scholarly journals Untargeted Metabolic Profiling of 4-Fluoro-Furanylfentanyl and Isobutyrylfentanyl in Mouse Hepatocytes and Urine by Means of LC-HRMS

Metabolites ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 97
Author(s):  
Camilla Montesano ◽  
Flaminia Vincenti ◽  
Federico Fanti ◽  
Matteo Marti ◽  
Sabrine Bilel ◽  
...  
Keyword(s):  
Metabolic Profiling ◽  
High Resolution Mass Spectrometry ◽  
Intraperitoneal Administration ◽  
New Psychoactive Substances ◽  
Metabolic Pattern ◽  
Mouse Hepatocytes ◽  
First Time

The diffusion of new psychoactive substances (NPS) is highly dynamic and the available substances change over time, resulting in forensic laboratories becoming highly engaged in NPS control. In order to manage NPS diffusion, efficient and innovative legal responses have been provided by several nations. Metabolic profiling is also part of the analytical fight against NPS, since it allows to identify the biomarkers of drug intake which are needed for the development of suitable analytical methods in biological samples. We have recently reported the characterization of two new analogs of fentanyl, i.e., 4-fluoro-furanylfentanyl (4F-FUF) and isobutyrylfentanyl (iBF), which were found for the first time in Italy in 2019; 4F-FUF was identified for the first time in Europe and was notified to the European Early Warning System. The goal of this study was the characterization of the main metabolites of both drugs by in vitro and in vivo experiments. To this end, incubation with mouse hepatocytes and intraperitoneal administration to mice were carried out. Samples were analyzed by means of liquid chromatography-high resolution mass spectrometry (LC–HRMS), followed by untargeted data evaluation using Compound Discoverer software with a specific workflow, designed for the identification of the whole metabolic pattern, including unexpected metabolites. Twenty metabolites were putatively annotated for 4F-FUF, with the dihydrodiol derivative appearing as the most abundant, whereas 22 metabolites were found for iBF, which was mainly excreted as nor-isobutyrylfentanyl. N-dealkylation of 4F-FUF dihydrodiol and oxidation to carbonyl metabolites for iBF were also major biotransformations. Despite some differences, in general there was a good agreement between in vitro and in vivo samples.

scholarly journals Identification and characterization of molecular interactions between mortalin/mtHsp70 and HSP60

2005 ◽  
Vol 391 (2) ◽  
pp. 185-190 ◽  
Author(s):  
Renu Wadhwa ◽  
Syuichi Takano ◽  
Kamaljit Kaur ◽  
Satoshi Aida ◽  
Tomoko Yaguchi ◽  
...  
Keyword(s):  
Heat Shock ◽  
Molecular Interactions ◽  
Heat Shock Protein 60 ◽  
Hsp60 Expression ◽  
Mitochondrial Hsp70 ◽  
Shock Proteins ◽  
First Time

Mortalin/mtHsp70 (mitochondrial Hsp70) and HSP60 (heat-shock protein 60) are heat-shock proteins that reside in multiple subcellular compartments, with mitochondria being the predominant one. In the present study, we demonstrate that the two proteins interact both in vivo and in vitro, and that the N-terminal region of mortalin is involved in these interactions. Suppression of HSP60 expression by shRNA (short hairpin RNA) plasmids caused the growth arrest of cancer cells similar to that obtained by suppression of mortalin expression by ribozymes. An overexpression of mortalin, but not of HSP60, extended the in vitro lifespan of normal fibroblasts (TIG-1). Taken together, this study for the first time delineates: (i) molecular interactions of HSP60 with mortalin; (ii) their co- and exclusive localizations in vivo; (iii) their involvement in tumorigenesis; and (iv) their functional distinction in pathways involved in senescence.

scholarly journals Bioconversion of Corticosterone into Corticosterone-Glucoside by Glucosyltransferase

Molecules ◽  
2018 ◽  
Vol 23 (7) ◽  
pp. 1783 ◽  
Author(s):  
Tokutaro Yamaguchi ◽  
Joo-Ho Lee ◽  
A-Rang Lim ◽  
Joon-Soo Sim ◽  
Eun-Ji Yu ◽  
...  
Keyword(s):  
High Resolution Mass Spectrometry ◽  
Hydroxyl Group ◽  
Oxygen Species ◽  
Neuroprotective Agents ◽  
Neuroprotective Effects ◽  
The Difference ◽  
First Time ◽  
Resolution Mass

Glucosylation of the 21-hydroxyl group of glucocorticoid changes its solubility into hydrophilicity from hydrophobicity and, as with glucocorticoid glucuronides as a moving object in vivo, it is conceivable that it exhibits the same behavior. Therefore, glucosylation to the 21-hydroxyl group while maintaining the 11β-hydroxyl group is particularly important, and glucosylation of corticosterone was confirmed by high-resolution mass spectrometry and 1D (1H and 13C) and 2D (COSY, ROESY, HSQC-DEPT and HMBC) NMR. Moreover, the difference in bioactivity between corticosterone and corticosterone 21-glucoside was investigated in vitro. Corticosterone 21-glucoside showed greater neuroprotective effects against H2O2-induced cell death and reactive oxygen species (ROS) compared with corticosterone. These results for the first time demonstrate that bioconversion of corticosterone through the region-selective glucosylation of a novel compound can present structural potential for developing new neuroprotective agents.

scholarly journals Novel Antiplasmodial Agents from Christia vespertilionis

2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Harish C. Upadhyay ◽  
Brijesh S. Sisodia ◽  
Harveer S. Cheema ◽  
Jyoti Agrawal ◽  
Anirban Pal ◽  
...  
Keyword(s):  
Antimalarial Activity ◽  
Complete Suppression ◽  
The Novel ◽  
Aqueous Methanol ◽  
Isolation And Characterization ◽  
First Time ◽  
Detailed Chemical

The roots, leaves and stems of Christia vespertilionis were separately and successively extracted with methanol and aqueous-methanol (1:4, v/v) and were evaluated in vitro for their antiplasmodial potential against Plasmodium falciparum NF-54. The aqueous-methanolic stem (AS) extract was the most active (IC50 7.5 μg/mL) followed by the methanolic leaf (ML) extract (IC50 32.0 μg/mL). The in vivo antimalarial activity of the combined plant extract of C. vespertilionis was also assessed in P. berghei infected mice, which showed 87.8% suppression of parasitaemia as compared with complete suppression by chloroquine on day 8. Finally, detailed chemical investigation of C. vespertilionis resulted in the isolation and characterization of fifteen compounds (1–15), of which two (1 and 4) are being reported for the first time from nature. The novel compound 1 possesses potent antiplasmodial activity (IC50 = 9.0 μg/mL).

scholarly journals Expression and Functional Characterization of the First Bacteriophage-Encoded Chaperonin

2012 ◽  
Vol 86 (18) ◽  
pp. 10103-10111 ◽  
Author(s):  
Lidia P. Kurochkina ◽  
Pavel I. Semenyuk ◽  
Victor N. Orlov ◽  
Johan Robben ◽  
Nina N. Sykilinda ◽  
...  
Keyword(s):  
Thermal Inactivation ◽  
Functional Characterization ◽  
Content Type ◽  
Chaperonin Groel ◽  
Phage Propagation ◽  
Physicochemical Methods ◽  
First Time

Chaperonins promote protein foldingin vivoand are ubiquitously found in bacteria, archaea, and eukaryotes. The first viral chaperonin GroEL ortholog, gene product 146 (gp146), whose gene was earlier identified in the genome of bacteriophage EL, has been shown to be synthesized during phage propagation inPseudomonas aeruginosacells. The recombinant gp146 has been expressed inEscherichia coliand characterized by different physicochemical methods for the first time. Using serum against the recombinant protein, gp146's native substrate, the phage endolysin gp188, has been immunoprecipitated from the lysate of EL-infected bacteria and identified by mass spectrometry.In vitroexperiments have shown that gp146 has a protective effect against endolysin thermal inactivation and aggregation, providing evidence of its chaperonin function. The phage chaperonin has been found to have the architecture and some properties similar to those of GroEL but not to require cochaperonin for its functional activity.

scholarly journals Metabolite Profiling of Ortho-, Meta- and Para-Fluorofentanyl by Hepatocytes and High-Resolution Mass Spectrometry

2019 ◽  
Vol 44 (2) ◽  
pp. 140-148 ◽  
Author(s):  
Per Ole M Gundersen ◽  
Anna Åstrand ◽  
Henrik Gréen ◽  
Martin Josefsson ◽  
Olav Spigset ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Urine Sample ◽  
High Resolution Mass Spectrometry ◽  
Parent Compound ◽  
Human Hepatocytes ◽  
New Psychoactive Substances ◽  
Metabolic Pattern ◽  
Drug Concentrations ◽  
Fentanyl Analogues

Abstract New psychoactive substances are emerging on the illegal drug market. Synthetic opioids including fentanyl analogues are of special concern due to their high potency. This indicates the possibility of low drug concentrations in vivo and calls for sensitive analytical methods and identification of the most appropriate analytical targets. In this study the in vitro metabolism of ortho-, meta- and para-fluorofentanyl, three fluorinated derivatives of fentanyl, has been investigated using human hepatocytes and compared to the results from an authentic human urine sample. Based on knowledge on the metabolism of similar fentanyl analogues N-dealkylation and hydroxylation was hypothesized to be the most central pathways. The three fluorofentanyl isomers were incubated with pooled human hepatocytes at 1, 3 and 5 h. Liquid chromatography quadrupole time of flight mass spectrometry operating in data-dependent mode was used to analyse the hepatocyte samples, as well as the hydrolysed and non-hydrolysed authentic urine sample. Data were analysed by a targeted approach with a database of potential metabolites. The major metabolite formed in vitro was the N-dealkylation product norfluorofentanyl. In addition various hydroxylated metabolites, a N-oxide, dihydrodiol metabolites and a hydroxymethoxy metabolite were found. In total, 14 different metabolites were identified for each fluorofentanyl isomer. In the authentic urine sample, three metabolites were detected in addition to the ortho-fluorofentanyl parent compound, with hydroxymethoxy metabolite having the highest abundance followed by norfluorofentanyl and a metabolite hydroxylated on the ethylphenyl ring. This in vitro study showed that the metabolic pattern for ortho-, meta-, and para-fluorofentanyl was close to those previously reported for other fentanyl analogues. We suggest that the hydroxymethoxy metabolite and the metabolite hydroxylated on the ethylphenyl ring should be the metabolites primarily investigated in further studies to determine the most appropriate marker for intake of fluorofentanyl derivatives in urine drug screening for human subjects.

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 IGF-II is regulated by microRNA-125b in skeletal myogenesis

2011 ◽  
Vol 192 (1) ◽  
pp. 69-81 ◽  
Author(s):  
Yejing Ge ◽  
Yuting Sun ◽  
Jie Chen
Keyword(s):  
Mammalian Target Of Rapamycin ◽  
Myoblast Differentiation ◽  
Skeletal Myogenesis ◽  
Dual Mechanism ◽  
First Time ◽  
Identification And Characterization ◽  
Major Target

MicroRNAs (miRNAs) have emerged as key regulators of skeletal myogenesis, but our knowledge of the identity of the myogenic miRNAs and their targets remains limited. In this study, we report the identification and characterization of a novel myogenic miRNA, miR-125b. We find that the levels of miR-125b decline during myogenesis and that miR-125b negatively modulates myoblast differentiation in culture and muscle regeneration in mice. Our results identify IGF-II (insulin-like growth factor 2), a critical regulator of skeletal myogenesis, as a direct and major target of miR-125b in both myocytes and regenerating muscles, revealing for the first time an miRNA mechanism controlling IGF-II expression. In addition, we provide evidence suggesting that miR-125b biogenesis is negatively controlled by kinase-independent mammalian target of rapamycin (mTOR) signaling both in vitro and in vivo as a part of a dual mechanism by which mTOR regulates the production of IGF-II, a master switch governing the initiation of skeletal myogenesis.

Cussonia arborea Hochst (Araliaceae): Ethnobotany, Pharmacology and Phytochemistry

2021 ◽  
Vol 14 (2) ◽  
pp. 6-10
Author(s):  
Modjinan Kayangar ◽  
N. Raymond Nono ◽  
T. Romuald Fouedjou ◽  
T. Billy Tchegnitegni ◽  
K. Beaudelaire Ponou ◽  
...  
Keyword(s):  
Short Review ◽  
Stem Bark ◽  
Published Data ◽  
Etoac Extract ◽  
Anticancer Properties ◽  
Isolation And Characterization ◽  
First Time

Background:  C. arborea belonging to Araliaceae family is used traditionally to cure many alien diseases including gonorrhoeae infection, diarrhea, malaria, and diabetes mellitus. The plant has been examined on the basis of scientific in vitro and in vivo evaluations possessing the major pharmacological activities including antimicrobial, antibacterial, antihyperglycemic,  antiplasmodial and anticancer properties. Aim of the study: In the present paper, we reported the isolation and characterization of secondary metabolites from the methanol extract of the stem bark of Cussonia arborea Hochst after a short review of the traditional and pharmacological studies done on this important medicinal plant. Materials and methods: MeOH extract of stem bark of C. arborea was suspended in water and successively extracted with EtOAc and n-BuOH. The EtOAc extract (18 g) was subjected to repeated column chromatography to yield seven (1-7) compounds. Their structures were determined by means of NMR, and published data. Results: The isolated compounds were identified as: protocatechuic acid (1), mixture of 3,23-dihydroxyolean-12-en-28-oic acid (2a) and 3,23-dihydroxyurs-12-en-28-oic acid (2b) in ratio 5/4, 3-O-β-D-xylopyranosylolean-12-en-28-oic acid (3), 3-O-α-L-arabinopyranosylolean-12-en-28-oic acid (4), β-resorcylic acid (5), mixture of 3-O-β-D-glucopyranosyl-23-hydroxyolean-12-en-28-oic acid (6a) and 3-O-β-D-glucopyranosyl-23-hydroxyurs-12-en-28-oic acid (6b) in ration 4/1, 3-O-β-D-glucopyranosyl-(1→2)-α-L-arabinopyranosyl-3β-hydroxyolean-12-en-28-oic acid (7). Compounds 3, 4, 5, 7, 2b and 6b are herein reported for the first time in this plant

scholarly journals Characterization of the Pseudomonas putida Mobile Genetic Element ISPpu10: an Occupant of Repetitive Extragenic Palindromic Sequences

2006 ◽  
Vol 188 (1) ◽  
pp. 37-44 ◽  
Author(s):  
María Isabel Ramos-González ◽  
María Jesús Campos ◽  
Juan Luis Ramos ◽  
Manuel Espinosa-Urgel
Keyword(s):  
Pseudomonas Putida ◽  
Insertion Element ◽  
Left And Right ◽  
First Time ◽  
Free Strain ◽  
Palindromic Sequences ◽  
Repetitive Extragenic Palindromic

ABSTRACT We have characterized the Pseudomonas putida KT2440 insertion element ISPpu10. This insertion sequence encodes a transposase which exhibits homology to the transposases and specific recombinases of the Piv/Moov family, and no inverted repeats are present at the borders of its left and right ends, thus constituting a new member of the atypical IS110/IS492 family. ISPpu10 was found in at least seven identical loci in the KT2440 genome, and variants were identified having an extra insertion at distinct loci. ISPpu10 always appeared within the core of specific repetitive extragenic palindromic (REP) sequences TCGCGGGTAAACCCG CT CCTAC, exhibiting high target stringency. One intragenic target was found associated with the truncation of a GGDEF/EAL domain protein. After active in vitro transposition to a plasmid-borne target, a duplication of the CT (underlined above) at the junction as a consequence of the ISPpu10 insertion was experimentally demonstrated for the first time in the IS110/IS492 family. The same duplication was observed after transposition of ISPpu10 from a plasmid to the chromosome of P. putida DOT-T1E, an ISPpu10-free strain with REPs similar to those of strain KT2440. Plasmid ISPpu10-mediated rearrangements were observed in vivo under laboratory conditions and in the plant rhizosphere.

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