis, xenograft transplantation was performed. Results CMT-1 tumor was a complex canine mammary carcinoma that stained negative to estrogen receptors (ER) and progesterone receptors (PR), but positive to human epidermal growth receptor-2 (HER2), defined as HER2-enriched subtype. In this study, a CMT-1 cell line obtained from CMT-1 tumor was immune-positive to vimentin, 伪-SMA, p63 and negative to E-cadherin (E-cad), indicating CMT-1 cells were myoepithelial cells. It was successfully cultured for more than 50 passages showing the same immunoreactivity to ER, PR, and HER2 as the primary canine tumor. The doubling time of CMT-1 cell line was 26.67聽h. The chromosome number of CMT-1 cells ranged from 31 to 64. A potential spontaneous epithelial to mesenchymal transition (EMT) was noticed during cell cultures. Potential EMT-induced CMT-1 cells showed no significance in migration rate compared to the original CMT-1 cells. CMT-1 cells was able to grow on a 3D culture and formed grape-like, solid, and cystic mammospheres at different time period. Inoculation of CMT-1 cells induced a complex HER2-enriched mammary tumor with metastasis in mice. Conclusions A canine cancerous HER2-enriched myoepithelial cell line was successfully established and a canine mammosphere developed from myoepithelial cells was documented in this study. We are expecting this novel cell line and its associated mammospheres could be used as a model to elucidate the role of myoepithelial cells in CMT carcinogensis in the future
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