scholarly journals Characterization of Membrane-Associated Progesterone Receptor Component-2 (MAPRC2) from Trichinella spiralis and Its Interaction with Progesterone and Mifepristone

Vaccines ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 934
Author(s):  
Muhammad Tahir Aleem ◽  
Jiawen Shi ◽  
Zhengqing Yu ◽  
Zhaohai Wen ◽  
Yang Zhang ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Adult Worm ◽  
Trichinella Spiralis ◽  
Protein Sequencing ◽  
Sequencing Analysis ◽  
Female Adult ◽  
Male Adult ◽  
Receptor Component ◽  
Female Adult Worm

Trichinellosis is a foodborne zoonotic disease caused by Trichinella spp., including Trichinella spiralis. In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized using protein sequencing analysis. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in vitro cell culture that showed different expression levels at all developmental stages (muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL), and newborn larvae (NBL)). Subsequently, the in vitro phenotypic effects of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later, the in vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The 300 ng/mL of P4 and 100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (p ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only 30 ng/mL P4 showed significant upregulation in F-AL (p ≤ 0.05). The current study provides new insights regarding the antihormone (P4 and RU486) drug design and vaccine therapy of recombinant (rTs-MAPRC2) protein as well as their combined effects to control T. spiralis infection.

scholarly journals Characterization of Membrane-associated Progesterone Receptor Component-2 (MAPRC2) From Trichinella spiralis and Its Interaction With Progesterone and Mifepristone

2021 ◽  
Author(s):  
Muhammad Tahir Aleem ◽  
Shi Jiawen ◽  
Zheng Qing Yu ◽  
Zhao hai Wen ◽  
Zhang Yang ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Adult Worm ◽  
Trichinella Spiralis ◽  
Protein Sequencing ◽  
Phenotypic Effect ◽  
Female Adult ◽  
Male Adult ◽  
Receptor Component ◽  
Female Adult Worm

Abstract Background: Trichinellosis is a food-borne zoonotic disease caused by nematode viz., Trichinella spiralis. Physiologically, the high progesterone (P4) doses cause new borne larvae (NBL) mortality in the parasite, while the low doses maintain pregnancy. In contrast, Mifepristone (RU486) works as an antagonist against the progesterone receptor (PR) and possesses abortifacient activities. Methods: In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized by protein sequencing. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in-vitro cell culture that showed different expression levels at all developmental stages [muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL) and new borne larvae (NBL)]. Subsequently, the in-vitro phenotypic effect of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later on, the in-vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Results: Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The P300 ng/mL of P4 and M100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (P ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only P30 ng/mL showed significant upregulation in F-AL (P ≤ 0.05). Conclusions: The current study provides new insights regarding the antihormone (P4 & RU486) drug design and vaccine therapy of recombinant (rTs-MAPRC2) protein as well as their combined effects to control T. spiralis infection.

scholarly journals Characterisation and preliminary functional analysis of N-acetyltransferase 13 from Schistosoma japonicum

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Yalan Tang ◽  
Kerou Zhou ◽  
Qingqing Guo ◽  
Cheng Chen ◽  
Jing Jia ◽  
...  
Keyword(s):  
Schistosoma Japonicum ◽  
Egg Production ◽  
Developmental Stages ◽  
Pcr Analysis ◽  
Female Adult ◽  
Male Adult ◽  
Transcript Levels ◽  
Reading Frame ◽  
Interfering Rna

Abstract Background N-acetyltransferase 13 (NAT13) is a probable catalytic component of the ARD1A-NARG1 complex possessing alpha (N-terminal) acetyltransferase activity. Results In this study, a full-length complementary DNA (cDNA) encoding Schistosoma japonicum NAT13 (SjNAT13) was isolated from schistosome cDNAs. The 621 bp open reading frame of SjNAT13 encodes a polypeptide of 206 amino acids. Real-time PCR analysis revealed SjNAT13 expression in all tested developmental stages. Transcript levels were highest in cercariae and 21-day-old worms, and higher in male adult worms than female adult worms. The rSjNAT13 protein induced high levels of anti-rSjNAT13 IgG antibodies. In two independent immunoprotection trials, rSjNAT13 induced 24.23% and 24.47% reductions in the numbers of eggs in liver. RNA interference (RNAi) results showed that small interfering RNA (siRNA) Sj-514 significantly reduced SjNAT13 transcript levels in worms and decreased egg production in vitro. Conclusions Thus, rSjNAT13 might play an important role in the development and reproduction of schistosomes.

scholarly journals Genotypic and Phenotypic Analyses of Hepatitis C Virus from Patients Treated with JTK-853 in a Three-Day Monotherapy

2012 ◽  
Vol 57 (1) ◽  
pp. 436-444 ◽  
Author(s):  
Naoki Ogura ◽  
Yukiyo Toyonaga ◽  
Izuru Ando ◽  
Kunihiro Hirahara ◽  
Tsutomu Shibata ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Amino Acid ◽  
Viral Resistance ◽  
Hcv Rna ◽  
Amino Acid Substitutions ◽  
Sequencing Analysis ◽  
Resistance Mutations ◽  
In The Wild

ABSTRACTJTK-853, a palm site-binding NS5B nonnucleoside polymerase inhibitor, shows antiviral activityin vitroand in hepatitis C virus (HCV)-infected patients. Here, we report the results of genotypic and phenotypic analyses of resistant variants in 24 HCV genotype 1-infected patients who received JTK-853 (800, 1,200, or 1,600 mg twice daily or 1,200 mg three times daily) in a 3-day monotherapy. Viral resistance in NS5B was investigated using HCV RNA isolated from serum specimens from the patients. At the end of treatment (EOT) with JTK-853, the amino acid substitutions M414T (methionine [M] in position 414 at baseline was replaced with threonine [T] at EOT), C445R (cysteine [C] in position 445 at baseline was replaced with arginine [R] at EOT), Y448C/H (tyrosine [Y] in position 448 at baseline was replaced with cysteine [C] or histidine [H] at EOT), and L466F (leucine [L] in position 466 at baseline was replaced with phenylalanine [F] at EOT), which are known to be typical resistant variants of nonnucleoside polymerase inhibitors, were observed in a clonal sequencing analysis. These substitutions were also selected by a treatment with JTK-853in vitro, and the 50% effective concentration of JTK-853 in the M414T-, C445F-, Y448H-, and L466V-harboring replicons attenuated the susceptibility by 44-, 5-, 6-, and 21-fold, respectively, compared with that in the wild-type replicon (Con1). These findings suggest that amino acid substitutions of M414T, C445R, Y448C/H, and L466F are thought to be viral resistance mutations in HCV-infected patients receiving JTK-853 in a 3-day monotherapy.

Induction of surface fluidity in Trichinella spiralis larvae during penetration of the host intestine: simulation by cyclic AMP in vitro

Parasitology ◽  
1997 ◽  
Vol 114 (1) ◽  
pp. 71-77 ◽  
Author(s):  
J. MODHA ◽  
M. C. ROBERTS ◽  
M. W. KENNEDY ◽  
J. R. KUSEL
Keyword(s):  
Cyclic Amp ◽  
Trichinella Spiralis ◽  
Lateral Diffusion ◽  
Epithelial Tissue ◽  
Intestinal Epithelial ◽  
Parasitic Helminths ◽  
Intracellular Parasites ◽  
Fluid Properties ◽  
Lipid Probe

The lateral diffusion (DL) properties of the fluorescent lipid probe 5-N (octadecanoyl) aminofluorescein (AF18) inserted into the surface of muscle-stage larvae of Trichinella spiralis were investigated by fluorescence recovery after photobleaching. AF18 was not free to diffuse laterally in dormant larvae, and this remained unchanged after larval activation in vitro with trypsin and bile. However, a significant increase in surface fluidity of the probe was demonstrated (%R = 74·5; DL = 11·5 × 10−9 cm2/sec) when larvae invaded intestinal epithelial tissue following oral infection of mice. Membrane-permeant photoactivatable caged cyclic AMP was used to analyse the putative mechanism responsible for this increase in lateral diffusion in the parasite surface. Although incubation of larvae with 1–50 μM caged cAMP had no effect on surface fluidity, incubation with 100 μM caged cAMP induced a substantial increase in the lateral mobility of AF18 (%R = 64·3; DL = 8·3 × 10−11 cm2/sec) immediately following photo-activation of the caged messenger. This induced fluidity, however, was transient and the larval surface reverted to immobility within 15 min. These observations constitute the first reported measurement of the fluid properties of the surface of intracellular parasites, the first demonstration of the parasite surface fluidity altering as a result of host cell invasion and the first indication of a mechanism underlying changes in surface fluidity in parasitic helminths.

In vitro and in vivo enhancement of progesterone binding to the uterine progesterone receptor by cortisol

Biochemistry ◽  
1977 ◽  
Vol 16 (13) ◽  
pp. 2829-2834 ◽  
Author(s):  
Syed Saiduddin ◽  
Hans Peter Zassenhaus
Keyword(s):  
Progesterone Receptor

Lack of Correlation between In vitro and In vivo Activation of Complement in Mice by Infective Trichinella spiralis Larvae

1989 ◽  
Vol 75 (4) ◽  
pp. 647 ◽  
Author(s):  
M. Stankiewicz ◽  
G. Sokolska ◽  
A. Pilarczyk ◽  
D. Sadowska ◽  
E. L. Jeska
Keyword(s):  
Trichinella Spiralis

Glucosiduronidation of androsterone by human breast tumors in vitro in relation to the progesterone receptor content

Steroids ◽  
1983 ◽  
Vol 42 (4) ◽  
pp. 449-455 ◽  
Author(s):  
Uma Raju ◽  
Mortimer Levitz ◽  
Alexander Kaganowicz ◽  
Ancel Blaustein
Keyword(s):  
Progesterone Receptor ◽  
Breast Tumors ◽  
Human Breast

scholarly journals In vitro effects of artesunate on the survival of worm pairs and egg production of Schistosoma mansoni

2009 ◽  
Vol 83 (1) ◽  
pp. 7-11 ◽  
Author(s):  
Y. Mitsui ◽  
M. Miura ◽  
Y. Aoki
Keyword(s):  
Schistosoma Mansoni ◽  
Adult Worm ◽  
Survival Time ◽  
Egg Production ◽  
Inhibitory Effect ◽  
Male And Female ◽  
Paired Male ◽  
In Vitro Effects ◽  
Paired Female

AbstractThe effect of artesunate (ART) on the survival time of adult worm pairs of Schistosoma mansoni and on their egg output during in vitro culture was assessed. ART significantly decreased the survival time of both paired male and female worms at concentrations of 5, 10, 20 and 40 mg l− 1 during in vitro cultivation. An inhibitory effect of ART on the daily egg output of paired female worms during in vitro cultivation was also observed.

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