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UNLABELLED: Cryptic conjugative plasmids lack antibiotic-resistance genes (ARGs). These plasmids can capture ARGs from the vast pool of the environmental metagenome, but the mechanism to recruit ARGs remains to be elucidated. To investigate the recruitment of ARGs by a cryptic plasmid, we sequenced and conducted mating experiments with Escherichia coli SW4848 (collected from a lake) that has a cryptic IncX (IncX4) plasmid and an IncF (IncFII/IncFIIB) plasmid with five genes that confer resistance to aminoglycosides (strA and strB), sulfonamides (sul2), tetracycline [tet(A)], and trimethoprim (dfrA5). In a conjugation experiment, a novel hybrid Tn21/Tn1721 transposon of 22,570 bp (designated Tn7714) carrying the five ARG mobilized spontaneously from the IncF plasmid to the cryptic IncX plasmid. The IncF plasmid was found to be conjugative when it was electroporated into E. coli DH10B (without the IncX plasmid). Two parallel conjugations with the IncF and the new IncX (carrying the novel Tn7714 transposon) plasmids in two separate E. coli DH10B as donors and E. coli J53 as the recipient revealed that the conjugation rate of the new IncX plasmid (with the novel Tn7714 transposon and five ARGs) is more than two orders of magnitude larger than the IncF plasmid. For the first time, this study shows experimental evidence that cryptic environmental plasmids can capture and transfer transposons with ARGs to other bacteria, creating novel multidrug-resistant conjugative plasmids with higher dispersion potential. IMPORTANCE: Cryptic conjugative plasmids are extrachromosomal DNA molecules without antibiotic-resistance genes (ARGs). Environmental bacteria carrying cryptic plasmids with a high conjugation rate threaten public health because they can capture clinically relevant ARGs and rapidly spread them to pathogenic bacteria. However, the mechanism to recruit ARG by cryptic conjugative plasmids in environmental bacteria has not been observed experimentally. Here, we document the first translocation of a transposon with multiple clinically relevant ARGs to a cryptic environmental conjugative plasmid. The new multidrug-resistant conjugative plasmid has a conjugation rate that is two orders of magnitude higher than the original plasmid that carries the ARG (i.e., the new plasmid from the environment can spread ARG more than two orders of magnitude faster). Our work illustrates the importance of studying the mobilization of ARGs in environmental bacteria. It sheds light on how cryptic conjugative plasmids recruit ARGs, a phenomenon at the root of the antibiotic crisis.

We determine which bipartite graphs embedded in a torus are move-reduced. In addition, we classify equivalence classes of such move-reduced graphs under square/spider moves. This extends the class of minimal graphs on a torus studied by Goncharov–Kenyon, and gives a toric analog of Postnikov’s and Thurston’s results on a disk.

A conjugation on a Hilbert space $ \mathcal {H} $ H means an antilinear bounded operator that the squares to the identity, which generalizes the traditional conjugation on complex Euclidean spaces. In this paper, with the help of normalized conjugation we introduce the notion of conjugate phase retrieval on general Hilbert spaces. We characterize frames that do conjugate phase retrieval; prove that every conjugate phase retrieval frame for $ \mathcal {H} $ H consisting of all real vectors has the complement property in $ \mathcal {H} $ H , and the converse is true if $ dim({\mathcal {H}})\leq ~2 $ dim (H) ≤ 2 ; and also prove that a small perturbation of conjugate phase retrieval frame still gives a conjugate phase retrieval frame if $ dim({\mathcal {H}}) \lt \infty $ dim (H) < ∞ , but it is false if $ dim({\mathcal H})=\infty $ dim (H) = ∞. [ABSTRACT FROM AUTHOR]

Bacterial conjugation, wherein DNA is transferred between cells through direct contact, is highly prevalent in complex microbial communities and is responsible for spreading myriad genes related to human and environmental health. Despite their importance, much remains unknown regarding the mechanisms driving the spread and persistence of these plasmids in situ. Studies have demonstrated that transferring, acquiring, and maintaining a plasmid imposes a significant metabolic burden on the host. Simultaneously, emerging evidence suggests that the presence of a conjugative plasmid can also provide both obvious and unexpected benefits to their host and local community. Combined, this highlights a continuous cost‐benefit tradeoff at the population level, likely contributing to overall plasmid abundance and long‐term persistence. Yet, while the metabolic burdens of plasmid conjugation, and their causes, are widely studied, their attendant potential advantages are less clear. Here, we summarize current perspectives on conjugative plasmids’ metabolic burden and then highlight the lesser‐appreciated yet critical benefits that plasmid‐mediated metabolic burdens may provide. We argue that this largely unexplored tradeoff is critical to both a fundamental theory of microbial populations and engineering applications and therefore warrants further detailed study. [ABSTRACT FROM AUTHOR]

Conjugative dissemination of mobile genetic elements (MGEs) among bacteria is initiated by assembly of the relaxosome at the MGE's origin‐of‐transfer (oriT) sequence. A critical but poorly defined step of relaxosome assembly involves recruitment of the catalytic relaxase to its DNA strand‐specific nicking site within oriT. Here, we present evidence by AlphaFold modeling, affinity pulldowns, and in vivo site‐directed photocrosslinking that the TraK Ribbon–Helix–Helix DNA‐binding protein recruits TraI to oriT through a dynamic interaction in which TraI's C‐terminal unstructured domain (TraICTD) wraps around TraK's C‐proximal tetramerization domain. Upon relaxosome assembly, conformational changes disrupt this contact, and TraICTD instead self‐associates as a prerequisite for relaxase catalytic functions or substrate engagement with the transfer channel. These findings delineate key early‐stage processing reactions required for conjugative dissemination of a model MGE. [ABSTRACT FROM AUTHOR]

A wide diversity of Mobile Genetic Elements (MGEs) are able to transfer host DNA. MGE-mediated 'sex' can be of significant influence on host evolution. Many instances of host DNA transfer are likely to be transient and non-adaptive. Evidence of MGE domestication suggests that host benefits of MGE-mediated sex could exist. The prokaryote world is replete with mobile genetic elements (MGEs) – self-replicating entities that can move within and between their hosts. Many MGEs not only transfer their own DNA to new hosts but also transfer host DNA located elsewhere on the chromosome in the process. This could potentially lead to indirect benefits to the host when the resulting increase in chromosomal variation results in more efficient natural selection. We review the diverse ways in which MGEs promote the transfer of host DNA and explore the benefits and costs to MGEs and hosts. In many cases, MGE-mediated transfer of host DNA might not be selected for because of a sex function, but evidence of MGE domestication suggests that there may be host benefits of MGE-mediated sex. [ABSTRACT FROM AUTHOR]

Horizontal gene transfer plays a key role in the global dissemination of antimicrobial resistance (AMR). AMR genes are often carried on self-transmissible plasmids, which are shared amongst bacteria primarily by conjugation. Antibiotic use has been a well-established driver of the emergence and spread of AMR. However, the impact of commonly used non-antibiotic compounds and environmental pollutants on AMR spread has been largely overlooked. Recent studies found common prescription and over-the-counter drugs, artificial sweeteners, food preservatives, and environmental pollutants, can increase the conjugative transfer of AMR plasmids. The potential mechanisms by which these compounds promote plasmid transmission include increased membrane permeability, upregulation of plasmid transfer genes, formation of reactive oxygen species, and SOS response gene induction. Many questions remain around the impact of most non-antibiotic compounds on AMR plasmid conjugation in clinical isolates and the long-term impact on AMR dissemination. By elucidating the role of routinely used pharmaceuticals, food additives, and pollutants in the dissemination of AMR, action can be taken to mitigate their impact by closely monitoring use and disposal. This review will discuss recent progress on understanding the influence of non-antibiotic compounds on plasmid transmission, the mechanisms by which they promote transfer, and the level of risk they pose. [ABSTRACT FROM AUTHOR]

Perfluoroalkyl sulfonamides (FASAs) and other FASA‐based per‐ and polyfluoroalkyl substances (PFASs) can transform into recalcitrant perfluoroalkyl sulfonates in vivo. We conducted high‐resolution mass spectrometry suspect screening of urine and tissues (kidney and liver) from mice dosed with an electrochemically fluorinated aqueous film‐forming foam (AFFF) to better understand the biological fate of AFFF‐associated precursors. The B6C3F1 mice were dosed at five levels (0, 0.05, 0.5, 1, and 5 mg kg−1 day−1) based on perfluorooctane sulfonate and perfluorooctanoate content of the AFFF mixture. Dosing continued for 10 days followed by a 6‐day depuration. Total oxidizable precursor assay of the AFFF suggested significant contributions from precursors with three to six perfluorinated carbons. We identified C4 to C6 FASAs and N‐glucuronidated FASAs (FASA‐N‐glus) excreted in urine collected throughout dosing and depuration. Based on normalized relative abundance, FASA‐N‐glus accounted for up to 33% of the total excreted FASAs in mouse urine, highlighting the importance of phase II metabolic conjugation as a route of excretion. High‐resolution mass spectrometry screening of liver and kidney tissue revealed accumulation of longer‐chain (C7 and C8) FASAs not detected in urine. Chain‐length–dependent conjugation of FASAs was also observed by incubating FASAs with mouse liver S9 fractions. Shorter‐chain (C4) FASAs conjugated to a much greater extent over a 120‐min incubation than longer‐chain (C8) FASAs. Overall, this study highlights the significance of N‐glucuronidation as an excretion mechanism for short‐chain FASAs and suggests that monitoring urine for FASA‐N‐glus could contribute to a better understanding of PFAS exposure, as FASAs and their conjugates are often overlooked by traditional biomonitoring studies. Environ Toxicol Chem 2024;43:2274–2284. © 2024 The Author(s). Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. [ABSTRACT FROM AUTHOR]

A delivery system was developed according to maillard reaction using mung bean protein and maltodextrin for encapsulation and sustain release of curcumin. The ultrasound assisted (150 W, 80 °C, 10 min) and classic wet heating (80 °C, 60 min), were used to prepare conjugates at three different ratios of Mung bean protein isolate to maltodextrin. Degree of conjugation was measured using OPA method Uv-visible spectroscopy was used to estimate the final products of maillard reaction. Different amounts of curcumin (0, 0.2, 0.4 and 0.6 mg. mL-1 was loaded using ethanol and change in pH. Primary analysis showed that the optimized samples were obtained when 0. 4 mg. mL-1 of curcumin was encapsulated using pH change method. FTIR spectra confirmed the conjugation of the MPI and MD and showed the electrostatic and hydrophobic interactions as well as hydrogen bonding are the main reasons of conjugates stability and curcumin encapsulation. The prepared curcumin containing conjugates under optimized method showed the higher DPPH radical scavenging activity. Our results showed that the release rate of encapsulated curcumin under simulated condition of gastrointestinal tract (GIT) was controlled and lower than that which encapsulated in mung bean protein. [ABSTRACT FROM AUTHOR]

Quaternion algebra ℍ is a noncommutative associative algebra. In recent years, quaternionic Fourier analysis has received increasing attention due to its applications in signal analysis and image processing. This paper addresses conjugate phase retrieval problem in the quaternion Euclidean space ℍ M with M ≥ 2 . Write ℂ η = { ξ : ξ = ξ 0 + β ⁢ η , ξ 0 , β ∈ ℝ } for η ∈ { i , j , k } . We remark that not only ℂ η M -vectors cannot allow traditional conjugate phase retrieval in ℍ M , but also ℂ i M ∪ ℂ j M -complex vectors cannot allow phase retrieval in ℍ M . We are devoted to conjugate phase retrieval of ℂ i M ∪ ℂ j M -complex vectors in ℍ M , where "conjugate" is not the traditional conjugate. We introduce the notions of conjugation, maximal commutative subset and conjugate phase retrieval. Using the phase lifting techniques, we present some sufficient conditions on complex vectors allowing conjugate phase retrieval. And some examples are also provided to illustrate the generality of our theory. [ABSTRACT FROM AUTHOR]

Optoelectronic properties of linear π‐conjugated polymers/oligomers are of great importance for the fabrication of organic photonic and electronic devices. To this end, the π‐conjugated polymers/oligomers need to meet both optoelectronic and key structural properties in order to fulfill their implementation as active components. In particular, they need to possess low bandgap and high thermal, conformational, and photochemical stabilities. So far, several strategies have been developed to attain such requirements including the covalent and non‐covalent rigidification concepts of the π‐conjugated systems. On the basis of these findings, we describe herein the theoretical studies of novel series of covalently bridged derivatives demonstrating the benefits of the strategy. Comparison of these derivatives with compounds previously described in the literature highlights enhanced optoelectronic properties and behaviors that would be beneficial for the construction and development of new linear π‐conjugated polymers. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the diversity of conjugative and chromosomally integrated mobile genetic elements (cciMGEs) within six oral streptococci species. cciMGEs, including integrative and conjugative elements (ICEs) and integrative and mobilizable elements (IMEs), are stably maintained on the host cell chromosome; however, under certain conditions, they are able to excise, form extrachromosomal circles, and transfer via a conjugation apparatus. Many cciMGEs encode "cargo" functions that aid survival in new niches and evolve new antimicrobial resistance or virulence properties, whereas others have been shown to influence host bacterial physiology. Here, using a workflow employing preexisting bioinformatics tools, we analyzed 551 genomes for the presence of cciMGEs across six common health- and disease-associated oral streptococci. We identified 486 cciMGEs, 173 of which were ICEs and 233 of which were IMEs. The cciMGEs were diverse in size, cargo genes, and relaxase types. We identified several novel relaxase proteins and a widespread IME carrying a small multidrug resistance transporter. Additionally, we provide evidence that several of the bioinformatically predicted cciMGEs encoded within various Streptococcus mutans strains are capable of excision and circularization, a critical step for cciMGE conjugative transfer. These findings highlight the significance and potential impact of MGEs in shaping the genetic landscape, pathogenicity, and antimicrobial resistance profiles of the oral microbiota. IMPORTANCE Oral streptococci are important players in the oral microbiome, influencing both health and disease states within dental bacterial communities. Evolutionary adaptation, shaped in a major part by the horizontal transfer of genes, is essential for their survival in the oral cavity and within new environments. Conjugation is a significant driver of horizontal gene transfer; however, there is limited information regarding this process in oral bacteria. This study utilizes publicly available genome sequences to identify conjugative and chromosomally integrated mobile genetic elements (cciMGEs) across several species of oral streptococci and presents the preliminary characterization of these elements. Our findings significantly enhance our understanding of the mobile genomic landscape of oral streptococci critical for human health, with valuable insights into how cciMGEs might influence the survival and pathogenesis of these bacteria in the oral microbiome. [ABSTRACT FROM AUTHOR]

Many multidrug-resistant (MDR) bacteria have evolved through the accumulation of antibiotic resistance genes (ARGs). Although the potential risk of probiotics as reservoirs of ARGs has been recognized, strategies for blocking the transfer of ARGs while using probiotics have rarely been explored. The probiotic Escherichia coli Nissle 1917 (EcN) has long been used for treating intestinal diseases. Here, we demonstrate frequent transfer of ARGs into EcN both in vitro and in vivo, raising concerns about its potential risk of accumulating antibiotic resistance. Given that no CRISPR-Cas system was found in natural EcN, we integrated the type I-E CRISPR-Cas3 system derived from E. coli BW25113 into EcN. The engineered EcN was able to efficiently cleave multiple ARGs [i.e., mcr-1, blaNDM-1, and tet(X)] encoding enzymes for degrading last-resort antibiotics. Through co-incubation of EcN expressing Cas3-Cascade and that expressing Cas9, we showed that the growth of the former strain outcompeted the latter strain, demonstrating a better clinical application prospect of EcN expressing the type I-E CRISPR-Cas3 system. In the intestine of a model animal (i.e., zebrafish), the engineered EcN exhibited immunity against the transfer of CRISPR-targeted ARGs. Our work equips EcN with immunity against the transfer of multiple ARGs by exploiting the exogenous type I-E CRISPR-Cas3 system, thereby reducing the risk of the spread of ARGs while using it as a probiotic chassis for generating living therapeutics. IMPORTANCE To reduce the development of antibiotic resistance, probiotics have been considered as a substitute for antibiotics. However, probiotics themselves are reservoirs of antibiotic resistance genes (ARGs). This study introduces a new strategy for limiting the spread of ARGs by engineering the typical probiotic strain Escherichia coli Nissle 1917 (EcN), which has been used for treating intestinal diseases and developed as living therapeutics. We also demonstrate that the type I CRISPR-Cas system imposes a lower growth burden than the type II CRISPR-Cas system, highlighting its promising clinical application potential. Our work not only provides a new strategy for restricting the transfer of ARGs while using probiotics but also enriches the genetic engineering toolbox of EcN, paving the way for the safe use and development of probiotics as living therapeutics. [ABSTRACT FROM AUTHOR]

Streptavidin is a tetrameric protein with high specificity and affinity for biotin. The interaction between avidin and biotin has become a valuable tool in nanotechnology. In recent years, the site‐specific biotin modification of proteins using biotin ligases, such as BirA, has attracted attention. This study established an in vivo method for achieving the complete biotinylation of target proteins using a single plasmid co‐expressing BirA and its target proteins. Specifically, a biotin‐modified protein was produced in Escherichia coli strain BL21(DE3) using a single plasmid containing genes encoding both BirA and a protein fused to BirA's substrate sequence, Avitag. This approach simplifies the production of biotinylated proteins in E. coli and allows the creation of various biotinylated protein types through gene replacement. Furthermore, the biotin modification rate of the obtained target protein could be evaluated using Native‐PAGE without performing complicated isolation operations of biotinylated proteins. In Native‐PAGE, biotin‐modified proteins and unmodified proteins were confirmed as clearly different bands, and it was possible to easily derive the modification rate from the respective band intensities. [ABSTRACT FROM AUTHOR]

Two regioisomeric extended tetrathiafulvalenes (TTFs) incorporating diindeno‐fused corannulene spacers were synthesized from suitable dione precursors using Horner‐Wadsworth‐Emmons olefination reactions. Both compounds are strong chromophores with a longest‐wavelength absorption that exhibited some degree of charge‐transfer character as inferred from solvatochromic behaviors and frontier orbital calculations. The compounds underwent two reversible one‐electron oxidations and a quasi‐reversible third oxidation during cyclic voltammetry conditions. The mono‐ and dications had characteristic NIR absorptions and both were EPR active. The dications with potential diradicaloid characters underwent reactions when generated in bulk. [ABSTRACT FROM AUTHOR]

Background/Objectives: Antibiotic resistance gene (ARG) spread is driven by horizontal gene transfer (HGT). Ciliated protozoa may contribute to this process, as their predation has been shown to facilitate HGT in certain bacteria. Here, this phenomenon was further investigated using A. salmonicida subsp. salmonicida. This fish pathogen bears an extensive and dynamic plasmidome, suggesting a high potential for HGT. Methods: A. salmonicida strains carrying one of three conjugative plasmids bearing ARGs (pSN254b, pRAS1b or pAsa4b) were cocultured with a recipient, either A. salmonicida, E. coli or A. hydrophila. Conjugation rates were assessed in the presence and absence of the ciliate Tetrahymena borealis. PCR genotyping confirmed the acquisition of the conjugative plasmids and was used to verify the mobilization of other plasmids. Results: The basal rate of conjugation observed was high. Under the conditions studied, ciliate predation did not appear to influence the conjugation rate, except at higher proportions of ciliates, which typically hampered conjugation. Microscopy revealed that most bacteria were digested in these conditions. PCR screening demonstrated that small mobilizable plasmids from A. salmonicida (pAsa1, pAsa2, pAsa3, and pAsal1) were acquired by the recipients along with the conjugative plasmids, with a slight effect of the ciliates in some donor/recipient cell combination. Conclusions: These results highlight how A. salmonicida can conjugate efficiently with different species and how complex its relationship with ciliates is. [ABSTRACT FROM AUTHOR]

Purpose: This study aims to enhance the targeting of erlotinib by grafting saccharides—galactose, pectin, and chitosan via oxalyl chloride-mediated processes. The objective is to evaluate the potential of these saccharide derivatives as ligands for targeted drug delivery, particularly to improve therapeutic efficacy in cancer treatments. Methods: Saccharides (galactose, pectin, and chitosan) were chemically grafted onto erlotinib, forming conjugates. Characterization was conducted using 1H NMR, differential scanning calorimetry (DSC), and Fourier-transform infrared spectroscopy (FTIR). The conjugates’ drug delivery efficiency was tested using A549 cell lines to assess their efficacy in targeting cancer cells. Results: Characterization confirmed the successful grafting of saccharides onto erlotinib, as evidenced by 1H NMR, DSC, and FTIR. The in vitro drug release study demonstrated enhanced targeting efficiency of the saccharide-erlotinib conjugates, with improved cell-specific distribution and potential therapeutic benefits. Conclusions: Grafting saccharides onto erlotinib offers a promising approach to enhance drug absorption, reduce degradation, and minimize side effects in cancer therapy. This targeted drug delivery strategy represents a significant advancement in biomedical engineering for optimizing cancer treatment efficacy. [ABSTRACT FROM AUTHOR]

AbstractAntimicrobial resistance (AMR) has been recognized as an important health crisis in the twenty first century. Type IV secretion systems (T4SSs) play key roles in the dissemination of AMR plasmids. Novel strategies that combat AMR problem by targeting T4SS sprung up in recent years. Here, we focus on the strategy of male-specific phages that could target and kill bacteria carrying conjugative AMR plasmids encoding T4SSs. We reviewed the recent advances in male-specific phages, including anti-conjugation mechanisms, clinical isolation and identification methods, classification and characteristics, in vitro and in vivo anti-conjugation efficacy and improving strategies. Male-specific phages constitute exciting candidates for developing sustainable anti-resistance biocontrol applications. [ABSTRACT FROM AUTHOR]

Introduction: Horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs) is one of the primary routes of antimicrobial resistance (AMR) dissemination. In the One Health context, tracking the spread of mobile genetic elements (MGEs) carrying ARGs in agri-food ecosystems is pivotal in understanding AMR diffusion and estimating potential risks for human health. So far, little attention has been devoted to plant niches; hence, this study aimed to evaluate the conjugal transfer of ARGs to the bacterial community associated with the plant rhizosphere, a hotspot for microbial abundance and activity in the soil. We simulated a source of AMR determinants that could enter the food chain via plants through irrigation. Methods: Among the bacterial strains isolated from treated wastewater, the strain Klebsiella variicola EEF15 was selected as an ARG donor because of the relevance of Enterobacteriaceae in the AMR context and the One Health framework. The strain ability to recolonize lettuce, chosen as a model for vegetables that were consumed raw, was assessed by a rifampicin resistant mutant. K. variicola EEF15 was genetically manipulated to track the conjugal transfer of the broad host range plasmid pKJK5 containing a fluorescent marker gene to the natural rhizosphere microbiome obtained from lettuce plants. Transconjugants were sorted by fluorescent protein expression and identified through 16S rRNA gene amplicon sequencing. Results and discussion: K. variicola EEF15 was able to colonize the lettuce rhizosphere and inhabit its leaf endosphere 7 days past bacterial administration. Fluorescence stereomicroscopy revealed plasmid transfer at a frequency of 10−3; cell sorting allowed the selection of the transconjugants. The conjugation rates and the strain’s ability to colonize the plant rhizosphere and leaf endosphere make strain EEF15::lacIq -pLpp-mCherry-gmR with pKJK5::Plac::gfp an interesting candidate to study ARG spread in the agri-food ecosystem. Future studies taking advantage of additional environmental donor strains could provide a comprehensive snapshot of AMR spread in the One Health context. [ABSTRACT FROM AUTHOR]

Objectives: Subinhibitory concentration of antibiotics in the environment is an important risk factor for the horizontal transmission of antibiotic resistance genes (ARGs). The signaling mechanism of resistance gene transmission remains unknown. The aim of this study was to investigate whether indole could be used as a molecular signal to help the spread of ARGs under the stress of subinhibitory concentrations of antibiotics. Methods: The effect of indole on conjugation frequency was investigated through a conjugation test, and its effect on the Type IV secretion system and pili gene expression of E. coli was observed. Meanwhile, we were investigating the trend of changes in indole regulatory factors ibpA, tnaA, and concentration pumps. Subsequently, we predicted the receptors that specifically bind to indole. Finally, our study focused on elucidating the regulatory mechanism of indole synthesis. Results: Conjugate frequency was significantly increased under 1/5MIC concentration cefotaxime stress. The transferred ARGs were blaCTX-M and foxA. The mobile plasmid was IncY or IncI2. Meanwhile, the concentration of endogenous indole was also significantly increased. And, surprisingly, inhibition of endogenous indole production resulted in a significant decrease in conjugate frequency. However, the conjugate frequency increased once again when the strains reacquired the exogenous indole. Furthermore, the fluctuation trends of indole‐regulated factor (ibpA, tnaA) mRNA and concentration pumps (acrEF, mtr) mRNA consistently with that of indole. Then, we found that the receptors of indole may be four targets of TCSs: CreC, PhoB, AtoC, and UhpA. More than that, when strains retrieved the exogenous indole again, the mRNA levels of T4SS (virB2, virB6, and virD4) and pppA (coding Pili precursor) genes significantly increased. This indicates that there is a close relationship between indole and conjugated channels, which are necessary for horizontal transfer of genetic material. And then, the trends of indole and tnaA mRNA were consistent with that of ibpA (one of SOS response). So, this result confirmed that indole was regulated by SOS response under subinhibitory concentrations of antibiotics. Conclusions: It is always known that subinhibitory concentrations of antibiotics stimulate an SOS response in E. coli, which helps in the horizontal spread of ARGs by modulating indole. [ABSTRACT FROM AUTHOR]

In recent decades, there has been a rapid increase in the prevalence of multidrug‐resistant pathogens, posing a challenge to modern antibiotic‐based medicine. This has highlighted the need for novel treatments that can specifically affect the target microorganism without disturbing other co‐inhabiting species, thus preventing the development of dysbiosis in treated patients. Moreover, there is a pressing demand for tools to effectively manipulate complex microbial populations. One of the approaches suggested to address both issues was to use conjugation as a tool to modify the microbiome by either editing the genome of specific bacterial species and/or the removal of certain taxonomic groups. Conjugation involves the transfer of DNA from one bacterium to another, which opens up the possibility of introducing, modifying or deleting specific genes in the recipient. In response to this proposal, there has been a significant increase in the number of studies using this method for gene delivery in bacterial populations. This MicroReview aims to provide a detailed overview on the use of conjugation for microbiome engineering, and at the same time, to initiate a discussion on the potential, limitations and possible future directions of this approach. [ABSTRACT FROM AUTHOR]

Simple Summary: The bacterium Yersinia ruckeri causes enteric redmouth disease, which affects salmonid fish, including farmed fish such as Atlantic salmon and rainbow trout, leading to significant commercial losses. In this study, we investigated a plasmid (an independently replicating DNA molecule) called pYR4 that is suspected of aiding Y. ruckeri in causing disease by producing adhesive structures known as type 4 pili. We identified a toxin–antitoxin locus on the plasmid, higBA, which we hypothesized might prevent loss of the plasmid from the bacterial cell through "addiction" to the antitoxin HigA. HigA is quickly degraded if the plasmid is lost, whereas the toxin HigB is stable and leads to cell death in the absence of HigA. We were able to demonstrate that HigBA is a functional toxin–antitoxin system and that stabilizes pYR4. We further showed that pYR4 can transfer between bacteria by a process known as conjugation. However, loss of the pYR4 plasmid did not have an effect in a simple insect larva infection model. Further work will have to be undertaken to evaluate the role of pYR4 in enteric redmouth disease. These results will help in understanding and ultimately managing a serious disease of commercially important fish. The bacterium Yersinia ruckeri causes enteric redmouth disease in salmonids and hence has substantial economic implications for the farmed fish industry. The Norwegian Y. ruckeri outbreak isolate NVH_3758 carries a relatively uncharacterized plasmid, pYR4, which encodes both type 4 pili and a type 4 secretion system. In this study, we demonstrate that pYR4 does not impose a growth burden on the Y. ruckeri host bacterium, nor does the plasmid contribute to twitching motility (an indicator of type 4 pilus function) or virulence in a Galleria mellonella larval model of infection. However, we show that pYR4 is conjugative. We also reveal, through mutagenesis, that pYR4 encodes a functional post-segregational killing system, HigBA, that is responsible for plasmid maintenance within Y. ruckeri. This is the first toxin–antitoxin system to be characterized for this organism. Whilst further work is needed to elucidate the virulence role of pYR4 and whether it contributes to bacterial disease under non-laboratory conditions, our results suggest that the plasmid possesses substantial stability and transfer mechanisms that imply importance within the organism. These results add to our understanding of the mobile genetic elements and evolutionary trajectory of Y. ruckeri as an important commercial pathogen, with consequences for human food production. [ABSTRACT FROM AUTHOR]

Summary: Plum protein isolate obtained from plum cake after oil extraction had protein content over 90% and amino acid composition of 36% which about 13% were essential amino acids. Evaluation of functional properties of conjugates obtained by interaction between plum protein isolate and caffeic acid, in three different concentrations, was carried out in this study. The conjugation of proteins with phenols forms complexes of higher molecular weights as a result of cross‐linking. The solubility of obtained conjugates was increased with the increase a concentration of caffeic acid at alkaline pH (8–10) for about 50%. Moreover, emulsion properties also were improved by conjugation with caffeic acid. In vitro digestion is considered to characterise and understand 'fate' of food components after passing gastrointestinal conditions. The obtained conjugates were digestible and have a good bioavailability of bioactive compounds. [ABSTRACT FROM AUTHOR]

Combinations of biological macromolecules can provide researchers with precise control and unique methods for regulating, studying, and manipulating cellular processes. For instance, combining the unmatched encodability afforded by nucleic acids with the diverse functionality of proteins has transformed our approach to solving several problems in chemical biology. Despite these benefits, there remains a need for new methods to site-specifically generate conjugates between different classes of biomolecules. Here we present a fully enzymatic strategy for combining nucleic acids and proteins using SNAP-tag and RNA-TAG (transglycosylation at guanosine) technologies via a bifunctional preQ1-benzylguanine small molecule probe. We demonstrate the robust ability of this technology to assemble site-specific SNAP-tag - RNA conjugates with RNAs of varying length and use our conjugation strategy to recruit an endonuclease to an RNA of interest for targeted degradation. We foresee that combining SNAP-tag and RNA-TAG will facilitate researchers to predictably engineer novel macromolecular complexes.

The purpose of this study was to investigate the effect of neutrophils and their antimicrobial factors, hydrogen peroxide and defensin α, on the biofilm biomass, the viability of bacteria in the biofilm and the efficiency of conjugative transfer of the pOX38:Cm plasmid from the E. coli N4i pOX38:Cm strain into different E. coli strains (commensal К12 TG1 and uropathogenic DL82, R32 and R45). The biofilm of the recipient E. coli TG1 with the donor E. coli N4i pOX38:Cm increased when 10⁵ cells/ml of neutrophils were added compared to the control, while the biofilm biomass of the uropathogenic E. coli recipient strains DL82/E. coli R45 with the donor E. coli N4i pOX38:Cm decreased when 10⁶/10⁴–10⁶ cells/ml of neutrophils were added, respectively. The survival of recipient E. coli TG1 cells and transconjugants in the biofilm was, compared to the control, higher when 10⁴, 10⁵, 10⁶ cells/ml of neutrophils were added. The addition of 0.1 mM H₂O₂ increased biofilm formation of E. coli DL82 and E. coli R45, and addition of 0.5 mM H₂O₂ reduced biofilm formation of E. coli DL82, while 0.5 mM or 2.5 mM reduced the E. coli R45 bacterial biofilm biomass in the conjugative mixture. The frequency of the pOX38:Cm conjugative transfer was lower in the presence of 2.5 mM H₂O₂ in the N4i pOX38:Cm × DL82 biofilm, and also in the presence of 0.5 and 2.5 mM H₂O₂ in the N4i pOX38:Cm × R45 biofilm, compared to the control. The frequency of pOX38:Cm conjugation from the donor E. coli N4i pOX38:Cm into E. coli DL82 decreased, when 5 or 25 ng/ml defensin α were added and the conjugation frequency in the mating mixture N4i pOX38:Cm×R45 decreased, when 5 ng/ml were added, while, when 25 ng/ml of defensin α were added it increased.

Mating assays are common laboratory experiments for measuring the conjugation frequency, i.e. efficiency at which a plasmid transfers from a population of donor cells to a population of recipient cells. Selective plating remains a widely used quantification method to enumerate transconjugants at the end of such assays. However, conjugation frequencies may be inaccurately estimated because plasmid transfer can occur on transconjugant-selective plates rather than only during the intended mating duration. We investigated the influence of cell density on this phenomenon. We conducted mating experiments with IncPα plasmid RP4 harbored in Escherichia coli at a fixed cell density and mating conditions, inoculated a serial dilution of the mating mixture on transconjugant-selective plates or in transconjugant-selective broth, and compared the results to a model of cell-to-cell distance distribution. Our findings suggest that irrespective of the mating mode (liquid vs solid), the enumeration of transconjugants becomes significantly biased if the plated cell density exceeds 28 Colony Forming Unit (CFU)/mm2 (or 1.68•105 CFU/standard 9 cm Petri dish). This threshold is determined with a 95% confidence interval of ± 4 CFU/mm2 (± 2.46•104 CFU/standard 9 cm Petri dish). Liquid mating assays were more sensitive to this bias because the conjugation frequency of RP4 is several orders of magnitude lower in suspension compared to surface mating. Therefore, if selective plating is used, we recommend to plate at this density threshold and that negative controls are performed where donors and recipients are briefly mixed before plating at the same dilutions as for the actual mating assay. As an alternative, a liquid enumeration method can be utilized to increase the signal-to-noise ratio and allow for more accurate enumeration of transconjugants. [ABSTRACT FROM AUTHOR]

In the past two to three decades, synthetic glycoconjugate vaccines have shown great potential in the prevention of severe infections and protection of high‐risk populations. Conjugation of synthetic oligosaccharide haptens to carrier proteins is the key step for the vaccine preparation. In this review, the conjugation methods currently used in the synthesis of glycoconjugate vaccines from synthetic/homogeneous oligosaccharide haptens are summarized with the focus on the reaction conditions (pH and sugar/protein ratio) and performance. This information can help researchers choose the appropriate conjugation methods. Further research directions toward site‐specific conjugations and fully homogeneous glycoconjugate vaccines are also discussed. [ABSTRACT FROM AUTHOR]

Recently, for the enhancement of the therapeutic qualities of peptides, proteins, small molecules, antibodies, or oligonucleotides, the conjugation technique has immerged as a new domain in drug delivery systems. Conjugated medications have a longer half-life, greater stability, fewer side effects, enhanced solubility, reduced immunogenicity, and a specific targeting approach. The present review discusses various conjugation methods, comparing cleavable and non-cleavable linkers and current advancements in linker technology. It focuses on the molecular characteristics of each component of drug conjugations and linker chemistry techniques. The correlation between in vitro and in vivo studies of these conjugated systems has also been discussed. [ABSTRACT FROM AUTHOR]

• Comparative analysis of the physicochemical stability of interchain cysteine ADCs that had matched DAR and identical linkers but differently charged auristatin payloads (neutral vs. acidic terminal group). • ADCs with negatively charged vcMMAF payloads showed higher aggregation than those with neutral but more hydrophobic vcMMAE payloads, both under shaking stress and after 2 weeks of storage at 40 °C. • Conjugation with vcMMAF decreased the pI, net charge, and electrostatic repulsion of the ADC compared with the parent antibody (mAb). • Surface pressure and apparent second virial coefficient data correlated with the aggregation propensities of the mAb, ADC-vcMMAE, and ADC-vcMMAF. Antibody-drug conjugates (ADCs) tend to be less stable than their parent antibodies, which is often attributed to the hydrophobic nature of their drug payloads. This study investigated how the payload charge affects ADC stability by comparing two interchain cysteine ADCs that had matched drug-to-antibody ratios and identical linkers but differently charged auristatin payloads, vcMMAE (neutral) and vcMMAF (negative). Both ADCs exhibited higher aggregation than their parent antibody under shaking stress and thermal stress conditions. However, conjugation with vcMMAF increased the aggregation rates to a greater extent than conjugation with uncharged but more hydrophobic vcMMAE. Consistent with the payload logD values, ADC-vcMMAE showed the greatest increase in hydrophobicity but minor changes in charge compared with the parent antibody, as indicated by hydrophobic interaction chromatography and capillary electrophoresis data. In contrast, ADC-vcMMAF showed a decrease in net charge and isoelectric point along with an increase in charge heterogeneity. This charge alteration likely contributed to a reduced electrostatic repulsion and increased surface activity in ADC-vcMMAF, thus affecting its aggregation propensity. These findings suggest that not only the hydrophobicity of the payload, but also its charge should be considered as a critical factor affecting the stability of ADCs. [ABSTRACT FROM AUTHOR]

Antibiotic-resistant microorganisms have become a serious threat to public health, resulting in hospital infections, the majority of which are caused by commonly used urinary tract catheters. Strategies for preventing bacterial adhesion to the catheters' surfaces have been potentially shown as effective methods, such as coating thesurface with antimicrobial biomolecules. Here, novel antimicrobial peptides (AMPs) were designed as potential biomolecules to prevent antibiotic-resistant bacteria from binding to catheter surfaces. Thiolated AMPs were synthesized using solid-phase peptide synthesis (SPPS), and prep-HPLC was used to obtain AMPs with purity greater than 90%. On the other side, the silicone catheter surface was activated by UV/ozone treatment, followed by functionalization with allyl moieties for conjugation to the free thiol group of cystein in AMPs using thiol-ene click chemistry. Peptide-immobilized surfaces were found to become more resistant to bacterial adhesion while remaining biocompatible with mammalian cells. The presence and site of conjugation of peptide molecules were investigated by immobilizing them to catheter surfaces from both ends (C-Pep and Pep-C). It was clearly demonstrated that AMPs conjugated to the surface via theirN terminus have a higher antimicrobial activity. This strategy stands out for its effective conjugation of AMPs to silicone-based implant surfaces for the elimination of bacterial infections. [ABSTRACT FROM AUTHOR]

The Vilsmeier–Haack reaction of ketones with DMF and POCl3- produced 3-chloroacrylcarbaldehydes, which were converted into the corresponding (Z)-4-[(Z)- or (Z)-4-[(E)-3-chloroallylidene)-2-phenyloxazol-5(4 H)-ones] (azlactones) or their isomeric mixtures when heated with hippuric acid in propionic anhydride. It was shown that the alcoholysis products of these compounds, resulting from the opening of the oxazolone ring, undergo copper-catalyzed intramolecular cross-coupling reactions with the formation of methyl 1 H -pyrrole-2-carboxylates. Thus, a one-pot method was developed for the synthesis of methyl 1 H -pyrrole-2-carboxylates starting from 4-(3-chloroallylidene)-2-phenyloxazol-5(4 H)-ones. [ABSTRACT FROM AUTHOR]

Emerging pollutants such as microplastics in water environments readily accumulate microorganisms on their surfaces, forming biofilms and concentrating antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). Consequently, microplastic biofilms have attracted the attention of researchers. Horizontal gene transfer (HGT) of ARGs is one of the primary ways that bacteria acquire antibiotic resistance. Most studies focus on the effects of nanomaterials on suspended bacteria, but microplastic biofilms as hotspots for horizontal gene transfer also warrant significant investigation. This study primarily explored and compared the effects of nano-titanium dioxide on the conjugation transfer frequency of ARGs in suspended bacteria and microplastic biofilms. Nano-titanium dioxide could promote ARG conjugation in both suspended bacteria and microplastic biofilms, with a greater effect on the former. The mechanism involved nano-titanium dioxide promoting the production of reactive oxygen species (ROS) in suspended and biofilm bacteria, increasing the synthesis of outer membrane proteins, enhancing the cell membrane permeability, and elevating the expression levels of conjugation-related genes, thereby facilitating the conjugation transfer of ARGs. Biofilm bacteria, being heavily encased and protected by extracellular polymeric substances (EPS), exhibit greater resistance to external environmental pressure, resulting in the weaker impact of nano-titanium dioxide on biofilm bacteria compared to suspended bacteria. This study reveals the risk of ARG conjugation transfer within microplastic biofilms induced by nanomaterials, providing valuable insights into the risks of microplastic and antibiotic resistance dissemination in water environments. [ABSTRACT FROM AUTHOR]

An operator T ∈ L (H) is said to be C-normal if there exists a conjugation C on H such that the commutator [ (C T) # , C T ] equals zero, where [ R , S ] : = R S - S R and R # is a Hermitian adjont operator of R as in (1). If there exists a conjugation C with respect to which T ∈ L (H) is C-normal, then T is called a conjugation-normal operator. In this paper, we study properties of conjugation-normal operator matrices. In particular, we focus on the conjugation-normality of the component operators of operator matrices which are conjugation-normal. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of low concentration of Wenyang Tonglin Decoction (WTD) on the binding conditions of R45 plasmid conjugative transfer under liquid phase conjugation and its mechanism. Methods: Escherichia coli CP9 (R45) and Staphylococcus aureus RN450RF were cultured in medium containing WTD, and their minimum inhibitory concentration (MIC) values were obtained. Using promoter fusion technology, E. coli CP9 (R45) containing a promoter fusion was obtained. β-Galactosidase activity of TrfAp and TrbBp was tested, and the mRNA expression of regulatory factors (TrbA, KorA, and KorB) was detected by real-time fluorescent quantitative polymerase chain reaction. Results: The MIC of E. coli CP9 (R45) was 400 g/L and that of S. aureus RN450RF was 200 g/L. When the drug concentration in the culture medium was 200 g/L, the highest number of conjugants was (3.47 ±0.20) × 107 CFU/mL At 90 h of conjugation, the maximum number of conjugants was (1.15 ±0.06) × 108 CFU/mL When the initial bacterial concentration was 108 CFU/mL, the maximum number of conjugants was (3.47 ± 0.20) × 107 CFU/mL. When the drug concentration was 200 g/L, the β-galactosidase activity of TrfAp and TrbBp significantly increased; the relative quantification of TrbA, KorA and KorB were significantly inhibited. Conclusion: Low concentration of WTD promoted the development of bacterial resistance by affecting promoters and inhibiting the expression of regulatory factors. [ABSTRACT FROM AUTHOR]

Investigations of the nature and degree of antiaromaticity of cycloheptatrienyl anion derivatives using both experimental and computational tools are presented. The ground state of cycloheptatrienyl anion in the gas phase is triplet, planar and Baird‐aromatic. In DMSO, it assumes a singlet distorted allylic form with a paratropic ring current. The other derivatives in both phases assume either allylic or diallylic conformations depending on the substituent pattern. A combination of experimental and computational methods was used to determine the pKa values of 16 derivatives in DMSO, which ranged from 36 to −10.7. We revealed that the stronger stabilization of the anionic system, which correlates with acidity, does not necessarily imply a lower degree of antiaromaticity in terms of magnetic properties. Conversely, the substitution pattern first affects the geometry of the ring through the bulkiness of the substituents and their better conjugation with a more distorted system. Consequently, the distortion reduces the cyclic conjugation in the π‐system and thereby decreases the paratropic current in a magnetic field, which manifests itself as a decrease in the NICS. The triplet‐state geometries and magnetic properties are nearly independent on the substitution pattern, which is typical for simple aromatic systems. [ABSTRACT FROM AUTHOR]

We introduce the C-decomposition property for reducible bounded linear operators on a Hilbert space, and prove that an arbitrary idempotent operator has the C-decomposition property with respect to a particular space decomposition, which is related to Halmos’ two projections theory. Using this, we obtain a general explicit description for all the conjugations C such that a given idempotent operator is a C-projection. We also present a characterization of the ranges of C-projections for any conjugation C. [ABSTRACT FROM AUTHOR]

4,16‐para‐substituted [2.2]paracylophanes with naphthalene (PCP‐NAP), anthracene (PCP‐ANTH), and tetraphenylethylene (PCP‐TPE), as new through‐space conjugated dimers, were prepared by the Suzuki‐Miyaura cross‐coupling reaction of 4,16‐diboryl[2.2]paracyclophane and the corresponding bromo derivative using Pd(PPh3)4 as a catalyst and KOH as a base. The synthesized compounds were fully characterized by NMR and HR‐MS, and their photophysical and electrochemical properties were studied. The photoluminescence quantum yield of PCP‐NAP, PCP‐ANTH were determined to be 0.21, 0.50, and for PCP‐TPE in aggregated state 0.31, respectively. PCP‐TPE exhibited aggregation‐induced emission characteristics when the water fraction was greater than 50 % in THF/water mixtures. PCP‐ANTH and PCP‐TPE were also characterized by X‐ray crystallography, PCP‐ANTH crystallizes in centrosymmetric monoclinic space group C2/c and PCP‐TPE crystallizes in the centrosymmetric triclinic space group P‐1 with one molecule residing at the inversion center. The observed properties of these π‐stacked dimers were compared with respect to through‐space conjugation and conjugation length in the structure. [ABSTRACT FROM AUTHOR]

Glycoconjugation is a well-established technology for vaccine development: linkage of the polysaccharide (PS) antigen to an appropriate carrier protein overcomes the limitations of PS T-independent antigens, making them effective in infants and providing immunological memory. Glycoconjugate vaccines have been successful in reducing the burden of different diseases globally. However, many pathogens still require a vaccine, and many of them display a variety of glycans on their surface that have been proposed as key antigens for the development of high-valency glycoconjugate vaccines. CDAP chemistry represents a generic conjugation strategy that is easily applied to PS with different structures. This chemistry utilizes common groups to a large range of PS and proteins, e.g., hydroxyl groups on the PS and amino groups on the protein. Here, new fast analytical tools to study CDAP reaction have been developed, and reaction conditions for PS activation and conjugation have been extensively investigated. Mathematical models have been built to identify reaction conditions to generate conjugates with wanted characteristics and successfully applied to a large number of bacterial PSs from different pathogens, e.g., Klebsiella pneumoniae, Salmonella Paratyphi A, Salmonella Enteritidis, Salmonella Typhimurium, Shighella sonnei and Shigella flexneri. Furthermore, using Salmonella Paratyphi A O-antigen and CRM197 as models, a design of experiment approach has been used to study the impact of conjugation conditions and conjugate features on immunogenicity in rabbits. The approach used can be rapidly extended to other PSs and accelerate the development of high-valency glycoconjugate vaccines. [ABSTRACT FROM AUTHOR]

The study delves into the use of the thiol-yne click reaction to enhance (bio)conjugation methodologies, particularly focusing on immobilizing biomolecules onto PLLA surfaces. The thiol-yne click reaction, known for its efficiency, selectivity, and versatility in forming carbon-sulfur bonds under mild conditions without transition metal catalysts, is explored for conjugating the fluorophore dansyl onto PLLA surfaces. This approach aims to broaden bioconjugation strategies beyond traditional methods like the Michael-type reaction, expanding their applicability to diverse biomolecules. Utilizing a photoinitiator and specific light for photo-immobilization, the thiol-yne click reaction offers spatial and temporal control, with the absence of transition metal catalysts mitigating concerns of cytotoxicity and metal contamination, rendering it suitable for biomedical applications. The objectives of this research encompass demonstrating the feasibility of the thiol-yne click reaction for surface functionalization and enriching bioconjugation strategies for tailoring PLLA surfaces, ultimately advancing biomedical technologies through precise control over surface properties and functionality. For this purpose, PLLA surfaces were activated through hydrolysis and amidation to introduce the activated alkyne moiety (PLLA-Alkyne), followed by photo-induced dansyl immobilization (PLLA-Dns) with Irgacure 651. Various surface characterization techniques, including SEM, WCA, XPS, ATR-FTIR, and fluorescence microscopy and spectroscopy, validated the successful conjugation. This metal-free method preserves the material's bulk properties while enabling thiol-containing molecule immobilization. [ABSTRACT FROM AUTHOR]

Public health faces daily challenges due to increasing reports of pathogenic microorganisms with new antimicrobial resistance. Klebsiella michiganensis, an emerging pathogen, poses difficulty in its identification using conventional techniques. This study presents the first documented case of NDM-1-producing K. michiganensis in Brazil, identified as the new ST418. Initially, the isolate from a tracheal secretion was misidentified as K. oxytoca. However, accurate identification was achieved through ANI analyses. Whole-genome sequencing was conducted to characterize the genetic context of the resistance genes, to identify virulence factors, and to construct a phylogenetic tree. The blaNDM-1 gene was found to be harbored on an IncFIB plasmid approximately 112 kb in length, which was transferable in conjugation assays. The detection of carbapenem resistance genes in this species highlights the importance of public health vigilance, as it may serve as a reservoir and disseminator of significant resistance genes. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance (AMR) is a major public health threat, exacerbated by the ability of bacteria to rapidly disseminate antimicrobial resistance genes (ARG). Since conjugative plasmids of the incompatibility group P (IncP) are ubiquitous mobile genetic elements that often carry ARG and are broad-host-range, they are important targets to prevent the dissemination of AMR. Plasmid-dependent phages infect plasmid-carrying bacteria by recognizing components of the conjugative secretion system as receptors. We sought to isolate plasmid-dependent phages from wastewater using an avirulent strain of Salmonella enterica carrying the conjugative IncP plasmid pKJK5. Irrespective of the site, we only obtained bacteriophages belonging to the genus Alphatectivirus. Eleven isolates were sequenced, their genomes analyzed, and their host range established using S. enterica, Escherichia coli, and Pseudomonas putida carrying diverse conjugative plasmids. We confirmed that Alphatectivirus are abundant in domestic and hospital wastewater using culture-dependent and culture-independent approaches. However, these results are not consistent with their low or undetectable occurrence in metagenomes. Therefore, overall, our results emphasize the importance of performing phage isolation to uncover diversity, especially considering the potential of plasmid-dependent phages to reduce the spread of ARG carried by conjugative plasmids, and to help combat the AMR crisis. [ABSTRACT FROM AUTHOR]

Maricaulales, the taxonomic Latin nomenclature for a particular bacterial order, have emerged as a prominent group among marine dimorphic prosthecate bacteria, primarily due to their distinctive cellular morphology. This study selected four representative strains of to devise a standardized gene knockout methodology. Through a systematic optimization of conjugation transfer factors, a dependable conjugation transfer technique was successfully established. Following this, the utilization of the optimized conjugation transfer protocol facilitated the successful knockout of the genes encoding the holdfast anchor protein in all four chosen Maricaulales strains. In conclusion, this research introduces a novel and efficient gene knockout tool specifically developed for engineering Maricaulales. [ABSTRACT FROM AUTHOR]

Can order markets lead participants towards price-taking equilibrium? Viewing market sessions as steps of iterative algorithms, this paper indicates positive prospects for convergence. Mathematical arguments turn on convolution, efficiency and generalized gradients. Economic arguments revolve around reservation costs, derived from indifference or threshold payments for quantities supplied or demanded. [ABSTRACT FROM AUTHOR]

Antimicrobial-resistance genes (ARGs) are spread among bacteria by horizontal gene transfer, however, the effect of environmental factors on the dynamics of the ARG in water environments has not been very well understood. In this systematic review, we employed the regression tree algorithm to identify the environmental factors that facilitate/inhibit the transfer of ARGs via conjugation in planktonic/biofilm-formed bacterial cells based on the results of past relevant research. Escherichia coli strains were the most studied genus for conjugation experiments as donor/recipient in the intra-genera category. Conversely, Pseudomonas spp. Acinetobacter spp. and Salmonella spp. were studied primarily as recipients across inter-genera bacteria. The conjugation efficiency (ce) was found to be highly dependent on the incubation period. Some antibiotics, such as nitrofurantoin (at ≥0.2 µg ml−1) and kanamycin (at ≥9.5 mg l−1) as well as metallic compounds like mercury (II) chloride (HgCl2, ≥3 µmol l−1), and vanadium (III) chloride (VCl3, ≥50 µmol l−1) had enhancing effect on conjugation. The highest ce value (−0.90 log10) was achieved at 15°C–19°C, with linoleic acid concentrations <8 mg l−1, a recognized conjugation inhibitor. Identifying critical environmental factors affecting ARG dissemination in aquatic environments will accelerate strategies to control their proliferation and combat antibiotic resistance. [ABSTRACT FROM AUTHOR]

Based on the properties of the conjugated p-electron system in cyclic polyenes, possible steric and electronic structures of transition states in thermal isomerizations of pyridine and phosphinine in an oxygen-free atmosphere have been determined by DFT calculations using B3LYP functional and 6-31G* basis set. The degree of stabilization of the transition states depends on the level of p-electron conjugation. Schemes of the thermal isomerizations of pyridine and phosphinine have been constructed, and potential barriers to these transformations have been calculated. The study of the reactivities of pyridine and phosphinine in the thermal isomerization processes has revealed lower aromaticity of phosphinine compared to pyridine. [ABSTRACT FROM AUTHOR]

Amanita phalloides is the primary species responsible for fatal mushroom poisoning, as its main toxin, α-amanitin, irreversibly and potently inhibits eukaryotic RNA polymerase II (RNAP II), leading to cell death. There is no specific antidote for α-amanitin, which hinders its clinical application. However, with the advancement of precision medicine in oncology, including the development of antibody–drug conjugates (ADCs), the potential value of various toxic small molecules has been explored. These ADCs ingeniously combine the targeting precision of antibodies with the cytotoxicity of small-molecule payloads to precisely kill tumor cells. We searched PubMed for studies in this area using these MeSH terms "Amanitins, Alpha-Amanitin, Therapeutic use, Immunotherapy, Immunoconjugates, Antibodies" and did not limit the time interval. Recent studies have conducted preclinical experiments on ADCs based on α-amanitin, showing promising therapeutic effects and good tolerance in primates. The current challenges include the not fully understood toxicological mechanism of α-amanitin and the lack of clinical studies to evaluate the therapeutic efficacy of ADCs developed based on α-amanitin. In this article, we will discuss the role and therapeutic efficacy of α-amanitin as an effective payload in ADCs for the treatment of various cancers, providing background information for the research and application strategies of current and future drugs. [ABSTRACT FROM AUTHOR]

Photocatalysis technology is an economical and effective new energy technology which depends on the conversion and storage of light energy through an energy transfer process or charge transfer process. Recently, organic semiconductor photocatalytic materials with the advantages of controllable structure, broad spectral response, designability, and flexibility have received wide attention. In particular, the organic polymeric materials containing poly-perylene diimides (PDI) show significant promise in the realm of photocatalysis due to their impressive catalytic capabilities and wide spectral reactivity. However, a poor charge separation and transportation (CST) process undermines their photocatalytic efficiency in most polymer photocatalysts, as well as in PDI photocatalysts. In this context, we propose a new strategy through regulating the monomer symmetry to construct highly efficient PDI photocatalysts. As proof-of-concept, a series of new PDI-based organic supramolecular photocatalytic materials with full visible spectral response from the perspectives of both the π-π conjugated structure and the symmetry of chain structure are successfully synthesized. Meanwhile, the structural compositions, morphology features, electrical properties, and photocatalytic performances of those obtained PDI photocatalysts were systematically studied. The results shown that the as-prepared PDI-1,5NDA exhibits 1.6-fold and 3.7-fold higher levels of photosynthesis of H2O2 activity than those of PDI-1,4NDA and PDI-PDA, respectively, which could be ascribe to its lower symmetry and large π-conjugate systems greatly enhances the separation of charge carriers. [ABSTRACT FROM AUTHOR]

The search for better energy storage systems that are less expensive, resource-abundant, and safer has sparked intense research into zinc ion batteries (ZIBs). Organic materials, especially quinones-based ZIBs, improved the rate performances by providing structural flexibility for the movement of zinc ions. In this work, a highly conjugated quinone molecule, tetrakis-lawsone (TLS), with multiple active sites, was used to enhance the capacity of the ZIBs. The non-planar geometry of TLS due to the different orientations of all four lawsone units of TLS provided a sufficient void for the Zn2+ movement, making it a suitable host cathode material for the ZIBs. TLS molecule consists of four LS units, which are aligned differently, thus, creating many empty void spaces in its matrix. Hence, it facilitates the Zn2+ ion movement within its lattice and thereby maximizes the utilization of TLS for energy storage. [ABSTRACT FROM AUTHOR]

The application of the amidohydrolase enzyme, L-asparaginase (ASNase), as a biocatalyst in the food and pharmaceutical industries has garnered significant interest. However, challenges such as hypersensitivity reactions, limited stability, and reusability under various operational conditions have hindered its cost-effective utilization. This paper introduces a novel nano-support for ASNase immobilization, namely the nanocomposite of iron oxide magnetic nanoparticles and amino acid-decorated graphene oxide (GO-Asp-Fe3O4). Characterization using FTIR spectroscopy, FE-SEM, and TEM microscopy revealed the homogeneous distribution of iron oxide nanoparticles on the surface of GO sheets. The effects of carrier functionalization and carrier-to-protein ratio on the immobilization of ASNase were studied to optimize the immobilization conditions. The magnetized nanocomposite of ASNase exhibited a 4.4-fold lower Michaelis-Menten constant (Km), suggesting an enhanced affinity for the substrate. The immobilized ASNase demonstrated two to eight times higher thermostability compared to the free enzyme and showed an extremely extended pH stability range. Furthermore, the immobilized enzyme retained over 80 % of its initial bioactivity after eight repeated reaction cycles. These findings suggest that the immobilization of ASNase on GO-Asp- Fe3O4 nanocomposite could be a viable option for industrial applications.

There is a growing demand for plant-based protein ingredients with improved functionality for use in diverse food and nutraceutical applications. In line with this, the current study aimed to investigate the effect of plant protein-prebiotic (polydextrose) conjugation on the techno-functional properties (emulsification, solubility, fat absorption and foaming) of pea proteins through wet heating Maillard reaction. Pea protein (PeP) was conjugated with polydextrose by incubating the mixture at various process conditions (pea protein: polydextrose mass ratios, temperature, and time). Response surface methodology coupled with Box–Behnken design was used to optimize multiple responses, including conjugation efficiency (CE), emulsifying activity (EAI), and foaming capacity (FC). The pea protein conjugate (optimized value) showed improved solubility throughout a wide pH (2–10) range and higher emulsification activity than pea protein alone. The development of conjugates (PeC) was validated through ultraviolet–visible spectroscopy, FTIR, and o-Phthaldialdehyde (OPA) assay. Browning index, FT-IR spectra, thermal properties, and scanning electron microscopy (SEM) micrographs were analyzed for the conjugate (PeC) obtained at optimized values. The FTIR spectra of the conjugates showed new peaks at 3100–3480 cm−1 and 1,000–1,166 cm−1 indicating conjugation. The Maillard conjugation increased the proportion of β-turn, random coil, accompanied by a decrease in α-helix, and β-sheet. These conformational changes were associated to the improved techno-functional properties of the pea protein upon conjugation, offering potential applications in the formulation of plant-based foods and beverages.