Intracellular H
S plays an important regulatory role in cell metabolism. The limited sensing materials and severe sensor passivation hinder its quantification. We functionalized conductive nanowires ...with MoS
and quercetin in a large-scale manner, developed single nanowire sensors with excellent electrocatalytic and anti-poisoning performance, and achieved the accurate quantification of H
S within single cells.
Doublesex (DSX) proteins are members of the Doublesex/mab-3-related (DMRT) protein family and play crucial roles in sex determination and differentiation among the animal kingdom. In the present ...study, we identified two Doublesex (Dsx)-like mRNA isoforms in the brine shrimp Artemia franciscana (Kellogg 1906), which are generated by the combination of alternative promoters, alternative splicing and alternative polyadenylation. The two transcripts exhibited sex-biased enrichment, which we termed AfrDsxM and AfrDsxF. They share a common region which encodes an identical N-terminal DNA-binding (DM) domain. RT-qPCR analyses showed that AfrDsxM is dominantly expressed in male Artemia while AfrDsxF is specifically expressed in females. Expression levels of both isoforms increased along with the developmental stages of their respective sexes. RNA interference with dsRNA showed that the knockdown of AfrDsxM in male larvae led to the appearance of female traits including an ovary-like structure in the original male reproductive system and an elevated expression of vitellogenin. However, silencing of AfrDsxF induced no clear phenotypic change in female Artemia. These results indicated that the male AfrDSXM may act as inhibiting regulator upon the default female developmental mode in Artemia. Furthermore, electrophoretic mobility shift assay analyses revealed that the unique DM domain of AfrDSXs can specifically bind to promoter segments of potential downstream target genes like AfrVtg. These data show that AfrDSXs play crucial roles in regulating sexual development in Artemia, and further provide insight into the evolution of sex determination/differentiation in sexual organisms.
Filtration of circulating tumor cells (CTCs) in peripheral blood is of proven importance for early cancer diagnosis, treatment monitoring, metastasis diagnosis, and prognostic evaluation. However, ...currently available strategies for enriching CTCs, such as magnetic activated cell sorting (MACS), face serious problems with purity due to nonspecific interactions between beads and leukocytes in the process of capturing. In the present study, the tumor-targeting molecule folic acid (FA) and magnetic nanoparticles (MNPs) were coated on the surface of red blood cells (RBCs) by hydrophobic interaction and chemical conjugation, respectively. The resulting engineered RBCs rapidly adhered to CTCs and the obtained CTC-RBC conjugates were isolated in a magnetic field. After treatment with RBC lysis buffer and centrifugation, CTCs were released and captured. The duration of the entire process was less than three hours. Cell counting showed that the capture efficiency was above 90% and the purity of the obtained CTCs was higher than 75%. The performance of the proposed method exceeded that of MACS® beads (80% for capture efficiency and 20% for purity) under the same conditions. The obtained CTCs could be successfully re-cultured and proliferated in vitro. Our engineered RBCs have provided a novel method for enriching rare cells in the physiological environment.
With the emergence of CRISPR/Cas9 technology, multiple gene editing procedures became available for the silkworm. Although binary transgene-based methods have been widely used to generate mutants, ...delivery of the CRISPR/Cas9 system via DNA-free ribonucleoproteins offers several advantages. However, the T7 promoter that is widely used in the ribonucleoprotein-based method for production of sgRNAs in vitro requires a 5' GG motif for efficient initiation. The resulting transcripts bear a 5' GG motif, which significantly constrains the number of targetable sites in the silkworm genome.
In this study, we used the T7 promoter to add two supernumerary G residues to the 5' end of conventional (perfectly matched) 20-nucleotide sgRNA targeting sequences. We then asked if sgRNAs with this structure can generate mutations even if the genomic target does not contain corresponding GG residues. As expected, 5' GG mismatches depress the mutagenic activity of sgRNAs, and a single 5' G mismatch has a relatively minor effect. However, tests involving six sgRNAs targeting two genes show that the mismatches do not eliminate mutagenesis in vivo, and the efficiencies remain at useable levels. One sgRNA with a 5' GG mismatch at its target performed mutagenesis more efficiently than a conventional sgRNA with 5' matched GG residues at a second target within the same gene. Mutations generated by sgRNAs with 5' GG mismatches are also heritable. We successfully obtained null mutants with detectable phenotypes from sib-mated mosaics after one generation.
In summary, our method improves the utility and flexibility of the ribonucleoprotein-based CRISPR/Cas9 system in silkworm.
Gut microbiota plays a critical role in the onset and development of depression, but the underlying molecular mechanisms are unclear. This study was conducted to explore the relationships between gut ...microbiota and host's metabolism in depression.
Chronic social defeat stress (CSDS) model of depression was established using C57BL/6 male mice. Fecal samples were collected from CSDS group and control group to measure gut microbiota and microbial metabolites. Meanwhile, tryptophan metabolism-related metabolites in hippocampus were also analyzed.
CSDS successfully induced depressive-like behaviors in CSDS group. The 24 differential bacterial taxa between the two groups were identified, and 14 (60.87%) differential bacterial taxa belonged to phylum Firmicutes. Functional analysis showed that tryptophan metabolism was significantly affected in CSDS mice. Meanwhile, 120 differential microbial metabolites were identified, and two key tryptophan metabolism-related metabolites (tryptophan and 5-hydroxytryptophan (5-HTP)) were significantly decreased in feces of CSDS mice. The correlation analysis found the significant relationships between tryptophan and differential bacterial taxa under Firmicutes, especially genus
(r=0.801, p=0.0002). In addition, the significantly decreased 5-hydroxytryptamine (5-HT) in hippocampus of depressed mice was also observed.
Our results showed that tryptophan metabolism might have an important role in the crosstalk between gut microbioa and brain in depression, and phylum Firmicutes, especially genus
, might be involved in the onset of depression through regulating tryptophan metabolism.
Spiroindoline‐3,7′‐pyrrolo1,2‐aazepines were conveniently synthesized in satisfactory yields and with high diastereoselectivity from three‐component reaction of L‐proline, isatins and chalcones and ...sequential reaction with dimethyl but‐2‐ynedioate or methyl propiolate in refluxing methanol. The ferrocenyl‐chalcones and dibenzylideneacetones were also successfully used in this one‐pot two‐step reaction to give novel ferrocenyl and styryl‐substituted spiro compounds. The reaction mechanism included domino 3+2 cycloadddtition reaction of azomethine ylide with chalcone and ring‐expansion reaction of in situ generated spiroindoline‐3,3′‐pyrrolizine with electron‐deficient alkynes.
Synthetic methods: Spiroindoline‐3,7′‐pyrrolo1,2‐aazepines were conveniently synthesized from domino 3+2 cycloadddtition reaction of azomethine ylide with chalcone and ring‐expansion reaction of in situ generated spiroindoline‐3,3′‐pyrrolizine.
Purpose
To evaluate the frequency and the degree of vital signs elevation, as well as to elucidate the risk factors for elevation of cardiopulmonary parameters.
Materials and methods
We ...retrospectively evaluated the medical records of 101 patients who received microwave ablation (MWA) under deep sedation with propofol. Univariate analysis followed by multivariate linear regression analysis was performed to determine the risk factors associated with the elevation of cardiopulmonary parameters.
Results
The heart rate (HR), mean blood pressure (BP) and respiratory rate (RR) were elevated in 53.5%, 45.5% and 30.7%. Hyperhemodynamic state (mean BP or HR increased > 30% of the baseline) and high RR (RR > 20 times/min) were detected in 23.8% and 13.9%. Age ≤ 50 years was signifiant for mean BP and HR elevation (
p
= 0.032;
p
= 0.027), ablation zone abutting the parietal peritoneum (
p
= 0.001;
p
= 0.001;
p
< 0.001) and the diaphragm (
p
= 0.001) were risk factors for BP and RR elevation.
Conclusions
Elevations in HR and BP are common. Risk factors for vital signs elevation include ablation zone abutting the parietal peritoneum and the diaphragm, as well as young age. These findings help devise strategies for anesthetic management.
•Quasi-brittle failure of flawed granite is evaluated in real time by integrated acousto-optic-mechanics (AOM) multi-physics field characterization methods.•Procedural sample-sized localization ...instabilities are forecasted and evaluated using the time‐reversed Omori law.•The physics relating to the failure predictability of flawed granite is revealed.
Forecasting quasibrittle failure in geomaterials is of paramount importance in the physics of fractures but rarely succeeds. To clarify the physics relating to the failure predictability in a quasibrittle regime, a combined AE (acoustic emission)-DIC (digital image correlation) technique is used in uniaxial compression experiments of flawed granite to trace its damage conditions in real time. The mechanistic correlations of the AE rate-process with the internal FPZ (fracture process zone) development are available. It is shown that the complete record of the AE activities does not meet a deterministic scaling law (e.g., typical time‐reversed Omori law), indicating a challenge in predicting ultimate failure via AE. Subsets of a power law acceleration registered much earlier than ultimate failure imply that the procedural sample-sized localization instability is predictable but still depends on the FPZ dynamic evolutions. More importantly, we find that the coexistence of FPZs and the intermittency of crack growth are the two main reasons for the difficulty of forecasting failure in flawed granite. This study sheds light on upcoming investigations in which the roles of FPZs, especially their spatiotemporal patterns, should be highlighted.
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