While CRISPR/Cas9 technology has proven to be a valuable system to generate gene-targeted modified animals in several species, this tool has been scarcely reported in farm animals. Myostatin is ...encoded by MSTN gene involved in the inhibition of muscle differentiation and growth. We determined the efficiency of the CRISPR/Cas9 system to edit MSTN in sheep and generate knock-out (KO) animals with the aim to promote muscle development and body growth. We generated CRISPR/Cas9 mRNAs specific for ovine MSTN and microinjected them into the cytoplasm of ovine zygotes. When embryo development of CRISPR/Cas9 microinjected zygotes (n = 216) was compared with buffer injected embryos (n = 183) and non microinjected embryos (n = 173), cleavage rate was lower for both microinjected groups (P<0.05) and neither was affected by CRISPR/Cas9 content in the injected medium. Embryo development to blastocyst was not affected by microinjection and was similar among the experimental groups. From 20 embryos analyzed by Sanger sequencing, ten were mutant (heterozygous or mosaic; 50% efficiency). To obtain live MSTN KO lambs, 53 blastocysts produced after zygote CRISPR/Cas9 microinjection were transferred to 29 recipient females resulting in 65.5% (19/29) of pregnant ewes and 41.5% (22/53) of newborns. From 22 born lambs analyzed by T7EI and Sanger sequencing, ten showed indel mutations at MSTN gene. Eight showed mutations in both alleles and five of them were homozygous for indels generating out-of frame mutations that resulted in premature stop codons. Western blot analysis of homozygous KO founders confirmed the absence of myostatin, showing heavier body weight than wild type counterparts. In conclusion, our results demonstrate that CRISPR/Cas9 system was a very efficient tool to generate gene KO sheep. This technology is quick and easy to perform and less expensive than previous techniques, and can be applied to obtain genetically modified animal models of interest for biomedicine and livestock.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The development of efficient and selective C−N bond‐forming reactions from abundant feedstock chemicals remains a central theme in organic chemistry owing to the key roles of amines in synthesis, ...drug discovery, and materials science. Herein, we present a dual catalytic system for the N‐alkylation of diverse aromatic carbocyclic and heterocyclic amines directly with carboxylic acids, by‐passing their preactivation as redox‐active esters. The reaction, which is enabled by visible‐light‐driven, acridine‐catalyzed decarboxylation, provides access to N‐alkylated secondary and tertiary anilines and N‐heterocycles. Additional examples, including double alkylation, the installation of metabolically robust deuterated methyl groups, and tandem ring formation, further demonstrate the potential of the direct decarboxylative alkylation (DDA) reaction.
No beating about the bush: (Hetero)aromatic amines underwent decarboxylative N‐alkylation directly with carboxylic acids with a dual copper/acridine photocatalytic system. The directional character of the acridine photocatalysis facilitates the challenging decarboxylation of unactivated carboxylic acids in the presence of more readily oxidizable anilines, thus enabling the use of a wide range of structurally diverse amine and acid substrates (see scheme).
The present work focused on the synthesis of novel ZnLaxFe2−xO4 catalysts (x = 0, 0.01, 0.03, 0.05) and their utilization for the photocatalytic degradation of Rhodamine B dye. Structurally, the band ...gap energy of the catalysts tended to decrease (1.94–1.70 eV) with increasing the amount of La3+ dopant. ZnLa0.05Fe1.95O4 had an average particle size (40 nm), high surface area (41.07 m2 g−1) and large pore volume (0.186 cm3 g−1). Moreover, the effect of doping ratio, reaction time, H2O2 concentration, catalyst loading on the treatment performance of La3+ substituted ZnFe2O4 nanocomposites was investigated. ZnLa0.05Fe1.95O4/H2O2 system exhibited the highest degradation efficiency of 99.5% and nonlinear pseudo first-order kinetic reaction rate (14.8 × 10−3 min−1) in the presence of visible light irradiation. The key role of reactive oxygen species involving •O2− and •OH radicals was well explained through the scavenger study. A plausible mechanism of the degradation of Rhodamine B dye was also proposed. Due to two advantageous points including high recyclability (up to 4 cycles) and stability, La3+ substituted ZnFe2O4 nanocomposites can be an effective and competitive catalyst for the visible light-driven photodegradation of toxic dyes in the real wastewaters.
•Facile synthesis of ZnFe2O4 nanocomposites with La3+ dopant (0–5%) was conducted.•Photocatalytic enhancement of ZnLaxFe2−xO4 was respect to pristine ZnFe2O4.•ZnLa0.05Fe1.95O4/H2O2 system obtained the highest photocatalytic efficiency of 99.5%.•The role of reactive oxygen species involving •O2− and •OH radicals was suggested
In this study, As-resistant endophytic bacteria (AEB) from four different ferns (i.e.,
Pteris vittata
,
Pityrogramma calomelanos, Blenchum orientale
, and
Nephrolepis exaltata
) collected from the ...polymetallic mine (Nui Phao) in Vietnam were isolated and characterized. The sampling locations were contaminated with As concentration within the range of 316–1606 mg kg
–1
. A total of 5 arsenite(III)- and 26 arsenate(V)-resistant endophytic bacterial strains that belonged to 13 different genera were obtained. The most predominant isolate (accounting for 36% of the total isolated AEB in root ferns) was identified as
Bacillus
sp. The isolates were more resistant to As(V) than to As(III). Among As(V)-resistant strains, 26 were resistant to As(V) at concentrations of 80–320 mM, whereas 5 As(III)-resistant strains were able to tolerate As(III) of up to 160 mM. Although all the isolates had the ability to produce indole acetic acid (IAA), only three strains (i.e.,
Sporosarcina luteola
R3.3.1,
Paenibacillus
sp. R3.8.3, and
Acinetobacter
sp. L2.5.1) had the phosphate-solubilizing ability. The pot experiments using
P.
vittata
and inoculation of the four selected AEB (i.e.,
Priestia megaterium
R2.5.2,
Micrococcus luteus
S3.4.1,
P. megaterium
R3.4.5, and
P. megaterium
L3.5.1) confirmed the important role of AEB in plant growth promotion and phytoremediation potential of As-contaminated soils.
Enterohemorrhagic Escherichia coli (EHEC) are major food-borne pathogens responsible for serious infections ranging from mild diarrhea to hemorrhagic colitis and life-threatening complications. Shiga ...toxins (Stxs) are the main virulence factor of EHEC. The antagonistic effect of a prophylactic treatment with the probiotic strain Saccharomyces cerevisiae against EHEC O157:H7 was investigated using complementary in vitro human colonic model and in vivo murine ileal loop assays. In vitro, the probiotic treatment had no effect on O157:H7 survival but favorably influenced gut microbiota activity through modulation of short-chain fatty acid production, increasing acetate production and decreasing that of butyrate. Both pathogen and probiotic strains had individual-dependent effects on human gut microbiota. For the first time, stx expression was followed in human colonic environment: at 9 and 12 h post EHEC infection, probiotic treatment significantly decreased stx mRNA levels. Besides, in murine ileal loops, the probiotic yeast specifically exerted a trophic effect on intestinal mucosa and inhibited O157:H7 interactions with Peyer’s patches and subsequent hemorrhagic lesions. Taken together, the results suggest that S. cerevisiae may be useful in the fight against EHEC infection and that host associated factors such as microbiota could influence clinical evolution of EHEC infection and the effectiveness of probiotics.
A cyclometalated iridium(
iii
) complex bearing a self-immolative quinolinium moiety was developed as a ratiometric substrate for transfer hydrogenation studies. This photoluminescent probe allowed ...the rapid screening of a variety of Ir catalysts using a microplate reader, offering a convenient method to assess activity using a minimum amount of catalyst sample.
A method for the simultaneous monitoring of multiple transfer hydrogenation reactions using a microplate reader is reported.
We present results from an analysis of all data taken by the BICEP2, Keck Array, and BICEP3 CMB polarization experiments up to and including the 2018 observing season. We add additional Keck Array ...observations at 220 GHz and BICEP3 observations at 95 GHz to the previous 95 / 150 / 220 GHz dataset. The Q / U maps now reach depths of 2.8, 2.8, and 8.8 μ KCMB arcmin at 95, 150, and 220 GHz, respectively, over an effective area of ≈ 600 square degrees at 95 GHz and ≈ 400 square degrees at 150 and 220 GHz. The 220 GHz maps now achieve a signal-to-noise ratio on polarized dust emission exceeding that of Planck at 353 GHz. We take auto- and cross-spectra between these maps and publicly available WMAP and Planck maps at frequencies from 23 to 353 GHz and evaluate the joint likelihood of the spectra versus a multicomponent model of lensed Λ CDM + r + dust + synchrotron + noise . The foreground model has seven parameters, and no longer requires a prior on the frequency spectral index of the dust emission taken from measurements on other regions of the sky. This model is an adequate description of the data at the current noise levels. The likelihood analysis yields the constraint r0.05 < 0.036 at 95% confidence. Running maximum likelihood search on simulations we obtain unbiased results and find that σ ( r ) = 0.009 . These are the strongest constraints to date on primordial gravitational waves.
Rabies is a fatal viral disease that causes an estimated 59,000 human deaths each year. The majority of these deaths occur in developing countries in Asia. Canine rabies is endemic to Vietnam, which ...is, however, moving towards the disease's elimination. Many countries, such as Vietnam, have invested tremendous resources in controlling rabies, highlighting the goal of regional and global elimination of this neglected disease. In Vietnam, rabies is recognised as one of five high-priority, zoonotic diseases by the Ministry of Health and the Ministry of Agriculture and Rural Development. Investment by the government and by international partners for rabies prevention and control has played a substantial role in reducing human rabies deaths from 404 cases in 1992 to 74 cases in 2017. The catalyst for this effort was the Prime Minister's creation of the National Rabies Program in 1996, which led to increased support and resources for rabies prevention and control. Interventions carried out since then include the expansion of post-exposure prophylaxis centres throughout the country, the introduction or revision of key legislation and guidelines, and improved multisectoral One Health collaboration. In addition, support from international partners, such as the World Organisation for Animal Health (OIE), the World Health Organization (WHO), the Food and Agriculture Organization of the United Nations (FAO), and the Centers for Disease Control and Prevention (CDC), has helped to increase awareness, manage dog populations more effectively, and improve Vietnam's surveillance and diagnostic capabilities. To pursue the goal of eliminating dog-mediated rabies in Vietnam, political commitment is crucial. Resources must be made available to enforce the regulations and guidelines that will enable Vietnam to achieve greater canine rabies vaccination coverage. In this paper, the authors provide an overview of the animal and human health systems in Vietnam, as well as past, current and future directions of rabies prevention and control.
This study aims to investigate the influence of near- and far-field earthquakes on the seismic performance of base-isolated Nuclear Power Plant (NPP) structures. Two earthquake motion groups of ...near-field and far-field characteristics are selected for fragility evaluation analysis. A base-isolated advanced reactor power 1400 (APR-1400) is employed for numerical analysis. A set of fragility curves are derived for various limit states based on the maximum likelihood estimation. The limit states are defined in terms of Lead Rubber Bearing (LRB) deformation capacity. The numerical results reveal that the median maximum deformations of LRBs were smaller for far-field ground motions than for near-field motions. Also, the comparison of fragility curves demonstrates that the probability of failure of base-isolated NPP structures is higher for near-field ground motions than far-field motions. It is crucial to select earthquake ground motions with both near- and far-field motions for the seismic evaluation of NPP structures.
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