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 Calpains Induce Egg Production Through Mirnas Secretion in Extracellular Vesicles Released From Paired Adult Worms of Schistosoma Japonicum

2021 ◽  
Author(s):  
Takashi Kumagai ◽  
Rieko Shimogawara ◽  
Koichiro Ichimura ◽  
Shiroh Iwanaga
Keyword(s):  
Electron Microscopy ◽  
Schistosoma Japonicum ◽  
Extracellular Vesicles ◽  
Egg Production ◽  
Developmental Stages ◽  
Calpain Inhibitor ◽  
Single Female ◽  
Female Specific ◽  
Male Specific

Abstract Extracellular vesicles (EVs) have been reported to be secreted from Schistosoma japonicum at all developmental stages. However, the reproduction and communication mechanisms between the paired adults through the EVs in dioecious Trematoda have not been reported. In this study, EVs containing many exosome-like vesicles and microvesicles were observed in the supernatants of paired adults cultured in vitro, and abundant selected miRNAs were contained in them. In particular, the female-specific miR-bantam was present only in vesicles and was hardly secreted outside the vesicles. In this study, we found that male-female pairing induced secretion of miR-3479 and miR-bantam in EVs, but not of male-specific miR-61. Furthermore, ingestion of mouse erythrocytes also increased the production of miRNAs in paired adult and single female worms. Vesicles were found in the teguments of females treated with erythrocytes under electron microscopy. After the paired worms were treated with several inhibitors against the secretion of EVs, only calpain inhibitor (calpeptin) significantly reduced the amount of miRNA in EVs. Furthermore, the worms treated with only calpeptin inhibited egg production in vitro. Together, these results indicate that qualitative miRNA production through EVs regulated by calpain plays a role in egg production in S. japonicum.

scholarly journals Gene Expression in Developmental Stages of Schistosoma japonicum Provides Further Insight into the Importance of the Schistosome Insulin-Like Peptide

2019 ◽  
Vol 20 (7) ◽  
pp. 1565 ◽  
Author(s):  
Xiaofeng Du ◽  
Malcolm Jones ◽  
Sujeevi Nawaratna ◽  
Shiwanthi Ranasinghe ◽  
Chunrong Xiong ◽  
...  
Keyword(s):  
Schistosoma Japonicum ◽  
Developmental Stages ◽  
Insulin Receptors ◽  
Glucose Consumption ◽  
Adenosine Diphosphate ◽  
Internal Cell ◽  
Parasite Survival ◽  
Survival And Growth ◽  
Insight Into

We showed previously that the Schistosoma japonicum insulin-like peptide (SjILP) binds the worm insulin receptors, thereby, activating the parasite’s insulin pathway and emphasizing its important role in regulating uptake of glucose, a nutrient essential for parasite survival. Here we show that SjILP is differentially expressed in the schistosome life cycle and is especially highly transcribed in eggs, miracidia, and adult female worms. RNA inference was employed to knockdown SjILP in adults in vitro, with suppression confirmed by significantly reduced protein production, declined adenosine diphosphate levels, and reduction in glucose consumption. Immunolocalization showed that SjILP is located to lateral gland cells of mature intra-ovular miracidia in the schistosome egg, and is distributed on the ciliated epithelium and internal cell masses of newly transformed miracidia. In schistosomula, SjILP is present on the tegument in two antero-lateral points, indicating highly polarized expression during cercarial transformation. Analysis of serum from S. japonicum-infected mice by ELISA using a recombinant form of SjILP as an antigen revealed IgG immunoreactivity to this molecule at 7 weeks post-infection indicating it is likely secreted from mature eggs into the host circulation. These findings provide further insights on ILP function in schistosomes and its essential roles in parasite survival and growth in different development stages.

Differentially Expressed and Novel Transcripts in Highly Purified Chronic Phase CML Stem Cells

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 193-193
Author(s):  
Yun Zhao ◽  
Allen Delaney ◽  
Afshin Raouf ◽  
Kamini Raghuram ◽  
Haiyan I Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Blood Cells ◽  
Molecular Mechanisms ◽  
Chronic Phase ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Direct Role ◽  
Hematopoietic Stem ◽  
Transcript Levels

Abstract The chronic phase of CML is sustained by rare BCR-ABL+ stem cells. These cells share many properties with normal pluripotent hematopoietic stem cells, but also differ in critical ways that alter their growth, drug responsiveness and genome stability. Understanding the molecular mechanisms underlying the biological differences between normal and CML stem cells is key to the development of more effective CML therapies. To obtain new insights into these mechanisms, we generated Long Serial Analysis of Gene Expression (SAGE) libraries from paired isolates of highly purified lin-CD34+CD45RA-CD36- CD71-CD7-CD38+ and lin-CD34+CD45RA-CD36-CD71-CD7-CD38- cells from 3 chronic phase CML patients (all with predominantly Ph+/BCR-ABL+ cells in both subsets) and from 3 control samples: a pool of 10 normal bone marrows (BMs), a single normal BM and a pool of G-CSF-mobilized blood cells from 9 donors. In vitro bioassays showed the CD34+CD38+ cells were enriched in CFCs (CML: 3–20% pure; normal: 4–19% pure) and the CD34+CD38- cells were enriched in LTC-ICs (CML: 0.2–26% pure; normal: 12–52% pure). Each of the 12 libraries was then sequenced to a depth of ~200,000 tags and tags from libraries prepared from like phenotypes were compared between genotypes using DiscoverySpace software and hierarchical clustering. 1687 (355 with clustering) and 1258 (316 with clustering) transcripts were thus identified as differentially expressed in the CML vs control CD34+CD38− and CD34+CD38+ subsets, respectively. 266 of these transcripts (11 with clustering) were differentially expressed in both subsets. The differential expression of 5 genes (GAS2, IGF2BP2, IL1R1, DUSP1 & SELL) was confirmed by real-time PCR analysis of lin-CD34+ cells isolated from an additional 5 normal BMs and 11 CMLs, and lin-CD34+CD38− cells from an additional 2 normal BMs and 2 CMLs (with dominant Ph+ cells). GAS2 and IL1R1 transcript levels were correlated with BCR-ABL transcript levels in both primitive subsets, and predicted differences in expression of IL1R1 and SELL were apparent within 3 days in CD34+ cord blood cells transduced with a lenti-BCR-ABL-IRES-GFP vs a control lenti-GFP vector (n=3). These findings support a direct role of BCR-ABL in perturbing the expression of these 3 genes. Further comparison of the meta CD34+CD38− and CD34+CD38+ CML cell libraries with most publicly accessible SAGE data revealed 69 novel tags in the CD34+ CML cells that correspond to unique but conserved genomic sequences. Nine of these were recovered by 5′- and 3′- RACE applied to cDNAs pooled from several human leukemic cell lines. These results illustrate the power of SAGE to reveal key components of the transcriptomes of rare human CML stem cell populations including transcripts of genes not previously known to exist. Continuing investigation of their biological roles in primary CML cells and primitive BCR-ABL-transduced human cells offer important strategies for delineating their potential as therapeutic targets.

Anoxia-induced transcriptional upregulation of sarp-19: cloning and characterization of a novel EF-hand containing gene expressed in hepatopancreas of Littorina littorea

2004 ◽  
Vol 82 (2) ◽  
pp. 285-293 ◽  
Author(s):  
Kevin Larade ◽  
Kenneth B Storey
Keyword(s):  
Time Course ◽  
Signal Sequence ◽  
Calcium Ionophore ◽  
Fold Increase ◽  
Feedback Mechanism ◽  
Littorina Littorea ◽  
Transcript Levels ◽  
Reading Frame ◽  
Ef Hand

Many marine molluscs have well-developed biochemical adaptations that allow them to live without oxygen for long periods of time, but very little is currently known about the molecular biology underlying these processes. Differential screening of a cDNA library derived from the hepatopancreas of the marine snail Littorina littorea revealed a novel anoxia-induced gene, sarp-19 (snail anoxia-responsive protein, 19 kDa). Examination of the sarp-19 transcript revealed an open reading frame that encoded a protein of 168 amino acids containing an N-terminal signal sequence and two putative EF-hand domains. Expression analysis of transcript levels established that sarp-19 accumulated over a time course of anoxia exposure, reaching a maximum 5.6-fold increase after 96 h compared with aerobic controls. However, transcript levels were reduced by 50% within 1 h when aerobic conditions were reestablished. Nuclear runoff assays confirmed transcriptional upregulation of sarp-19 during anoxia exposure, and organ explant experiments showed that the gene was also responsive to anoxia exposure in vitro. sarp-19 transcripts were also elevated in response to freezing, suggesting that the protein may have a role in the physiological responses of this intertidal snail to both aerial exposure and winter freezing. Hepatopancreas explants treated with a calcium ionophore showed increased levels of the sarp-19 transcript, suggesting a possible feedback mechanism regulated by levels of intracellular calcium. Expression was also responsive to tissue incubation with cyclic GMP and phorbol 12-myristate 13-acetate but was not affected by cyclic AMP, implicating involvement of protein kinases G and C but not protein kinase A in the expression of sarp-19. The SARP-19 protein may play a role in calcium-activated signaling during anoxia exposure in L. littorea.Key words: anoxia, mollusc, gastropod, calcium, EF hand.

scholarly journals Molecular cloning, expression and characterization of Bmserpin-2 gene from Bombyx mori.

2009 ◽  
Vol 56 (4) ◽  
Author(s):  
Ye Pan ◽  
Hengchuan Xia ◽  
Peng Lü ◽  
KePing Chen ◽  
Qin Yao ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Developmental Stages ◽  
Pcr Analysis ◽  
Post Inoculation ◽  
Amino Acid Residues ◽  
Gene Encoding ◽  
Reading Frame ◽  
E Coli ◽  
Real Time Quantitative Pcr ◽  
Localization Analysis

Serpins are a broadly distributed family of protease inhibitors. In this study, the gene encoding Bombyx mori serpin-2 (Bmserpin-2) was cloned and expressed in E. coli. The Bmserpin-2 cDNA contains a 1125 bp open reading frame (ORF). The deduced protein has 374 amino-acid residues, contains a conserved SERPIN domain and shares extensive homology with other invertebrate serpins. RT-PCR analysis showed that Bmserpin-2 was expressed in all developmental stages of B. mori larvae and various larval tissues. Subcellular localization analysis indicated that Bmserpin-2 protein was located in the cytoplasm. Interestingly, real-time quantitative PCR revealed that the expression of Bmserpin-2 in the midgut of susceptible B. mori strain 306 significantly increased at 72 hours post inoculation (hpi) when infected with BmNPV. However, there was no significant increase of the Bmserpin-2 expression in resistant strain NB infected with BmNPV. Thus, our data indicates that Bmserpin-2 may be involved in B. mori antiviral response.

scholarly journals Functional analysis of the Frzb2 gene in Schistosoma japonicum

2019 ◽  
Vol 50 (1) ◽  
Author(s):  
Guifeng Cheng ◽  
Xiaochun Li ◽  
Fanglin Qin ◽  
Rong Xu ◽  
Yuanyuan Zhang ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Morphological Structure ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Single Sex ◽  
Reproductive Ability ◽  
Protein Receptor ◽  
Rapid Amplification ◽  
Novel Target ◽  
Interfering Rna

AbstractSchistosomiasis is a globally important helminthic disease of humans and animals, and it is the second most common parasitic disease after malaria. Eggs produced by mature females are responsible for the disease’s occurrence and spread. Frzb2, a secreted frizzled-related protein, can inhibit Wnt signalling by competitive binding to the specific frizzled protein receptor. In this study, the complete gene sequence of SjFrzb2 was obtained by using 3′-rapid amplification of cDNA ends technology. SjFrzb2 transcript levels at different stages of S. japonicum maturation were evaluated by quantitative real-time RT-PCR analysis. SjFrzb2 was expressed at all developmental stages examined and exhibited the highest transcription level in 7-day-old worms, then gradually decreased during the growth and developmental stages to reach the lowest level at 18 days post-infection. SjFrzb2 gene expression was higher in female worms than in male worms and was significantly higher in female worms from a single-sex infection than in female worms from a bisexual infection. The functions of SjFrzb2 were explored via a small interfering RNA-based gene silencing approach and the soaking method. The results showed that SjFrzb2 gene knockdown impaired the growth and development of S. japonicum in mice, affecting not only the survival and morphological structure of the worms but also their reproductive ability and the viability of the produced eggs. Collectively, these observations imply that Frzb2 may be a novel target for the development of immuno- and/or small molecule-based therapeutics to control schistosomiasis fecundity and transmission.

scholarly journals Ameliorative effects of Schisandrin B on Schistosoma mansoni-induced hepatic fibrosis in vivo

2021 ◽  
Vol 15 (6) ◽  
pp. e0009554
Author(s):  
Ho Yin Pekkle Lam ◽  
Ting-Ruei Liang ◽  
Shih-Yi Peng
Keyword(s):  
Liver Fibrosis ◽  
Schistosoma Mansoni ◽  
Egg Production ◽  
Treatment Options ◽  
Parasite Burden ◽  
Inflammasome Activation ◽  
Schisandrin B ◽  
Male Adult

Schistosomiasis is second only to malaria as the most devastating parasitic disease in the world. It is caused by the helminths Schistosoma mansoni (S. mansoni), S. haematobium, or S. japonicum. Typically, patients with schistosomiasis suffer from symptoms of liver fibrosis and hepatosplenomegaly. Currently, patients were treated with praziquantel. Although praziquantel effectively kills the worm, it cannot prevent re-infection or resolve liver fibrosis. Also, current treatment options are not ample to completely cure liver fibrosis and splenic damages. Moreover, resistance of praziquantel has been reported in vivo and in vitro studies. Therefore, finding new effective treatment agents is urgently needed. Schisandrin B (Sch B) of Schisandra chinensis has been shown to protect against different liver injuries including fatty liver disease, hepatotoxicity, fibrosis, and hepatoma. We herein investigate the potential of using Sch B to treat S. mansoni-induced liver fibrosis. Results from the present study demonstrate that Sch B is beneficial in treating S. mansoni-induced liver fibrosis and splenic damages, through inhibition of inflammasome activation and apoptosis; and aside from that regulates host immune responses. Besides, Sch B treatment damages male adult worm in the mice, consequently helps to reduce egg production and lessen the parasite burden.

scholarly journals Molecular Cloning and Characterization of an Anthocyanidin Synthase Gene in Prunus persica (L.) Batsch

2017 ◽  
Vol 45 (1) ◽  
pp. 28-35 ◽  
Author(s):  
Jiabao YE ◽  
Feng XU ◽  
Guiyuan WANG ◽  
Qiangwen CHEN ◽  
Tingting TAO ◽  
...  
Keyword(s):  
Prunus Persica ◽  
Transcript Level ◽  
Pcr Analysis ◽  
Anthocyanin Content ◽  
Anthocyanin Accumulation ◽  
Anthocyanidin Synthase ◽  
Enzyme Activity Assay ◽  
Reading Frame ◽  
Fruit Skin

To elucidate the effect of anthocyanidin synthase (ANS) gene on anthocyanin accumulation in fruit skin of Prunus persica (L.) Batsch cv. 'Chunmei', this study cloned and characterized an ANS gene (PpANS) from peach. PpANS (GenBank accession No. KX760117) was encoded by a 1074 bp-long open reading frame (ORF) corresponding to a polypeptide consisting of 358 amino acids with a molecular mass of 40.45 kD and an isoelectric point of 5.46. PpANS contains a conserved 2-oxoglutarate- and iron-dependent dioxygenases and non-haem dioxygenase binding regions. PpANS shared high similarities to angiosperm ANS and displayed the closest genetic relationship to Prunus domestica. Real-time PCR analysis indicated that PpANS was highly expressed in fruit skin, flesh and flowers, and peach fruit skin showed the highest transcript level of PpANS. Anthocyanin accumulation analysis indicated that it was highly accumulated in fruit skin and flesh of peach. Changes in the transcript level were highly correlated with anthocyanin content in the different tissues of peach. Prokaryotic expression analysis showed PpANS that protein can be expressed correctly in E. coli, and the size of PpANS recombinant protein was consistent with its predicted size. In vitro enzyme activity assay revealed that recombinant PpANS protein could catalyze the formation the cyanidin from leucocyanidin. These results indicated that PpANS was responsible for anthocyanin accumulation in P. persica.

scholarly journals Dose-dependent Inhibition of Gynecophoral Canal Protein Gene Expression in Vitro in the Schistosome (Schistosoma japonicum) by RNA Interference

2005 ◽  
Vol 37 (6) ◽  
pp. 386-390 ◽  
Author(s):  
Guo-Feng Cheng ◽  
Jiao-Jiao Lin ◽  
Yi Shi ◽  
You-Xin Jin ◽  
Zhi-Qiang Fu ◽  
...  
Keyword(s):  
Schistosoma Japonicum ◽  
Protein Gene ◽  
Female Worm ◽  
Transcript Level ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Male Worm ◽  
Dependent Inhibition ◽  
Dose Dependent

Abstract The gynecophoral canal protein gene SjGCP of Schistosoma japonicum that is necessary for the pairing between the male and female worms is specifically expressed in the adult male worm. This protein is widely distributed in the adult female worm after pairing. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence were employed to analyze the relationship between the RNAi effect and dsRNA dosage in the parasites. The results revealed that the inhibition of SjGCP expression by siRNA is dose-dependent. RT-PCR analysis showed that the SjGCP transcript level was reduced by 75% when 100 nM dsRNA was applied.

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.

Sign in / Sign up

Export Citation Format

Share Document