The effect of albendazole sulfoxide on the expression of miR-61 and let-7 in different in vitro developmental stages of Echinococcus granulosus

Background:: The dog tapeworm, Echinococcus granulosus, is a zoonotic parasite affecting human and livestock across the globe. Basic research on the molecular biology and genetics of E. granulosus improves our understanding of the biology and potential drug targets in various developmental stages of E. granulosus in both definitive and intermediate hosts. There has been increasing interest in identification of microRNAs in parasitic organisms. The purpose of the current study was to compare the activity of a selected profile of miRNAs in different developmental stages of E. granulosus. Methods:: Different developmental stages of the parasite were obtained from ex vivo as well as in vitro cultured E. granu-losus. MicroRNAs were extracted from the ex vivo germinal layer and invaginated protoscoleces as well as the in vitro gen-erated microcysts, evaginated protoscoleces and strobilated worms. Expression of the selected miRNAs was evaluated by RT-qPCR for each stage. Results:: Four out of five miRNAs were present and active in different developmental stages of E. granulosus. A significant over-expression of miR-61 was observed in germinal layer and during the protoscolex transformation into the microcysts, however miR-10 was more expressed in the mature strobilated forms than the other stages. Let-7 and miR-3489 showed a high expression in germinal layer. Conclusion:: Differential expression of four miRNAs among different in vitro and ex vivo developmental stages of E. granu-losus was documented in the present study. Further experimental investigations are required to elucidate the probable role of the miRNAs in bi-directional differentiation of protoscoleces either into the strobilated worm or to a secondary hydatid cyst and the potential of these miRNAs as drug targets.

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Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010005161x ◽

1974 ◽

Vol 32 ◽

pp. 320-321

Author(s):

J. P. Revel

Keyword(s):

Developmental Stages ◽

Three Dimensional ◽

Cell Movement ◽

Cellular Interactions ◽

Serial Sections ◽

Cell Sheets ◽

Freeze Etching ◽

Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

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In vitro immunoregulatory activity and anti-inflammatory effect of Echinococcus granulosus laminated layer

Acta Tropica ◽

10.1016/j.actatropica.2021.105886 ◽

2021 ◽

Vol 218 ◽

pp. 105886

Author(s):

Sara Benazzouz ◽

Manel Amri ◽

Junhua Wang ◽

Samia Bouaziz ◽

Fahima Ameur ◽

...

Keyword(s):

Echinococcus Granulosus ◽

Anti Inflammatory ◽

Laminated Layer ◽

Inflammatory Effect

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Genome-Wide Transcriptomic Identification and Functional Insight of Lily WRKY Genes Responding to Botrytis Fungal Disease

Plants ◽

10.3390/plants10040776 ◽

2021 ◽

Vol 10 (4) ◽

pp. 776

Author(s):

Shipra Kumari ◽

Bashistha Kumar Kanth ◽

Ju young Ahn ◽

Jong Hwa Kim ◽

Geung-Joo Lee

Keyword(s):

Abiotic Stress ◽

Biotic Stress ◽

Developmental Stages ◽

Expression Patterns ◽

Lilium Longiflorum ◽

Biotic And Abiotic Stress ◽

Biotic Stresses ◽

Genome Wide

Genome-wide transcriptome analysis using RNA-Seq of Lilium longiflorum revealed valuable genes responding to biotic stresses. WRKY transcription factors are regulatory proteins playing essential roles in defense processes under environmental stresses, causing considerable losses in flower quality and production. Thirty-eight WRKY genes were identified from the transcriptomic profile from lily genotypes, exhibiting leaf blight caused by Botrytis elliptica. Lily WRKYs have a highly conserved motif, WRKYGQK, with a common variant, WRKYGKK. Phylogeny of LlWRKYs with homologous genes from other representative plant species classified them into three groups- I, II, and III consisting of seven, 22, and nine genes, respectively. Base on functional annotation, 22 LlWRKY genes were associated with biotic stress, nine with abiotic stress, and seven with others. Sixteen unique LlWRKY were studied to investigate responses to stress conditions using gene expression under biotic and abiotic stress treatments. Five genes—LlWRKY3, LlWRKY4, LlWRKY5, LlWRKY10, and LlWRKY12—were substantially upregulated, proving to be biotic stress-responsive genes in vivo and in vitro conditions. Moreover, the expression patterns of LlWRKY genes varied in response to drought, heat, cold, and different developmental stages or tissues. Overall, our study provides structural and molecular insights into LlWRKY genes for use in the genetic engineering in Lilium against Botrytis disease.

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Zika Virus Infection Leads to Demyelination and Axonal Injury in Mature CNS Cultures

Viruses ◽

10.3390/v13010091 ◽

2021 ◽

Vol 13 (1) ◽

pp. 91

Author(s):

Verena Schultz ◽

Stephanie L. Cumberworth ◽

Quan Gu ◽

Natasha Johnson ◽

Claire L. Donald ◽

...

Keyword(s):

Nervous System ◽

Neural Development ◽

Zika Virus ◽

Developmental Stages ◽

Cell Types ◽

Neural Cells ◽

Zikv Infection ◽

Axonal Pathology ◽

Time Frames

Understanding how Zika virus (Flaviviridae; ZIKV) affects neural cells is paramount in comprehending pathologies associated with infection. Whilst the effects of ZIKV in neural development are well documented, impact on the adult nervous system remains obscure. Here, we investigated the effects of ZIKV infection in established mature myelinated central nervous system (CNS) cultures. Infection incurred damage to myelinated fibers, with ZIKV-positive cells appearing when myelin damage was first detected as well as axonal pathology, suggesting the latter was a consequence of oligodendroglia infection. Transcriptome analysis revealed host factors that were upregulated during ZIKV infection. One such factor, CCL5, was validated in vitro as inhibiting myelination. Transferred UV-inactivated media from infected cultures did not damage myelin and axons, suggesting that viral replication is necessary to induce the observed effects. These data show that ZIKV infection affects CNS cells even after myelination—which is critical for saltatory conduction and neuronal function—has taken place. Understanding the targets of this virus across developmental stages including the mature CNS, and the subsequent effects of infection of cell types, is necessary to understand effective time frames for therapeutic intervention.

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In vitro scolicidal effects of Sideritis perfoliata extract against Echinococcus granulosus

International Journal of Clinical Practice ◽

10.1111/ijcp.14498 ◽

2021 ◽

Author(s):

Tuncay Çelik ◽

Muhittin Önderci ◽

Mustafa Pehlivan ◽

Önder Yumrutaş ◽

Fatih Üçkardeş

Keyword(s):

Echinococcus Granulosus

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Design, Synthesis and Antiparasitic Evaluation of Click Phospholipids

Molecules ◽

10.3390/molecules26144204 ◽

2021 ◽

Vol 26 (14) ◽

pp. 4204

Author(s):

George E. Magoulas ◽

Pantelis Afroudakis ◽

Kalliopi Georgikopoulou ◽

Marina Roussaki ◽

Chiara Borsari ◽

...

Keyword(s):

Leishmania Donovani ◽

Developmental Stages ◽

Neglected Tropical Diseases ◽

Heterocyclic Ring ◽

Head Group ◽

Ether Phospholipid ◽

Design Synthesis ◽

Antiparasitic Activity ◽

Intracellular Amastigotes

A library of seventeen novel ether phospholipid analogues, containing 5-membered heterocyclic rings (1,2,3-triazolyl, isoxazolyl, 1,3,4-oxadiazolyl and 1,2,4-oxadiazolyl) in the lipid portion were designed and synthesized aiming to identify optimised miltefosine analogues. The compounds were evaluated for their in vitro antiparasitic activity against Leishmania infantum and Leishmania donovani intracellular amastigotes, against Trypanosoma brucei brucei and against different developmental stages of Trypanosoma cruzi. The nature of the substituents of the heterocyclic ring (tail) and the oligomethylene spacer between the head group and the heterocyclic ring was found to affect the activity and toxicity of these compounds leading to a significantly improved understanding of their structure–activity relationships. The early ADMET profile of the new derivatives did not reveal major liabilities for the potent compounds. The 1,2,3-triazole derivative 27 substituted by a decyl tail, an undecyl spacer and a choline head group exhibited broad spectrum antiparasitic activity. It possessed low micromolar activity against the intracellular amastigotes of two L. infantum strains and T. cruzi Y strain epimastigotes, intracellular amastigotes and trypomastigotes, while its cytotoxicity concentration (CC50) against THP-1 macrophages ranged between 50 and 100 μM. Altogether, our work paves the way for the development of improved ether phospholipid derivatives to control neglected tropical diseases.

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Development of an embryo germination protocol for shy-seeded grape (Vitis vinifera L.)

Plant Genetic Resources ◽

10.1017/s1479262121000307 ◽

2021 ◽

pp. 1-9

Author(s):

Ashwini P. Benke ◽

Ram Krishna ◽

Roshni R. Samarth ◽

Shweta S. Dhumal ◽

Waquar A. Ansari ◽

...

Keyword(s):

Developmental Stages ◽

Germination Rate ◽

Germination Percentage ◽

Vitis Vinifera L ◽

Growth Media ◽

Embryo Germination ◽

Indole Butyric Acid ◽

Grape Berries ◽

Benzyl Amino Purine

Abstract Acquisition and germination of seeds are the most desired targets for the improvement of vegetatively propagated crops. In the present study, we developed a potential embryo germination protocol for the Red Globe grape cultivar having a low seed germination rate. Three grape berries at different developmental stages, viz. 50, 60 and 70 days after flowering (DAF), were selected for in-vitro embryo germination. Three growth media, namely Emershad and Ramming (ER), Nitsch and Nitsch (NN) and Murashige and Skoog (MS), and plant growth regulators (benzyl amino purine (BA), 0.5, 0.7 and 0.9 mg/l; indole butyric acid (IBA), 1.0, 1.5 and 2.0 mg/l; and gibberellic acid (GA), 0.1, 0.3 and 0.9 mg/l) were screened individually in different combinations with three amino acids, namely cysteine, glutamine and proline (2.0 μmol/l each). The maximum embryos germination percentage recorded at 70 DAF was 63.33, 47.78 and 45.56% in ER, NN and MS media, respectively, supplemented with 0.9 mg/l BA, 2.0 mg/l IBA, 0.9 mg/l GA and 2.0 μmol glutamine. Glutamine was found to have the most significant impact, and it improved the rescued embryos germination. The present study provides a potential recipe for a medium that can facilitate efficient germination of grape embryos.

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Characterization of isolated bovine preantral follicles based on morphology, diameter and cell number

Zygote ◽

10.1017/s0967199419000832 ◽

2020 ◽

Vol 28 (2) ◽

pp. 154-159

Author(s):

Juliana I. Candelaria ◽

Anna C. Denicol

Keyword(s):

Granulosa Cells ◽

Developmental Stages ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Cell Number ◽

Culture Conditions ◽

Preantral Follicles ◽

Secondary Stage

SummaryPreantral follicles are a potential reservoir of oocytes to be used in assisted reproductive technologies. With the increasing interest in developing techniques to grow preantral follicles in vitro, and as the bovine emerges as an appropriate model species to understand human folliculogenesis, the establishment of an accurate classification of developmental stages is needed. Classification of bovine preantral follicles has been mostly based on histological analysis and estimation models, which may not translate well to correctly characterize preantral follicles isolated from the ovary. In this study, we classified bovine preantral follicles by morphology upon isolation, determined diameter and number of granulosa cells by direct counting, and compared our results with previous studies reporting bovine preantral follicle classification. Follicles were isolated via homogenization of ovary tissue and classified into primary, early secondary and secondary stage based on morphology and number of layers of granulosa cells. Diameter was individually measured and Hoechst 33342 was used as a nuclear stain to count granulosa cells. We found that follicles classified by morphology into primary, early secondary, and secondary had different mean diameter and cell number (P < 0.01); cell number and diameter were positively correlated, as were cell density and cell number in each developmental stage (P < 0.01). Results obtained here were mostly in agreement with previous classifications based on histological sections and on isolated follicles, with some discrepancies. The present data add accuracy to classification of bovine preantral follicles that is critical to optimize culture conditions to produce developmentally competent oocytes.

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In Vitro Susceptibilities of the MicrosporidiaEncephalitozoon cuniculi, Encephalitozoon hellem, and Encephalitozoon intestinalis to Albendazole and Its Sulfoxide and Sulfone Metabolites

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.42.12.3301 ◽

1998 ◽

Vol 42 (12) ◽

pp. 3301-3303 ◽

Cited By ~ 15

Author(s):

Olivier Ridoux ◽

Michel Drancourt

Keyword(s):

Encephalitozoon Cuniculi ◽

Albendazole Sulfoxide ◽

Encephalitozoon Intestinalis ◽

Degree Of Oxidation ◽

Albendazole Sulfone

ABSTRACT In vitro comparisons demonstrated that the efficacy of albendazole, albendazole-sulfoxide, and albendazole-sulfone against pathogenicEncephalitozoon species was proportional to the degree of oxidation at a concentration of >10−3 μg/ml. Furthermore, at a concentration of <10−2 μg/ml, benzimidazoles were more effective against Encephalitozoon cuniculi and Encephalitozoon hellem than againstEncephalitozoon intestinalis.

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