scholarly journals Shifted-excitation Raman difference spectroscopy for in vitro and in vivo biological samples analysis

2010 ◽  
Vol 1 (2) ◽  
pp. 617 ◽  
Author(s):  
Mário Augusto da Silva Martins ◽  
Dayana Gonçalves Ribeiro ◽  
Edson Aparecido Pereira dos Santos ◽  
Airton Abrahão Martin ◽  
Adriana Fontes ◽  
...  
Keyword(s):  
Biological Samples ◽  
Difference Spectroscopy

scholarly journals Ghrelin Octanoylation Is Completely Stabilized in Biological Samples by Alkyl Fluorophosphonates

Endocrinology ◽  
2016 ◽  
Vol 157 (11) ◽  
pp. 4330-4338 ◽  
Author(s):  
Kayleigh R. McGovern-Gooch ◽  
Trevor Rodrigues ◽  
Joseph E. Darling ◽  
Michelle A. Sieburg ◽  
Alfonso Abizaid ◽  
...  
Keyword(s):  
Blood Serum ◽  
Biological Samples ◽  
Energy Homeostasis ◽  
Peptide Hormone ◽  
Microsomal Protein ◽  
Ghrelin Concentration ◽  
Acyltransferase Activity ◽  
Desacyl Ghrelin

Ghrelin is a peptide hormone involved in multiple physiological processes related to energy homeostasis. This hormone features a unique posttranslational serine octanoylation modification catalyzed by the enzyme ghrelin O-acyltransferase, with serine octanoylation essential for ghrelin to bind and activate its cognate receptor. Ghrelin deacylation rapidly occurs in circulation, with both ghrelin and desacyl ghrelin playing important roles in biological signaling. Understanding the regulation and physiological impact of ghrelin signaling requires the ability to rapidly protect ghrelin from deacylation in biological samples such as blood serum or cell lysates to preserve the relative concentrations of ghrelin and desacyl ghrelin. In in vitro ghrelin O-acyltransferase activity assays using insect microsomal protein fractions and mammalian cell lysate and blood serum, we demonstrate that alkyl fluorophosphonate treatment provides rapid, complete, and long-lasting protection of ghrelin acylation against serine ester hydrolysis without interference in enzyme assay or ELISA analysis. Our results support alkyl fluorophosphonate treatment as a general tool for stabilizing ghrelin and improving measurement of ghrelin and desacyl ghrelin concentrations in biochemical and clinical investigations and suggest current estimates for active ghrelin concentration and the ghrelin to desacyl ghrelin ratio in circulation may underestimate in vivo conditions.

scholarly journals A novel coumarin-based colorimetric and fluorescent probe for detecting increasing concentrations of Hg2+in vitro and in vivo

RSC Advances ◽  
2021 ◽  
Vol 11 (38) ◽  
pp. 23597-23606
Author(s):  
Li Huang ◽  
Wenlong Sheng ◽  
Lizhen Wang ◽  
Xia Meng ◽  
Hongdong Duan ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Biological Samples ◽  
Biological Toxicity ◽  
Environmental And Biological Samples

Mercury has complex biological toxicity and can cause a variety of physiological diseases and even death, so it is of great importance to develop novel strategies for detecting trace mercury in environmental and biological samples.

Lipid Peroxidation Due to In Vitro and In Vivo Exposure of Biological Samples to Nanoparticles

2013 ◽  
pp. 155-164 ◽  
Author(s):  
Anca Dinischiotu ◽  
Loredana Stanca ◽  
Daniela Gradinaru ◽  
Sorina Nicoleta Petrache ◽  
Mihaela Radu ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Biological Samples

scholarly journals Nanotherapeutics With Anthracyclines: Methods of Determination and Quantification of Anthracyclines in Biological Samples

2015 ◽  
pp. S1-S10
Author(s):  
E. KOZIOLOVA ◽  
O. JANOUSKOVA ◽  
P. CHYTIL ◽  
M. STUDENOVSKY ◽  
L. KOSTKA ◽  
...  
Keyword(s):  
Side Effects ◽  
Biological Samples ◽  
Chromatographic Analysis ◽  
Accurate Method ◽  
Cellular Level ◽  
Cytostatic Agents ◽  
Antitumor Therapy ◽  
Microscopy Techniques

Anthracyclines, e.g. doxorubicin, pirarubicin, are widely used as cytostatic agents in the polymer nanotherapeutics designed for the highly effective antitumor therapy with reduced side effects. However, their precise dosage scheme needs to be optimized, which requires an accurate method for their quantification on the cellular level in vitro during nanocarrier development and in body fluids and tissues during testing in vivo. Various methods detecting the anthracycline content in biological samples have already been designed. Most of them are highly demanding and they differ in exactness and reproducibility. The cellular uptake and localization is predominantly observed and determined by microscopy techniques, the anthracycline content is usually quantified by chromatographic analysis using fluorescence detection. We reviewed and compared published methods concerning the detection of anthracycline nanocarriers.

scholarly journals Metabolism of Genipin in Rat and Identification of Metabolites by Using Ultraperformance Liquid Chromatography/Quadrupole Time-of-Flight Tandem Mass Spectrometry

2013 ◽  
Vol 2013 ◽  
pp. 1-13 ◽  
Author(s):  
Yue Ding ◽  
Jian-Wei Hou ◽  
Yong Zhang ◽  
Li-Ying Zhang ◽  
Tong Zhang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Biological Samples ◽  
Time Of Flight ◽  
Tandem Mass ◽  
Enzymatic Study ◽  
Fragmentation Patterns

Thein vivoandin vitrometabolism of genipin was systematically investigated in the present study. Urine, plasma, feces, and bile were collected from rats after oral administration of genipin at a dose of 50 mg/kg body weight. A rapid and sensitive method using ultraperformance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UPLC-Q/TOF MS) was developed for analysis of metabolic profile of genipin in rat biological samples (urine, plasma, feces, and bile). A total of ten metabolites were detected and identified by comparing their fragmentation patterns with that of genipin using MetaboLynx software tools. On the basis of the chromatographic peak area, the sulfated and glucuronidated conjugates of genipin were identified as major metabolites. And the existence of major metabolites G1 and G2 was confirmed by thein vitroenzymatic study further. Then, metabolite G1 was isolated from rat bile by semipreparative HPLC. Its structure was unambiguously identified as genipin-1-o-glucuronic acid by comparison of its UV, IR, ESI-MS,1H-NMR, and13C-NMR spectra with conference. In general, genipin was a very active compound that would transform immediately, and the parent form of genipin could not be observed in rats biological samples. The biotransformation pathways of genipin involved demethylated, ring-opened, cysteine-conjugated, hydroformylated, glucuronidated, and sulfated transformations.

scholarly journals Inactivation and Elimination of SARS-CoV-2 in Biosamples Using Simple Fixatives and Ultrafiltration

2021 ◽  
Vol 4 (1) ◽  
pp. 18
Author(s):  
Ranjeet Kumar ◽  
Afsal Kolloli ◽  
Selvakumar Subbian
Keyword(s):  
Biological Samples ◽  
Culture Media ◽  
Ex Vivo ◽  
Host Cells ◽  
Laboratory Research ◽  
Health Economy ◽  
Safety Level ◽  
In Vivo Models

The Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) causes Coronavirus disease-2019 (COVID-19), which is an ongoing pandemic that has significantly affected the health, economy, and socio-economic status of individuals worldwide. Laboratory research using in vitro, ex vivo and in vivo models has been accelerated to understand the pathogenesis of SARS-CoV-2 infection. However, such experimental research involving SARS-CoV-2 is restricted to biocontainment/safety level-3 (BSL-3) settings, due to the high pathogenicity of this virus. Since many of the downstream analyses of SARS-CoV-2-infected biological samples need to be conducted in a non-BSL3 setting, it is important to ensure that the samples are fully decontaminated and safe for subsequent analysis. Here, we report the effectiveness of standard procedures used to fix cells and tissues for pathological analysis, including 2% or 4% paraformaldehyde, 50%–70% ethanol, 10% neutral buffered formalin and ultrafiltration using membranes with a molecular weight cut-off (MWCO) ranging from 3 to 30 kDa, for inactivating or eliminating SARS-CoV-2. We validated these methods in experimental laboratory samples, such as viral inoculum in cell culture media, SARS-CoV-2 infected host cells and animal tissue lysates. We found that 15 minutes’ treatment of viral inoculum (105 plaque-forming units; PFU) or SARS-CoV-2 infected cells with paraformaldehyde or 70% ethanol resulted in complete inactivation of the virus. The treatment of infected hamster lung tissues with 10% neutral buffered formalin also fully inactivated the virus. However, only 3 kDa ultracentrifuge filter was effective in eliminating the virus to an undetectable limit in the filtrate. Our validated methods are useful for decontaminating biological samples to reduce infection risk and safe handling in BSL2 facilities.

Induction and Differentiation of Dental Epithelium in Vivo and in Vitro

1982 ◽  
Vol 40 ◽  
pp. 142-145
Author(s):  
E. J. Kollar
Keyword(s):  
Connective Tissue ◽  
Oral Mucosa ◽  
Basal Lamina ◽  
Functional Derivatives ◽  
Specific Source ◽  
Dental Epithelium ◽  
Connective Tissue Stroma

The differentiation and maintenance of many specialized epithelial structures are dependent on the underlying connective tissue stroma and on an intact basal lamina. These requirements are especially stringent in the development and maintenance of the skin and oral mucosa. The keratinization patterns of thin or thick cornified layers as well as the appearance of specialized functional derivatives such as hair and teeth can be correlated with the specific source of stroma which supports these differentiated expressions.

Ultrastructural Correlations between Clonal Human Tumor Colonies and in Vivo Neoplasms

1982 ◽  
Vol 40 ◽  
pp. 210-211
Author(s):  
M.J. Murphy ◽  
R.R. Price ◽  
J.C. Sloman
Keyword(s):  
Cultured Cells ◽  
Human Tumor ◽  
Cell Type ◽  
Tumor Mass ◽  
Initial Cell ◽  
Cloning Assay ◽  
Human Tumor Cloning ◽  
The Relationship

The in vitro human tumor cloning assay originally described by Salmon and Hamburger has been applied recently to the investigation of differential anti-tumor drug sensitivities over a broad range of human neoplasms. A major problem in the acceptance of this technique has been the question of the relationship between the cultured cells and the original patient tumor, i.e., whether the colonies that develop derive from the neoplasm or from some other cell type within the initial cell population. A study of the ultrastructural morphology of the cultured cells vs. patient tumor has therefore been undertaken to resolve this question. Direct correlation was assured by division of a common tumor mass at surgical resection, one biopsy being fixed for TEM studies, the second being rapidly transported to the laboratory for culture.

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