scholarly journals Proteome Profiling of PMJ2-R and Primary Peritoneal Macrophages

2021 ◽  
Vol 22 (12) ◽  
pp. 6323
Author(s):  
Alexander L. Rusanov ◽  
Peter M. Kozhin ◽  
Olga V. Tikhonova ◽  
Victor G. Zgoda ◽  
Dmitry S. Loginov ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Living Organism ◽  
Peritoneal Macrophages ◽  
In Vitro Models ◽  
Individual Characteristics ◽  
Essential Proteins ◽  
Comparative Proteomic Analysis ◽  
Immortalized Cell ◽  
The Individual

In vitro models are often used for studying macrophage functions, including the process of phagocytosis. The application of primary macrophages has limitations associated with the individual characteristics of animals, which can lead to insufficient standardization and higher variability of the obtained results. Immortalized cell lines do not have these disadvantages, but their responses to various signals can differ from those of the living organism. In the present study, a comparative proteomic analysis of immortalized PMJ2-R cell line and primary peritoneal macrophages isolated from C57BL/6 mice was performed. A total of 4005 proteins were identified, of which 797 were quantified. Obtained results indicate significant differences in the abundances of many proteins, including essential proteins associated with the process of phagocytosis, such as Elmo1, Gsn, Hspa8, Itgb1, Ncf2, Rac2, Rack1, Sirpa, Sod1, C3, and Msr1. These findings indicate that outcomes of studies utilizing PMJ2-R cells as a model of peritoneal macrophages should be carefully validated. All MS data are deposited in ProteomeXchange with the identifier PXD022133.

scholarly journals Comparative proteomic analysis of the mouse ovary in response to GnIH and GnRH: An in vitro approach

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Anushree Dave ◽  
Michael D. Powell ◽  
Rajagopala Sridaran ◽  
Kazuyoshi Tsutsui ◽  
Amitabh Krishna
Keyword(s):  
Proteomic Analysis ◽  
Mouse Ovary ◽  
Comparative Proteomic Analysis

Evaluation of the Antioxidant and Antidiabetic Potential of the Poly Herbal Formulation: Identification of Bioactive Factors

2020 ◽  
Vol 18 (2) ◽  
pp. 111-123
Author(s):  
V.V. Sathibabu Uddandrao ◽  
Parim Brahmanaidu ◽  
Saravanan Ganapathy
Keyword(s):  
Plant Extracts ◽  
Antioxidant Properties ◽  
In Vitro Models ◽  
Piper Nigrum ◽  
Individual Plant ◽  
Glucose Lowering ◽  
Methanolic Extracts ◽  
Herbal Formulation ◽  
The Individual

Background & Objectives: The present investigation is intended to prepare a Poly Herbal Formulation (PHF) with Piper nigrum (fruits), Terminalia paniculata (bark) and Bauhinia purpurea (bark) and assess their antioxidant and glucose-lowering effects utilizing in vitro models. Methods: The individual plant methanolic extracts and PHF are exposed to phytochemical examination and to distinguish the bioactive factors by GC-MS. We assessed the antioxidant properties of individual plant extracts and the PHF by using the DPPH scavenging method, H2O2 scavenging assay, TBARS assay and total antioxidant estimation. Likewise, the anti-diabetic activity was assessed by ɑ-amylase and α-glucosidase enzyme inhibition and glucose diffusion inhibitory techniques. Results: We found that PHF contains a high measure of total phenolics, total flavonoids and tannin compared to individual plant extracts. The GC-MS identified the bioactive components. We also found that PHF had significantly higher antioxidant and glucose-lowering effects than the individual plant concentrates. Conclusion: In conclusion, it could be reasoned that due to the nearness of antioxidant components, the PHF has good potential in the administration of hyperglycemia, diabetes and the related state of oxidative stress. This study shows that PHF is superior to individual plant extracts, supporting the conventional PHF concept.

Effects of Colchicine on the Golgi Complex and Gerl of Cultured Rat Peritoneal Macrophages and Epiphyseal Chondrocytes

1980 ◽  
Vol 45 (1) ◽  
pp. 41-58
Author(s):  
JOHAN THYBERG ◽  
ANDRZEJ PIASEK ◽  
STANISLAW MOSKALEWSKI
Keyword(s):  
Golgi Complex ◽  
Structural Integrity ◽  
Cell Types ◽  
Peritoneal Macrophages ◽  
Electron Microscopic ◽  
Thiamine Pyrophosphatase ◽  
Cytoplasmic Microtubules ◽  
The Individual ◽  
Functionally Diverse

Thioglycollate-elicited rat peritoneal macrophages and epiphyseal chondrocytes were cultured in vitro, treated with colchicine, and then studied by electron-microscopic and cytochemical techniques. Colchicine, but not lumicolchicine, caused disappearance of cytoplasmic microtubules and breakup of the Golgi complex with spreading of its dictyosomes from a well defined juxta nuclear area throughout the cytoplasm. There was also an altered distribution of lysosomes, which oriented themselves close to the dictyosomes both in controland colchicine-treated cells. Further, the structure of the individual dictyosomes was changed, especially in the chondrocytes. GERL equivalents were observed in control cells but were difficult to detect after exposure to colchicine. Reaction product for thiamine pyrophosphatase was found in narrow cisternae on the inner side of the dictyosomes in control cells but invacuole-like structures in colchicine-treated cells. Reaction product for acid phosphatase was present in GERL equivalents and lysosomes in control cells but mainly in lysosomes incolchicine-treated cells. Nevertheless, the total specific activities of these enzymes as well as of 5′-nucleotidase, a plasma membrane marker, remained unaffected by the drug treatment. These observations show that cytoplasmic microtubules play an important and, in many respects similar, cytoskeletal role in two so functionally diverse cell types as macrophages and chondrocytes. They are particularly important for the structural integrity of the Golgi complex, which in both cells is normally organized in the area around the centrioles, from which numerous microtubules radiate into the cytoplasm. The observations further suggest that GERL is an integrated part of the Golgi complex in these cells.

scholarly journals Comparative Proteomic Analysis of Buffalo Oocytes Matured in vitro Using iTRAQ Technique

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Lingsheng Chen ◽  
Linhui Zhai ◽  
Chunfeng Qu ◽  
Chengpu Zhang ◽  
Sheng Li ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Comparative Proteomic Analysis

62 COMPARATIVE PROTEOMIC ANALYSIS OF LIVER MITOCHONDRIA DERIVED FROM DECEASED NEWBORN CLONED CALVES AND ADULT CLONES BY TWO-DIMENSIONAL DIFFERENTIAL GEL ELECTROPHORESIS

2011 ◽  
Vol 23 (1) ◽  
pp. 137
Author(s):  
K. Takeda ◽  
M. Tasai ◽  
S. Akagi ◽  
S. Watanabe ◽  
M. Oe ◽  
...  
Keyword(s):  
Protein Expression ◽  
Somatic Cell ◽  
Proteomic Analysis ◽  
Gel Electrophoresis ◽  
Mitochondrial Protein ◽  
Two Dimensional ◽  
Cell Nuclei ◽  
Comparative Proteomic Analysis ◽  
Mitochondrial Fractions

Aberrant reprogramming of donor somatic cell nuclei may result in many severe problems in animal cloning. The inability to establish functional interactions between donor nucleus and recipient mitochondria is also likely responsible for developmental deficiency. However, an understanding of the expressed proteins in cattle is lacking. In the present study, alterations in mitochondrial protein levels between somatic cell nuclear transferred (SCNT) and control animals (mostly produced by AI) were investigated. Nuclear transfer was performed using donor cells prepared from cumulus cells (B1), ear skin, or skeletal muscle from adult Japanese Black cattle, and enucleated in vitro matured oocytes (Holstein or Japanese Black) as previously reported (Akagi et al. 2003). Liver samples were collected from postmortem SCNT calves (CB1-3; 0, 1, and 9 days postnatally) and adult SCNT cattle (CA1-4; 6, 6, 6, and 5 years of age) produced from the same cell line (B1) and preserved at –80°C. Mitochondrial fractions were prepared from the frozen–thawed liver samples by mechanical homogenization and differential centrifugation, and subjected to two-dimensional difference in gel electrophoresis (2D-DIGE) using CyDye™ dyes (Cy2, Cy3, Cy5; GE Healthcare) for specific labelling. Protein expression changes were confirmed by ImageMaster 2D Platinum software with a volume ratio greater than 2.0 (Student’s t-test; P < 0.05). The expression of 5 proteins were up-regulated in SCNT calves compared to control calves (n = 6; Day 250 fetus, 0, 4, 8, 8, and 8 days after birth; P < 0.05). Expressed protein patterning compared to control groups was different among SCNT calves. The protein spots of CB-1 showed great differences compared with other SCNT calves; 13 spots were up-regulated, and 18 spots were down-regulated. In adult SCNT cattle, the concentrations of 3 proteins were higher when compared to control cattle (n = 4; 2, 2, 6, and 8 years of age; P < 0.05). Protein expression was different among individual SCNT animals even if they were produced from the same donor cell source. For example, 9 spots were up-regulated and 7 spots were down-regulated in CA-1. In contrast, no differences were detected in 2 of the SCNT cattle (CA-3 and 4; P < 0.05). Novel proteins were not identified in any of the SCNT cattle or calves. In conclusion, alteration of mitochondrial protein expression levels were observed in non-viable neonatal SCNT calves and varied among SCNT individuals; suggesting that mitochondrial related gene expression may be implicated in early losses. Comparative proteomic analysis represents an important tool for further studies on SCNT animals. We thank Dr. C. A. Pinkert (Auburn Univ.) and Dr. Somfai (NARO) for their assistance. This work was supported by a grant from the NARO, Japan.

In-silico Subtractive Proteomic Analysis Approach for Therapeutic Targets in MDR Salmonella enterica subsp. enterica serovar Typhi str. CT18

2019 ◽  
Vol 19 (29) ◽  
pp. 2708-2717 ◽  
Author(s):  
Noor Rahman ◽  
Ijaz Muhammad ◽  
Gul E. Nayab ◽  
Haroon Khan ◽  
Rosanna Filosa ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Metabolic Pathways ◽  
Salmonella Enterica ◽  
Drug Targets ◽  
Protein Sequences ◽  
Analysis Approach ◽  
Essential Proteins ◽  
Pathways Analysis

Objective: In the present study, an attempt has been made for subtractive proteomic analysis approach for novel drug targets in Salmonella enterica subsp. enterica serover Typhi str.CT18 using computational tools. Method: Paralogous, redundant and less than 100 amino acid protein sequences were removed by using CD-HIT. Further detection of bacterial proteins which are non-homologous to host and are essential for the survival of pathogens by using BLASTp against host proteome and DEG`s, respectively. Comparative Metabolic pathways analysis was performed to find unique and common metabolic pathways. The non-redundant, non-homologous and essential proteins were BLAST against approved drug targets for drug targets while Psortb and CELLO were used to predict subcellular localization. Result: There were 4473 protein sequences present in NCBI Database for Salmonella enterica subsp. enterica serover Typhi str. CT18 out of these 327 were essential proteins which were non-homologous to human. Among these essential proteins, 124 proteins were involved in 19 unique metabolic pathways. These proteins were further BLAST against approved drug targets in which 7 cytoplasmic proteins showed druggability and can be used as a therapeutic target. Conclusion: Drug targets identification is the prime step towards drug discovery. We identified 7 cytoplasmic druggable proteins which are essential for the pathogen survival and non-homologous to human proteome. Further in vitro and in vivo validation is needed for the evaluation of these targets to combat against salmonellosis.

scholarly journals Distinct Effects of Inflammation on Cytochrome P450 Regulation and Drug Metabolism: Lessons from Experimental Models and a Potential Role for Pharmacogenetics

Genes ◽  
2020 ◽  
Vol 11 (12) ◽  
pp. 1509
Author(s):  
Laura M. de Jong ◽  
Wim Jiskoot ◽  
Jesse J. Swen ◽  
Martijn L. Manson
Keyword(s):  
Cytochrome P450 ◽  
Drug Metabolism ◽  
Drug Response ◽  
Genetic Factors ◽  
Potential Role ◽  
In Vitro Models ◽  
Experimental Models ◽  
Individual Variability ◽  
The Individual

Personalized medicine strives to optimize drug treatment for the individual patient by taking into account both genetic and non-genetic factors for drug response. Inflammation is one of the non-genetic factors that has been shown to greatly affect the metabolism of drugs—primarily through inhibition of cytochrome P450 (CYP450) drug-metabolizing enzymes—and hence contribute to the mismatch between the genotype predicted drug response and the actual phenotype, a phenomenon called phenoconversion. This review focuses on inflammation-induced drug metabolism alterations. In particular, we discuss the evidence assembled through human in-vitro models on the effect of inflammatory mediators on clinically relevant CYP450 isoform levels and their metabolizing capacity. We also present an overview of the current understanding of the mechanistic pathways via which inflammation in hepatocytes may modulate hepatic functions that are critical for drug metabolism. Furthermore, since large inter-individual variability in response to inflammation is observed in human in-vitro models and clinical studies, we evaluate the potential role of pharmacogenetic variability in the inflammatory signaling cascade and how this can modulate the outcome of inflammation on drug metabolism and response.

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