Search Results (59)

The two probiotic bacteria Lactobacillus helveticus MIMLh5 and L. acidophilus NCFM exhibit homology, are both equipped with an S-layer made up of highly homologous proteins and are capable of stimulating Th1-inducing signals in dendritic cells. In this study, we aimed to compare the two strains as regards the thickness of the S-layer and their capacity to induce the production of the two Th1-inducing cytokines IL-12 and IFN-β. For both bacteria, stimulation with an increasing number of bacteria led to the higher and prompter production of IL-12 and IFN-β, but at all MOIs tested, the IL-12 response induced by NCFM was always the strongest. For both bacteria, the induction of IL-12 peaked at a multiplicity of infection (MOI) of 2–5, while IL-10, known to inhibit the induction of IL-12 cytokines, was induced more slowly and continued to increase at a higher MOI. By employing specific inhibitors, MIMLh5 and NCFM were also shown to activate different MAP kinase pathways. Endocytosed MIMLh5 showed higher survival in the DCs compared to NCFM. In the presence of mannan, previously shown to accelerate endosomal killing of Gram-positive bacteria, the survival of MIMLh5 was strongly decreased, and IL-12 increased to a level close to that induced by NCFM without the addition of mannan, indicating the importance of rapid endosomal degradation for a strong IL-12 response. When measuring the S-layer thickness, MIMLh5’s S-layer appeared to be more than twice the thickness of NCFM and exhibited an elastic modulus approximately twice as high, which is a measure of a cell’s resistance to an applied mechanic stress. When the two strains were depleted of S-layer protein, the elastic modulus was comparable. Together, our data suggests that the thicker S-layer of MIMLh5 compared to NCFM may contribute to its endosomal survival, thus reducing its capacity to induce IL-12. This may constitute an important parameter in the selection of probiotic bacteria for specific purposes. Full article

This review summarizes the current knowledge on the probiotic characteristics of dairy propionibacteria, represented by Propionibacterium freudenreichii and some Acidipropionibacterium species commonly consumed through raw milk cheese. For example, in Swiss-type cheeses, P. freudenreichii is added as a starter culture. Some strains of P. freudenreichii have been included in mixed probiotic commercial preparations or used to produce tablets from fermented culture media containing bioactive substances such as short-chain fatty acids (SCFAs), bifidogenic molecules, and vitamins. Acidipropionibacterium acidipropionici and A. jensenii strains have mainly been evaluated as health and productivity promoters in farm animals. For P. freudenreichii, the molecular mechanisms behind its probiotic action have been well elucidated, and recently, novel potential applications have been demonstrated in animal models. P. freudenreichii strains have been shown to mitigate inflammatory bowel diseases (IBDs) and mucositis and prevent necrotizing enterocolitis (NEC) in newborns. Their immunomodulation capacity has alleviated symptoms of food allergies, obesity, diabetes, colorectal cancer (CRC), and infections. Moreover, P. freudenreichii inhibited osteoclastogenesis in a rheumatoid arthritis model. Most observed effects are mediated by proteins on the cell surface or contained in extracellular vesicles (EVs) such as the surface layer (S-layer) protein SlpB, DlaT, and GroEL. No safety issues have been reported for these bacteria. However, investigations into transferable antibiotic resistance traits are still needed, and clinical trials are required to evaluate their effectiveness as probiotics for humans. Full article

Background: The 7^th Asia Dengue Summit (ADS), titled “Road Map to Zero Dengue Death”, was held in Malaysia from 5 to 7 June 2024. The summit was co-organized by Asia Dengue Voice and Action (ADVA); Global Dengue and Aedes-Transmitted Diseases Consortium (GDAC); Southeast Asian Ministers of Education Tropical Medicine and Public Health Network (SEAMEO TROPMED); Fondation Mérieux (FMx); and the International Society for Neglected Tropical Diseases (ISNTD). Objectives: Dengue experts from academia and research, as well as representatives from the Ministries of Health, Regional and Global World Health Organization (WHO), and International Vaccine Institute (IVI), came together to highlight the crucial need for an integrated approach for dengue control and achieve the target of zero dengue deaths. Methods: With more than 50 speakers and delegates from over 28 countries, twelve symposiums, and three full days, the 7^th ADS highlighted approaches to curb the growing danger of dengue. The summit included topics ranging from emerging dengue trends, insights from dengue human infection models, the immunology of dengue, and vaccine updates to antivirals and host-directed therapeutics. Conclusions: The 7^th Asia Dengue Summit reinforced the importance of an integrated, collaborative approach to dengue prevention and control. By bringing together diverse stakeholders and launching innovative initiatives such as the Dengue Slayers Challenge, the summit advanced the regional and global agenda to achieve zero dengue deaths. The exchange of knowledge and strategies at the summit is expected to contribute significantly to improved dengue management and community engagement in affected regions. Full article

Although rare, the ability to produce surface S-layer proteins is beneficially associated with particular Lactobacillus strains being investigated as probiotics. Therefore, this work aimed to study specific probiotic functionalities of selected Levilactobacillus brevis strains MB1, MB2, MB13 and MB20, isolated from human milk microbiota, and to assess the contribution of S-proteins. Firstly, Rapid Annotation using Subsystem Technology revealed that cell wall-related genes were abundant in analysed L. brevis genomes. Furthermore, the results demonstrated that S-proteins mediate aggregation capacity and competitive exclusion of selected pathogens by L. brevis strains. The improvement of Caco-2 epithelial monolayer barrier function was demonstrated by the increase in JAM-A and occludin expressions when L. brevis strains or S-proteins were added, with the effect being most pronounced after treatment with MB2 and S-proteins of MB1. L. brevis strains, especially MB20, exerted the potential to adhere to recombinant human ZG16. Strain MB2 and MB20-S-proteins improved the barrier function of HT29 epithelial monolayer, as evidenced by increased ZG16 expression. Analysed L. brevis strains and S-proteins differentially affected the protein expression of IL-1β, IL-6 and IL-8, and IL-10 cytokines. The most prominent effect was observed by S-proteins of MB20, since IL-1β production was decreased while IL-10 production was significantly increased. Full article

The potential of neuromorphic (NM) solutions often lies in their low-SWaP (Size, Weight, and Power) capabilities, which often drive their application to domains that could benefit from this. Nevertheless, spiking neural networks (SNNs), with their inherent time-based nature, present an attractive alternative also for areas where data features are present in the time dimension, such as time series forecasting. Time series data, characterized by seasonality and trends, can benefit from the unique processing capabilities of SNNs, which offer a novel approach for this type of task. Additionally, time series data can serve as a benchmark for evaluating SNN performance, providing a valuable alternative to traditional datasets. However, the challenge lies in the real-valued nature of time series data, which is not inherently suited for SNN processing. In this work, we propose a novel spike-encoding mechanism and two loss functions to address this challenge. Our encoding system, inspired by NM event-based sensors, converts the derivative of a signal into spikes, enhancing interoperability with the NM technology and also making the data suitable for SNN processing. Our loss functions then optimize the learning of subsequent spikes by the SNN. We train a simple SNN using SLAYER as a learning rule and conduct experiments using two electricity load forecasting datasets. Our results demonstrate that SNNs can effectively learn from encoded data, and our proposed DecodingLoss function consistently outperforms SLAYER’s SpikeTime loss function. This underscores the potential of SNNs for time series forecasting and sets the stage for further research in this promising area of research. Full article

In 2009, Seth Grahame-Smith published Pride and Prejudice and Zombies, sparking a subgenre that situates itself within multiple genres. I draw from the rebellious nature of nineteenth-century proto-feminists who tried to reclaim the female monster as an initial methodology to analyze Grahame-Smith’s Elizabeth Bennet. I argue that the (white) women in this horror rewriting inadvertently become the oppressors alongside contextualized zombie theory. This article also explores Grahame-Smith’s Charlotte Lucas as a complex female monster, as she is bitten and turned into a zombie, which reflects in part Jane Austen’s Charlotte’s social status and (potential) spinsterdom. It is the mythos of the zombie that makes Grahame-Smith’s Elizabeth Bennet’s feminist subversion less remarkable. And it is Charlotte’s embodiment of both the rhetorical and the religio-mythic monster that merges two narratives: the Americanized appropriated zombie and the oppressed woman. Grahame-Smith’s characters try to embody the resistance of twenty-first feminist sensibilities but fail due to the racial undertones of the zombie tangentially present in Pride and Prejudice and Zombies. Full article

Lactobacillus helveticus 34.9 was isolated from a sample of Romanian home-made fermented milk, producing both surface layer proteins and a class III bacteriocin. The present study aimed to investigate the biological and functional role of the S-layer in correlation with its probiotic properties. The presence of S-layer proteins resulted in various degrees of co-aggregation of L. helveticus 34.9 with pathogens and with other lactic acid bacteria, but the removal of these proteins reduced the co-aggregation with all the tested strains. Moreover, the S-layer proved to be involved in cell wall hydrophobicity and cellular protection during freeze-drying. In the simulated passage through the gastrointestinal tract, S-layer depleted cells exhibited increased vulnerability, with greater viability loss in low pH and pepsin treatment compared to control cells. Subsequently, in the small intestine simulation, these cells lost all viability, underscoring the vital role of extracellular proteins for cell protection. The morphological effects of these treatments were observed by scanning electron microscopy. Severe structural damage was noticed when the S-layer was absent, including loss of cell shape and integrity as well as many ghost cells emptied of their content. Finally, the elimination of surface proteins reduced the interaction between L. helveticus 34.9 and mammalian cells. Full article

Previously, the protective role of the S-layer protein 2 (Slp2) of the vaginal Lactobacillus crispatus 2029 (LC2029) strain against foodborne pathogens Campylobacter jejuni, Salmonella enterica serovar Enteritidis, and Escherichia coli O157:H was demonstrated. We demonstrate the new roles of the Slp2-positive LC2029 strain and soluble Slp2 against C. albicans infections. We show that LC2029 bacteria can adhere to the surface of the cervical epithelial HeLa cells, prevent their contact with C. albicans, and block yeast transition to a pathogenic hyphal form. Surface-bound Slp2 provides the ability for LC2029 to co-aggregate with various C. albicans strains, including clinical isolates. C. albicans-induced necrotizing epithelial damage is reduced by colonization with the Slp2-positive LC2029 strain. Slp2 inhibits the adhesion of various strains of C. albicans to different human epithelial cells, blocks yeast transition to a pathogenic hyphal form, and prevents the colonization and pathogenic infiltration of mucosal barriers. Only Slp2 and LC2029 bacteria stimulate the production of protective human β-defensin 3 in various epithelial cells. These findings support the anti-Candida albicans potential of the probiotic LC2029 strain and Slp2 and form the basis for further research on their ability to prevent and manage invasive Candida infections. Full article

In this review, we discuss the progress in the investigation of macromolecular crystals obtained through the use of atomic force microscopy (AFM), a powerful tool for imaging surfaces and specimens at high resolution. AFM enables the visualization of soft samples at the nanoscale and can provide precise visual details over a wide size range, from the molecular level up to hundreds of micrometers. The nonperturbative nature, the ability to scan in a liquid environment, and the lack of need for freezing, fixing, or staining make AFM a well-suited tool for studying fragile samples such as macromolecular crystals. Starting from the first morphological investigations revealing the surface morphology of protein crystals, this review discusses the achievements of AFM in understanding the crystal growth processes, both at the micro- and nanoscale. The capability of AFM to investigate the sample structure at the single molecular level is analyzed considering in-depth the structure of S-layers. Lastly, high-speed atomic force microscopy (HS-AFM) is discussed as the evolution to overcome the limitations of low imaging speed, allowing for the observation of molecular dynamics and weakly adsorbed, diffusing molecules. HS-AFM has provided intuitive views and directly visualized phenomena that were previously described indirectly, answering questions that were challenging to address using other characterization methods. Full article

Biological treatment methods overcome many of the drawbacks of physicochemical strategies and play a significant role in removing dye contamination for environmental sustainability. Numerous microorganisms have been investigated as promising dye-degrading candidates because of their high metabolic potential. However, few can be applied on a large scale because of the extremely harsh conditions in effluents polluted with multiple dyes, such as alkaline pH, high salinity/heavy metals/dye concentration, high temperature, and oxidative stress. Therefore, extremophilic microorganisms offer enormous opportunities for practical biodegradation processes as they are naturally adapted to multi-stress conditions due to the special structure of their cell wall, capsule, S-layer proteins, extracellular polymer substances (EPS), and siderophores structural and functional properties such as poly-enzymes produced. This review provides scientific information for a broader understanding of general dyes, their toxicity, and their harmful effects. The advantages and disadvantages of physicochemical methods are also highlighted and compared to those of microbial strategies. New techniques and methodologies used in recent studies are briefly summarized and discussed. In particular, this study addresses the key adaptation mechanisms, whole-cell, enzymatic degradation, and non-enzymatic pathways in aerobic, anaerobic, and combination conditions of extremophiles in dye degradation and decolorization. Furthermore, they have special metabolic pathways and protein frameworks that contribute significantly to the complete mineralization and decolorization of the dye when all functions are turned on. The high potential efficiency of microbial degradation by unculturable and multi-enzyme-producing extremophiles remains a question that needs to be answered in practical research. Full article

Deer hunting has a long history in the US. It is supported by hunting cultures, described as necessary for protecting forest/plant biodiversity and ecosystems, but opposed by animal welfare and rights advocates as cruel. Using multiple literature sources, we examine the trade-off between protecting deer and ecosystems from harm in the context of contemporary America. We examine various approaches for exploring harms affecting nonhuman animal populations found in the green criminological, environmental sociology, wildlife conservation and management, and ecological literature. We argue that making sense of these opposing positions requires examining the extent of deer hunting to quantify those harms in some way. Here, we examine reported deer kills for US states for the period 1999–2020. These data indicate that nearly 7 million deer are taken annually in the US. We also examined some hypothesized correlates of deer harvesting across states. While these data tell us something about the number of deer killed, these data alone are insufficient. We argue no clear conclusion about the acceptability of deer hunting can be reached given the difficulty rectifying opposing moral/philosophical positions on deer hunting, opposing deer management objectives, and scientific evidence on the ecological impacts of deer populations in the US under contemporary conditions that include shrinking forest ecosystems and impaired ecosystem stability. Full article

The control of spoilage microorganisms and foodborne pathogens in meat and meat products is a challenge for food producers, which potentially can be overcome through the combined use of biopreservatives, in the form of a mix of various microbial hurdles. The objective of this work is to systematically review the available knowledge to reveal whether various microbial hurdles applied in combination can pose an effective decontamination strategy for meat and meat products. PubMed, Web of Science, and Scopus were utilized to identify and evaluate studies through February 2023. Search results yielded 45 articles that met the inclusion criteria. The most common meat biopreservatives were combinations of various starter cultures (24 studies), and the use of mixtures of non-starter protective cultures (13 studies). In addition, studies evaluating antimicrobial combinations of bacteriocins with other bacteriocins, BLIS (bacteriocin-like inhibitory substance), non-starter protective cultures, reuterin, and S-layer protein were included in the review (7 studies). In one study, a biopreservative mixture comprised antifungal protein PgAFP and protective cultures. The literature search revealed a positive effect, in most of the included studies, of the combination of various bacterial antimicrobials in inhibiting the growth of pathogenic and spoilage bacteria in meat products. The main advantages of the synergistic effect achieved were: (1) the induction of a stronger antimicrobial effect, (2) the extension of the spectrum of antibacterial action, and (3) the prevention of the regrowth of undesirable microorganisms. Although further research is required in this area, the combination of various microbial hurdles can pose a green and valuable biopreservation approach for maintaining the safety and quality of meat products. Full article

Clostridioides difficile, a nosocomial pathogen, is an emerging gut pathobiont causing antibiotic-associated diarrhea. C. difficile infection involves gut colonization and disruption of the gut epithelial barrier, leading to the induction of inflammatory/immune responses. The expression of two major exotoxins, TcdA and TcdB is the major cause of C. difficile pathogenicity. Attachment of bacterial abundant cell wall proteins or surface S-layer proteins (SLPs) such as SlpA with host epithelial cells is critical for virulence. In addition to being toxins, these surface components have been shown to be highly immunogenic. Recent studies indicate that C. difficile SLPs play important roles in the adhesion of the bacteria to the intestinal epithelial cells, disruption of tight junctions, and modulation of the immune response of the host cells. These proteins might serve as new targets for vaccines and new therapeutic agents. This review summarizes our current understanding of the immunological role of SLPs in inducing host immunity and their use in the development of vaccines and novel therapeutics to combat C. difficile infection. Full article

S-layers are involved in the adaptation of bacteria to the outside environment and in pathogenesis, often representing special virulence factors. Vegetative cells of the entomopathogenic bacterium Brevibacillus laterosporus are characterized by an overproduction of extracellular surface layers that are released in the medium during growth. The purpose of this study was to characterize cell wall proteins of this bacterium and to investigate their involvement in pathogenesis. Electron microscopy observations documented the presence of multiple S-layers, including an outermost (OW) and a middle (MW) layer, in addition to the peptidoglycan layer covering the plasma membrane. After identifying these proteins (OWP and MWP) by mass spectrometry analyses, and determining their gene sequences, the cell wall multilayer-released fraction was successfully isolated and used in insect bioassays alone and in combination with bacterial spores. This study confirmed a central role of spores in bacterial pathogenicity to insects but also detected a significant virulence associated with fractions containing released cell wall multilayer proteins. Taken together, S-layer proteins appear to be part of the toxins and virulence factors complex of this microbial control agent of invertebrate pests. Full article

Human milk not only provides a perfect balance of nutrients to meet all the needs of the infant in the first months of life but also contains a variety of bacteria that play a key role in tailoring the neonatal faecal microbiome. Microbiome analysis of human milk and infant faeces from mother-breastfed infant pairs was performed by sequencing the V1–V3 region of the 16S rRNA gene using the Illumina MiSeq platform. According to the results, there is a connection in the composition of the microbiome in each mother-breastfed infant pair, supporting the hypothesis that the infant’s gut is colonised with bacteria from human milk. MiSeq sequencing also revealed high biodiversity of the human milk microbiome and the infant faecal microbiome, whose composition changes during lactation and infant development, respectively. A total of 28 genetically distinct strains were selected by hierarchical cluster analysis of RAPD-PCR (Random Amplified Polymorphic DNA-Polymerase Chain Reaction) electrophoresis profiles of 100 strains isolated from human milk and identified by 16S RNA sequencing. Since certain cellular molecules may support their use as probiotics, the next focus was to detect (S)-layer proteins, bacteriocins and exopolysaccharides (EPSs) that have potential as therapeutic biomolecules. SDS-PAGE (Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis) coupled with LC-MS (liquid chromatography-mass spectrometry) analysis revealed that four Levilactobacillus brevis strains expressed S-layer proteins, which were identified for the first time in strains isolated from human milk. The potential biosynthesis of plantaricin was detected in six Lactiplantibacillus plantarum strains by PCR analysis and in vitro antibacterial studies. ¹H NMR (Proton Nuclear Magnetic Resonance) analysis confirmed EPS production in only one strain, Limosilactobacillus fermentum MC1. The overall microbiome analysis suggests that human milk contributes to the establishment of the intestinal microbiota of infants. In addition, it is a promising source of novel Lactobacillus strains expressing specific functional biomolecules. Full article