scholarly journals ANÁLISE DO EFEITO OVICIDA DO dsRNA QUITINA SINTASE NO MOSQUITO Aedes aegypti (DIPTERA: CULICIDAE)

2021 ◽  
Author(s):  
Brenda Martins Vasconcelos ◽  
Victor Guimarães Ribeiro ◽  
Tiago Souza Salles ◽  
Thayane da Encarnação Sá Guimarães ◽  
Mônica Ferreira Moreira
Keyword(s):  
Escherichia Coli ◽  
Aedes Aegypti ◽  
Gfp Gene

Introdução: O mosquito Aedes aegypti é o principal vetor das arboviroses dengue, chikungunya, Zika e febre amarela. Para que ocorra a produção e a maturação de ovos, as fêmeas do mosquito realizam o repasto sanguíneo, quando ocorre a infecção e transmissão viral. A melhor maneira de evitar a propagação destas doenças é através do controle vetorial. Inseticidas químicos são utilizados no controle, entretanto, ao longo do tempo, os mosquitos adquirem resistência a esses compostos. Neste contexto, o RNA de interferência (RNAi) tem sido uma grande alternativa para o controle. Os genes de quitina sintase (CHSA e CHSB) são considerados excelentes alvos de silenciamento, pois, não são transcritos em vertebrados e plantas. Objetivos: Analisar os efeitos do silenciamento dos genes CHSA e CHSB na oviposição dos mosquitos tratados com dsRNACHS em baixas concentrações. Materiaisemétodos: A produção de dsRNA foi feita in vivo através da transformação de Escherichia coli, cepa HT115 com o plasmídeo L4440, contendo o inserto para a região do sítio catalítico de CHSA e CHSB e GFP (gene não-relacionado para mosquitos), usado como controle. Estas culturas foram lisadas com clorexidina 0,5%. O experimento foi feito em triplicata biológica, utilizando 10 larvas de 4° instar por grupo em 2 mL de água. Foi utilizado, em ambas condições, a concentração 2x10-5 µg/mL de células contendo dsRNACHS ou dsRNAGFP. Os mosquitos adultos sobreviventes foram alimentados com sangue de aves para testar a capacidade de oviposição e a viabilidade dos ovos postos. Resultados: As fêmeas adultas oriundas das larvas tratadas com dsRNACHS tiveram menor postura de ovos, e estes apresentaram fenótipo de ressecamento e deformidades, acarretando baixa eclosão, quando comparados com o controle. Conclusões: Concluímos que o dsRNACHS administrado afeta a produção e a qualidade dos ovos das fêmeas de Ae. aegypti, sendo uma boa estratégia de controle para este inseto.

Безопасность соединений из группы терпенов ментанового ряда in vitro и in vivo

2020 ◽  
Vol 83 (4) ◽  
pp. 24-30
Author(s):  
Ирина Владимировна Акулина ◽  
Светлана Ивановна Павлова ◽  
Ирина Семеновна Степаненко ◽  
Назира Сунагатовна Карамова ◽  
Александр Владиславович Сергеев ◽  
...  
Keyword(s):  
Escherichia Coli

Проведено токсикологическое исследование соединений с антибактериальными свойствами из группы терпенов ментанового ряда в условиях in vitro и in vivo: лимонена (B34), его производного (+)-1,2-оксида лимонена (B60) и серосодержащего монотерпенового соединения 2-(1’-гидрокси-4’-изопренил-1’-метилциклогексил-2’-тио)метилэтаноата (B65). В условиях in vitro (культура опухолевых клеток HeLa) изучаемые монотерпены в диапазоне концентраций 2 – 200 мкг/мл обладали цитотоксичностью. Ингибирующая концентрация (ИК50) для B34 составила 231 (167 – 295) мкг/мл, для B60 – 181 (105 – 257) мкг/мл, ИК50 B65 – 229 (150 – 308) мкг/мл. Исследование генотоксичности показало, что B34 и B65 в диапазоне концентраций 50 – 1000 мкг/мл не индуцируют SOS мутагенез в клетках Escherichia coli PQ37, тогда как B60 в концентрациях 500 и 1000 мкг/мл проявляет генотоксичность. In vivo в остром эксперименте на беспородных мышах установлена низкая токсичность B34 и его производных при различных путях введения. Наименьший показатель острой токсичности имеет B65, в связи с чем дополнительно на крысах проведено изучение его хронической токсичности. Ежедневное внутрижелудочное введение B65 в разовых дозах, составляющих 1/10 и 1/20 ЛД50 (1000 мг/кг и 500 мг/кг), в течение 1 мес не вызывало гибели животных, значимых нарушений общего состояния, изменения динамики массы тела, морфопатологических изменений. Внутрижелудочное введение B65 крысам в высокой токсической дозе 2000 мг/кг (1/5 ЛД50) в течение месяца вызывает патоморфологические изменения структуры печени.

Garlic Extract (Allium sativum L.) Effectively Inhibits Staphylococcus aureus and Escherichia coli by Invitro Test

2020 ◽  
Vol 2 (2) ◽  
pp. 61-68
Author(s):  
Agnina Listya Anggraini ◽  
Ratih Dewi Dwiyanti ◽  
Anny Thuraidah
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Bacterial Infections ◽  
Allium Sativum ◽  
Antibacterial Properties ◽  
Herbal Medicines ◽  
Garlic Extract ◽  
Dilution Method ◽  
Initial Stage

Infection is a disease caused by the presence of pathogenic microbes, including Staphylococcus aureus and Escherichia coli. Garlic (Allium sativum L.) has chemical contents such as allicin, alkaloids, flavonoids, saponins, tannins, and steroids, which can function as an antibacterial against Staphylococcus aureus and Escherichia coli. This study aims to determine the antibacterial properties of garlic extract powder against Staphylococcus aureus and Escherichia coli. This research is the initial stage of the development of herbal medicines to treat Staphylococcus aureus and Escherichia coli infections. The antibacterial activity test was carried out by the liquid dilution method. The concentrations used were 30 mg/mL, 40 mg/mL, 50 mg/mL, 60 mg/mL and 70 mg/mL. The results showed that the Minimum Inhibitory Concentration (MIC) against Staphylococcus aureus and Escherichia coli was 40 mg/mL and 50 mg / mL. Minimum Bactericidal Concentration (MBC) results for Staphylococcus aureus and Escherichia coli are 50 mg/mL and 70 mg/mL. Based on the Simple Linear Regression test, the R2 value of Staphylococcus aureus and Escherichia coli is 0.545 and 0.785, so it can be concluded that there is an effect of garlic extract powder on the growth of Staphylococcus aureus and Escherichia coli by 54.5% and 78.5%. Garlic (Allium sativum L.) extract powder has potential as herbal medicine against bacterial infections but requires further research to determine its effect in vivo.

scholarly journals The Lethal(2)-Essential-for-Life [L(2)EFL] Gene Family Modulates Dengue Virus Infection in Aedes aegypti

2020 ◽  
Vol 21 (20) ◽  
pp. 7520
Author(s):  
Lucky R. Runtuwene ◽  
Shuichi Kawashima ◽  
Victor D. Pijoh ◽  
Josef S. B. Tuda ◽  
Kyoko Hayashida ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Cell Line ◽  
Dengue Infection ◽  
Fruit Fly ◽  
Initiation Factor ◽  
Poly I:C ◽  
Gene Products ◽  
Vector Mosquito

Efforts to determine the mosquito genes that affect dengue virus replication have identified a number of candidates that positively or negatively modify amplification in the invertebrate host. We used deep sequencing to compare the differential transcript abundances in Aedes aegypti 14 days post dengue infection to those of uninfected A. aegypti. The gene lethal(2)-essential-for-life [l(2)efl], which encodes a member of the heat shock 20 protein (HSP20) family, was upregulated following dengue virus type 2 (DENV-2) infection in vivo. The transcripts of this gene did not exhibit differential accumulation in mosquitoes exposed to insecticides or pollutants. The induction and overexpression of l(2)efl gene products using poly(I:C) resulted in decreased DENV-2 replication in the cell line. In contrast, the RNAi-mediated suppression of l(2)efl gene products resulted in enhanced DENV-2 replication, but this enhancement occurred only if multiple l(2)efl genes were suppressed. l(2)efl homologs induce the phosphorylation of eukaryotic initiation factor 2α (eIF2α) in the fruit fly Drosophila melanogaster, and we confirmed this finding in the cell line. However, the mechanism by which l(2)efl phosphorylates eIF2α remains unclear. We conclude that l(2)efl encodes a potential anti-dengue protein in the vector mosquito.

scholarly journals Characterization and in Vivo Cloning of prlC, a Suppressor of Signal Sequence Mutations in Escherichia coli K12

Genetics ◽  
1987 ◽  
Vol 116 (4) ◽  
pp. 513-521
Author(s):  
Nancy J Trun ◽  
Thomas J Silhavy
Keyword(s):  
Escherichia Coli ◽  
Genetic Mapping ◽  
Signal Sequence ◽  
Illegitimate Recombination ◽  
Mutant Phenotype ◽  
E Coli ◽  
Physical Location ◽  
Sequence Deletion ◽  
New Alleles

ABSTRACT The prlC gene of E. coli was originally identified as an allele, prlC1, which suppresses certain signal sequence mutations in the genes for several exported proteins. We have isolated six new alleles of prlC that also confer this phenotype. These mutations can be placed into three classes based on the degree to which they suppress the lamBsignal sequence deletion, lamBs78. Genetic mapping reveals that the physical location of the mutations in prlC correlates with the strength of the suppression, suggesting that different regions of the gene can be altered to yield a suppressor phenotype. We also describe an in vivo cloning procedure using λplacMu9H. The procedure relies on transposition and illegitimate recombination to generate a specialized transducing phage that carries prlC1. This method should be applicable to any gene for which there is a mutant phenotype.

scholarly journals An infection-induced RhoB-Beclin 1-Hsp90 complex enhances clearance of uropathogenic Escherichia coli

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chunhui Miao ◽  
Mingyu Yu ◽  
Geng Pei ◽  
Zhenyi Ma ◽  
Lisong Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Treatment Strategies ◽  
Uropathogenic Escherichia Coli ◽  
Beclin 1 ◽  
Host Cells ◽  
Infection Model ◽  
Intracellular Bacteria ◽  
Bladder Epithelium ◽  
Binding Residue

AbstractHost cells use several anti-bacterial pathways to defend against pathogens. Here, using a uropathogenic Escherichia coli (UPEC) infection model, we demonstrate that bacterial infection upregulates RhoB, which subsequently promotes intracellular bacteria clearance by inducing LC3 lipidation and autophagosome formation. RhoB binds with Beclin 1 through its residues at 118 to 140 and the Beclin 1 CCD domain, with RhoB Arg133 being the key binding residue. Binding of RhoB to Beclin 1 enhances the Hsp90-Beclin 1 interaction, preventing Beclin 1 degradation. RhoB also directly interacts with Hsp90, maintaining RhoB levels. UPEC infections increase RhoB, Beclin 1 and LC3 levels in bladder epithelium in vivo, whereas Beclin 1 and LC3 levels as well as UPEC clearance are substantially reduced in RhoB+/− and RhoB−/− mice upon infection. We conclude that when stimulated by UPEC infections, host cells promote UPEC clearance through the RhoB-Beclin 1-HSP90 complex, indicating RhoB may be a useful target when developing UPEC treatment strategies.

scholarly journals Thermostability Engineering of a Class II Pyruvate Aldolase from Escherichia coli by in Vivo Folding Interference

2021 ◽  
Vol 9 (15) ◽  
pp. 5430-5436
Author(s):  
Sandra Bosch ◽  
Esther Sanchez-Freire ◽  
María Luisa del Pozo ◽  
Morana C̆esnik ◽  
Jaime Quesada ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Class Ii
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