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 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 Regulation of Sex-Specific Selection offruitless 5′ Splice Sites by transformerand transformer-2

1998 ◽  
Vol 18 (1) ◽  
pp. 450-458 ◽  
Author(s):  
Volker Heinrichs ◽  
Lisa C. Ryner ◽  
Bruce S. Baker
Keyword(s):  
Splice Site ◽  
Courtship Behavior ◽  
Specific Pattern ◽  
Splice Sites ◽  
Repeat Elements ◽  
Specific Alternative ◽  
Female Specific ◽  
Male Specific ◽  
Splicing Enhancer

ABSTRACT In Drosophila melanogaster, the fruitless(fru) gene controls essentially all aspects of male courtship behavior. It does this through sex-specific alternative splicing of the fru pre-mRNA, leading to the production of male-specific fru mRNAs capable of expressing male-specificfru proteins. Sex-specific fru splicing involves the choice between alternative 5′ splice sites, one used exclusively in males and the other used only in females. Here we report that the Drosophila sex determination genestransformer (tra) and transformer-2(tra-2) switch fru splicing from the male-specific pattern to the female-specific pattern through activation of the female-specific fru 5′ splice site. Activation of female-specific fru splicing requirescis-acting tra and tra-2 repeat elements that are part of an exonic splicing enhancer located immediately upstream of the female-specific fru 5′ splice site and are recognized by the TRA and TRA-2 proteins in vitro. Thisfru splicing enhancer is sufficient to promote the activation by tra and tra-2 of both a 5′ splice site and the female-specific doublesex (dsx) 3′ splice site, suggesting that the mechanisms of 5′ splice site activation and 3′ splice site activation may be similar.

scholarly journals Molecular Sex Identification in the Hardy Rubber Tree (Eucommia ulmoides Oliver) via ddRAD Markers

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Wencai Wang ◽  
Guoqian Yang ◽  
Xin Deng ◽  
Fengqing Shao ◽  
Yongquan Li ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Polymerase Chain Reaction Amplification ◽  
Rubber Tree ◽  
Sex Identification ◽  
Morphological Observation ◽  
Eucommia Ulmoides ◽  
Sex Recognition ◽  
Sex Determination System ◽  
Female Specific ◽  
Male Specific

Eucommia ulmoides, also known as the industrially and medicinally important hardy rubber tree, is the sole species of Eucommiaceae. Nevertheless, its dioecious property hinders sex recognition by traditional morphological observation at very early developmental stages, thus inhibiting breeding and economic cropping. In this study, double-digest restriction site-associated DNA sequencing (ddRAD-seq) was applied to screen sex-linked molecular markers for sex identification and investigation of the sex determination system in 20 male and female E. ulmoides individual plants, respectively. In consequence, five candidate male-specific loci but no female-specific loci were predicated among the 183,752 male and 147,122 female catalogue loci by bioinformatics analysis. Subsequent PCR (polymerase chain reaction) amplification and Sanger sequencing examinations were performed on another 24 individuals, 12 for each sex, from a separate population. One ideal sex-linked locus, MSL4, was identified among the five putative male-specific loci that were found using ddRAD data. MSL4 is 479 bp in length and highly conserved in all the male individuals, suggesting its feature of being stable and repeatable. Our results also indicated that the sex of E. ulmoides is likely determined genetically. In short, this study provides a consistent and reproducible ddRAD marker (MSL4) that is able to discriminate male from female seedlings in E. ulmoides, which will be valuable for rapid breeding practice and better commercial production of this economically important tree.

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.

scholarly journals Radicicol, a Novel Lead Compound against the Migratory-Stage Schistosomula of Schistosoma japonicum

2020 ◽  
Vol 65 (3) ◽  
Author(s):  
Liangxiong Xu ◽  
Qiuli Liu ◽  
Qingren Zeng ◽  
Ping Wu ◽  
Quan Yu ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Schistosoma Japonicum ◽  
Primary Treatment ◽  
Parasitic Disease ◽  
Lead Compound ◽  
Content Type ◽  
Acid Lactone ◽  
Scanning Electron

ABSTRACT Schistosomiasis poses a serious threat to human health and remains a major tropical and parasitic disease in more than 70 countries. Praziquantel (PZQ) has been the primary treatment for schistosomiasis for nearly 4 decades. However, its efficacy against migratory-stage schistosomula is limited. Radicicol (RAD), a β-resorcylic acid lactone derived from Paecilomyces sp. strain SC0924, was investigated as an alternative treatment for Schistosoma japonicum. In vitro tests showed that within 72 h, RAD (10 μmol/liter) completely killed schistosomula of both skin and liver stages with an efficacy significantly higher than that of PZQ, although it was less potent against adult worms than PZQ. In vivo, RAD reduced worm burdens and liver eggs by 91.18% and 86.01%, respectively, by killing migratory-stage schistosomula. Optical microscopy and scanning electron microscopy revealed that RAD damaged the epiderm and tegument morphology of S. japonicum worms at various stages and altered their motility to different degrees. RAD exhibited schistosomicidal effects at different stages in vitro and in vivo, especially at the migratory stage, implying that its mechanism could be different from that of PZQ. Collectively, these results showed that RAD is promising as a lead for the development of drugs to control the migratory-stage schistosomula of S. japonicum.

scholarly journals Synergistic effect of combination chemotherapy with praziquantel and DW-3-15 for Schistosoma japonicum in vitro and in vivo

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Zi-Yin Yang ◽  
Zi-Hao Liu ◽  
Ya-Nan Zhang ◽  
Chen Li ◽  
Lei Liu ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Synergistic Effect ◽  
Schistosoma Japonicum ◽  
Combination Chemotherapy ◽  
Combination Index ◽  
First Choice ◽  
Scanning Electron

Abstract Background Schistosomiasis is a debilitating and neglected tropical disease for which praziquantel (PZQ) remains the first-choice drug for treatment and control of the disease. In our previous studies, we found that the patented compound DW-3-15 (patent no. ZL201110142538.2) displayed significant and stabilized antiparasitic activity through a mechanism that might be distinct from PZQ. Here, we investigated the antischistosomal efficacy of PZQ combined with DW-3-15 against schistosomula and adult worms of Schistosoma japonicum in vitro and in vivo, to verify whether there was a synergistic effect of the two compounds. Methods The antischistosomal efficacy of PZQ combined with DW-3-15 in comparison with an untreated control and monotherapy group against schistosomula and adult worms was assessed both in vitro and in vivo. Parasitological studies, scanning electron microscopy, combination index, and histopathological analysis were used for the assessment. Results The results showed significantly reduced viability of schistosomes, achieving 100% viability reduction for juveniles and males by combination chemotherapy using PZQ together with DW-3-15 in vitro. The combination index was 0.28, 0.27, and 0.53 at the higher concentration of PZQ combined with DW-3-15 against juveniles, males, and females, respectively, indicating that the two compounds display strong synergism. Scanning electron microscopy observations also demonstrated that the compound combination induced more severe and extensive alterations to the tegument and subtegument of S. japonicum than those with each compound alone. In vivo, compared with the single-compound-treated group, the group treated with the higher-dose combination demonstrated the best schistosomicidal efficacy, with significantly reduced worm burden, egg burden, and granuloma count and area, which was evident against schistosomula and adult worms. Conclusions Our study provides a potential novel chemotherapy for schistosomiasis caused by S. japonicum. It would improve the antischistosomal effect on schistosomula and adult worms of S. japonicum, and decrease individual dosages. Graphical Abstract

scholarly journals Synergic effects of decellularized bone matrix, hydroxyapatite, and extracellular vesicles on repairing of the rabbit mandibular bone defect model

2020 ◽  
Author(s):  
Asrin Emami ◽  
Tahereh Talaei-Khozani ◽  
Saeid Tavanafar ◽  
Nehleh Zareifard ◽  
Negar Azarpira ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Bone Repair ◽  
Bone Matrix ◽  
Bone Cell ◽  
Histochemical Staining ◽  
Beneficial Effects ◽  
Mg63 Cells

Abstract Background: Extracellular vesicles (ECV) and bone extracellular matrix (ECM) have beneficial effects on the treatment of some pathological conditions. The purpose of this study was to find the synergic effects of decellularized bone (DB) ECM and ECVs on the repair of rabbit. Methods: The quality of decellularized sheep bones was confirmed by H&E, Hoechst, DNA quantification, immunohistochemistry, histochemical staining, and scanning electron microscopy (SEM). Osteoblast-derived ECVs were evaluated by internalization test, Transmission electron microscopy, Dynamic light scattering, and flow cytometry for CD9, CD63, CD81 markers. The hydrogel containing DB and hydroxyapatite (HA) with or without ECVs was evaluated for osteoblast functions and bone repair both in vitro and in vivo. Results: The data indicated ECM preservation after decellularization as well as cell depletion. In vitro assessments revealed that mineralization and alkaline phosphatase activity did not improve after treatment of MG63 cells by ECVs, while in vivo morphomatrical estimations showed synergic effects of ECVs and DB+HA hydrogels on increasing the number of bone-specific cells and vessel and bone area compared to the control, DB+HA and ECV-treated groups. Conclusions: The DB enriched with ECVs can be an ideal scaffold for bone tissue engineering and may provide a suitable niche for bone cell migration and differentiation.

Schistosoma japonicum: immunolocalization of paramyosin during development

Parasitology ◽  
1997 ◽  
Vol 114 (1) ◽  
pp. 45-52 ◽  
Author(s):  
G. N. GOBERT ◽  
D. J. STENZEL ◽  
M. K. JONES ◽  
D. E. ALLEN ◽  
D. P. McMANUS
Keyword(s):  
Electron Microscopy ◽  
Schistosoma Japonicum ◽  
Polyclonal Antibody ◽  
Developmental Stages ◽  
Muscle Layer ◽  
Protective Properties ◽  
Vaccine Target ◽  
Target Molecule

This paper describes the localization of paramyosin immunoreactivity in Schistosoma japonicum and represents the first comparative immunolocalization study among schistosome adult, cercariae and lung schistosomula by electron microscopy. A polyclonal antibody was utilized to immunolabel paramyosin or paramyosin-like proteins. Paramyosin was localized within the muscle layer of all 3 developmental stages. Furthermore, paramyosin was localized within granules of the post-acetabular glands of cercariae, and within the tegument matrix and surface of lung schistosomules. Adults and cercariae did not display any detectable paramyosin on the surface or within the tegument. The possible functions of paramyosin within S. japonicum and the relevance of these findings in relation to the reported protective properties of paramyosin as an anti-schistosome vaccine target molecule are discussed.

scholarly journals CSIG-11. CENTRAL NERVOUS SYSTEM TUMOR PATIENTS HAVE ELEVATED LEVELS OF CIRCULATING EXTRACELLULAR VESICLES

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi46-vi46
Author(s):  
Franz Ricklefs ◽  
Cecile Maire ◽  
Katharina Kolbe ◽  
Mareike Holz ◽  
Rudolph Reimer ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Molecular Subtype ◽  
Cell Types ◽  
Central Nervous System Tumor ◽  
Healthy Controls ◽  
Double Positive ◽  
Tumor Patients ◽  
System Tumor

Abstract BACKGROUND We previously demonstrated that extracellular vesicles (EV) from CNS tumors reflect the molecular subtype of the original tumor and mediate an exchange of pro-oncogenic signals. EVs are commonly characterized by nanoparticle analysis (NTA), electron microscopy and tetraspanin markers, including CD9, CD81 and CD63. It is unclear, however, to what extent circulating tumor EVs are utilizable for diagnostic purposes and how their marker profile overlaps with EVs derived from other cell types. Aiming to define markers that allow distinction and enrichment of glioma EVs from patient blood, we utilized Imaging Flow Cytometry (IFC) to discriminate single EVs via multiple surface markers. METHODS EVs were isolated from blood of patients suffering from glioblastoma (n=24), anaplastic astrocytoma (n=8), brain metastasis (n=7), meningioma (n=12), pituitary gland tumor (n=11), epilepsy (n=11) and from healthy controls (n=18) and were analyzed by IFC, immunoblotting, electron microscopy and NTA. In addition, circulating EVs from PALM-GFP-GL261 and PALM-GFP-CT2A tumor-bearing mice (n=5) as well as from glioblastoma stem-like (GSC) cultures (n=4), neural stem cells (NSC), cerebral endothelial cells (cEC) and T-cells (n=4) were characterized. RESULTS CNS tumor patients have significantly elevated levels of circulating EVs (P < 0.001), as measured by NTA and IFC. In particular, the proportion of double positive CD9+/CD81+, CD9+/CD63+and CD63+/CD81+EVs is increased in glioblastoma patients (p=0.018) compared with healthy controls[L1]. In accordance, cultured GSCs secrete increased levels of CD9+/CD81+EVs in vitro. In the two syngeneic murine PALM-GFP glioma models, only 0.1-0.01% of circulating plasma EVs were found to be derived from intracranial tumors, underlining the need to identify markers that can enrich tumor-specific EVs for molecular profiling. CONCLUSION Glioma patients display increased levels of circulating plasma EVs that can be profiled by IFC, which is a unique and novel technique that facilitates discrimination of different EV subpopulations.

scholarly journals Synergic effects of decellularized bone matrix, hydroxyapatite, and extracellular vesicles on repairing of the rabbit mandibular bone defect model

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Asrin Emami ◽  
Tahereh Talaei-Khozani ◽  
Saeid Tavanafar ◽  
Nehleh Zareifard ◽  
Negar Azarpira ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Bone Repair ◽  
Bone Matrix ◽  
Bone Cell ◽  
Histochemical Staining ◽  
Beneficial Effects ◽  
Mg63 Cells

Abstract Background Extracellular vesicles (ECV) and bone extracellular matrix (ECM) have beneficial effects on the treatment of some pathological conditions. The purpose of this study was to find the synergic effects of decellularized bone (DB) ECM and ECVs on the repair of rabbit. Methods The quality of decellularized sheep bones was confirmed by H&E, Hoechst, DNA quantification, immunohistochemistry, histochemical staining, and scanning electron microscopy (SEM). Osteoblast-derived ECVs were evaluated by internalization test, Transmission electron microscopy, Dynamic light scattering, and flow cytometry for CD9, CD63, CD81 markers. The hydrogel containing DB and hydroxyapatite (HA) with or without ECVs was evaluated for osteoblast functions and bone repair both in vitro and in vivo. Results The data indicated ECM preservation after decellularization as well as cell depletion. In vitro assessments revealed that mineralization and alkaline phosphatase activity did not improve after treatment of MG63 cells by ECVs, while in vivo morphomatrical estimations showed synergic effects of ECVs and DB + HA hydrogels on increasing the number of bone-specific cells and vessel and bone area compared to the control, DB + HA and ECV-treated groups. Conclusions The DB enriched with ECVs can be an ideal scaffold for bone tissue engineering and may provide a suitable niche for bone cell migration and differentiation.

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