The usefulness of post-genomics tools for characterization of the amine cross-talk in mammalian cells

2007 ◽  
Vol 35 (2) ◽  
pp. 381-385 ◽  
Author(s):  
F. Sánchez-Jiménez ◽  
R. Montañez ◽  
F. Correa-Fiz ◽  
P. Chaves ◽  
C. Rodríguez-Caso ◽  
...  
Keyword(s):  
Systems Biology ◽  
Mathematical Models ◽  
Mammalian Cells ◽  
State Of The Art ◽  
Molecular Approaches ◽  
Amine Metabolism ◽  
Genomics Tools ◽  
Stored Information

Evidence is growing in favour of a relationship between cancer and chronic inflammation, and particularly of the role of a polyamine and histamine metabolic interplay involved in these physiopathological problems, which are indeed highly complex biological systems. Decodification of the complex inter- and intra-cellular signalling mechanisms that control these effects is not an easy task, which must be helped by systems biology technologies, including new tools for location and integration of database-stored information and predictive mathematical models, as well as functional genomics and other experimental molecular approaches necessary for hypothesis validation. We review the state of the art and present our latest efforts in this area, focused on the amine metabolism field.

scholarly journals Characterization of symbiotic and associated bacteria from entomopathogenic nematode Heterorhabditis sp. (nematode: Heterorhabditidae) isolated from India

2020 ◽  
Vol 30 (1) ◽  
Author(s):  
Rashid Pervez ◽  
Showkat Ahmad Lone ◽  
Sasmita Pattnaik
Keyword(s):  
Insect Pests ◽  
Entomopathogenic Nematode ◽  
Alcaligenes Faecalis ◽  
Symbiotic Bacteria ◽  
Rrna Gene ◽  
Main Body ◽  
Associated Bacteria ◽  
Molecular Approaches

Abstract Background Entomopathogenic nematodes (EPNs) harboring symbiotic bacteria are one of the safest alternatives to the chemical insecticides for the control of various insect pests. Infective juveniles of EPNs locate a target insect, enter through the openings, and reach the hemocoel, where they release the symbiotic bacteria and the target gets killed by the virulence factors of the bacteria. Photorhabdus with Heterorhabditis spp. are well documented; little is known about the associated bacteria. Main body In this study, we explored the presence of symbiotic and associated bacteria from Heterorhabditis sp. (IISR-EPN 09) and characterized by phenotypic, biochemical, and molecular approaches. Six bacterial isolates, belonging to four different genera, were recovered and identified as follows: Photorhabdus luminescens, one each strain of Providencia vermicola, Pseudomonas entomophila, Alcaligenes aquatilis, and two strains of Alcaligenes faecalis based on the phenotypic, biochemical criteria and the sequencing of 16S rRNA gene. Conclusion P. luminescens is symbiotically associated with Heterorhabditis sp. (IISR-EPN 09), whereas P. vermicola, P. entomophila, A. aquatilis, and A. faecalis are the associated bacteria. Further studies are needed to determine the exact role of the bacterial associates with the Heterorhabditis sp.

The growing role of structural mass spectrometry in the discovery and development of therapeutic antibodies

The Analyst ◽  
2018 ◽  
Vol 143 (11) ◽  
pp. 2459-2468 ◽  
Author(s):  
Yuwei Tian ◽  
Brandon T. Ruotolo
Keyword(s):  
Mass Spectrometry ◽  
State Of The Art ◽  
Therapeutic Antibodies ◽  
Critical Importance ◽  
Current State ◽  
Measurement Science ◽  
Structural Mass Spectrometry ◽  
Structural Mass

The comprehensive structural characterization of therapeutic antibodies is of critical importance for the successful discovery and development of such biopharmaceuticals, yet poses many challenges to modern measurement science. Here, we review the current state-of-the-art mass spectrometry technologies focusing on the characterization of antibody-based therapeutics.

scholarly journals 3213 Unraveling the role of Phospholamban (PLN) in humans via the characterization of Induced Pluripotent Stem Cell (iPSC) Cardiomyocytes (CM) derived from carriers of a lethal PLN mutation

2019 ◽  
Vol 3 (s1) ◽  
pp. 26-26
Author(s):  
Maria Giovanna Trivieri ◽  
Francesca Stillitano ◽  
Delaine Ceholski ◽  
Irene Turnbull ◽  
Kevin Costa ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
State Of The Art ◽  
Induced Pluripotent Stem Cell ◽  
Relevant Model ◽  
Study Population ◽  
Induced Pluripotent ◽  
Ca Homeostasis

OBJECTIVES/SPECIFIC AIMS: To study the biology of Phosholamban (PLN) in a human relevant model. METHODS/STUDY POPULATION: State of the art stem-cell technologies using iPSC-CMs derived from carriers of a lethal PLN mutation. RESULTS/ANTICIPATED RESULTS: Our preliminary data demonstrate that this particular PLN mutation (L39) results in reduced expression and mis-localization of PLN as well as increased incidence of early after depolarization in isolated iPSC-CMs. DISCUSSION/SIGNIFICANCE OF IMPACT: Phospholamban (PLN) is a critical regulator of Ca++ homeostasis yet many uncertainties still remain regarding its role in humans. Our study will provide unique insights into the pathophysiology of this protein in HF.

Molecular mechanisms of iron uptake in eukaryotes

1996 ◽  
Vol 76 (1) ◽  
pp. 31-47 ◽  
Author(s):  
D. M. de Silva ◽  
C. C. Askwith ◽  
J. Kaplan
Keyword(s):  
Iron Uptake ◽  
Mammalian Cells ◽  
Iron Homeostasis ◽  
Molecular Mechanisms ◽  
Iron Acquisition ◽  
Genetic Disorders ◽  
Essential Element ◽  
Recent Developments

Iron serves essential functions in both prokaryotes and eukaryotes, and cells have highly specialized mechanisms for acquiring and handling this metal. The primary mechanism by which the concentration of iron in biologic systems is controlled is through the regulation of iron uptake. Although the role of transferrin in mammalian iron homeostasis has been well characterized, the study of genetic disorders of iron metabolism has revealed other, transferrin-independent, mechanisms by which cells can acquire iron. In an attempt to understand how eukaryotic systems take up this essential element, investigators have begun studying the simple eukaryote Saccharomyces cerevisiae. Several genes have been identified and cloned that act in concert to allow iron acquisition from the environment. Some of these genes appear to have functional homologues in human systems. This review focuses on the recent developments in understanding eukaryotic iron uptake with an emphasis on the genetic and molecular characterization of these systems in both cultured mammalian cells and S. cerevisiae. An unexpected connection between iron and copper homeostasis has been revealed by recent genetic studies, which confirm biologic observations made several decades ago.

scholarly journals Generation and Characterization of a CRISPR/Cas9—Induced 3-mst Deficient Zebrafish

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 317 ◽  
Author(s):  
Antonia Katsouda ◽  
Maria Peleli ◽  
Antonia Asimakopoulou ◽  
Andreas Papapetropoulos ◽  
Dimitris Beis
Keyword(s):  
Mammalian Cells ◽  
Redox Homeostasis ◽  
Biological Processes ◽  
Oxygen Species ◽  
Regenerative Capacity ◽  
Mercaptopyruvate Sulfurtransferase ◽  
Bioinformatic Approach ◽  
Ubiquitous Presence

3-mercaptopyruvate sulfurtransferase (3-MST) is an enzyme capable of synthesizing hydrogen sulfide (H2S) and polysulfides. In spite of its ubiquitous presence in mammalian cells, very few studies have investigated its contribution to homeostasis and disease development, thus the role of 3-MST remains largely unexplored. Here, we present a clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR–associated protein-9 (Cas9) induced 3-mst mutant zebrafish line, which will allow the study of 3-MST’s role in several biological processes. The 3-mst zebrafish orthologue was identified using a bioinformatic approach and verified by its ability to produce H2S in the presence of 3-mercaptopyruvate (3-MP). Its expression pattern was analyzed during zebrafish early development, indicating predominantly an expression in the heart and central nervous system. As expected, no detectable levels of 3-Mst protein were observed in homozygous mutant larvae. In line with this, H2S levels were reduced in 3-mst−/− zebrafish. Although the mutants showed no obvious morphological deficiencies, they exhibited increased lethality under oxidative stress conditions. The elevated levels of reactive oxygen species, detected following 3-mst deletion, are likely to drive this phenotype. In line with the increased ROS, we observed accelerated fin regenerative capacity in 3-mst deficient zebrafish. Overall, we provide evidence for the expression of 3-mst in zebrafish, confirm its important role in redox homeostasis and indicate the enzyme’s possible involvement in the regeneration processes.

scholarly journals Thyroid Ultrasound: State of the Art Part 1 – Thyroid Ultrasound reporting and Diffuse Thyroid Diseases

2017 ◽  
Vol 19 (1) ◽  
pp. 79 ◽  
Author(s):  
Manjiri Dighe ◽  
Richard Barr ◽  
Jörg Bojunga ◽  
Vito Cantisani ◽  
Maria Cristina Chammas ◽  
...  
Keyword(s):  
Thyroid Nodules ◽  
State Of The Art ◽  
Thyroid Diseases ◽  
World Federation ◽  
Thyroid Ultrasound ◽  
Therapeutic Work ◽  
Work Up ◽  
Thyroid Abnormalities

Accurate differentiation of focal thyroid nodules (FTL) and thyroid abnormalities is pivotal for proper diagnostic and therapeutic work-up. In these two part articles, the role of ultrasound techniques in the characterization of FTL and evaluation of diffuse thyroid diseases is described to expand on the recently published World Federation in Ultrasound and Medicine (WFUMB) thyroid elastography guidelines and review how this guideline fits into a complete thyroid ultrasound exam.

scholarly journals Thyroid Ultrasound: State of the Art. Part 2 – Focal Thyroid Lesions

2017 ◽  
Vol 19 (2) ◽  
pp. 195 ◽  
Author(s):  
Manjiri Dighe ◽  
Richard Barr ◽  
Jörg Bojunga ◽  
Vito Cantisani ◽  
Maria Cristina Chammas ◽  
...  
Keyword(s):  
Thyroid Nodules ◽  
State Of The Art ◽  
World Federation ◽  
Thyroid Ultrasound ◽  
Therapeutic Work ◽  
Work Up ◽  
Thyroid Abnormalities ◽  
Thyroid Lesions

Accurate differentiation of focal thyroid nodules (FTL) and thyroid abnormalities is pivotal for proper diagnostic and therapeutic work-up. In these two part articles, the role of ultrasound techniques in the characterization of FTL and evaluation of diffuse thyroid diseases is described to expand on the recently published World Federation in Ultrasound and Medicine (WFUMB) thyroid elastography guidelines and review how this guideline fits into a complete thyroid ultrasound exam.

Inflamm-Aging, Cytokines and Aging: State of the Art, New Hypotheses on the Role of Mitochondria and New Perspectives from Systems Biology

2006 ◽  
Vol 12 (24) ◽  
pp. 3161-3171 ◽  
Author(s):  
S. Salvioli ◽  
M. Capri ◽  
S. Valensin ◽  
P. Tieri ◽  
D. Monti ◽  
...  
Keyword(s):  
Systems Biology ◽  
State Of The Art

scholarly journals Expression and Functional Characterization of Bluetongue Virus VP5 Protein: Role in Cellular Permeabilization

2001 ◽  
Vol 75 (18) ◽  
pp. 8356-8367 ◽  
Author(s):  
S. H. Hassan ◽  
C. Wirblich ◽  
M. Forzan ◽  
P. Roy
Keyword(s):  
Bluetongue Virus ◽  
Mammalian Cells ◽  
Functional Characterization ◽  
Recombinant Baculovirus ◽  
Structural Features ◽  
Amino Terminal ◽  
Amphipathic Helices ◽  
Outer Capsid

ABSTRACT Segment 5 of bluetongue virus (BTV) serotype 10, which encodes the outer capsid protein VP5, was tagged with glutathioneS-transferase and expressed by a recombinant baculovirus. The recombinant protein was subsequently purified to homogeneity, and its possible biological role in virus infection was investigated. Purified VP5 was able to bind mammalian cells but was not internalized, which indicates it is not involved in receptor-mediated endocytosis. The purified VP5 protein was shown to be able to permeabilize mammalian and Culicoides insect cells, inducing cytotoxicity. Sequence analysis revealed that VP5 possesses characteristic structural features (including two amino-terminal amphipathic helices) compatible with virus penetration activity. To assess the role of each feature in the observed cytotoxicity, a series of deleted VP5 molecules were generated, and their expression and biological activity was compared with the parental molecule. VP5 derivatives that included the two amphipathic helices exhibited cytotoxicity, while those that omitted these sequences did not. To confirm their role in membrane destabilization two synthetic peptides (amino acids [aa] 1 to 20 and aa 22 to 41) encompassing the two helices and an additional peptide representing the adjacent downstream sequences were also assessed for their effect on the cell membrane. Both helices, but not the downstream VP5 sequence, exhibited cytotoxicity with the most-amino-terminal helix (aa 1 to 20) showing a higher activity than the adjacent peptide (aa 22 to 41). Purified VP5 was shown to readily form trimers in solution, a feature of many proteins involved in membrane penetration. Taken together, these data support a role for VP5 in virus-cell penetration consistent with its revelation in the entry vesicle subsequent to cell binding and endocytosis.

scholarly journals Crystal structure of the inclusion complex of cholesterol in β-cyclodextrin and molecular dynamics studies

2018 ◽  
Vol 14 ◽  
pp. 838-848 ◽  
Author(s):  
Elias Christoforides ◽  
Andreas Papaioannou ◽  
Kostas Bethanis
Keyword(s):  
Crystal Structure ◽  
Inclusion Complex ◽  
Mammalian Cells ◽  
Inclusion Compound ◽  
Physicochemical Characterization ◽  
Md Simulations ◽  
Cholesterol Removal ◽  
Triclinic Space Group

The role of beta-cyclodextrin (β-CD) in cholesterol removal primarily from mammalian cells and secondly from dairy products has been studied thoroughly in recent years. Although the physicochemical characterization of the inclusion compound of cholesterol in β-CD has been achieved by various methods, no crystal structure has been determined so far. We report here the crystal structure of the inclusion compound of cholesterol in β-CD. The inclusion complex crystallizes in the triclinic space group P1 forming head-to-head dimers which are stacked along the c-axis. One well-defined cholesterol molecule ‘axially’ encapsulated inside the β-CD dimer and 22 water molecules that stabilize the complexes in the crystalline state comprise the asymmetric unit of the structure. The dimers are arranged in an intermediate (IM) channel packing mode in the crystal. Moreover, MD simulations, at 300 and 340 K, based on the crystallographically determined coordinates of the complex show that the formed cholesterol/β-CD inclusion compound remains very stable in aqueous solution at both temperatures.

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