Cross-talk between the gut microbiota and monocyte-like macrophages mediates an inflammatory response to promote colitis-associated tumourigenesis.

OBJECTIVE: Macrophages are among the most abundant cells in the colon tumour microenvironment, and there is a close relationship among monocytes, macrophages and the gut microbiota. Alterations in the gut microbiota are involved in tumour development, but the underlying mechanisms remain unclear. We aim to elucidate the temporal changes in macrophage subsets and functions, and how these dynamics are regulated by microbial cues in the initiation of colitis-associated cancer. DESIGN: A mouse model of colitis-associated tumourigenesis was established to determine macrophage dynamics. The role of monocyte-like macrophage (MLM) was confirmed by targeting its chemotaxis. The effects of the gut microbiota were assessed by antibiotic treatment and faecal microbiota transplantation. RESULTS: A selective increase in MLMs was observed in the initial stages of colitis-associated cancer, with an enhanced secretion of inflammatory cytokines. MLM accumulation was regulated by CCL2 expression of colonic epithelial cells, which was influenced by bacteria-derived lipopolysaccharide (LPS). LPS further stimulated interleukin 1ß production from MLMs, inducing interleukin-17-producing T-helper cell activation to promote inflammation. These observations were also supported by altered microbial composition associated with human colitis and colorectal cancer, evolving transcriptional signature and immune response during human colitis-associated tumourigenesis. CONCLUSIONS: The gut microbiota uses LPS as a trigger to regulate MLM accumulation in a chemokine-dependent manner and generate a precancerous inflammatory milieu to facilitate tumourigenesis.

Synthesis and biological evaluation of novel structure-related hGHRH agonistic analogs.

Activity and half-life play key roles in the application of GHRH analogues. The GHRH monomers produced in a solid synthesizer were incubated, respectively, in NH4OH solution and lyophilized to obtain their dimers. The activities, specificities, and receptor affinities of the GHRH dimers were evaluated in rGH release/inhibition, rACTH/LH/PRL release, pituitary homogenate binding, and fluorescent staining. Compared to hGHRH(1-44)NH2 (S), PP-hGHRH(1-44)-GGC-CGG-hGHRH(44-1)-PP (2D), P-hGHRH(1-44)-GGC-CGG-hGHRH(44-1)-P (2E), (1)P-hGHRH(2-44)-GGC-CGG-hGHRH(44-2)-(1)P (2F), or hGHRH(1-44)-GGC-CGG-hGHRH(44-1) (2Y) had potency of 104 ± 16.7%, 94 ± 32.6%, 114 ± 16.6%, or 122 ± 14.5% and similar specificities. The inhibition effect of GHIH on rGH stimulated by GHRH dimer was in dose-/time-dependent manner. The staining of FITC-labeled dimer showed cytomembrane distribution and the binding ranking was 2F>2D>2Y>2E>S. 2F presents the strongest activity and the highest affinity to pituitary cells. The dimer with (1)Pro-GHRH stimulates stronger rGH release than that with (1)Tyr-GHRH and the N-terminal single cyclic amino acid is required for the stimulation.

Multi-tailed vision transformer for efficient inference.

Recently, Vision Transformer (ViT) has achieved promising performance in image recognition and gradually serves as a powerful backbone in various vision tasks. To satisfy the sequential input of Transformer, the tail of ViT first splits each image into a sequence of visual tokens with a fixed length. Then, the following self-attention layers construct the global relationship between tokens to produce useful representation for the downstream tasks. Empirically, representing the image with more tokens leads to better performance, yet the quadratic computational complexity of self-attention layer to the number of tokens could seriously influence the efficiency of ViT's inference. For computational reduction, a few pruning methods progressively prune uninformative tokens in the Transformer encoder, while leaving the number of tokens before the Transformer untouched. In fact, fewer tokens as the input for the Transformer encoder can directly reduce the following computational cost. In this spirit, we propose a Multi-Tailed Vision Transformer (MT-ViT) in the paper. MT-ViT adopts multiple tails to produce visual sequences of different lengths for the following Transformer encoder. A tail predictor is introduced to decide which tail is the most efficient for the image to produce accurate prediction. Both modules are optimized in an end-to-end fashion, with the Gumbel-Softmax trick. Experiments on ImageNet-1K demonstrate that MT-ViT can achieve a significant reduction on FLOPs with no degradation of the accuracy and outperform compared methods in both accuracy and FLOPs.

Intracellular ATP delivery to in vitro expanded mouse CD27- γδ T cells.

Intracellular ATP supports the function of γδT17 cells in mice. Here, we present a protocol for intracellular ATP delivery to in vitro expanded mouse CD27- γδ T cells. We describe steps for pre-coating well plates, preparing lymphocytes, culturing CD27- γδ T cells, and ATP delivery. We then detail functional evaluation of γδ T cells by flow cytometry. Appropriate concentrations of control and ATP vesicles are detailed for intracellular ATP delivery, which can also be applied to other immune cells. For complete details on the use and execution of this protocol, please refer to Wang et al. (2023).1.

Transcription factor c-Maf-targeted cancer immunotherapy.

In innate immune cells, the transcription factor cellular musculoaponeurotic fibrosarcoma (c-Maf) influences cell fate and function through molecular and metabolic programming, thereby influencing immune homeostasis and antitumor immunity. We discuss recent c-Maf landmark discoveries with a focus on the immunosuppressive tumor microenvironment (TME) and provide a new perspective on c-Maf-targeted cancer immunotherapy.

Natural γδT17 cell development and functional acquisition is governed by the mTORC2-c-Maf-controlled mitochondrial fission pathway.

Natural IL-17-producing γδ T cells (γδT17 cells) are unconventional innate-like T cells that undergo functional programming in the fetal thymus. However, the intrinsic metabolic mechanisms of γδT17 cell development remain undefined. Here, we demonstrate that mTORC2, not mTORC1, selectively controls the functional fate commitment of γδT17 cells through regulating transcription factor c-Maf expression. scRNA-seq data suggest that fetal and adult γδT17 cells predominately utilize mitochondrial metabolism. mTORC2 deficiency results in impaired Drp1-mediated mitochondrial fission and mitochondrial dysfunction characterized by mitochondrial membrane potential (ΔΨm) loss, reduced oxidative phosphorylation (OXPHOS), and subsequent ATP depletion. Treatment with the Drp1 inhibitor Mdivi-1 alleviates imiquimod-induced skin inflammation. Reconstitution of intracellular ATP levels by ATP-encapsulated liposome completely rescues γδT17 defect caused by mTORC2 deficiency, revealing the fundamental role of metabolite ATP in γδT17 development. These results provide an in-depth insight into the intrinsic link between the mitochondrial OXPHOS pathway and γδT17 thymic programming and functional acquisition.

Temporal dynamics of competitive fertilization in social groups of red junglefowl (Gallus gallus) shed new light on avian sperm competition.

Studies of birds have made a fundamental contribution to elucidating sperm competition processes, experimentally demonstrating the role of individual mechanisms in competitive fertilization. However, the relative importance of these mechanisms and the way in which they interact under natural conditions remain largely unexplored. Here, we conduct a detailed behavioural study of freely mating replicate groups of red junglefowl, Gallus gallus, to predict the probability that competing males fertilize individual eggs over the course of 10-day trials. Remating frequently with a female and mating last increased a male's probability of fertilization, but only for eggs ovulated in the last days of a trial. Conversely, older males, and those mating with more polyandrous females, had consistently lower fertilization success. Similarly, resistance to a male's mating attempts, particularly by younger females, reduced fertilization probability. After considering these factors, male social status, partner relatedness and the estimated state of male extragonadal sperm reserves did not predict sperm competition outcomes. These results shed new light on sperm competition dynamics in taxa such as birds, with prolonged female sperm storage and staggered fertilizations. This article is part of the theme issue 'Fifty years of sperm competition'.

Kinetic Study of DNA Topoisomerases by Supercoiling-Dependent Fluorescence Quenching.

DNA topoisomerases are essential enzymes for all living organisms and important targets for anticancer drugs and antibiotics. Although DNA topoisomerases have been studied extensively, steady-state kinetics has not been systematically investigated because of the lack of an appropriate assay. Previously, we demonstrated that newly synthesized, fluorescently labeled plasmids pAB1_FL905 and pAB1_FL924 can be used to study DNA topoisomerase-catalyzed reactions by fluorescence resonance energy transfer (FRET) or supercoiling-dependent fluorescence quenching (SDFQ). With the FRET or SDFQ method, we performed steady-state kinetic studies for six different DNA topoisomerases including two type IA enzymes (Escherichia coli and Mycobacterium smegmatis DNA topoisomerase I), two type IB enzymes (human and variola DNA topoisomerase I), and two type IIA enzymes (E. coli DNA gyrase and human DNA topoisomerase IIα). Our results show that all DNA topoisomerases follow the classical Michaelis-Menten kinetics and have unique steady-state kinetic parameters, K M, V max, and k cat. We found that k cat for all topoisomerases are rather low and that such low values may stem from the tight binding of topoisomerases to DNA. Additionally, we confirmed that novobiocin is a competitive inhibitor for adenosine 5'-triphosphate binding to E. coli DNA gyrase, demonstrating the utility of our assay for studying topoisomerase inhibitors.

[Study on the mechanism of energy transfer between fluorescein sodium and tetrabromofluorescein sodium in micelles of cationic surface active agent].

In the present paper, the fluorescence reaction of cationic surface-active agents (CSAA) with Tetrabromofluorescein sodium (TBF) in aqueous solution was investigated. It was found that the fluorescence quenching of TBF appears when it reacts with the cation monomer of a CSAA and a new stronger fluorescence is obtained when the ion-associates react with the micellate of CSAA. The authors investigated the condition of energy transfer between acidic fluorescent dyes in micelles of cetyl trimethyl ammonium bromide or hexadecyl pyridinium bromide. It was indicated that in the micelles formed by cationic surface active agent with dyes embedded (cationic surface active agent and dyes are charge opposite), the effective energy transfer between anionic dyes could occur. When the concentration of cationic surface active agent reached certain value, the energy transfer could occur. With the value of two thirds of critical micelle concentration, the efficiency of energy transfer reached the maximum; when the concentration of cationic surface active agent increased, the efficiency of energy transfer would be decreased. The authors also deduced the model of energy transfer between dyes in micelles and laws of it.

[Study on mechanism of lanthanide sensitized fluorescence energy transfer between terbium(III)-ciprofloxacin and rhodamine B].

A complex composed of ciprofloxacin and terbium (Tb3+) in the solution of acetic acid-sodium acetate buffer (pH 6.2) was studied by fluorescence spectroscopy and ultra-violet absorption spectroscopy. The emission peak of Tb3+ appeared at 490, 545, 590 nm, (the sensitized fluorescence was enforced mightily) and the intensity of 545 nm emission peak was increased obviously. In its acute emission spectrum, the strongest emission peak of Tb3+ was at 545 nm, which was close to the wavelength of the biggest absorption peak of RB, 552 nm. Therefore, as the basic dye rhodamine B(RB) was added, the fluorescence intensity of 545 nm emission peak decreased regularly, indicating that there was a great quenching process. The result showed that the course was statistic. Based on the mechanism of the Förster energy transfer, the efficiency of energy transfer and the distance between the acceptor RB and the complex were found. Thereby, it was indicated that the course of action was single static quenching and the mechanism of quenching was based on energy transfer.

One step synthesis of high-efficiency AgBr-Br-g-C3N4 composite catalysts for photocatalytic H2O2 production via two channel pathway.

In this work, a two-component modified AgBr-Br-g-C3N4 composite catalyst with outstanding photocatalytic H2O2 production ability is synthesized. XRD, UV-Vis, N2 adsorption, TEM, XPS, EPR and PL were used to characterize the obtained catalysts. The as-prepared AgBr-Br-g-C3N4 composite catalyst shows the highest H2O2 equilibrium concentration of 3.9 mmol L-1, which is 7.8 and 19.5 times higher than that of GCN and AgBr. A "two channel pathway" is proposed for this reaction system which causes the remarkably promoted H2O2 production ability. In addition, compared with another two-component modified catalyst, Ag-AgBr-g-C3N4, AgBr-Br-g-C3N4 composite catalyst displays the higher photocatalytic H2O2 production ability and stability.

Targeted delivery and release of doxorubicin using a pH-responsive and self-assembling copolymer.

Developing a smart cancer drug delivery carrier with enhanced cancer-targeting and effective drug release in tumors is critical for efficient cancer chemotherapy. Herein, we designed a pH-responsive copolymer (PEG-ELP[VH4-70]) that entraps DOX into a hydrophobic core and self-assembles into smart DOX-loaded nanoparticles. The DOX-loaded nanoparticles were stabilized by Zn2+ and disrupted as the pH drops from 7.4 to 5.6. An in vitro study demonstrated that the DOX-loaded nanoparticle system exhibited efficient internalization, triggered the release of DOX into the cytoplasm and enabled the inhibition of tumors effectively. When used to deliver chemotherapeutics to a murine cancer model, PEG-ELP[VH4-70]/DOX accomplished a 4.8-fold suppressed effect relative to the free drug after intravenous injection. This simple strategy can promote preeminent stability for targeting hydrophobic drugs to tumor tissues.

Relationship between hypnosis and personality trait in participants with high or low hypnotic susceptibility.

BACKGROUND: The relationship between normal personality and hypnotic susceptibility is important for understanding mental processing and mental disorders, but it is less consistent in normal people or in patients with a psychiatric disorder. We have hypothesized that the correlation exists but varies in individuals with different levels of hypnotizability. PARTICIPANTS AND METHODS: We invited 72 individuals with high (HIGH group) and 47 individuals with low (LOW group) hypnotic susceptibilities to undertake tests of NEO-PI-R and the Stanford Hypnotic Susceptibility Scale, Form C (SHSSC). RESULTS: The HIGH group scored significantly higher than the LOW group did on openness to experience and its facet openness to feelings. In the LOW group, SHSSC total was positively predicted by openness to ideas; age regression was positively predicted by openness to experience and negatively predicted by extraversion; anosmia to ammonia was negatively predicted by agreeableness; and negative visual hallucination was positively predicted by openness to experience. In the HIGH group, hallucinated voice was positively predicted by openness to experience and negatively predicted by agreeableness, and posthypnotic amnesia was positively predicted by extraversion and negatively predicted by openness to experience. CONCLUSION: The associations between normal personality traits and hypnotic susceptibility items were weak and different in the two groups, which imply that managing mental or somatoform disorders might be through adjusting hypnotizability and mobilizing personality functions.

Investigation of mtDNA control region sequences in a Tibetan population sample from China.

Mitochondrial hypervariable region sequences including HVI and HVII (15,751-520) were investigated from 174 unrelated Tibetan individuals living in Tibet Autonomous Region in People's Republic of China. The resulted sequences were aligned and compared with revised Cambridge sequence (rCRS). This sequence variability rendered a high gene diversity value (0.9940 ± 0.0021) and a high random match probability (0.0118) was determined with PIC of 0.9882. Among a total of 174 samples, 217 polymorphic sites were identified, which defined 135 haplotypes. A total of 135 different haplotypes were detected, 113 of them were unique and 22 were shared. The most common haplogroup was M9a1a1c1b1 (16.09%), followed by A11 (6.32%), A (5.17%), R (4.60%), A15 (4.60%), and G3a1 (3.45). The proportions of macro-haplogroups M, N, and L were 54.60%, 42.53%, and 2.87%, respectively. By principal component analysis (PCA), there was no special cluster between Tibetans and other populations except that the structure of Tibetans closely resembled that of Uygur in component 2.