In Vivo and In Vitro Characterization of the ARR11 Response Regulator Implicated in the His-to-Asp Phosphorelay Signal Transduction in Arabidopsis thaliana

The δ-opioid receptor (δOR) holds great potential as a therapeutic target. Yet, clinical drug development, which has focused on δOR agonists that mimic the potent and selective tool compound SNC80 have largely failed. It has increasingly become apparent that the SNC80 scaffold carries with it potent and efficacious β-arrestin recruitment. Here, we screened a relatively small (5120 molecules) physical drug library to identify δOR agonists that underrecruit β-arrestin, as it has been suggested that compounds that efficaciously recruit β-arrestin are proconvulsant. The screen identified a hit compound and further characterization using cellular binding and signaling assays revealed that this molecule (R995045, compound 1) exhibited ten-fold selectivity over µ- and κ-opioid receptors. Compound 1 represents a novel chemotype at the δOR. A subsequent characterization of fourteen analogs of compound 1, however did not identify a more potent δOR agonist. Computational modeling and in vitro characterization of compound 1 in the presence of the endogenous agonist leu-enkephalin suggest compound 1 may also bind allosterically and negatively modulate the potency of Leu-enkephalin to inhibit cAMP, acting as a ‘NAM-agonist’ in this assay. The potential physiological utility of such a class of compounds will need to be assessed in future in vivo assays.

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In vivo and in vitro characterization of rifampicine suppositories

Journal of Drug Delivery Science and Technology ◽

10.1016/s1773-2247(09)50039-1 ◽

2009 ◽

Vol 19 (3) ◽

pp. 217-221

Author(s):

H.M. El-Nahas ◽

F.S. Ghazy ◽

H.A. El-Ghamry ◽

A.M. El-Wsaby

Keyword(s):

In Vitro Characterization

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In vivo and in vitro characterization of the B1 and B2 zinc-binding domains from the acute promyelocytic leukemia protooncoprotein PML.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.93.4.1601 ◽

1996 ◽

Vol 93 (4) ◽

pp. 1601-1606 ◽

Cited By ~ 75

Author(s):

K. L. Borden ◽

J. M. Lally ◽

S. R. Martin ◽

N. J. O'Reilly ◽

E. Solomon ◽

...

Keyword(s):

Acute Promyelocytic Leukemia ◽

Promyelocytic Leukemia ◽

Zinc Binding ◽

Binding Domains ◽

In Vitro Characterization

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In vivo and in vitro trimethadione oxidation activity of the liver from various animal species including mouse, hamster, rat, rabbit, dog, monkey and human

Human & Experimental Toxicology ◽

10.1177/096032719901800102 ◽

1999 ◽

Vol 18 (1) ◽

pp. 12-16 ◽

Cited By ~ 2

Author(s):

E Tanaka ◽

A Ishikawa ◽

T Horie

Keyword(s):

Animal Species ◽

In Vitro Study ◽

Metabolic Clearance ◽

Oxidation Activity ◽

In Vitro Characterization ◽

Demethylase Activity ◽

Total Body

Trimethadione (TMO) has the properties required of a probe drug for the evaluation of hepatic drug-oxidizing capacity and, in this study, we have summarized the in vivo and in vitro metabolism of TMO in various animal species including mouse, hamster, rat, rabbit, dog, monkey and human. In the in vivo study, the plasma TMO level was measured after intravenous or oral (human) administration of TMO at a dose of 4 mg/kg to various animal species. The rate of TMO metabolic clearance in these animal species in vivo was in the order mouse > hamster >rat>rabbit>dog>monkey>human. In the in vitro study, species differences were observed in the cytochrome P450 (P450) content and drug-oxidizing enzyme activity. The content of P450 was monkey> mouse>dog>rabbit>hamster>rat>human. On the other hand, TMO N-demethylation was in the order mouse >hamster >rat >rabbit>dog>monkey>human. There was a good correlation between the mean total body clearance of TMO ( in vivo)andthemeanTMON-demethylase activity ( in vitro) (y=1.7×+0.11, r=0.965, P<0.001). These results show that TMO is a probe agent with metabolic and pharmacokinetic characteristics making it attractive for the in vivo and in vitro characterization of metabolic activity in various animal species.

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Functional Gene Network of Prenyltransferases in Arabidopsis thaliana

Molecules ◽

10.3390/molecules24244556 ◽

2019 ◽

Vol 24 (24) ◽

pp. 4556 ◽

Cited By ~ 1

Author(s):

Diana Kopcsayová ◽

Eva Vranová

Keyword(s):

Arabidopsis Thaliana ◽

Amino Acid Level ◽

Chain Elongation ◽

Pathway Network ◽

Isoprenoid Pathway ◽

Prenyl Diphosphate ◽

Mutant Complementation

Prenyltransferases (PTs) are enzymes that catalyze prenyl chain elongation. Some are highly similar to each other at the amino acid level. Therefore, it is difficult to assign their function based solely on their sequence homology to functional orthologs. Other experiments, such as in vitro enzymatic assay, mutant analysis, and mutant complementation are necessary to assign their precise function. Moreover, subcellular localization can also influence the functionality of the enzymes within the pathway network, because different isoprenoid end products are synthesized in the cytosol, mitochondria, or plastids from prenyl diphosphate (prenyl-PP) substrates. In addition to in vivo functional experiments, in silico approaches, such as co-expression analysis, can provide information about the topology of PTs within the isoprenoid pathway network. There has been huge progress in the last few years in the characterization of individual Arabidopsis PTs, resulting in better understanding of their function and their topology within the isoprenoid pathway. Here, we summarize these findings and present the updated topological model of PTs in the Arabidopsis thaliana isoprenoid pathway.

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In vitro characterization of PRRSV isolates with different in vivo virulence using monocyte-derived macrophages

Veterinary Microbiology ◽

10.1016/j.vetmic.2019.03.008 ◽

2019 ◽

Vol 231 ◽

pp. 139-146 ◽

Cited By ~ 3

Author(s):

Giulia Ogno ◽

Carmen A. Sautter ◽

Elena Canelli ◽

Obdulio García-Nicolás ◽

Tomasz Stadejek ◽

...

Keyword(s):

In Vitro Characterization

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Invertebrate Gonadotropin-Releasing Hormone Receptor Signaling and Its Relevant Biological Actions

International Journal of Molecular Sciences ◽

10.3390/ijms21228544 ◽

2020 ◽

Vol 21 (22) ◽

pp. 8544

Author(s):

Tsubasa Sakai ◽

Tatsuya Yamamoto ◽

Shin Matsubara ◽

Tsuyoshi Kawada ◽

Honoo Satake

Keyword(s):

Signal Transduction ◽

Biological Functions ◽

Hpg Axis ◽

Intracellular Calcium Mobilization ◽

Releasing Factors ◽

Species Specific ◽

Gonadotropin Releasing Hormone Receptor

Gonadotropin-releasing hormones (GnRHs) play pivotal roles in reproduction via the hypothalamus-pituitary-gonad axis (HPG axis) in vertebrates. GnRHs and their receptors (GnRHRs) are also conserved in invertebrates lacking the HPG axis, indicating that invertebrate GnRHs do not serve as “gonadotropin-releasing factors” but, rather, function as neuropeptides that directly regulate target tissues. All vertebrate and urochordate GnRHs comprise 10 amino acids, whereas amphioxus, echinoderm, and protostome GnRH-like peptides are 11- or 12-residue peptides. Intracellular calcium mobilization is the major second messenger for GnRH signaling in cephalochordates, echinoderms, and protostomes, while urochordate GnRHRs also stimulate cAMP production pathways. Moreover, the ligand-specific modulation of signal transduction via heterodimerization between GnRHR paralogs indicates species-specific evolution in Ciona intestinalis. The characterization of authentic or putative invertebrate GnRHRs in various tissues and their in vitro and in vivo activities indicate that invertebrate GnRHs are responsible for the regulation of both reproductive and nonreproductive functions. In this review, we examine our current understanding of and perspectives on the primary sequences, tissue distribution of mRNA expression, signal transduction, and biological functions of invertebrate GnRHs and their receptors.

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