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2022 ◽  
Vol 15 (1) ◽  
Author(s):  
Rui-xin Shen ◽  
Yi-ting Wang ◽  
Jia-hong Wu ◽  
Ning Zhang ◽  
Heng-duan Zhang ◽  
...  

Abstract Background Odorant-binding proteins (OBPs) play important roles in many physiological processes of mosquitoes. Previous high-throughput sequencing studies have revealed that some OBPs of Culex quinquefasciatus might be involved in the development of resistance to insecticides. Methods Based on the results of sequencing analyses, the OBP28 gene was selected for evaluation in this study. Three laboratory strains of Cx. quinquefasciatus [susceptible strain (SS), deltamethrin-resistant strain 1 (HN) and deltamethrin-resistant strain 2 (RR)] were first examined by using the Centers for Disease Control and Prevention bottle bioassay, after which the expression level of the OBP28 gene in the susceptible and deltamethrin-resistant strains was determined by real-time quantitative polymerase chain reaction. The OBP28 gene in deltamethrin-resistant strain RR was silenced using RNA interference technology. The expression level of OBP28 and the resistance level were tested in the silenced strain and control strain after microinjection of double-stranded RNA for a 48-h interference period. Four field-collected strains (henceforth ‘field strains’) of Cx. quinquefasciatus were also examined for their resistance to deltamethrin and levels of OBP28 expression. Finally, a correlation analysis between deltamethrin resistance and gene expression was carried out for all seven strains, i.e. the four field strains and the three laboratory strains. Results In the bioassay, the mortality of SS, HN and RR was 100%, 21.33% and 1.67%, respectively. The relative expression levels of OBP28 in strains HN and RR were 6.30- and 6.86-fold higher, respectively, than that of strain SS. After silencing of the OBP28 gene, the mortality of strain RR was 72.20% and that of the control strain 26.32%. The mortality of strain RR increased significantly after interference compared to that of the control strain. There was a negative correlation between OBP28 gene expression and mortality in adult mosquitoes after exposure to deltamethrin. Conclusions To our knowledge, this study shows for the first time a correlation between the expression of a gene coding for OBP and insecticide resistance in mosquitoes. The potential resistance mechanism that was elucidated provides a new target gene for the surveillance of resistance in mosquitoes. Graphical abstract


2021 ◽  
Vol 15 ◽  
Author(s):  
Sarah Kim ◽  
Stephanie A. Gacek ◽  
Madaline M. Mocchi ◽  
Eva E. Redei

Genetic predisposition and environmental stress are known etiologies of stress-related psychiatric disorders. Environmental stress during adolescence is assumed to be particularly detrimental for adult affective behaviors. To investigate how genetic stress-reactivity differences modify the effects of stress during adolescence on adult affective behaviors we employed two inbred strains with differing stress reactivity. The Wistar Kyoto More Immobile (WMI) rat strain show increased stress-reactivity and despair-like behaviors as well as passive coping compared to the nearly isogenic control strain, the Wistar Kyoto Less Immobile (WLI). Males and females of these strains were exposed to contextual fear conditioning (CFC) during early adolescence (EA), between 32 and 34 postnatal days (PND), and were tested for the consequences of this mild EA stress in adulthood. Early adolescent stress significantly decreased anxiety-like behavior, measured in the open field test (OFT) and increased social interaction and recognition in adult males of both strains compared to controls. In contrast, no significant effects of EA stress were observed in adult females in these behaviors. Both males and females of the genetically less stress-reactive WLI strain showed significantly increased immobility in the forced swim test (FST) after EA stress compared to controls. In contrast, immobility was significantly attenuated by EA stress in adult WMI females compared to controls. Transcriptomic changes of the glucocorticoid receptor (Nr3c1, GR) and the brain-derived neurotrophic factor (Bdnf) illuminate primarily strain and stress-dependent changes, respectively, in the prefrontal cortex and hippocampus of adults. These results suggest that contrary to expectations, limited adolescent stress is beneficial to males thru decreasing anxiety and enhancing social behaviors, and to the stress more-reactive WMI females by way of decreasing passive coping.


AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Takashi Koyanagi ◽  
Ayumi Hara ◽  
Kanako Kobayashi ◽  
Yuji Habara ◽  
Akira Nakagawa ◽  
...  

AbstractPeptidyl-prolyl cis-trans isomerase (PPIase, EC 5.2.1.8) catalyzes the racemization reaction of proline residues on a polypeptide chain. This enzyme is also known to function as a molecular chaperon to stabilize protein conformation during the folding process. In this study, we noted FK506 binding protein (FKBP)-type PPIase from a hyperthemophilic archaeon Thermococcus sp. strain KS-1 (PPIase KS−1) to improve the solubility of Pseudomonas putida aromatic amino acid decarboxylase (AADC) that is an indispensable enzyme for fermentative production of plant isoquinoline alkaloids. AADC fused N-terminally with the PPIase KS−1 (PPIase KS−1-AADC), which was synthesized utilizing Escherichia coli host, showed improved solubility and, consequently, the cell-free extract from the recombinant strain exhibited 2.6- to 3.4-fold elevated AADC activity than that from the control strain that expressed the AADC gene without PPIase KS−1. On the other hand, its thermostability was slightly decreased by fusing PPIase KS−1. The recombinant E. coli cells expressing the PPIase KS−1-AADC gene produced dopamine and phenylethylamine from L-dopa and phenylalanine by two- and threefold faster, respectively, as compared with the control strain. We further demonstrated that the efficacy of PPIase KS−1-AADC in solubility and activity enhancement was a little but obviously higher than that of AADC fused N-terminally with NusA protein, which has been assumed to be the most effective protein solubilizer. These results suggest that PPIase KS−1 can be used as one of the best choices for producing heterologous proteins as active forms in E. coli.


Molecules ◽  
2021 ◽  
Vol 26 (22) ◽  
pp. 6794
Author(s):  
Aleksandrina S. Volobueva ◽  
Olga I. Yarovaya ◽  
Marina V. Kireeva ◽  
Sophia S. Borisevich ◽  
Kseniya S. Kovaleva ◽  
...  

A number of framework amides with a ginsenol backbone have been synthesized using the Ritter reaction. We named the acetamide as Ginsamide. A method was developed for the synthesis of the corresponding amine and thioacetamide. The new compounds revealed a high activity against H1N1 influenza, which was confirmed using an animal model. Biological experiments were performed to determine the mechanism of action of the new agents, a ginsamide-resistant strain of influenza virus was obtained, and the pathogenicity of the resistant strain and the control strain was studied. It was shown that the emergence of resistance to Ginsamide was accompanied by a reduction in the pathogenicity of the influenza virus.


2021 ◽  
Vol 12 (4) ◽  
pp. 664-669
Author(s):  
P. A. Shumilova ◽  
N. A. Sennikova ◽  
E. A. Silivanova ◽  
M. A. Levchenko

Insect populations exist under acute and chronic exposures to lethal and sublethal insecticide concentrations. Among the sublethal effects of insecticides on insects are reductions in life span, development rates, population growth, fertility, fecundity, changes in sex ratio, deformities, changes in behaviour, feeding, searching, and oviposition. These effects may differ depending on the modes of action of insecticides, their doses, and developmental stage of application. This study evaluated the life-history parameters in Musca domestica Linnaeus, 1758 (Diptera:Muscidae) strains that were exposed to two insecticides (fipronil and chlorfenapyr) with different modes of action at sublethal concentrations in each generation up to tenth. Two approaches to each insecticide’s exposure were used in this study, particularly in one approach, only adults M. domestica were exposed to fipronil or chlorfenapyr and in another approach, only larvae were exposed to each insecticide. The susceptibility of adult flies to these insecticides was assessed by resistance ratio based on results of non-choice feeding bioassays. Fipronil exposure at the sublethal concentration in each generation did not affect the susceptibility of adult M. domestica (in the tenth generation) to fipronil. The resistance ratio values revealed tolerance to chlorfenapyr in adults of M. domestica strains that were exposed to this insecticide, independent of the approach used to insecticide exposure. Most of the life-history parameters (such as durations of separate developmental stages, the emergence duration, the adult emergence ratio, the female ratio, and the number of eggs per female per day) of the second, fourth, sixth, eighth, and tenth generations of the insecticide-exposed strains were similar to those of the control strain. At the same time, the sublethal effects of both insecticides fipronil and chlorfenapyr prolonged the larval duration (1.63–2.22 times) and the number of days from egg to adult (1.18–1.39 times) compared to the control strain. Further studies are needed to investigate a possible genetic variability in M. domestica in response to exposure of parental generation to sublethal doses of fipronil and chlorfenapyr.


Plant Disease ◽  
2021 ◽  
Author(s):  
Juan Moral Moral ◽  
MARÍA TERESA GARCÍA LOPEZ ◽  
Ana Gordon ◽  
Alejandro Ortega-Beltran ◽  
Ryan D Puckett ◽  
...  

Aflatoxin contamination of almond kernels, caused by Aspergillus flavus and A. parasiticus, is a severe concern for growers due to its high toxicity. In California, the global leader of almond production, aflatoxin can be managed by applying the biological control strain AF36 of A. flavus and selecting resistant cultivars. Here, we classified the almond genotypes by K-Means cluster analysis into three groups [Susceptible (S), Moderately Susceptible (MS), or Resistant (R)] based on aflatoxin content of inoculated kernels. The protective effects of the shell and seedcoat in preventing aflatoxin contamination were also examined. The presence of intact shells reduced aflatoxin contamination over 100-fold. The seedcoat provided a layer of protection, but not complete. In kernel inoculation assays, none of the studied almond genotypes showed a total resistance to the pathogen. However, nine traditional cultivars and four advanced selections were classified as R. Because these advanced selections contained germplasm derived from peach, we compared the kernel resistance of three peach cultivars to that shown by kernels of a R (‘Sonora’) and a S (‘Carmel’) almond cultivar and five pistachio cultivars. Overall, peach kernels were significantly more resistant to the pathogen than almond kernels, which were more resistant than pistachio kernels. Finally, we studied the combined effect of the cultivar resistance and the biocontrol strain AF36 in limiting aflatoxin contamination. For this, we co-inoculated almond kernels of R ‘Sonora’ and S ‘Carmel’ with AF36 72 h before or 48 h after inoculating with an aflatoxin-producing strain of A. flavus. The percentage of aflatoxin reduction by AF36 strain was greater in kernels of ‘Carmel’ kernels (98%) than in those of ‘Sonora’ (83%). Cultivar resistance also affected the kernel colonization by the biological control strain. AF36 strain limited aflatoxin contamination in almond kernels even when applied 48 h after the aflatoxin-producing strain. Our results show that biocontrol combined with the use of cultivars with resistance to aflatoxin contamination can result in a more robust protection strategy than the use of either practices in isolation.


Author(s):  
Brianna M. Schick ◽  
Hunter Dlugas ◽  
Teresa L. Czeiszperger ◽  
Alexandra R. Matus ◽  
Melissa J. Bukowski ◽  
...  

Author(s):  
Zhijian Ni ◽  
Jinyong Wu ◽  
Zhongkui Li ◽  
Lixia Yuan ◽  
Yu Wang ◽  
...  

Abstract 3-fucosyllactose (3-FL) and 2′-fucosyllactose (2′-FL), are two important fucosylated oligosaccharides in human milk. Extensive studies on 2'-FL enabled its official approval for use in infant formula. However, commercialization of 3-FL seems to be a bit sluggish due to its low content in human milk and poor yield in enlarged production. Here, an α-1,3-fucosyltransferase mutant was expressed in an engineered E. coli able to produce GDP-L-fucose, and gave a promising 3-FL titer in a 5.0-L bioreactor. To increase the availability of cofactors (NADPH and GTP) for optimized 3-FL production, zwf, pntAB, and gsk genes were successively overexpressed, finally resulting in a high 3-FL level with a titer of 35.72 g/L, a yield of 0.82 mol 3-FL/mol lactose, and a specific productivity of 0.46 g/L*h. Unexpectedly, the deletion of pfkA gene led to a much lower performance of 3-FL production than the control strain. Taken together, our production strategy finally achieved the highest 3-FL level in E. coli to date.


2021 ◽  
Vol 38 (2) ◽  
pp. 120-125
Author(s):  
V. I. Sergevnin ◽  
O. V. Tukacheva ◽  
О. R. Sibiryakova ◽  
E. V. Sarmometov ◽  
P. B. Azanov

Objective. To study the sensitivity to chemical disinfectants against Mycobacterium tuberculosis isolated from HIV-infected individuals and patients with monotuberculosis in comparison to the reference strain of Mycobacterium terrae. Materials and methods. The sensitivity of 12 M. tuberculosis strains isolated from patients with HIV-associated tuberculosis (n = 6) and monotuberculosis (n = 6) to chlorine-containing and quaternary ammonium compound (QAC) disinfectants was studied. The M. terrae strain was used as a reference. Seedings of microbial suspensions treated with disinfectants on the culture medium Levenshtein-Jensen were carried out and were monitored for 3 months. Results. One, two, three months following the treatment of microorganisms with QAC-containing disinfectant in the absence of growth of the control strain, there was noted growth of M. tuberculosis, particularly intense in the strains isolated from HIV-infected persons. Conclusions. Thus, the resistance of tuberculosis pathogens to disinfectants can be acquired. M. tuberculosis strains isolated from patients with HIV-associated tuberculosis are detected to be more resistant to disinfectants than the strains isolated from patients with monotubercular infection.


2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Simon Boecker ◽  
Björn-Johannes Harder ◽  
Regina Kutscha ◽  
Stefan Pflügl ◽  
Steffen Klamt

Abstract Background The alcohol 2,3-butanediol (2,3-BDO) is an important chemical and an Escherichia coli producer strain was recently engineered for bio-based production of 2,3-BDO. However, further improvements are required for realistic applications. Results Here we report that enforced ATP wasting, implemented by overexpressing the genes of the ATP-hydrolyzing F1-part of the ATPase, leads to significant increases of yield and especially of productivity of 2,3-BDO synthesis in an E. coli producer strain under various cultivation conditions. We studied aerobic and microaerobic conditions as well as growth-coupled and growth-decoupled production scenarios. In all these cases, the specific substrate uptake and 2,3-BDO synthesis rate (up to sixfold and tenfold higher, respectively) were markedly improved in the ATPase strain compared to a control strain. However, aerobic conditions generally enable higher productivities only with reduced 2,3-BDO yields while high product yields under microaerobic conditions are accompanied with low productivities. Based on these findings we finally designed and validated a three-stage process for optimal conversion of glucose to 2,3-BDO, which enables a high productivity in combination with relatively high yield. The ATPase strain showed again superior performance and finished the process twice as fast as the control strain and with higher 2,3-BDO yield. Conclusions Our results demonstrate the high potential of enforced ATP wasting as a generic metabolic engineering strategy and we expect more applications to come in the future.


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