scholarly journals Cancerous Tissue Diagnosis by LIF Spectroscopy Derived From Body-Compatible Fluorophores

2021 ◽  
Vol 12 (1) ◽  
pp. e10-e10
Author(s):  
Atefeh Asghari Moghaddam ◽  
Batool Sajad ◽  
Fariba Mehrad Nia ◽  
Seyed Hamid Madani
Keyword(s):  
Diagnostic Method ◽  
Rhodamine 6G ◽  
Spectral Shift ◽  
Clinical Applications ◽  
Intensity Difference ◽  
Sodium Fluorescein ◽  
Dye Molecules ◽  
Lif Spectroscopy ◽  
Tissue Specimens

Introduction: The laser-induced fluorescence (LIF) method as molecular emission spectroscopy is used to diagnose cancerous tissues. According to the previous reports, the red-shift in the fluorescence spectrum from Rhodamine 6G (Rd6G)-stained cancerous tissues compared to healthy ones impregnated with the same dye provides the feasibility for diagnosis. In this paper, we have employed the LIF emissions as a diagnostic method to distinguish between cancerous and healthy tissues infiltrated by a body-compatible fluorophore to avoid the toxicity and hazard of Rd6G dye. Methods: Biological tissue specimens are stained with sodium fluorescein (NaFl) dye and then irradiated by the blue CW diode laser (405 nm) to examine the spectral properties that are effective in detecting cancerous tissues. Results: The spectral shift and the intensity difference of fluorescence are keys to diagnosing in vitro cancerous breast, colon, and thyroid tissues for clinical applications. The notable tubular densities in the breast and colon tissues and the space between the papillae in the thyroid ones cause the cancerous tissues to be prominently heterogeneous, providing numerous micro-cavities and thus more room for dye molecules. Conclusion: Here, we have assessed the spectral shift and intensity difference of fluorescence as a diagnostic method to distinguish between cancerous and healthy tissues for clinical applications.

RNA interference and its clinical applications

2007 ◽  
Vol 148 (47) ◽  
pp. 2235-2240 ◽  
Author(s):  
Gyöngyi Munkácsy ◽  
Zsolt Tulassay ◽  
Balázs Győrffy
Keyword(s):  
Rna Interference ◽  
Clinical Applications

Az RNS-interferencia a poszttranszkripciós génelcsendesítés olyan formája, amelynek során rövid, specifikusan RNS-molekulák elnyomják a gének kifejeződésében kulcsszerepet játszó hírvivő RNS-ek működését. A sejtbe juttatott dupla szálú vagy rövid interferáló RNS-molekulák aktiválják az RNS-indukált elcsendesítő komplexet, amely a célgén hírvivő RNS-ét lebontja. A sejtek saját szabályozó mikro-RNS-molekulákkal is rendelkeznek, amelyeknek hírvivő RNS-e képes önmagával hajtűt képezni, amit a sejt dupla szálú RNS-ként értelmez. Az RNS-interferencia élettani működései közé tartozik a vírusok és a transzpozonok elleni védekezés, valamint a génkifejeződés szabályozása. Az RNS-interferencia nemcsak in vitro alkalmazható az egyes gének működésének vizsgálatára, hanem klinikai alkalmazásainak lehetőségei is megjelentek. Eddig vírusfertőzésekben, az időskori makuladegeneráció gátlására, a vér koleszterinszint-csökkentésére, daganatellenes és neurodegeneratív betegségek kezelésében alkalmazták. Az RNS-interferencia alkalmazását azonban nehezíti, hogy a megfelelő rövid interferáló RNS-molekulák tervezéséhez szükséges bioinformatikai algoritmusok nem tökéletesek; a szervezet szöveteibe való bejuttatásuk nehéz; illetve csak olyan esetekben alkalmazható, amelyekben átmeneti antagonista génelcsendesítő hatás és nem hosszú távú kezelés szükséges. Az alkalmazás legnagyobb előnye a jelentős specificitás, ami miatt mellékhatása is kevés. Az RNS-interferencia alapú kezelések megjelenése már a közeli jövőben várható.

scholarly journals Lentiviral Vectors for Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1288
Author(s):  
Wendy Dong ◽  
Boris Kantor
Keyword(s):  
Large Scale ◽  
Lentiviral Vector ◽  
Clinical Applications ◽  
Scale Production ◽  
Base Editing ◽  
Epigenome Editing ◽  
Vector Systems ◽  
Dividing Cells

CRISPR/Cas technology has revolutionized the fields of the genome- and epigenome-editing by supplying unparalleled control over genomic sequences and expression. Lentiviral vector (LV) systems are one of the main delivery vehicles for the CRISPR/Cas systems due to (i) its ability to carry bulky and complex transgenes and (ii) sustain robust and long-term expression in a broad range of dividing and non-dividing cells in vitro and in vivo. It is thus reasonable that substantial effort has been allocated towards the development of the improved and optimized LV systems for effective and accurate gene-to-cell transfer of CRISPR/Cas tools. The main effort on that end has been put towards the improvement and optimization of the vector’s expression, development of integrase-deficient lentiviral vector (IDLV), aiming to minimize the risk of oncogenicity, toxicity, and pathogenicity, and enhancing manufacturing protocols for clinical applications required large-scale production. In this review, we will devote attention to (i) the basic biology of lentiviruses, and (ii) recent advances in the development of safer and more efficient CRISPR/Cas vector systems towards their use in preclinical and clinical applications. In addition, we will discuss in detail the recent progress in the repurposing of CRISPR/Cas systems related to base-editing and prime-editing applications.

Clinical applications of techniques used in human in vitro fertilization research

1983 ◽  
Vol 146 (5) ◽  
pp. 477-481 ◽  
Author(s):  
Richard P. Marrs ◽  
Joyce M. Vargyas ◽  
Hidekazu Saito ◽  
William E. Gibbons ◽  
Trish Berger ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Clinical Applications ◽  
Vitro Fertilization ◽  
Human In Vitro Fertilization

Development of temporal spectroscopic properties for Xerogel matrices doped with Rhodamine 6G dye

Open Physics ◽  
2006 ◽  
Vol 4 (3) ◽  
Author(s):  
Abbas Al-Wattar ◽  
Baha Chiad ◽  
Wesam Twej ◽  
Sarmed Al-Awadi
Keyword(s):  
Fluorescence Spectra ◽  
Rhodamine 6G ◽  
Sol Gel ◽  
Blue Shift ◽  
Spectroscopic Properties ◽  
Dye Molecules ◽  
Absorption And Fluorescence Spectra ◽  
Silica Network ◽  
Sol Gel Process ◽  
Two Parameters

AbstractThe solid host of a laser dye modifies its spectroscopic properties with respect to its liquid host. During the Sol-Gel process the dye molecules suffer from changing their environment. Two parameters affect this matter, the change in the concentration due to the evaporation of the solvent (drying) and the caging of dye molecules inside the pores or attachment to the silica network. Rhodamine 6G absorption and fluorescence spectra with different concentrations, during Sol-Gel time processing, have been studied. Both, absorption and fluorescence spectra of the dye in the solid host, for different concentrations, show a blue-shift relative to its liquid phase.

scholarly journals Studies on Fluorescence Efficiency and Photodegradation of Rhodamine 6G Doped PMMA Using a Dual Beam Thermal Lens Technique

2002 ◽  
Vol 20 (2-4) ◽  
pp. 99-110 ◽  
Author(s):  
Achamma Kurian ◽  
Nibu A. George ◽  
Binoy Paul ◽  
V. P. N. Nampoori ◽  
C. P. G. Vallabhan
Keyword(s):  
Thermal Lens ◽  
Fluorescence Quantum Yield ◽  
Laser Excitation ◽  
Rhodamine 6G ◽  
Fluorescence Emission ◽  
Concentration Quenching ◽  
Dye Molecules ◽  
Dual Beam ◽  
Fluorescence Efficiency ◽  
Absolute Fluorescence

In this paper we report the use of the dual beam thermal lens technique as a quantitative method to determine absolute fluorescence quantum efficiency and concentration quenching of fluorescence emission from rhodamine 6G doped Poly(methyl methacrylate) (PMMA), prepared with different concentrations of the dye. A comparison of the present data with that reported in the literature indicates that the observed variation of fluorescence quantum yield with respect to the dye concentration follows a similar profile as in the earlier reported observations on rhodamine 6G in solution. The photodegradation of the dye molecules under cw laser excitation is also studied using the present method.

scholarly journals Expression of PRDX6 Correlates with Migration and Invasiveness of Colorectal Cancer Cells

2018 ◽  
Vol 51 (6) ◽  
pp. 2616-2630 ◽  
Author(s):  
Wen-Shih Huang ◽  
Cheng-Yi Huang ◽  
Meng-Chiao Hsieh ◽  
Yi-Hung Kuo ◽  
Shui-Yi Tung ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Transcriptional Activation ◽  
Colorectal Adenomas ◽  
Tissue Array ◽  
Boyden Chamber ◽  
Node Positive ◽  
Proliferation Capacity ◽  
Twist 1 ◽  
Tissue Specimens

Background/Aims: Colorectal cancer (CRC) is the third most common type of cancer and the second leading cause of cancer-related deaths worldwide. PRDXs are antioxidant enzymes that play an important role in cell differentiation, proliferation and apoptosis and have diverse functions in malignancy development. However, the mechanism of aberrant overexpression of PRDX6 in CRC remains unclear. Methods: Boyden chamber assay, flow cytometry and a lentiviral shRNA targeting PRDX6 and transient transfection with pCMV-6-PRDX6 plasmid were used to examine the role of PRDX6 in the proliferation capacity and invasiveness of CRC cells. Immunohistochemistry (IHC) with tissue array containing 40 paraffin- embedded CRC tissue specimens and Western blot assays were used to detect target proteins. Results: PRDX6 was significantly up-expressed in different comparisons of metastasis of colorectal adenomas in node-positive CRC (P = 0.03). In in vitro HCT-116, PRDX6 silencing markedly suppressed CRC cell migration and invasiveness while also inducing cell cycle arrest as well as the generation of reactive oxygen species (ROS); specific overexpression of PRDX6 had the opposite effect. Mechanistically, the PRDX6 inactivation displayed decreased levels of PRDX6, N-cadherin, β-catenin, Vimentin, Slug, Snail and Twist-1 through the activation of the PI3K/ AKT/p38/p50 pathways, but they were also significantly inhibited by PRDX6 transfectants. There was also increased transcriptional activation of dimethylation of histone H3 lysine 4 (H3K4me3) of PRDX6 promoter via the activation of the PI3K/Akt/NFkB pathways. Conclusion: Our findings demonstrated that PRDX6 expression plays a characteristic growth-promoting role in CRC metastasis. This study suggests that PRDX6 may serve as a biomarker of node-positive status and may have a role as an important endogenous regulator of cancer cell tumorigenicity in CRC. PRDX6 may also be an effective therapeutic target.

scholarly journals LncRNA PVT1 Knockdown Ameliorates Myocardial Ischemia Reperfusion Damage via Suppressing Gasdermin D-Mediated Pyroptosis in Cardiomyocytes

2021 ◽  
Vol 8 ◽  
Author(s):  
Cuizhi Li ◽  
Huafeng Song ◽  
Chunlin Chen ◽  
Shaoxian Chen ◽  
Qiyu Zhang ◽  
...  
Keyword(s):  
Myocardial Ischemia ◽  
Cell Viability ◽  
Ischemia Reperfusion ◽  
Inflammatory Factors ◽  
H9c2 Cells ◽  
Myocardial Ischemia Reperfusion ◽  
Tissue Specimens ◽  
Masson Staining

Objective: Myocardial ischemia reperfusion (I/R) damage is a life-threatening vascular emergency after myocardial infarction. Here, we observed the cardioprotective effect of long non-coding RNA (lncRNA) PVT1 knockdown against myocardial I/R damage.Methods: This study constructed a myocardial I/R-induced mouse model and a hypoxia/reoxygenation (H/R)-treated H9C2 cells. PVT1 expression was examined via RT-qPCR. After silencing PVT1 via shRNA against PVT1, H&E, and Masson staining was performed to observe myocardial I/R damage. Indicators of myocardial injury including cTnI, LDH, BNP, and CK-MB were examined by ELISA. Inflammatory factors (TNF-α, IL-1β, and IL-6), Gasdermin D (GSDMD), and Caspase1 were detected via RT-qPCR, western blot, immunohistochemistry, or immunofluorescence. Furthermore, CCK-8 and flow cytometry were presented for detecting cell viability and apoptosis.Results: LncRNA PVT1 was markedly up-regulated in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Silencing PVT1 significantly lowered serum levels of cTnI, LDH, BNP, and CK-MB in myocardial I/R mice. H&E and Masson staining showed that silencing PVT1 alleviated myocardial I/R injury. PVT1 knockdown significantly lowered the production and release of inflammatory factors as well as inhibited the expression of GSDMD-N and Caspase1 in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Moreover, silencing PVT1 facilitated cell viability and induced apoptosis of H/R-treated H9C2 cells.Conclusion: Our findings demonstrated that silencing PVT1 could alleviate myocardial I/R damage through suppressing GSDMD-mediated pyroptosis in vivo and in vitro. Thus, PVT1 knockdown may offer an alternative therapeutic strategy against myocardial I/R damage.

scholarly journals Association of CD44−/CD24− Breast Cancer Cells with Late Stage Tumor Recurrence

2020 ◽  
Author(s):  
Xinbo Qiao ◽  
Yixiao Zhang ◽  
Lisha Sun ◽  
Qingtian Ma ◽  
Jie Yang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Cells ◽  
Tumor Metastasis ◽  
Cancer Prognosis ◽  
Molecular Events ◽  
Cell Conversion ◽  
Tissue Specimens ◽  
Distant Tumor

AbstractTumor metastasis remains the main cause of breast cancer-related deaths, especially the later breast cancer distant metastasis. This study assessed CD44−/CD24− tumor cells in 576 tissue specimens for associations with clinicopathological features and metastasis and then investigated the underlying molecular events. The data showed that level of CD44−/CD24− cells was associated with later postoperative distant tumor metastasis. Furthermore, CD44−/CD24− triple negative cells could spontaneously convert into CD44+/CD24− cancer stem cells (CSCs) with properties similar to CD44+/CD24− CSCs from parental MDA-MB-231 cells in terms of gene expression, tumor cell xenograft formation, and lung metastasis in vitro and in vivo. Single-cell RNA sequencing identified RHBDL2 as a regulator that enhanced spontaneous CD44+/CD24− CSC conversion, whereas knockdown of RHBDL2 expression inhibited YAP/NF-κB signaling and blocked spontaneous CD44−/CD24− cell conversion to CSCs. These data suggested that the level of CD44−/CD24− tumor cells could predict breast cancer prognosis, metastasis, and response to adjuvant therapy.

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