scholarly journals Innovative approach for the application of MTT, LDH and bis-ANS

2021 ◽  
Author(s):  
Emese Szilvia Jánosi-Mózes
Keyword(s):  
Quantitative Determination ◽  
Hippocampal Slice ◽  
Ex Vivo ◽  
Hippocampal Slices ◽  
Fluorescent Imaging ◽  
Tissue Slices ◽  
Hippocampus Slice ◽  
Acute Hippocampal Slices

The acute hippocampus slice-based models can serve as a feasible tool to gain deeper understanding on how pathological events in AD, in particular Aβ oligomerization, influence hippocampal functionality and therefore paving the way to develop and test related hypothesis but also to screen potential protective agents or validate results seen in vivo or predicted in silico. To overcome the shortcomings of the in vitro and in vivo methods, we developed a cost effective and simple apparatus for maintaining the viability of acute tissue slices, named: ExViS (Ex Vivo System; Universal Mini-Chamber Tube System for Acute Tissue Slices). The system allowed us to further develop methods that use acute (ex vivo) hippocampal slices to model AD-related patomechanisms. Over the course of my PhD studies we have developed the following techniques and ex vivo models based on the features of acute hippocampal slices: 1. Rapid, reliable and quantitative determination of Aβ1-42 toxicity in ageing (OGD) acute hippocampal slice model using MTT and LDH assays. 2. Quantitative determination of zinc-induced Aβ1-42 oligomerization toxicity in acute hippocampal slice model using MTT assay. 3. Fluorescent imaging of neurite cross-sections and detailed imaging of neurite structures in acute hippocampal slices via a novel application of bis-ANS and its co-staining variations. 4. Modelling the effect of Zn2+ released in glutamergic synaptic cleft in the hippocampus on Aβ1-42 aggregation and its consequences on cell viability and synaptic functionality, learning and memory (LTP).

Validation of Doxepin Quantitative Determination Methods for their Application to In Vitro, Ex Vivo and In Vivo Studies

2015 ◽  
Vol 11 (4) ◽  
pp. 269-277 ◽  
Author(s):  
Roser Sanz ◽  
Beatriz Clares ◽  
Mireia Mallandrich ◽  
Isidre Casals ◽  
David Bellido ◽  
...  
Keyword(s):  
Quantitative Determination ◽  
Ex Vivo ◽  
Determination Methods

scholarly journals Performance of a novel protease-activated fluorescent imaging system for intraoperative detection of residual breast cancer during breast conserving surgery

2021 ◽  
Vol 187 (1) ◽  
pp. 145-153
Author(s):  
Conor R. Lanahan ◽  
Bridget N. Kelly ◽  
Michele A. Gadd ◽  
Michelle C. Specht ◽  
Carson L. Brown ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Imaging System ◽  
Ex Vivo ◽  
Menopausal Status ◽  
Fluorescent Imaging ◽  
Cancer Subtypes ◽  
Surgical Workflow ◽  
Tumor Types

Abstract Purpose Safe breast cancer lumpectomies require microscopically clear margins. Real-time margin assessment options are limited, and 20–40% of lumpectomies have positive margins requiring re-excision. The LUM Imaging System previously showed excellent sensitivity and specificity for tumor detection during lumpectomy surgery. We explored its impact on surgical workflow and performance across patient and tumor types. Methods We performed IRB-approved, prospective, non-randomized studies in breast cancer lumpectomy procedures. The LUM Imaging System uses LUM015, a protease-activated fluorescent imaging agent that identifies residual tumor in the surgical cavity walls. Fluorescent cavity images were collected in real-time and analyzed using system software. Results Cavity and specimen images were obtained in 55 patients injected with LUM015 at 0.5 or 1.0 mg/kg and in 5 patients who did not receive LUM015. All tumor types were distinguished from normal tissue, with mean tumor:normal (T:N) signal ratios of 3.81–5.69. T:N ratios were 4.45 in non-dense and 4.00 in dense breasts (p = 0.59) and 3.52 in premenopausal and 4.59 in postmenopausal women (p = 0.19). Histopathology and tumor receptor testing were not affected by LUM015. Falsely positive readings were more likely when tumor was present < 2 mm from the adjacent specimen margin. LUM015 signal was stable in vivo at least 6.5 h post injection, and ex vivo at least 4 h post excision. Conclusions Intraoperative use of the LUM Imaging System detected all breast cancer subtypes with robust performance independent of menopausal status and breast density. There was no significant impact on histopathology or receptor evaluation.

scholarly journals Insight into Glutamatergic Involvement in Rewarding Effects of Mephedrone in Rats: In Vivo and Ex Vivo Study

2021 ◽  
Author(s):  
Olga Wronikowska ◽  
Maria Zykubek ◽  
Agnieszka Michalak ◽  
Anna Pankowska ◽  
Paulina Kozioł ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Interdisciplinary Approach ◽  
Nmda Antagonist ◽  
Exchange Chromatography ◽  
Resonance Spectroscopy ◽  
Monoamine Transporters ◽  
Ex Vivo Study ◽  
Insight Into

AbstractMephedrone is a widely used drug of abuse, exerting its effects by interacting with monoamine transporters. Although this mechanism has been widely studied heretofore, little is known about the involvement of glutamatergic transmission in mephedrone effects. In this study, we comprehensively evaluated glutamatergic involvement in rewarding effects of mephedrone using an interdisciplinary approach including (1) behavioural study on effects of memantine (non-selective NMDA antagonist) on expression of mephedrone-induced conditioned place preference (CPP) in rats; (2) evaluation of glutamate concentrations in the hippocampus of rats following 6 days of mephedrone administration, using in vivo magnetic resonance spectroscopy (MRS); and (3) determination of glutamate levels in the hippocampus of rats treated with mephedrone and subjected to MRS, using ion-exchange chromatography. In the presented research, we confirmed priorly reported mephedrone-induced rewarding effects in the CPP paradigm and showed that memantine (5 mg/kg) was able to reverse the expression of this effect. MRS study showed that subchronic mephedrone administration increased glutamate level in the hippocampus when measured in vivo 24 h (5 mg/kg, 10 mg/kg and 20 mg/kg) and 2 weeks (5 mg/kg and 20 mg/kg) after last injection. Ex vivo chromatographic analysis did not show significant changes in hippocampal glutamate concentrations; however, it showed similar results as obtained in the MRS study proving its validity. Taken together, the presented study provides new insight into glutamatergic involvement in rewarding properties of mephedrone.

scholarly journals Comparison of the Resistance to Bending Forces of the 4.5 LCP Plate-rod Construct and of 4.5 LCP Alone Applied to Segmental Femoral Defects in Miniature Pigs

2010 ◽  
Vol 79 (4) ◽  
pp. 613-620 ◽  
Author(s):  
Lucie Urbanová ◽  
Robert Srnec ◽  
Pavel Proks ◽  
Ladislav Stehlík ◽  
Zdeněk Florian ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Femoral Diaphysis ◽  
Miniature Pig ◽  
Femoral Bone ◽  
Bone Implant ◽  
Miniature Pigs ◽  
Bone Defect Healing ◽  
Considerable Strain

The study deals with the determination of mechanical properties, namely resistance to bending forces, of flexible buttress osteosynthesis using two different bone-implant constructs stabilizing experimental segmental femoral bone defects (segmental ostectomy) in a miniature pig ex vivo model using 4.5 mm titanium LCP and a 3 mm intramedullary pin (“plate and rod” construct) (PR-LCP), versus the 4.5 mm titanium LCP alone (A-LCP). The “plate and rod” fixation (PR-LCP) of the segmental femoral defect is significantly more resistant (p < 0.05) to bending forces (200 N, 300 N, and 500 N) than LCP alone (A-LCP). Stabilisation of experimental segmental lesions of the femoral diaphysis in miniature pigs by flexible bridging osteosynthesis 4.5 mm LCP in combination with the “plate and rod” construct appears to be a suitable fixation of non-reducible fractures where considerable strain of the implants by bending forces can be assumed. These findings will be used in upcoming in vivo experiments in the miniature pig to investigate bone defect healing after transplantation of mesenchymal stem cells in combination with biocompatible scaffolds.

Pegylated magnetic nanocarriers for doxorubicin delivery: A quantitative determination of stealthiness in vitro and in vivo

2012 ◽  
Vol 81 (3) ◽  
pp. 498-505 ◽  
Author(s):  
E. Allard-Vannier ◽  
S. Cohen-Jonathan ◽  
J. Gautier ◽  
K. Hervé-Aubert ◽  
E. Munnier ◽  
...  
Keyword(s):  
Quantitative Determination ◽  
Doxorubicin Delivery ◽  
Magnetic Nanocarriers

Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices

1993 ◽  
Vol 50 (3) ◽  
pp. 341-351 ◽  
Author(s):  
Patrizia Casaccia-Bonnefil ◽  
Eirikur Benedikz ◽  
Hong Shen ◽  
Armin Stelzer ◽  
Diane Edelstein ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Hippocampal Slice ◽  
Hippocampal Slices ◽  
Hippocampal Slice Cultures ◽  
Acute Hippocampal Slices ◽  
Organotypic Hippocampal Slice Cultures

scholarly journals Development and Optimization of a Fluorescent Imaging System to Detect Amyloid-β Proteins: Phantom Study

2018 ◽  
Vol 9 ◽  
pp. 117959721878108 ◽  
Author(s):  
David Tes ◽  
Karl Kratkiewicz ◽  
Ahmed Aber ◽  
Luke Horton ◽  
Mohsin Zafar ◽  
...  
Keyword(s):  
Alzheimer Disease ◽  
Imaging System ◽  
Ex Vivo ◽  
Amyloid Β ◽  
The United States ◽  
Fluorescent Imaging ◽  
Fluorescent Properties ◽  
In Vivo Experiments ◽  
The Brain

Alzheimer disease is the most common form of dementia, affecting more than 5 million people in the United States. During the progression of Alzheimer disease, a particular protein begins to accumulate in the brain and also in extensions of the brain, ie, the retina. This protein, amyloid-β (Aβ), exhibits fluorescent properties. The purpose of this research article is to explore the implications of designing a fluorescent imaging system able to detect Aβ proteins in the retina. We designed and implemented a fluorescent imaging system with a range of applications that can be reconfigured on a fluorophore to fluorophore basis and tested its feasibility and capabilities using Cy5 and CRANAD-2 imaging probes. The results indicate a promising potential for the imaging system to be used to study the Aβ biomarker. A performance evaluation involving ex vivo and in vivo experiments is planned for future study.

Electronic determination of root canal working length in primary molar teeth: an in vivo and ex vivo study

2011 ◽  
Vol 44 (5) ◽  
pp. 402-406 ◽  
Author(s):  
A. P. C. A. Beltrame ◽  
T. C. Triches ◽  
N. Sartori ◽  
M. Bolan
Keyword(s):  
Root Canal ◽  
Ex Vivo ◽  
Primary Molar ◽  
Working Length ◽  
Molar Teeth ◽  
Ex Vivo Study

scholarly journals TMOD-31. ADAPTING ENGINEERED CELL THERAPIES TO UNDERSTAND AND OVERCOME GLIOBLASTOMA RESISTANCE USING INTEGRATED IN VIVO AND EX VIVO MODELS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi269-vi269
Author(s):  
Andrew Satterlee ◽  
Denise Dunn ◽  
Scott Floyd ◽  
Shawn Hingtgen
Keyword(s):  
Primary Tumor ◽  
Ex Vivo ◽  
Tumor Burden ◽  
In Vivo Studies ◽  
Pcr Analysis ◽  
Tissue Slices ◽  
Live Animal ◽  
Invasive Tumor ◽  
Recurrent Tumors

Abstract Genetically engineered neural stem cells (NSCs) are a promising therapy for the highly aggressive brain cancer glioblastoma (GBM), yet treatment durability remains a major challenge. We sought to define the events that contribute to dynamic adaption of GBM during NSC treatment and develop strategies to convert initial tumor kill into sustained GBM suppression. Using a unique hybrid tumor model treated with human skin-derived induced NSCs (iNSCs) releasing the pro-apoptotic agent TRAIL, we investigated how spatial distribution of tumor and iNSCs affects GBM adaption throughout recurrence. Serial bioluminescent imaging (BLI) was used to track tumor volumes in vivo, while a subset of mice were sacrificed 6, 13, and 20 days post-treatment to harvest brains and generate living ex vivo tissue slices. Live animal imaging showed iNSC-TRAIL treatment rapidly decreased tumor volumes when delivered into the primary tumor mass; however, minimal impact on tumor growth was observed when cells were delivered into distal regions of the brain. In contrast, high-resolution imaging of living brain sections showed extensive impacts of iNSC-TRAIL therapy that could not be visualized with BLI. The living slices showed iNSC-TRAIL treatment into the primary tumor decreased the solid, but not the invasive, tumor burden. Treatment into the lateral ventricles did impact tumor kill and was more effective at treating the invasive tumor burden and maintaining inhibition than treatment into the contralateral parenchyma. We next utilized the living tissue slices to explore the sensitivity of the recurrent tumors to TRAIL. When therapy was applied to slices harboring recurrent tumor, treatment again significantly reduced tumor volumes, suggesting that tumors had not acquired TRAIL resistance. These results informed an additional in vivo survival study and subsequent PCR analysis of untreated and recurrent tumors, and combine the fidelity of in vivo studies with the speed and spatial resolution of living brain slice technology.

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