Real-Time Visualization and Quantification of Oncolytic M1 Virus In Vitro and In Vivo

2021 ◽  
Vol 32 (3-4) ◽  
pp. 158-165
Author(s):  
Wenbo Zhu ◽  
Jiankai Liang ◽  
Jingyi Tan ◽  
Li Guo ◽  
Jing Cai ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Visualization

scholarly journals Near Infra-Red Labelling and Tracking of Transplanted Corneal Endothelial Graft

2021 ◽  
Author(s):  
Maninder Bhogal ◽  
Heng-Pei Ang ◽  
Shu-Jun Lin ◽  
Lwin Chan ◽  
Khadijah Adnan ◽  
...  
Keyword(s):  
Real Time ◽  
Corneal Transplantation ◽  
Specular Microscopy ◽  
Post Transplantation ◽  
In Vivo Evaluation ◽  
Non Invasive ◽  
Real Time Visualization ◽  
Donor Graft

Abstract Following corneal transplantation, there is an initial decline in corneal endothelial cells (CECs) following graft preparation and surgery. Monitoring post-transplantation is only possible months after surgery by specular microscopy, which has a limited field of view. We have developed a labelling approach using 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindotricarbocyanine iodide (DIR) dye solution, that enabled tracking of labelled CECs in vivo for at least one month. Initial in vitro optimization of dye concentration, cellular toxicity and real-time cell migration was assessed using propagated primary CECs. Subsequent in vivo evaluation of cellular labelling was assessed within a rabbit wound healing model. Finally, real-time visualization of human cadaver donor tissue incubated in DIR transplanted into rabbits was achieved using the Heidelberg Spectralis. Results revealed detectable fluorescence increased with concentration to a plateau of 100µg/ml, with no toxicity of CECs at any concentration evaluated. DIR-labelled CECs were detectable in vivo upto 1 month, and transplanted labelled donor graft could be visualized and were trackable in vivo. Acute endothelial rejection in 1 rabbit was evidenced by detectable DIR positive cells within the anterior chamber. DIR imaging allowed for detailed imaging of the transplanted corneal endothelium, and enabled non-invasive observation of the corneal endothelial morphology following transplantation.

scholarly journals Transient ICD malfunctions during oncologic radiotherapy: a multi-centre, randomized, in-vitro study

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
E Di Girolamo ◽  
M Appignani ◽  
N Furia ◽  
M Marini ◽  
P De Filippo ◽  
...  
Keyword(s):  
Real Time ◽  
Treatment Plan ◽  
Low Energy ◽  
In Vivo Dosimetry ◽  
Implantable Cardioverter Defibrillators ◽  
Shock Delivery ◽  
Transient Electromagnetic ◽  
Direct Exposure

Abstract Background Direct exposure of implantable cardioverter-defibrillators (ICDs) during radiotherapy is still considered potentially harmful, or even unsafe, by manufacturers and current recommendations. The effects of photon beams on ICDs are unpredictable, depending on multiple factors, and malfunctions may present during exposure. Purpose To evaluate transient ICD malfunctions by direct exposure to doses up to 10 Gy during low-energy RT, forty-three contemporary wireless-enabled ICDs, with at least 4 months to elective replacement indicator (ERI) were evaluated in a real-time in-vitro session in three different centres. Methods All ICDs had baseline interrogation. Single chamber devices were programmed to the VVI/40 mode and dual or triple chamber devices were programmed to the DDD/40 mode. Rate response function and antitachycardia therapies were disabled, with the ventricular tachycardia (VT)/ventricular fibrillation (VF) detection windows still active. A centring computed tomography was performed to build the corresponding treatment plan and the ICDs were blinded randomized to receive either 2-, 5- or 10-Gy exposure by a low photon-energy linear accelerator (6MV) in a homemade water phantom (600 MU/min). The effective dose received by the ICDs was randomly assessed by an in-vivo dosimetry. During radiotherapy, the ICDs were observed in a real-time session using manufacturer specific programmer, and device function (pacing, sensing, programmed parameters, arrhythmia detections) was recorder by the video camera in the bunker throughout the entire photon exposure. All ICDs had an interrogation session immediately after exposure. Results During radiotherapy course, almost all ICDs (93%) recorded major or minor transient electromagnetic interferences. On detail, sixteen ICDs (37.2%) reported atrial and/or ventricular oversensing, with base-rate-pacing inhibition and VT/VF detection. Twenty-four ICDs (55.8%) recorded non clinically relevant noise, and no detections were observed. Only three ICDs (7%) reported neither transient malfunction nor minor noise, withstanding direct radiation exposure. At immediate post-exposure interrogation, the ICDs that recorded major real-time malfunctions had VT/VF detections stored in the device memory. In none of the ICDs spontaneous changes in parameter settings were reported. Malfunctions occurred regardless of either 2-, 5- or 10-Gy photon beam exposure. Conclusions Transient electromagnetic interferences were observed in most of the contemporary ICDs during radiotherapy course, regardless of photon dose. To avoid potentially life-threatening ICD malfunctions such as pacing inhibition or inappropriate shock delivery, magnet application on the pocket site or ICD reprogramming to the asynchronous mode are still suggested in ICD patients ongoing even low energy radiotherapy exposure. Funding Acknowledgement Type of funding source: None

Facile solvothermal synthesis of ultrathin spinel ZnMn2O4 nanospheres: An efficient electrocatalyst for in vivo and in vitro real time monitoring of H2O2

2021 ◽  
Vol 900 ◽  
pp. 115674
Author(s):  
Muthaiah Annalakshmi ◽  
Sakthivel Kumaravel ◽  
T.S.T. Balamurugan ◽  
Shen-Ming Chen ◽  
Ju-Liang He
Keyword(s):  
Real Time ◽  
Solvothermal Synthesis ◽  
Real Time Monitoring

scholarly journals Non-invasive, real-time reporting drug release in vitro and in vivo

2015 ◽  
Vol 51 (32) ◽  
pp. 6948-6951 ◽  
Author(s):  
Yanfeng Zhang ◽  
Qian Yin ◽  
Jonathan Yen ◽  
Joanne Li ◽  
Hanze Ying ◽  
...  
Keyword(s):  
Drug Release ◽  
Real Time ◽  
Near Infrared ◽  
Reporting System ◽  
Real Time Monitoring ◽  
Near Infrared Fluorescence ◽  
Non Invasive ◽  
Fluorescence Dye

Anin vitroandin vivodrug-reporting system is developed for real-time monitoring of drug release via the analysis of the concurrently released near-infrared fluorescence dye.

scholarly journals Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion

mBio ◽  
2011 ◽  
Vol 2 (3) ◽  
Author(s):  
Christina D. Orrú ◽  
Jason M. Wilham ◽  
Lynne D. Raymond ◽  
Franziska Kuhn ◽  
Björn Schroeder ◽  
...  
Keyword(s):  
Real Time ◽  
Blood Test ◽  
Prion Diseases ◽  
Proteinase K ◽  
Transmissible Spongiform Encephalopathies ◽  
Serum Samples ◽  
Spongiform Encephalopathies ◽  
Jakob Disease

ABSTRACT A key challenge in managing transmissible spongiform encephalopathies (TSEs) or prion diseases in medicine, agriculture, and wildlife biology is the development of practical tests for prions that are at or below infectious levels. Of particular interest are tests capable of detecting prions in blood components such as plasma, but blood typically has extremely low prion concentrations and contains inhibitors of the most sensitive prion tests. One of the latter tests is quaking-induced conversion (QuIC), which can be as sensitive as in vivo bioassays, but much more rapid, higher throughput, and less expensive. Now we have integrated antibody 15B3-based immunoprecipitation with QuIC reactions to increase sensitivity and isolate prions from inhibitors such as those in plasma samples. Coupling of immunoprecipitation and an improved real-time QuIC reaction dramatically enhanced detection of variant Creutzfeldt-Jakob disease (vCJD) brain tissue diluted into human plasma. Dilutions of 1014-fold, containing ~2 attogram (ag) per ml of proteinase K-resistant prion protein, were readily detected, indicating ~10,000-fold greater sensitivity for vCJD brain than has previously been reported. We also discriminated between plasma and serum samples from scrapie-infected and uninfected hamsters, even in early preclinical stages. This combined assay, which we call “enhanced QuIC” (eQuIC), markedly improves prospects for routine detection of low levels of prions in tissues, fluids, or environmental samples. IMPORTANCE Transmissible spongiform encephalopathies (TSEs) are largely untreatable and are difficult to diagnose definitively prior to irreversible clinical decline or death. The transmissibility of TSEs within and between species highlights the need for practical tests for even the smallest amounts of infectivity. A few sufficiently sensitive in vitro methods have been reported, but most have major limitations that would preclude their use in routine diagnostic or screening applications. Our new assay improves the outlook for such critical applications. We focused initially on blood plasma because a practical blood test for prions would be especially valuable for TSE diagnostics and risk reduction. Variant Creutzfeldt-Jakob disease (vCJD) in particular has been transmitted between humans via blood transfusions. Enhanced real-time quaking-induced conversion (eRTQ) provides by far the most sensitive detection of vCJD to date. The 15B3 antibody binds prions of multiple species, suggesting that our assay may be useful for clinical and fundamental studies of a variety of TSEs of humans and animals.

scholarly journals The circRAB3IP Mediated by eIF4A3 and LEF1 Contributes to Enzalutamide Resistance in Prostate Cancer by Targeting miR-133a-3p/miR-133b/SGK1 Pathway

2021 ◽  
Vol 11 ◽  
Author(s):  
Dong Chen ◽  
Yaqin Wang ◽  
Feiya Yang ◽  
Adili Keranmu ◽  
Qingxin Zhao ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Real Time ◽  
Expression Pattern ◽  
Real Time Pcr ◽  
Bioinformatic Analysis ◽  
Biological Functions ◽  
Quantitative Real Time Pcr ◽  
Regulatory Effects

An increasing number of studies have shown that circRNAs are closely related to the carcinogenesis and development of prostate cancer (PCa). However, little is known about the effect of the biological functions of circRNAs on the enzalutamide resistance of PCa. Through bioinformatic analysis and experiments, we investigated the expression pattern of circRNAs in enzalutamide-resistant PCa cells. Quantitative real-time PCR was used to detect the expression of circRAB3IP, and plasmids that knock down or overexpress circRAB3IP were used to evaluate its effect on the enzalutamide sensitivity of PCa cells. Mechanistically, we explored the potential regulatory effects of eIF4A3 and LEF1 on the biogenesis of circRAB3IP. Our in vivo and in vitro data indicated that increased expression of circRAB3IP was found in enzalutamide-resistant PCa, and knockdown of circRAB3IP significantly enhanced enzalutamide sensitivity in PCa cells. However, upregulation of circRAB3IP resulted in the opposite effects. Further mechanistic research demonstrated that circRAB3IP could regulate the expression of serum and glucocorticoid-regulated kinase 1 (SGK1) by serving as a sponge that directly targets miR-133a-3p/miR-133b. Then, we showed that circRAB3IP partially exerted its biological functions via SGK1 signaling. Furthermore, we discovered that eIF4A3 and LEF1 might increase circRAB3IP expression in PCa.

Real time imaging reveals a peroxisomal reticulum in living cells

2000 ◽  
Vol 113 (20) ◽  
pp. 3663-3671 ◽  
Author(s):  
M. Schrader ◽  
S.J. King ◽  
T.A. Stroh ◽  
T.A. Schroer
Keyword(s):  
Real Time ◽  
Cytoplasmic Dynein ◽  
Living Cells ◽  
Real Time Imaging ◽  
Peroxisomal Targeting ◽  
Microtubule Motor ◽  
Targeting Signal ◽  
Peroxisomal Targeting Signal

We have directly imaged the dynamic behavior of a variety of morphologically different peroxisomal structures in HepG2 and COS-7 cells transfected with a construct encoding GFP bearing the C-terminal peroxisomal targeting signal 1. Real time imaging revealed that moving peroxisomes interacted with each other and were engaged in transient contacts, and at higher magnification, tubular peroxisomes appeared to form a peroxisomal reticulum. Local remodeling of these structures could be observed involving the formation and detachment of tubular processes that interconnected adjacent organelles. Inhibition of cytoplasmic dynein based motility by overexpression of the dynactin subunit, dynamitin (p50), inhibited the movement of peroxisomes in vivo and interfered with the reestablishment of a uniform distribution of peroxisomes after recovery from nocodazole treatment. Isolated peroxisomes moved in vitro along microtubules in the presence of a microtubule motor fraction. Our data reveal that peroxisomal behavior in vivo is significantly more dynamic and interactive than previously thought and suggest a role for the dynein/dynactin motor in peroxisome motility.

Analyzing Gene Expression through Real Time PCR while Neo-tissue Regeneration using Developed Tissue Constructs

2020 ◽  
pp. 15-34
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Tissue Regeneration ◽  
Real Time Pcr ◽  
Molecular Level ◽  
Northern Blotting ◽  
Tissue Constructs ◽  
Low Sensitivity

Real-time PCR offers a wide area of application to analyze the role of gene activity in various biological aspects at the molecular level with higher specificity, sensitivity and the potential to troubleshoot with post-PCR processing and difficulties. With the recent advancement in the development of functional tissue graft for the regeneration of damaged/diseased tissue, it is effective to analyze the cell behaviour and differentiation over tissue construct toward specific lineage through analyzing the expression of an array of specific genes. With the ability to collect data in the exponential phase, the application of Real-Time PCR has been expanded into various fields such as tissue engineering ranging from absolute quantification of gene expression to determine neo-tissue regeneration and its maturation. In addition to its usage as a research tool, numerous advancements in molecular diagnostics have been achieved, including microbial quantification, determination of gene dose and cancer research. Also, in order to consistently quantify mRNA levels, Northern blotting and in situ hybridization (ISH) methods are less preferred due to low sensitivity, poor precision in detecting gene expression at a low level. An amplification step is thus frequently required to quantify mRNA amounts from engineered tissues of limited size. When analyzing tissue-engineered constructs or studying biomaterials–cells interactions, it is pertinent to quantify the performance of such constructs in terms of extracellular matrix formation while in vitro and in vivo examination, provide clues regarding the performance of various tissue constructs at the molecular level. In this chapter, our focus is on Basics of qPCR, an overview of technical aspects of Real-time PCR; recent Protocol used in the lab, primer designing, detection methods and troubleshooting of the experimental problems.

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