Quantitative Analysis of Helical Flow with Accuracy Using Ultrasound Speckle Image Velocimetry: In Vitro and in Vivo Feasibility Studies

A quantitative analysis of activity-related calcium dynamics was performed in motoneurons of the nucleus hypoglossus in the brain stem slice preparation from mouse by simultaneous patch-clamp and microfluorometric calcium measurements. Motoneurons were analyzed under in vitro conditions that kept them in a functionally intact state represented by rhythmic, inspiratory-related bursts of excitatory postsynaptic currents and associated action potential discharges. Bursts of electrical activity were paralleled by somatic calcium transients resulting from calcium influx through voltage-activated calcium channels, where each action potential accounted for a calcium-mediated charge influx around 2 pC into the somatic compartment. Under in vivo conditions, rhythmic-respiratory activity in young mice occurred at frequencies up to 5 Hz, demonstrating the necessity for rapid calcium elevation and recovery in respiratory-related neurons. The quantitative analysis of hypoglossal calcium homeostasis identified an average extrusion rate, but an exceptionally low endogenous calcium binding capacity as cellular parameters accounting for rapid calcium signaling. Our results suggest that dynamics of somatic calcium transients 1) define an upper limit for the maximum frequency of respiratory-related burst discharges and 2) represent a potentially dangerous determinant of intracellular calcium profiles during pathophysiological and/or excitotoxic conditions.

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In Vitro and Preliminary In Vivo Validation of Echo Particle Image Velocimetry in Carotid Vascular Imaging

Ultrasound in Medicine & Biology ◽

10.1016/j.ultrasmedbio.2010.11.017 ◽

2011 ◽

Vol 37 (3) ◽

pp. 450-464 ◽

Cited By ~ 64

Author(s):

Fuxing Zhang ◽

Craig Lanning ◽

Luciano Mazzaro ◽

Alex J. Barker ◽

Phillip E. Gates ◽

...

Keyword(s):

Particle Image Velocimetry ◽

Particle Image ◽

Vascular Imaging ◽

Image Velocimetry ◽

In Vivo Validation

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In Vivo and In Vitro Feasibility Studies of Intraocular Use of Polyethylene Glycol–Based Synthetic Sealant to Close Retinal Breaks in Porcine and Rabbit Eyes

Investigative Opthalmology & Visual Science ◽

10.1167/iovs.14-15349 ◽

2015 ◽

Vol 56 (8) ◽

pp. 4705 ◽

Cited By ~ 13

Author(s):

Sujin Hoshi ◽

Fumiki Okamoto ◽

Mikki Arai ◽

Tatsuo Hirose ◽

Yoshimi Sugiura ◽

...

Keyword(s):

Polyethylene Glycol ◽

Feasibility Studies ◽

Retinal Breaks

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Variations in modifications of sugar residues in hamster zona pellucida after in vivo fertilization and in vitro egg activation

Reproduction ◽

10.1530/rep.0.1230671 ◽

2002 ◽

pp. 671-682 ◽

Cited By ~ 5

Author(s):

M El-Mestrah ◽

FW Kan

Keyword(s):

Quantitative Analysis ◽

Zona Pellucida ◽

Hydrolytic Enzymes ◽

Outer Region ◽

Egg Activation ◽

Cortical Granules ◽

Neuraminidase Treatment ◽

Thin Sections

Lectins were used as probes in conjunction with quantitative analysis to investigate the distribution of different carbohydrate residues in hamster zona pellucida and their possible modification patterns after in vivo fertilization and in vitro egg activation. Several lectins including HPA, WGA, RCA-I, PNA, DSA, BSAIB(4), DBA, AAA and MAA were used to label the zona pellucida of both unfertilized and fertilized eggs. With the exception of PNA and BSAIB(4), the same lectins were also used to label the zona pellucida of oocytes activated in vitro. A multicomparison quantitative analysis of the density of labelling in the inner and outer regions of the zona pellucida before and after fertilization in vivo, as well as after in vitro egg activation, was performed. Of all the lectins studied, preferential localization of labelling by RCA-I and DSA to the inner zona pellucida of unfertilized eggs was observed. After in vivo fertilization, there was an increase in labelling in the inner region of the zona pellucida when thin sections of fertilized oocytes were incubated with HPA, BSAIB(4) and AAA. Although increased labelling by RCA-I was observed, a significant decrease in labelling intensity was obtained with WGA and the sequence Neu-WGA in both the inner and outer zona pellucida of fertilized oocytes. A significant increase in the density of labelling with WGA was also observed after digestion with neuraminidase. In parallel, when hamster oocytes activated in vitro were compared with those fertilized in vivo, a difference in lectin-gold labelling was observed in both the inner and outer region of the zona pellucida. Labelling with HPA, WGA, DSA and MAA increased significantly in both the inner and outer regions of the zona pellucida, whereas labelling by DBA significantly decreased in the inner portion of the zona pellucida. After neuraminidase treatment, a significant increase in labelling density was observed when thin sections of in vitro-activated oocytes were incubated with WGA. These results demonstrate: (i) the post-fertilization modifications of major saccharidic determinants that may play a role in the sperm-egg interaction process of fertilization in vivo; and (ii) that the modified properties of zonae pellucidae of fertilized and in vitro-activated eggs resulting from the action of hydrolytic enzymes, as well as glycoproteins released through exocytosis of cortical granules, are not identical.

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A Review on Chromatography—Mass Spectrometry Applications on Anthocyanin and Ellagitannin Metabolites of Blackberries and Raspberries

Foods ◽

10.3390/foods10092150 ◽

2021 ◽

Vol 10 (9) ◽

pp. 2150

Author(s):

Dilip K. Rai ◽

Katerina Tzima

Keyword(s):

Mass Spectrometry ◽

Quantitative Analysis ◽

Health Effects ◽

Analytical Techniques ◽

In Vitro Models ◽

Qualitative And Quantitative Analysis ◽

Qualitative And Quantitative ◽

Chromatography Mass Spectrometry

Berries have been widely assessed for their beneficial health effects, predominately due to their high (poly)phenol content of anthocyanins and ellagitannins. After ellagitannins and ellagic acid are metabolized by the gut microbiome, a class of compounds known as urolithins are produced, which exert potential advantageous health effects. Anthocyanins, on the other hand, undergo a complex metabolic pathway after their interaction with microbial and endogenous enzymes, forming a broad range of metabolites and catabolic products. In most cases, in vitro models and cell lines are used to generate metabolites, whereas their assessment in vivo is currently limited. Thus far, several analytical methods have been developed for the qualitative and quantitative analysis of phenolic metabolites in berries, including liquid chromatography, mass spectrometry, and other hyphenated techniques, and have been undoubtedly valuable tools for the detailed metabolite characterization and profiling. In this review, a compilation of studies providing information on the qualitative and quantitative analysis of (poly)phenol metabolites in blackberries and raspberries after the utilization of in vitro and in vivo methods is presented. The different analytical techniques employed are assessed, focusing on the fate of the produced metabolic compounds in order to provide evidence on their characteristics, formation, and beneficial effects.

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The stentable in vitro artery: an instrumented platform for endovascular device development and optimization

Journal of The Royal Society Interface ◽

10.1098/rsif.2016.0834 ◽

2016 ◽

Vol 13 (125) ◽

pp. 20160834 ◽

Cited By ~ 3

Author(s):

Elizabeth E. Antoine ◽

François P. Cornat ◽

Abdul I. Barakat

Keyword(s):

Animal Studies ◽

Shear Stresses ◽

Detailed Measurement ◽

Collagen Hydrogel ◽

Image Velocimetry ◽

Broad Array ◽

Wide Scale ◽

Endovascular Device

Although vascular disease is a leading cause of mortality, in vitro tools for controlled, quantitative studies of vascular biological processes in an environment that reflects physiological complexity remain limited. We developed a novel in vitro artery that exhibits a number of unique features distinguishing it from tissue-engineered or organ-on-a-chip constructs, most notably that it allows deployment of endovascular devices including stents, quantitative real-time tracking of cellular responses and detailed measurement of flow velocity and lumenal shear stress using particle image velocimetry. The wall of the stentable in vitro artery consists of an annular collagen hydrogel containing smooth muscle cells (SMCs) and whose lumenal surface is lined with a monolayer of endothelial cells (ECs). The system has in vivo dimensions and physiological flow conditions and allows automated high-resolution live imaging of both SMCs and ECs. To demonstrate proof-of-concept, we imaged and quantified EC wound healing, SMC motility and altered shear stresses on the endothelium after deployment of a coronary stent. The stentable in vitro artery provides a unique platform suited for a broad array of research applications. Wide-scale adoption of this system promises to enhance our understanding of important biological events affecting endovascular device performance and to reduce dependence on animal studies.

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Phytochemical Screening, In-vitro and In-vivo Anti-diabetic Activity of Nelumbo nucifera Leaves against Alloxan-Induced Diabetic Rabbits

Indian Journal of Animal Research ◽

10.18805/ijar.b-1181 ◽

2019 ◽

Author(s):

Shahid Maqbool ◽

Najeeb Ullah ◽

Aqal Zaman ◽

Atif Akbar ◽

Saima Saeed ◽

...

Keyword(s):

Body Weight ◽

Quantitative Analysis ◽

Cold Water ◽

Nelumbo Nucifera ◽

Plant Leaves ◽

Standard Drug ◽

Antihyperglycemic Activity ◽

Butanol Fraction

A recent survey has demonstrated that traditional healers and herbalists frequently use Nelumbo nucifera plant to treat diabetes. The plant investigated here has been previously reported as natural folk medicine, but until now no scientific investigation of potential anti-diabetic effects have been reported. Plant leaves of Nelumbo nucifera were screened out for quantitative analysis of phytochemicals, in-vitro anti-diabetic activity (Alpha-amylase inhibitory assay) and In-vivo anti-diabetic assay of animals (rabbits). Quantitative analysis indicates that different phytochemicals were present in plant leaves extract/fractions. In-vitro antidiabetic analysis results poses that butanol fraction is significantly more active than other fractions or extract in antihyperglycemic activity with minimum IC50 value of 8.56µg/ml. For In-vivo anti-diabetic assay, animals (rabbits) were examined and animals were divided into 10 groups and except blank group all were treated with standard and plant extracts/fractions doses (25mg and 50mg per Kg body weight) as an aqueous suspension. Their blood sugar level was thoroughly recorded with a glucometer before the administration of sample and standard drug. Alloxan Monohydrate was administrated orally (as an ice-cold water solution) at dose rate 150mg/kg body weight for one week to make animals hyperglycemic. In-vivo analysis indicates that butanol fraction is the most active fraction in antihyperglycemic activity. So Leaves of N. nucifera can be used as antidiabetic remedy. So, it is strongly recommended that N. nucifera leaves should be elucidated further for isolation of active phytochemicals.

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