To investigate the investigation of gut microbial communities to chicken egg production performance during different laying stages, we conducted a comparative study of the diversity, composition, and function of microbial communities in the fecal and cecal contents of six hens at the early stage, 25 at the peak stage, and 15 at the late stage. We obtained clean data averaging 13.40 and 13.79 Gb for each of the fecal and cecal content samples. The metagenomic analysis revealed significant differences in fecal and cecal content microbial diversity during different laying stages, especially during the peak stage, and the microbiota structure of fecal contents was more stable compared to that of cecal contents. The dominant microflora in the fecal contents during the peak stage were Firmicutes (74.84%) and Lactobacillus (28.13%). The dominant microflora of cecal contents during different stages were basically the same at the phylum level and genus level. During the peak stage, the dominant bacteria shared by microorganisms in the fecal and cecal contents were Lactobacillus. Functional analyses of the gut microbiome indicate that fecal and cecal content microbes have different functional capacities at different egg‐laying stages. We therefore hypothesized that the gut microbiome would vary with laying stage and have a non‐negligible effect on egg production performance. These results improved our ability to provide some theoretical basis for feeding management of laying hens during different laying stages and provided insights into the influence of the gut microbiota on the laying performance of chickens.

AimsPorphyromonas gingivalis is a keystone pathogen in periodontitis that releases bacterial extracellular vesicles (bEVs) with a content derived from the membrane and cytosol of the bacteria itself. The bEVs are taken up by host tissue cells, such as oral fibroblasts, yet their effects on human cells remain incompletely understood.MethodsThe aim of this study was to investigate the impact of bEVs on human oral fibroblasts. Targeted liquid chromatography–mass spectrometry was used to assess changes in the metabolome of these cells following exposure to bEVs from three strains of P. gingivalis (ATCC 33277, A7A1‐28, and W83). Metabolite content of the bEVs from each strain was also characterized.ResultMetabolomic analyses revealed both common and strain‐specific metabolites in the bEVs from these strains. Additionally, pathways mainly associated with amino acid metabolism were enriched for all. Following exposure to bEVs, the fibroblasts exhibited a metabolic shift in energy metabolism. In particular, fibroblasts incubated with bEVs from ATCC 33277 had a unique profile of enriched pathways after 10 min. However, after 24 h, this profile changed and became more similar to the profile in fibroblasts incubated with bEVs from the two more pathogenic strains. Pathway analysis revealed enrichment of glycerophospholipid and butanoate metabolism for fibroblasts exposed to all the bEVs. Additionally, altered sphingolipid metabolism was observed in fibroblasts following exposure to bEVs from ATCC 33277.ConclusionOur findings demonstrate that bEVs derived from P. gingivalis strains can induce changes in cellular processes in fibroblasts. This suggests a role of bEVs in the pathogenesis of periodontitis.

To explore the peculiarities of neutrophil motility, two models of chemoattraction were created: a horizontal model, where a container with bacterial chemoattractant was attached laterally to the endotheliocyte monolayer, and a vertical model, simulating a pyemic focus in the lower part of the modified Boyden chamber. Low‐molecular weight product secretion and/or degradation of Enterococcus faecalis caused “disorientations” of neutrophil migration with hyperproduction reactive oxygen species (ROS) by immune cells, while Proteus mirabilis inhibited both migration of most neutrophils and the production of ROS by them, while the activity of the remaining uninhibited neutrophils increased. Neutrophils generated ROS during migration, especially actively in the case of a large number of mobile cells (under stimulation with low‐molecular weight product secretion and/or degradation of Enterococcus faecalis and Escherichia coli). Using high‐resolution microscopy, it was shown that low‐activity neutrophils cause changes in the morphology of endothelial cells during migration more than high‐activity neutrophils. In the vertical migration model, the morphology of endothelial cells significantly changed during neutrophils diapedesis. It was observed that space between endothelial cells was increased (especially in the case of neutrophil swarming).

The prevalence of low back pain (LBP) due to lumbar disc herniation (LDH) was recorded as 31.9% in the year 2022. Studies carried out around the world have failed to confirm the primary cause of disc herniation. Among the multiple hypothesized contributing factors, a low‐grade bacterial infection has been identified as one of the major causes of LDH. Researchers have reported that Propionibacterium acnes (P. acnes) is the predominant bacterial species isolated using culture‐derived methods. However, biofilm formation leads to a low bacterial yield in culture methods. Although culture methods remain the gold standard for the identification of bacterial species, there is a growing need for the usage of advanced techniques that are more sensitive, reliable, less time‐consuming, and precise. Advancement of high‐throughput sequencing tools allows thorough mining of complete bacterial profiles, even for bacteria that are challenging to cultivate in conventional laboratory settings. Currently, both high‐throughput sequencing and omics have opened a new avenue, providing clear evidence for addressing queries related to bacterial contamination that have been frequently addressed in culture isolates of herniated discs over the past few decades. This review evaluates how advanced techniques in microbial identification have revolutionized our understanding of bacteria in disc health. Traditional methods confirmed the existence of known bacteria, but advanced techniques revealed a vast, previously unseen diversity, challenging the output of culture‐based methods. This new information has even overturned the understanding of the role of P. acnes in evaluating disc health. Advanced techniques have opened a window to the hidden world of microbes and have been attributed to altered views on bacterial communities in healthy and herniated discs.

Proteases degrade proteins and peptides, recycling materials and preventing unnecessary buildup within the cell. They can also be secreted and act in extracellular space. Bacterial proteases are often secreted and function as virulence factors. In the context of the microbiome, they can contribute to host–microbe interactions to facilitate colonization and disease pathogenesis. Thus, proteolytic activity is often found to be upregulated in patient cohorts. In this minireview, we describe how bacterial proteases in the microbiome can display various bioactivities such as disruption of barrier function, degradation of host defense compounds, modulating inflammatory responses, and allowing for microbial movement. We focus on the gut, skin, vaginal, and urinary microbiomes and describe how specific bacterial organisms have proteolytic activities that can exacerbate or lead to human diseases.

The epidermal growth factor receptor (EGFR), a protein located on the cell surface, belongs to the tyrosine kinase family and plays a crucial role in cell development and proliferation. Abnormal expression or mutations in the EGFR gene can lead to non–small cell lung cancer. Although established EGFR inhibitors have been effective in the treatment of cancer, they are associated with several side effects. As a result, there is an urgent need to develop novel EGFR inhibitors that can effectively target the receptor while causing no adverse side effects. In this study, a series of bis(6‐amino‐1,3‐dimethylpyrimidine‐2,4(1H,3H)‐dione derivatives was synthesized in water at room temperature without the use of any catalyst, and pharmaceutical properties are investigated. Computational methods, including density functional theory (DFT), molecular docking, and molecular dynamics (MD), were utilized to investigate the chemical properties, drug‐like characteristics, and anticancer potential of the molecule. Quantum chemical calculations indicated that the molecules are relatively stable and exhibit significant electrophilic properties. The analysis of HOMO‐LUMO contour maps was conducted to illustrate charge density distributions that may be associated with biological activity. Docking studies with EGFR enzymes indicated that all compounds demonstrated favorable binding affinities, with docking scores ranging from −4.412 to −6.158 kcal/mol. Particularly, Compound 3f, with an energy of −6.158 kcal/mol, showed the best binding affinity, outperforming the native ligand, which had a docking score of −5.076 kcal/mol. The stability of the EGFR‐3f complex is significantly enhanced by the formation of five conventional hydrogen bonds and one carbon–hydrogen bond in ligand–protein interactions. MD simulations, which included analyses such as root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration (rGyr), molecular surface area (MolSA), and polar surface area (PSA), were conducted on the EGFR‐3f complex. It was found that the EGFR‐3f complex is stable, and the results show that Compound 3f has a strong interaction with the target enzyme.

The alveolins are a family of intermediate filament‐like proteins that form cytoskeletal structures in both free‐living and parasitic members of the alveolate kingdom. Despite their important functions, the alveolins’ biochemical properties and organizing principles are still poorly understood. Here, we characterize four alveolins of Plasmodium falciparum, the deadliest malaria parasite, to understand how alveolin domains mediate protein–protein interactions and highly specific recruitment to substructures of the cytoskeleton. Unexpectedly, we uncover variable dependence on alveolin domains for each substructure rather than an overarching mechanism. While PfIMC1e requires 1f to be sequentially recruited to the basal complex, PfIMC1c and PfIMC1g do not require interactions with each other to localize properly to the inner membrane complex. Moreover, alveolin domains are not interchangeable—they contain unique signatures for specialized localization. Finally, we identify a region outside the alveolin domain of PfIMC1e that is important for basal complex recruitment. These results provide direct evidence that alveolin domains mediate both alveolin–alveolin interactions and compartment‐specific localization.

Trachelomonas hispida var. coronata is one of the most widely reported varieties of T. hispida from water bodies worldwide. The specimens of this variety, apart from their species‐specific features, such as an ellipsoidal lorica covered with strong, sharp spines, have a crown consisting of long spines surrounding the apical pore opening. The process of lorica formation is poorly understood, and in the few studies dealing with this topic, results indicate that these taxa and the entire species can produce lorica completely devoid of spines, a diacritic feature of the species. In our study, we observed in detail the formation process of the lorica in this taxon under different chemical conditions in relation to the concentration of the basic elements, Fe and Mn, which are saturated in trachelomonad lorica. The results showed that in the Fe‐enriched medium, monads formed delicate, porous, spiny envelopes, whereas in the Mn‐enriched medium, the loricae were more solid and less porous and had weaker developed spines; rather, they were in the form of short papillae. Differences were also observed in the structure surrounding the apical pore, which was developed differently in the two sets of media modifications (Fe‐ or Mn‐supplemented media). We also observed different elemental compositions and colouration of loricae of cells growing in different media. This revealed that the features considered during the process of species identification are very unstable making the entire exercise highly complicated. Our research also shows that a broad discussion of the problem should be undertaken, and modern methods must be developed to unravel the complexities not only within the species but also within the entire Trachelomonas.

Bacillales is an order of Gram‐positive bacteria comprising well‐known genera such as Staphylococcus, Listeria, and Bacillus. Staphylococcus aureus has been shown to cause various types of tissue and systemic infections in humans and livestock. Some isolates can be highly resistant to antibiotics hence the need to identify better drugs and drug targets. Previous studies have shown that (E)‐N‐(4‐fluorophenyl)‐2‐phenylethene sulfonamide (FPSS) could inhibit activities of a bacteria‐specific transcription factor sigma B. Sigma B regulons in Bacillus subtilis and in Listeria monocytogenes, including virulence factor genes in L. monocytogenes, were decreased. Since sigma B is also presented in S. aureus, we initially postulated the use of FPSS to target S. aureus sigma B activity and to attenuate its virulence gene expression. Surprisingly, qRT‐PCR results revealed that FPSS induced the expression of sigma B‐dependent gene asp23. RNAseq results showed 39 S. aureus genes were affected by FPSS (37 genes were upregulated and two genes were downregulated). FPSS had no effect on expression of sigB operon in S. aureus. Therefore, we hypothesized that the effects of FPSS on sigma B regulons in L. monocytogenes, B. subtilis, and S. aureus were different. FPSS may target the expression of upstream regulators of sigma B (Rsbs), particularly the stressosome proteins which are lacking in S. aureus. Indeed, in L. monocytogenes, FPSS significantly increased the rsbR expression and could, thereby, dampen the sigma B activity and downregulate the expression of sigma B‐dependent virulence genes in L. monocytogenes. This study proposes a narrower spectrum of FPSS application to listerial infections, and not staphylococcal infections.

Apilactobacillus kosoi (A. kosoi) 10H is a fructophilic lactic acid bacterium found in vegetable sugar fermentation liquid. It has been shown to possess high intestinal immunostimulatory activity. In this study, we investigated the potential for A. kosoi 10H heat‐killed cells and their components to convert the phenotype of immunosuppressive M2 macrophages to immunostimulatory M1 macrophages. We further investigated the ability of induction of apoptosis of cancer cell lines by macrophage culture medium in the presence of A. kosoi 10H heat‐killed cells. After induction of macrophage differentiation by phorbol 12‐myristate 13‐acetate in a THP‐1 human monocyte–derived cell line, immunosuppressive M2 macrophages were induced by interleukin‐6 (IL‐6). When A. kosoi 10H heat‐killed cells were brought into contact with IL‐6‐induced M2 macrophages, the expression of CD163 and CD209, which are M2 macrophage markers, markedly decreased, and the expression of CD80, human leukocyte antigen‐DR isotype (HLA‐DR), and tumor necrosis factor‐α (TNF‐α), markers of M1 macrophages, markedly increased. A similar effect was observed with a water‐soluble extract and a hydrophobic liquid extract from A. kosoi 10H heat‐killed cells. In addition, when M2 macrophage culture medium that had been conditioned with A. kosoi 10H heat‐killed cells was added to the MCF‐7 breast cancer cell line, we found that apoptosis was induced. TNF‐α in the medium conditioned with macrophages cultured in the presence of A. kosoi 10H heat‐killed cells was found to be partially responsible for MCF‐7 cell death. This study presents basic data on the potential to use lactic acid bacteria to improve the cancer microenvironment.