Bioinformatic Comparison of Kunitz Protease Inhibitors in Echinococcus Granulosus and E. Multilocularis and the Genes Expressed in Different Developmental Stages of E. Granulosus

2021 ◽  
Author(s):  
Hui Zhang ◽  
Mengxiao Tian ◽  
Wenjing Qi ◽  
Juan Wu ◽  
Huajun Zheng ◽  
...  
Keyword(s):  
Protease Inhibitors ◽  
Echinococcus Granulosus ◽  
Alveolar Echinococcosis ◽  
Developmental Stages ◽  
Zoonotic Diseases ◽  
Neighbor Joining ◽  
Transcriptome Data ◽  
Kunitz Domain ◽  
Kunitz Type ◽  
Echinococcus Spp

Abstract Background: Cystic and alveolar echinococcosis caused by the tapeworms Echinococcus granulosus and E. multilocularis, respectively, are important zoonotic diseases. Protease inhibitors are crucial for the survival of both Echinococcus spp. Kunitz-type inhibitors play a regulatory role in the control of protease activity. In this study，we identified all the Kunitz-type protease inhibitors present in the genomes of these two tapeworms and analyzed the gene sequences using computational, structural bioinformatics and phylogenetic approaches to evaluate the evolutionary relationships of these genes. Results: A total of 19 genes from E. multilocularis and 23 genes from E. granulosus contained single or multiple Kunitz-domains. A neighbor-joining phylogenetic tree indicated that the E. granulosus and E. multilocularis Kunitz domain peptides were divided into three branches containing 9 clusters. Based on available transcriptome data, we analyzed the expression of these Kunitz-domain protease inhibitors in four major developmental stages of E. granulosus and found they were differentially expressed.Conclusion: We identified 19 and 23 Kunitz protease inhibitors in E. multilocularis and E. granulosus respectively; the majority of these genes were expressed in one or four stages of E. granulosus with some being highly expressed in adult worms indicating that these genes likely play different roles in the different developmental stages.

scholarly journals Bioinformatic comparison of Kunitz protease inhibitors in Echinococcus granulosus sensu stricto and E. multilocularis and the genes expressed in different developmental stages of E. granulosus s.s.

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Hui Zhang ◽  
Mengxiao Tian ◽  
Wenjing Qi ◽  
Juan Wu ◽  
Huajun Zheng ◽  
...  
Keyword(s):  
Adult Worm ◽  
Protease Inhibitors ◽  
Echinococcus Granulosus ◽  
Developmental Stages ◽  
Sensu Stricto ◽  
Likelihood Method ◽  
Homologous Gene ◽  
Transcription Analysis ◽  
Kunitz Domain ◽  
Kunitz Type

Abstract Background Cystic and alveolar echinococcosis caused by the tapeworms Echinococcus granulosus sensu stricto (s.s.) and E. multilocularis, respectively, are important zoonotic diseases. Protease inhibitors are crucial for the survival of both Echinococcus spp. Kunitz-type inhibitors play a regulatory role in the control of protease activity. In this study,we identified Kunitz-type domain protease inhibitors(KDPIs) present in the genomes of these two tapeworms and analyzed the gene sequences using computational, structural bioinformatics and phylogenetic approaches to evaluate the evolutionary relationships of these genes. Hi-seq transcriptome analysis showed that E. granulosuss.s. KDPIs were differentially expressed in the different developmental stages. We validated some of the genes expressed in adult worm, protoscolex and cyst germinal membrane of E. granulosuss.s. and E. multilocularis by quantitative PCR. Results A total of 19 genes from E. multilocularis and 23 genes from E. granulosuss.s. were predicted to be KDPIs with the most containing a single Kunitz-domain. A maximum likelihood method phylogenetic tree indicated that the E. granulosuss.s. and E. multilocularis Kunitz domain peptides were divided into three branches containing 9 clusters. The ratio of positively charged residues and neutral residues are different between E. multilocularis and E. granulosuss.s. KDPIs. We also found that E. multilocularis had higher percentage of sequences containing signal peptides (17/19, 89.47%) than that of E. granulosuss.s. (14/23, 60.87%). Transcript analysis showed all the E. granulosuss.s. KDPI genes were expressed differentially in four developmental stages of the worm. Transcription analysis showed that 9 KDPIs (including EG_07244,EGR_08716 and EGR_10096) were highly upregulated in adult worm, and 2 KDPIs (EG_09268 and EG_09490) were highly expressed in the cyst germinal membrane. Quantitative gene expression analysis(qPCR) of four genes confirmed the expression of these genes. EGR_08716 and its homologous gene (EmuJ_001137000) were highly and specifically expressed in adult worms of the two worms. Conclusions A total 19 and 23 KDPIs were identified in the genomes of E. multilocularis and E. granulosus s.s. , respectively. The differential expression of these KDPIs in different stages may indicate their different roles in the different hosts. The difference in characterization of KDPIs may be associated with the different pathology of metacestode stage of these two parasites.

scholarly journals Cloning and Characterization of Two Potent Kunitz Type Protease Inhibitors from Echinococcus granulosus

2015 ◽  
Vol 9 (12) ◽  
pp. e0004268 ◽  
Author(s):  
Shiwanthi L. Ranasinghe ◽  
Katja Fischer ◽  
Wenbao Zhang ◽  
Geoffrey N. Gobert ◽  
Donald P. McManus
Keyword(s):  
Protease Inhibitors ◽  
Echinococcus Granulosus ◽  
Kunitz Type

scholarly journals Echinococcus spp.: Tapeworms that Pose a Danger to Both Animals and Humans – a Review

2017 ◽  
Vol 48 (4) ◽  
pp. 193-201 ◽  
Author(s):  
A. Brožová ◽  
I. Jankovská ◽  
V. Bejček ◽  
S. Nechybová ◽  
P. Peřinková ◽  
...  
Keyword(s):  
Small Intestine ◽  
Life Cycle ◽  
Echinococcus Multilocularis ◽  
Alveolar Echinococcosis ◽  
Sensu Stricto ◽  
Mammal Species ◽  
Current State ◽  
Wide Range ◽  
The World ◽  
Echinococcus Spp

Abstract Species of the genus Echinococcus (Cestoda; Taeniidae) are minute tapeworms of carnivores. Their larvae are known as hydatids (metacestode), which proliferate asexually in various mammals. Like the majority of cestodes, Echinococcus spp. require two different host species to complete their life cycle. Definitive hosts harbouring the adult cestodes in the small intestine are exclusively carnivores of the Canidae and Felidae families. A wide range of mammal species including humans is susceptible to infection by the metacestode of Echinococcus spp., which develops in their viscera. The disease, caused by species of the genus Echinococcus, is called echinococcosis, and it is one of the most dangerous zoonoses in the world. The traditional species Echinococcus granulosus and Echinococcus multilocularis are agents of significant diseases due to the high number of cases and the wide geographical species range. The taxonomy of the genus is controversial; in the current state of ongoing complex revisions, the agent of cystic echinococcosis E. granulosus sensu lato is divided into five species (E. granulosus sensu stricto, E. felidis, E. equinus, E. ortleppi, E. canadensis), in addition to the agents of alveolar echinococcosis (E. multilocularis, E. shiquicus) and polycystic/unicystic echinococcosis (E. vogeli, E. oligarthrus). Here we provide an overview of the current situation, which continues to develop.

scholarly journals Cystic Echinococcosis: Clinical, Immunological, and Biomolecular Evaluation of Patients from Sardinia (Italy)

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 907
Author(s):  
Cinzia Santucciu ◽  
Piero Bonelli ◽  
Angela Peruzzu ◽  
Alessandro Fancellu ◽  
Vincenzo Marras ◽  
...  
Keyword(s):  
Molecular Biology ◽  
Echinococcus Granulosus ◽  
Cystic Echinococcosis ◽  
Public Health Problem ◽  
Sensu Stricto ◽  
Surgical Removal ◽  
Neighbor Joining ◽  
Intermediate Hosts ◽  
High Prevalence ◽  
Echinococcus Granulosus Sensu Lato

Cystic echinococcosis (CE), a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus sensu lato (s.l.), is a worldwide public health problem. Echinococcus granulosus sensu stricto (s.s.), associated with G1 and G3 genotypes, is endemic with high prevalence in the Mediterranean basin. The parasite’s life cycle comprises definitive hosts (canids) and intermediate hosts (ruminants) and can occasionally involve humans. The main aim of this research was to confirm the diagnosis of 13 patients suspected of CE who presented different complications and needed the surgical removal of the cysts. We also wanted to understand and clarify more the diagnosis of echinococcosis in humans. For this purpose, the patients first underwent cyst evaluation by ultrasound (US), immunological analysis, and then total pericystectomy, followed by parasitological, histopathological, and molecular biology examinations of the cysts. US stadiated one CE1, one CE2, eight CE3b, one CE4, and two CE5; immunology evidenced nine positives; histopathology confirmed 11 CE cysts, of which 8 fertile presenting protoscoleces were identified as E. granulosus s.s. by molecular biology, genotyped as three G1 and four G3 by neighbor-joining (NJ) phylogenetic tree. In conclusion, the results showed that 11 patients were affected by E. granulosus s.s. G1 orG3, and 2 cystic neoformations were of non-parasitic origin.

scholarly journals The complex world of plant protease inhibitors: Insights into a Kunitz-type cysteine protease inhibitor of Arabidopsis thaliana

2017 ◽  
Vol 11 (1) ◽  
pp. e1368599 ◽  
Author(s):  
Sachin Rustgi ◽  
Edouard Boex-Fontvieille ◽  
Christiane Reinbothe ◽  
Diter von Wettstein ◽  
Steffen Reinbothe
Keyword(s):  
Arabidopsis Thaliana ◽  
Protease Inhibitor ◽  
Protease Inhibitors ◽  
Cysteine Protease ◽  
Cysteine Protease Inhibitor ◽  
Complex World ◽  
Plant Protease ◽  
Kunitz Type

Evaluation of reactivity to Echinococcus spp. among rural inhabitants in Poland

2015 ◽  
Vol 60 (3) ◽  
Author(s):  
Ewa Cisak ◽  
Jacek Sroka ◽  
Angelina Wójcik-Fatla ◽  
Violetta Zając ◽  
Jacek Dutkiewicz
Keyword(s):  
Western Blot ◽  
Echinococcus Granulosus ◽  
Cross Reactivity ◽  
Farm Animals ◽  
Control Group ◽  
Laboratory Diagnostics ◽  
Group 14 ◽  
Serum Samples ◽  
The City ◽  
Echinococcus Spp

AbstractA group of 172 rural inhabitants from eastern Poland (68 males and 104 females, mean age 49.0 ± 12.0 years) was examined for the presence of antibodies against Echinococcus granulosus and Echinococcus multilocularis. A population of 38 healthy urban dwellers from the city of Lublin (17 males and 21 females, mean age 36.2 ± 9.6 years) were examined as a control group. Sera of 22 rural inhabitants (12.8%) reacted positively to Echinococcus granulosus hydatid fluid antigen in the screening test. A cross-reactivity was observed with two serum samples that tested positive in ELISA for E. granulosus. Three serum samples were tested positive for E. multilocularis using the Em2plus ELISA assay and also positive for Western blot. None of the members of control group showed the presence of a seropositive reaction to Echinococcus spp. The reactivity to Echinococcus spp. among rural inhabitants decreased with age and this correlation was statistically significant (R = -0.197151, p = 0.009535). The percentage of positive findings was the highest (50.0%) in the youngest age group (14-20). No significant correlations were found between responses to interview questions (possession of domestic and farm animals, contact with wild animals, eating unwashed berries, drinking unboiled water) and the presence of seropositive reactions to Echinococcus spp. The presented results seem to indicate that echinococcosis is still a current problem in Poland that should not be neglected and, moreover, indicates the need for improvement in the routine laboratory diagnostics of Echinococcus spp. by standardizing the ELISA and Western blot tests.

scholarly journals Transcriptome Analysis Identities Key Candidate Genes Mediating Purple Ovary Coloration in Asiatic Hybrid Lilies

2016 ◽  
Author(s):  
Leifeng Xu ◽  
Panpan Yang ◽  
Suxia Yuan ◽  
Yayan Feng ◽  
Hua Xu ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Developmental Stages ◽  
Anthocyanin Biosynthesis ◽  
Differential Expression Analysis ◽  
Transcriptome Data ◽  
Comparative Transcriptome ◽  
Myb Gene ◽  
Short Lifespan ◽  
H Gene ◽  
Pigment Accumulation

Lily tepals have a short lifespan. Once the tepals senesce, the ornamental value of the flower is lost. Some cultivars have attractive purple ovaries and fruits which greatly enhance the ornamental value of Asiatic hybrid lilies. However, little is known about the molecular mechanisms of anthocyanin biosynthesis in Asiatic hybrid lily ovaries. To investigate the transcriptional network that governs purple ovary coloration in Asiatic hybrid lilies, we obtained transcriptome data from green ovaries (S1) and purple ovaries (S2) of Asiatic ‘Tiny Padhye’. Comparative transcriptome analysis revealed 4228 differentially expressed genes. Differential expression analysis revealed that nine unigenes including four CHS genes, one CHI gene, one F3H gene, one F3′H gene, one DFR gene, and one UFGT gene were significantly up-regulated in purple ovaries. One MYB gene, LhMYB12-Lat, was identified as a key transcription factor determining the distribution of anthocyanins in Asiatic hybrid lily ovaries. Further qPCR results showed unigenes related to anthocyanin biosynthesis were highly expressed in purple ovaries of Asiatic ‘Tiny Padhye’ at stages 2 and 3, while they showed an extremely low level of expression in ovaries of Asiatic ‘Yellow Pixels’ during all developmental stages. In addition, shading treatment significantly decreased pigment accumulation by suppressing the expression of several unigenes related to anthocyanin biosynthesis in ovaries of Asiatic ‘Tiny Padhye’. These results could further our understanding of the molecular mechanisms of anthocyanin biosynthesis in Asiatic hybrid lily ovaries.

scholarly journals Kunitz type protease inhibitor EgKI-1 from the canine tapeworm Echinococcus granulosus as a promising anti-cancer therapeutic

2018 ◽  
Author(s):  
Shiwanthi L Ranasinghe ◽  
Glen M Boyle ◽  
Katja Fischer ◽  
Jeremy Potriquet ◽  
Jason P Mulvenna ◽  
...  
Keyword(s):  
Cell Growth ◽  
Protease Inhibitor ◽  
Hydatid Cyst ◽  
Echinococcus Granulosus ◽  
Cancer Cell ◽  
Dependent Manner ◽  
Anti Cancer ◽  
Kunitz Type ◽  
Kunitz Type Protease Inhibitor

AbstractEgKI-1, a member of the Kunitz type protease inhibitor family, is highly expressed by the oncosphere of the canine tapeworm Echinococcus granulosus, the stage that is infectious to humans and ungulates, giving rise to a hydatid cyst localized to the liver and other organs. Larval protoscoleces, which develop within the hydatid cyst, have been shown to possess anti-cancer properties, although the precise molecules involved have not been identified. We show that recombinant EgKI-1 inhibits the growth and migration of a range of human cancers including breast, melanoma and cervical cancer cell lines in a dose-dependent manner in vitro without affecting normal cell growth. Furthermore, EgKI-1 treatment arrested the cancer cell growth by disrupting the cell cycle and induced apoptosis of cancer cells in vitro. An in vivo model of triple negative breast cancer (MDA-MB-231) in BALB/c nude mice showed significant tumor growth reduction in EgKI-1-treated mice compared with controls. These findings indicate that EgKI-1 shows promise for future development as an anti-cancer therapeutic.

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