scholarly journals Mesenchymal Stromal Cell Characteristics and Regenerative Potential in Cardiovascular Disease

2018 ◽  
Vol 27 (5) ◽  
pp. 765-785 ◽  
Author(s):  
F. C. C. van Rhijn-Brouwer ◽  
H. Gremmels ◽  
J. O. Fledderus ◽  
M. C. Verhaar
Keyword(s):  
Cardiovascular Disease ◽  
Clinical Success ◽  
Predictive Ability ◽  
In Vitro Assays ◽  
Future Research ◽  
Promising Strategy

Administration of mesenchymal stromal cells (MSCs) is a promising strategy to treat cardiovascular disease (CVD). As progenitor cells may be negatively affected by both age and comorbidity, characterization of MSC function is important to guide decisions regarding use of allogeneic or autologous cells. Definitive answers on which factors affect MSC function can also aid in selecting which MSC donors would yield the most therapeutically efficacious MSCs. Here we provide a narrative review of MSC function in CVD based on a systematic search. A total of 41 studies examining CVD-related MSC (dys)function were identified. These data show that MSC characteristics and regenerative potential are often affected by CVD. However, studies presented conflicting results, and directed assessment of MSC parameters relevant to regenerative medicine applications was lacking in many studies. The predictive ability of in vitro assays for in vivo efficacy was rarely assessed. There was no correlation between quality of study reporting and study findings. Age mismatch was also not associated with study findings or effect size. Future research should focus on assays that assess regenerative potential in MSCs and parameters that relate to clinical success.

scholarly journals Functional Characterization of the mazEF Toxin-Antitoxin System in the Pathogenic Bacterium Agrobacterium tumefaciens

2021 ◽  
Vol 9 (5) ◽  
pp. 1107
Author(s):  
Wonho Choi ◽  
Yoshihiro Yamaguchi ◽  
Ji-Young Park ◽  
Sang-Hyun Park ◽  
Hyeok-Won Lee ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Physiological Function ◽  
Functional Characterization ◽  
Site Directed Mutagenesis ◽  
In Vitro Assays ◽  
Cell Growth Inhibition ◽  
The Right

Agrobacterium tumefaciens is a pathogen of various plants which transfers its own DNA (T-DNA) to the host plants. It is used for producing genetically modified plants with this ability. To control T-DNA transfer to the right place, toxin-antitoxin (TA) systems of A. tumefaciens were used to control the target site of transfer without any unintentional targeting. Here, we describe a toxin-antitoxin system, Atu0939 (mazE-at) and Atu0940 (mazF-at), in the chromosome of Agrobacterium tumefaciens. The toxin in the TA system has 33.3% identity and 45.5% similarity with MazF in Escherichia coli. The expression of MazF-at caused cell growth inhibition, while cells with MazF-at co-expressed with MazE-at grew normally. In vivo and in vitro assays revealed that MazF-at inhibited protein synthesis by decreasing the cellular mRNA stability. Moreover, the catalytic residue of MazF-at was determined to be the 24th glutamic acid using site-directed mutagenesis. From the results, we concluded that MazF-at is a type II toxin-antitoxin system and a ribosome-independent endoribonuclease. Here, we characterized a TA system in A. tumefaciens whose understanding might help to find its physiological function and to develop further applications.

scholarly journals Method combining BAC film and positive staining for the characterization of DNA intermediates by dark-field electron microscopy

2020 ◽  
Vol 5 (1) ◽  
Author(s):  
Yann Benureau ◽  
Eliana Moreira Tavares ◽  
Ali-Akbar Muhammad ◽  
Sonia Baconnais ◽  
Eric Le Cam ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Dark Field ◽  
In Vitro Assays ◽  
Positive Staining ◽  
Synthetic Dna ◽  
Dark Field Imaging ◽  
Alkyl Ammonium

Abstract DNA intermediate structures are formed in all major pathways of DNA metabolism. Transmission electron microscopy (TEM) is a tool of choice to study their choreography and has led to major advances in the understanding of these mechanisms, particularly those of homologous recombination (HR) and replication. In this article, we describe specific TEM procedures dedicated to the structural characterization of DNA intermediates formed during these processes. These particular DNA species contain single-stranded DNA regions and/or branched structures, which require controlling both the DNA molecules spreading and their staining for subsequent visualization using dark-field imaging mode. Combining BAC (benzyl dimethyl alkyl ammonium chloride) film hyperphase with positive staining and dark-field TEM allows characterizing synthetic DNA substrates, joint molecules formed during not only in vitro assays mimicking HR, but also in vivo DNA intermediates.

scholarly journals RBP-L, a transcription factor related to RBP-Jkappa.

1997 ◽  
Vol 17 (5) ◽  
pp. 2679-2687 ◽  
Author(s):  
S Minoguchi ◽  
Y Taniguchi ◽  
H Kato ◽  
T Okazaki ◽  
L J Strobl ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Notch Receptor ◽  
In Vitro Assays ◽  
Functional Interaction ◽  
Protein Protein Interaction ◽  
Isolation And Characterization ◽  
Signalling Molecule

RBP-Jkappa is a sequence-specific DNA binding protein which plays a central role in signalling downstream of the Notch receptor by physically interacting with its intracellular region. Although at least four Notch genes exist in mammals, it is unknown whether each Notch requires a specific downstream signalling molecule. Here we report isolation and characterization of a mouse RBP-Jkappa-related gene named RBP-L that is expressed almost exclusively in lung, in contrast to the ubiquitous expression of RBP-Jkappa. For simplicity, we propose to call RBP-Jkappa RBP-J. The RBP-L protein bound to a DNA sequence almost identical to that of RBP-J. Surprisingly, RBP-L did not interact with any of the known four mouse Notch proteins. Although we found that RBP-L and EBNA-2 cooperated in transcriptional activation, they did not show significantly strong protein-protein interaction that can be detected by several in vivo and in vitro assays. This is again in contrast to physical association of RBP-J with EBNA-2. Several models to explain functional interaction between RBP-L and EBNA-2 are discussed.

scholarly journals Use of in vivo and in vitro assays for the characterization of mammalian excision repair and isolation of repair proteins

1990 ◽  
Vol 236 (2-3) ◽  
pp. 223-238 ◽  
Author(s):  
J.H.J. Hoeijmakers ◽  
A.P.M. Eker ◽  
R.D. Wood ◽  
P. Robins
Keyword(s):  
Excision Repair ◽  
In Vitro Assays

scholarly journals A comparative study of GT27 Family N-acetylgalactosaminyltransferases

2021 ◽  
Author(s):  
Biprajit Sanyal
Keyword(s):  
Half Life ◽  
Homo Sapiens ◽  
Effective Means ◽  
Therapeutic Proteins ◽  
In Vitro Assays ◽  
Future Research ◽  
E Coli ◽  
Glycosylation Sites

Therapeutic proteins face short half lives in vivo. Their high costs and associated toxicity effects of increasing dosage warrant exploration of methods to increase serum half-life. These proteins can be produced with native or engineered glycosylation sites, which has been shown to be an effective means of prolonging serum half-life. Engineered E. coli represents an economical route of production. I have been able to produce, purify and test the activity of three N-acetylgalactosaminyltransferase isoform 2 in Escherichia coli and show glycosylation on peptides derived from Interleukin 29. I followed the activity of these enzymes on three candidate therapeutic proteins via lectin blotting. Data suggest the Homo sapiens orthologue of GalNAcT2 is the most efficient enzyme in the in vitro assays with all candidate therapeutic protein substrates displaying the Tn-antigen. Future research should investigate continuous assays for precise results as well as assaying native peptides as opposed to non-native ones.

scholarly journals Pharmacological characterization of repeated administration of the first generation abused synthetic cannabinoid CP47,497

2016 ◽  
Vol 27 (3) ◽  
Author(s):  
Travis W. Grim ◽  
Kimberly L. Samano ◽  
Bogna Ignatowska-Jankowska ◽  
Qing Tao ◽  
Laura J. Sim-Selly ◽  
...  
Keyword(s):  
G Proteins ◽  
First Generation ◽  
Repeated Administration ◽  
Synthetic Cannabinoid ◽  
In Vitro Assays ◽  
Pharmacological Effects ◽  
Pharmacological Characterization

AbstractA series of in vivo and in vitro assays were conducted to characterize the pharmacological effects of the first generation abused synthetic cannabinoid CP47,497, a racemic bicyclic cannabinoid that is similar in structure to the potent, high-efficacy synthetic cannabinoid CP55,940. CP47,497 was less efficacious than CP55,940 in activating G-proteins and dose-dependently produced common CB

Natural Products as Potential Agents Against SARS-CoV and SARS-CoV-2

2021 ◽  
Vol 28 ◽  
Author(s):  
Joanda Paolla Raimundo e Silva ◽  
Chonny Alexander Herrera Acevedo ◽  
Thalisson Amorim de Souza ◽  
Renata Priscila Barros de Menezes ◽  
Zoe L. Sessions ◽  
...  
Keyword(s):  
Natural Products ◽  
In Silico ◽  
Principal Component ◽  
In Vivo Studies ◽  
In Vitro Assays ◽  
Future Research ◽  
Natural Substances ◽  
Effective Drugs

Background: Natural products are useful agents for the discovery of new lead-compounds and effective drugs to combat coronaviruses (CoV). Objective: The present work provides an overview of natural substances, plant extracts, and essential oils as potential antiSARS-CoV agents. In addition, this work evaluates their drug-like properties which are essential in the selection of compounds in order to accelerate the drug development process. Methods: The search was carried out using PubMed, ScienceDirect and SciFinder. Articles addressing plant-based natural products as potential SARS-CoV or SARS-CoV-2 agents within the last seventeen years were analyzed and selected. The descriptors for Chemometrics analyzes were obtained in alvaDesc and the principal component analyzes (PCA) were carried out in SIMCA version 13.0. Results: Based on in vitro assays and computational analyzes, this review covers twenty nine medicinal plant species and more than 300 isolated substances as potential anti-coronavirus agents. Among them, flavonoids and terpenes were the most promising compound classes. In silico analyses of drug-like properties corroborate these findings and indicate promising candidates for in vitro and in vivo studies to validate their activity. Conclusion: This paper highlights the role of ethnopharmacology in drug discovery and simulates the use of integrative (in silico/ in vitro) and chemocentric approaches to strengthen current studies and guide future research in the field of antivirals agents.

scholarly journals PPIP5K2 and PCSK1 are Candidate Genetic Contributors to Familial Keratoconus

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Mariam Lofty Khaled ◽  
Yelena Bykhovskaya ◽  
Chunfang Gu ◽  
Alice Liu ◽  
Michelle D. Drewry ◽  
...  
Keyword(s):  
Gene Trap ◽  
Whole Exome ◽  
The Us ◽  
Related Quality ◽  
First Time

AbstractKeratoconus (KC) is the most common corneal ectatic disorder affecting >300,000 people in the US. KC normally has its onset in adolescence, progressively worsening through the third to fourth decades of life. KC patients report significant impaired vision-related quality of life. Genetic factors play an important role in KC pathogenesis. To identify novel genes in familial KC patients, we performed whole exome and genome sequencing in a four-generation family. We identified potential variants in the PPIP5K2 and PCSK1 genes. Using in vitro cellular model and in vivo gene-trap mouse model, we found critical evidence to support the role of PPIP5K2 in normal corneal function and KC pathogenesis. The gene-trap mouse showed irregular corneal surfaces and pathological corneal thinning resembling KC. For the first time, we have integrated corneal tomography and pachymetry mapping into characterization of mouse corneal phenotypes which could be widely implemented in basic and translational research for KC diagnosis and therapy in the future.

scholarly journals Virulence parameters in the characterization of strains of Entamoeba histolytica

1997 ◽  
Vol 39 (2) ◽  
pp. 65-70 ◽  
Author(s):  
Maria A GOMES ◽  
Maria N. MELO ◽  
Gil P.M. PENA ◽  
Edward F. SILVA
Keyword(s):  
Entamoeba Histolytica ◽  
Cytopathic Effect ◽  
Axenic Culture ◽  
Vero Cells ◽  
In Vitro Assays ◽  
Biological Assays ◽  
Restriction Fragment Pattern

Differences in virulence of strains of Entamoeba histolytica have long been detected by various experimental assays, both in vivo and in vitro. Discrepancies in the strains characterization have been arisen when different biological assays are compared. In order to evaluate different parameters of virulence in the strains characterization, five strains of E. histolytica, kept under axenic culture, were characterized in respect to their, capability to induce hamster liver abscess, erythrophagocytosis rate and cytopathic effect upon VERO cells. It was found significant correlation between in vitro biological assays, but not between in vivo and in vitro assays. Good correlation was found between cytopathic effect and the mean number of uptaken erythrocytes, but not with percentage of phagocytic amoebae, showing that great variability can be observed in the same assay, according to the variable chosen. It was not possible to correlate isoenzyme and restriction fragment pattern with virulence indexes since all studied strains presented pathogenic patterns. The discordant results observed in different virulence assays suggests that virulence itself may not the directly assessed. What is in fact assessed are different biological characteristics or functions of the parasite more than virulence itself. These characteristics or functions may be related or not with pathogenic mechanisms occurring in the development of invasive amoebic disease

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