Cell Wall Surface Research Articles

Alginate‑carbon dot nanocomposite: A green approach towards designing turn-on aptasenor for Candida albicans fungus

Candida albicans fungus, an opportunistic microorganism that becomes a pathogen in its host, can often cause superficial and severe infections in human body. In this work, a turn-on fluorescent aptasenor based on Alginate‑carbon dot nanocomposite has been designed through a green approach. Carbon dots (CDs) with average size of 4 ± 1 nm were synthesized hydrothermally using Philodendron bipinnatifidum plant extract as precursor. Then, mixture of carbon dot and alginate was cross-linked by Ca2+ ions to make Alginate-CD nanocomposite with negligible effect on fluorescence intensity of CDs (λex = 360 nm, λem = 400 nm). The fluorescence quantum yield and lifetime were found as 14.1 % and 3.15 ns, respectively. The nanocomposite was characterize using different spectroscopic (Photoluminescence, UV–visible, XPS, FT-IR) and microscopic (TEM and SEM) analytical techniques. An aptamer was used to selectively target (1 → 3)-β-D-glucan on the surface of the Candida albicans cell wall. It was found that the fluorescence energy resonance transfer between nanocomposite and aptamer quenches the fluorescence intensity of nanocomposite, making it suitable for designing turn-on fluorescent probe. It was found that the fluorescence could be recovered in the presence of Candida albicans fungus (in <30 min), owing to higher affinity between aptamer and (1 → 3)-β-D-glucan on the surface of Candida albicans fungus, where there were two linear ranges: low concentration of 50–200 cells/mL and high concentration of 1000–6000 cells/mL (R2 ≥ 0.989 for both linear ranges), with limit of detection of 40 cells/mL. The selectivity of the designed probe was examined against a mixture of microbial agents (Staphylococcus aureus and Escherichia coli) and Curdlan polysaccharide (1→3)-b-D-Glucan (L). The designed biosensor showed an excellent ability to directly detect Candida albicans fungus via a simple and fast (<30 min) approach. The results of this experimental study show that Alginate-CD-aptamer nanocomposite is a bright prospect in clinical applications and a suitable alternative to traditional methods of Candida albicans diagnosis.

Alcoholysis-induced changes in cell wall surfaces: Structural insights for the effective delignification of lignocellulosic biomass

Alcoholysis (organosolv delignification)-induced changes in cell wall surfaces were investigated to verify whether structural assessments are required for effective delignification. Softwood blocks of Cryptomeria japonica were subjected to alcoholysis at 100–150 °C, which gradually decreased their lignin content. Scanning electron microscopy revealed the emergence of amorphous mesh structures on the intercellular side and their transformation into spherical particles with increasing temperature. In addition, warty layers changed from uneven structures into spherical particles on the lumen side of tracheids. These particles produced in cell walls under harsh alcoholysis conditions, damaging the cell wall layers on both sides. Confocal laser scanning microscopy identified that they were mainly lignin eluted by alcoholysis. Alcoholysis at 130 °C providing the largest specific surface area showed intermediate stages of growth into spherical particles but allowed complete delignification when combined with NaClO2 bleaching. Therefore, the role of the spherical particles, which has so far been debatable, was clarified as causing damage rather than a bleaching accelerant. Focusing only on compositional changes while ignoring structural ones leads to the incorrect identification of optimal conditions that remove lignin but damage the cell walls. Our findings demonstrate that structural considerations are required for effective and noninvasive delignification.

GPI anchoring controls cell wall integrity, immune evasion and surface localization of ChFEM1 for infection of Cochlibolus heterostrophus1

Glycosylphosphatidylinositol (GPI) anchoring is one of the common post-translational modifications in eukaryotic cells. In fungi, it exerts a wide range of biological functions by targeting proteins to the cell wall, but only few studies focus on the roles of GPI anchoring in plant pathogenic fungi. Here, we reveal a role of GPI anchoring in the maize fungal pathogen Cochlibolus heterostrophus. We found that GPI-anchored proteins were widely accumulated in hyphae, appressorium and infection hyphae of C. heterostrophus. Deletion of ChGPI7, which encodes a key enzyme involved in the biosynthesis of GPI anchors, resulted in significant reduction of vegetative growth and conidiation, as well as virulence due to impairment of appressorium formation and invasive growth. The ΔChgpi7 mutants also showed severe defects in cell wall integrity, resulting in a significant reduction of stress resistance. Deletion of ChGPI7 and hydrofluoric acid (HF) pyridine treatment both led to removal of cell wall GPI-anchored proteins and exposure of chitin, the results suggested that GPI anchored proteins could protect chitin from host immune recognition. A total of 124 proteins were predicted to be GPI anchored proteins in C. heterostrophus, including a putative cell wall glycoprotein ChFEM1. Deletion of ChFEM1 also resulted in significant reduction in virulence and defects in infection structures, as well as cell wall integrity. We further found that cell wall localization and protein abundance of ChFEM1 were affected by ChGPI7. Our results showed that GPI anchoring regulates cell wall integrity and immune evasion for infection of C. heterostrophus.

Effect of O-Polysaccharide Modifications on Successful Plant Colonization by Bacteria

O-polysaccharides of gram-negative bacteria are a highly variable component of the lipopolysaccharide molecules located at the cell wall surface and involved in microbial interaction with plant and animal cells. Activity of prophage genes often results in various non-stoichiometric modifications (methylation, acetylation, etc.) of glycans at bacterial cell surface. The share of modified O-polysaccharides increases during the stationary growth phase and results in increased hydrophobicity of microbial surface. Bacterial cells with different hydrophobicity showed difference in attachment to plant roots. Increased cell hydrophobicity index was found to result in a significant increase in the number of adsorbed microorganisms per unit root length. Thus, acetyl transferase and methyl transferase genes of viral origin may be indirectly involved in successful colonization of plant roots by rhizosphere bacteria.

Strain-Dependent Adhesion Variations of Shouchella clausii Isolated from Healthy Human Volunteers: A Study on Cell Surface Properties and Potential Probiotic Benefits.

The probiotic potential of Shouchella clausii is widely recognized, but little is known about its adhesive properties. Hence, this study aims to investigate the adhesion potential and cell surface properties of four human-origin S. clausii strains (B619/R, B603/Nb, B106, and B637/Nm). We evaluated epithelial adhesion, Extracellular Matrix (ECM) binding, aggregation ability, and cell surface hydrophobicity and used genome analysis for validation. Our results demonstrate that adhesion capability is a strain-specific attribute, with significant variations observed among the four strains. B619/R, B603/Nb, and B106 displayed stronger adhesion properties than B637/Nm. Supplementary adhesion assays showed that B637/Nm displayed high hydrophobicity, significant auto-aggregation, and significant mucin-binding abilities. Conversely, B619/R, B603/Nb, and B106 had mildly hydrophobic surfaces and low aggregation abilities. Genome annotation revealed the presence of various adhesion proteins in four strains. Notably, the reduced adhesion potential of B637/Nm was supported by the absence of the cell wall surface anchor family protein (LPxTG motif), which is crucial for interactions with intestinal epithelial cells or mucus components. Further, docking studies provided insights into the interaction of adhesion proteins with gut mucins. These findings contribute to a better understanding of how S. clausii strains interact with the gut environment, facilitating the development of probiotic formulations tailored for improved gut health and well-being.

Immunopeptidomics Mapping of Listeria monocytogenes T Cell Epitopes in Mice

Listeria monocytogenes is a foodborne intracellular bacterial model pathogen. Protective immunity against Listeria depends on an effective CD8+ T cell response, but very few T cell epitopes are known in mice as a common animal infection model for listeriosis. To identify epitopes, we screened for Listeria immunopeptides presented in the spleen of infected mice by mass spectrometry-based immunopeptidomics. We mapped more than 6000mouse self-peptides presented on MHC class I molecules, including 12 high confident Listeria peptides from 12 different bacterial proteins. Bacterial immunopeptides with confirmed fragmentation spectra were further tested for their potential to activate CD8+ T cells, revealing VTYNYINI from the putative cell wall surface anchor family protein LMON_0576 as a novel bona fide peptide epitope. The epitope showed high biological potency in a prime boost model and can be used as a research tool to probe CD8+ T cell responses in the mouse models of Listeria infection. Together, our results demonstrate the power of immunopeptidomics for bacterial antigen identification.

Caffeine Protects Keratinocytes from Trichophyton mentagrophytes Infection and Behaves as an Antidermatophytic Agent.

Caffeine affords several beneficial effects on human health, acting as an antioxidant, anti-inflammatory agent, and analgesic. Caffeine is widely used in cosmetics, but its antimicrobial activity has been scarcely explored, namely against skin infection agents. Dermatophytes are the most common fungal agents of human infection, mainly of skin infections. This work describes the in vitro effect of caffeine during keratinocyte infection by Trichophyton mentagrophytes, one of the most common dermatophytes. The results show that caffeine was endowed with antidermatophytic activity with a MIC, determined following the EUCAST standards, of 8 mM. Caffeine triggered a modification of the levels of two major components of the fungal cell wall, β-(1,3)-glucan and chitin. Caffeine also disturbed the ultrastructure of the fungal cells, particularly the cell wall surface and mitochondria, and autophagic-like structures were observed. During dermatophyte-human keratinocyte interactions, caffeine prevented the loss of viability of keratinocytes and delayed spore germination. Overall, this indicates that caffeine can act as a therapeutic and prophylactic agent for dermatophytosis.

Insight into the role of Streptococcus suis zinc metalloprotease C from the new serotype causing meningitis in piglets

Streptococcus suis (S. suis) is an important gram-positive pathogen and an emerging zoonotic pathogen that causes meningitis in swine and humans. Although several virulence factors have been characterized in S. suis, the underlying mechanisms of pathogenesis are not fully understood. In this study, we identified Zinc metalloproteinase C (ZmpC) probably as a critical virulence factor widely distributed in S. suis strains. ZmpC was identified as a critical facilitator in the development of bacterial meningitis, as evidenced by the detection of increased expression of TNF-α, IL-8, and matrix metalloprotease 9 (MMP-9). Subcellular localization analysis further revealed that ZmpC was localized to the cell wall surface and gelatin zymography analysis showed that ZmpC could cleave human MMP-9. Mice challenge demonstrated that ZmpC provided protection against S. suis CZ130302 (serotype Chz) and ZY05719 (serotype 2) infection. In conclusion, these results reveal that ZmpC plays an important role in promoting CZ130302 to cause mouse meningitis and may be a potential candidate for a S. suis CZ130302 vaccine.

Size/Shape-Controllable Carbonized Wood Electrodes Enabled by an MXene Shell with Spatial Confinement and a Traction Effect on the Wood Cell Wall for Shape-Customizable Energy Storage Devices.

The shrinkage and collapse of wood cell walls during carbonization make it challenging to control the size and shape of carbonized wood (CW) through pre- or postprocessing (e.g., sawing, cutting, and milling). Herein, a shape-adaptive MXene shell (MS) is created on the surface of the wood cell walls. The MS limits the deformation of wood cell walls by spatial confinement and traction effects, which is supported by the inherent dimensional stability of the MS and the formation of new C-O-Ti covalent bonds between the wood cell wall and MS. Consequently, the volumetric shrinkage ratio of CW encapsulated by the MS (CW-MS) is significantly reduced from 54.8% for CW to 2.6% for CW-MS even at 800 °C. The harnessing of this collapse enables the production of CW-MS with prolonged stability and high electric conductivity (384 S m-1). These properties make CW-MS suitable for energy storage devices with various designed shapes, matching the increasingly compact and complex structures of electronic devices.

Halenaquinol Blocks Staphylococcal Protein A Anchoring on Cell Wall Surface by Inhibiting Sortase A in Staphylococcus aureus.

Sortase A (SrtA) is a cysteine transpeptidase that binds to the periplasmic membrane and plays a crucial role in attaching surface proteins, including staphylococcal protein A (SpA), to the peptidoglycan cell wall. Six pentacyclic polyketides (1-6) were isolated from the marine sponge Xestospongia sp., and their structures were elucidated using spectroscopic techniques and by comparing them to previously reported data. Among them, halenaquinol (2) was found to be the most potent SrtA inhibitor, with an IC50 of 13.94 μM (4.66 μg/mL). Semi-quantitative reverse transcription PCR data suggest that halenaquinol does not inhibit the transcription of srtA and spA, while Western blot analysis and immunofluorescence microscopy images suggest that it blocks the cell wall surface anchoring of SpA by inhibiting the activity of SrtA. The onset and magnitude of the inhibition of SpA anchoring on the cell wall surface in S. aureus that has been treated with halenaquinol at a value 8× that of the IC50 of SrtA are comparable to those for an srtA-deletion mutant. These findings contribute to the understanding of the mechanism by which marine-derived pentacyclic polyketides inhibit SrtA, highlighting their potential as anti-infective agents targeting S. aureus virulence.

Desorption of rare earth elements biosorbed on Euglena mutabilis suspensions and biofilms

AbstractIncreasing demand for rare earth elements (REEs) has stimulated research on novel recovery methods from sources like industrial effluents. Our recent study demonstrated that algal biofilms are good candidates for the adsorption of REEs and that the extracellular polymeric substances (EPS) within the biofilm were a key accumulator of REEs. In this work, we measured the desorption kinetics, extents, and selectivity of REEs from the algae Euglena mutabilis in suspensions and biofilms using aqueous solutions of HCl at pH between 1.5 and 4, and Na2EDTA at pH 5. We examined the potential for reusing suspended and biofilm biomass for repeated adsorption–desorption cycles. The desorption kinetics were similar for suspensions and biofilms, with equilibrium being reached within 20 min. The extent of desorption was higher in biofilms with 86% and 95% desorption, compared to suspensions which had 72% and 74% desorption using HCl and Na2EDTA, respectively. We postulate that the difference between suspensions and biofilms is attributed to the binding sites in the EPS matrix of the biofilm being more readily protonated than those at the algal cell wall surface. Heavy REEs were found to preferentially desorb at pH 4 over lighter REEs. After 3 repeated adsorption–desorption cycles, the adsorption capacity of biofilms increased by 280% and 80% using HCl and Na2EDTA, respectively, compared to an 80% decrease in biosorption capacity for the suspension. The increase in biofilm biosorption capacity was attributed to the simultaneous desorption of divalent cations alongside REEs from the EPS increasing the number of available binding sites.

The unusual mode of action of the polyketide glycoside antibiotic cervimycin C.

Cervimycins A-D are bis-glycosylated polyketide antibiotics produced by Streptomyces tendae HKI 0179 with bactericidal activity against Gram-positive bacteria. In this study, cervimycin C (CmC) treatment caused a spaghetti-like phenotype in Bacillus subtilis 168, with elongated curved cells, which stayed joined after cell division, and exhibited a chromosome segregation defect, resulting in ghost cells without DNA. Electron microscopy of CmC-treated Staphylococcus aureus (3 × MIC) revealed swollen cells, misshapen septa, cell wall thickening, and a rough cell wall surface. Incorporation tests in B. subtilis indicated an effect on DNA biosynthesis at high cervimycin concentrations. Indeed, artificial downregulation of the DNA gyrase subunit B gene (gyrB) increased the activity of cervimycin in agar diffusion tests, and, in high concentrations (starting at 62.5 × MIC), the antibiotic inhibited S. aureus DNA gyrase supercoiling activity in vitro. To obtain a more global view on the mode of action of CmC, transcriptomics and proteomics of cervimycin treated versus untreated S. aureus cells were performed. Interestingly, 3 × MIC of cervimycin did not induce characteristic responses, which would indicate disturbance of the DNA gyrase activity in vivo. Instead, cervimycin induced the expression of the CtsR/HrcA heat shock operon and the expression of autolysins, exhibiting similarity to the ribosome-targeting antibiotic gentamicin. In summary, we identified the DNA gyrase as a target, but at low concentrations, electron microscopy and omics data revealed a more complex mode of action of cervimycin, which comprised induction of the heat shock response, indicating protein stress in the cell.IMPORTANCEAntibiotic resistance of Gram-positive bacteria is an emerging problem in modern medicine, and new antibiotics with novel modes of action are urgently needed. Secondary metabolites from Streptomyces species are an important source of antibiotics, like the cervimycin complex produced by Streptomyces tendae HKI 0179. The phenotypic response of Bacillus subtilis and Staphylococcus aureus toward cervimycin C indicated a chromosome segregation and septum formation defect. This effect was at first attributed to an interaction between cervimycin C and the DNA gyrase. However, omics data of cervimycin treated versus untreated S. aureus cells indicated a different mode of action, because the stress response did not include the SOS response but resembled the response toward antibiotics that induce mistranslation or premature chain termination and cause protein stress. In summary, these results point toward a possibly novel mechanism that generates protein stress in the cells and subsequently leads to defects in cell and chromosome segregation.

Characteristics of Laboratory-Derived Vancomycin-Intermediate Staphylococcus aureus Strains

Four clinical vancomycin susceptible Staphylococcus aureus (VSSA) isolates were subjected to selection for the vancomycin-intermediate S. aureus (VISA) phenotype using increasing concentrations of vancomycin. The vancomycin MIC achieved for VISA strains was 7 µg/mL. Population analysis profiles of the bacteria revealed that almost 100% of population growing at 4 µg/mL of vancomycin while some subpopulations were found to be resistant to concentrations ranging from 6 to 9 µg/mL of vancomycin. The results correlated with the AUC ratios for VISA, which ranged from 3.02-3.3 in this study. In the absence of vancomycin in the assay buffer, all the laboratory-derived VISA strains exhibited reduced whole cell autolysis compared to that of their VSSA counterparts. Examination of cell wall morphologies revealed that almost all the laboratory-derived VISA strains had thicker and rougher cell wall surfaces compared to their parental strains. Vancomycin had no remarkable effects on the thickness and roughness of the cell wall of all the laboratory-derived VISA derivatives.

Glycosyltransferases: Unraveling Molecular Insights and Biotechnological Implications

Glycosyltransferases (GTs) are present in almost all living organisms; plants, animals and microorganisms. GTs transfer sugar molecule from nucleotide sugars to a wide range of molecules including hormones, secondary metabolites, biotic and abiotic chemicals. When glycosyltransferases add a sugar moiety in any molecule, the hydrophilicity of that molecule changes and thus alter the chemical properties of the molecule. This phenomenon is vital for appropriate working of living organisms. For the first time, X-ray structure of bacteriophage T4-glucosyltransferase was reported in 1994. In bacteria, GTs play essential roles in various biological processes such as cell wall biosynthesis, surface glycosylation and virulence factor production. The point mutation as well as the domain-swapping has been reported in Bacteria. The sequence change as well as the whole cells has been engineered in bacteria too. GTs play very important role in survival, growth, development, metabolism, detoxification, insecticide resistance, chitin formation, chemosensation, defense and immunity, involved in various signaling pathways, etc. In plants, glycosyltransferase enzymes play essential role in biosynthesis of cell wall components, secondary metabolites, and signaling molecules. GTs are involved in the transfer of sugar moieties from activated donor molecules to specific acceptor molecules, leading to the formation of glycosidic bonds. GTs modify flavonoids, alkaloids and terpenoids, etc. with sugar residues and alter the solubility, stability, and bioactivity of these compounds and regulate the plant defense mechanism and interaction with insects, microorganisms and other organisms. GTs have direct impact on plant homeostasis. Site directed mutagenesis (SDM) in UGTs or GTs cause a change in substrate specificity and produce increased or total loss in the catalytic activity GTs. This kind of change demonstrated that a change in substrate specificity could cause better glycosylation and perked up anticancer activity of UGTs. GTs are also involved in glycosylaton of phytohormones and regulate their metabolism and signaling pathways. GTs are involved in the activity, stability, and transport of these hormones and influence the plant growth, development, and responses to various environmental stimuli. Four UGT families encoding 200 genes are reported in humans which regulate cell signaling, protein folding, immune response, growth and development, detoxification, metabolism and elimination of drugs, DNA methylation and histone modifications, transcriptional regulation, post-transcriptional regulation and post-translational regulation, synthesis of human blood group antigens A and B and recently GTs are also reported as linked with COVID-19-related loss of smell or taste. Various bioinformatics tools have been developed which would help analyse in silico, the structure of the GTs using any reference enzyme. The activity and the ordered structures along with various stability assays can be performed before to conduct in vitro analyses such as mutagenesis. Targeted mutagenesis have been reported through site directed mutagenesis (SDM) or domain-swapping. Standard protocols of molecular biology i.e. transformation, protein expression, extraction and purification followed by mass spectrometric analysis has been described. This molecular technique would direct future endeavours to engineer more glycosyltransferases to augment their activity with different substrates and provide a basis for more exploration of GTs as an active compound for potential anti-cancer therapeutics. Additionally, the role of GTs in medicine, food industry, pharmaceutical industry and agriculture is discussed. More research work is needed for the better understanding of the biological processes and the mechanisms of glycosyltransferases involved in cancer, tumor, drug metabolism etc. New era of engineering is awaited to engineer these GT enzymes in vitro to get them boost in industry as well as to help cure cancer and other diseases as well.

Tracing the entry process of submicrometre plastics in soybean sprouts by leaf-derived fluorescent carbon dots

As a global emerging contaminant, microplastics (MPs) in water or soil can accumulate in vegetables, making them easily ingested through the diet. With excellent and tunable optical properties, carbon dots (CDs) are highly advantageous for tracing the entry process of MPs. Originally, long-wavelength CDs were synthesized from leaf-derived extracts, and fluorescent submicrometer plastics (CDs-MPs) with clean surfaces and concentrated particle sizes were obtained by soap-free microemulsion polymerization. The concentration of CDs-MPs exhibits a significant linear relationship with long-wavelength fluorescence intensity (λEx/λEm: 415/676 nm). Soybean sprouts (SBS), as an important type of food, are susceptible to contamination of MPs due to their soft epidermis and rapidly growing biomass. The results showed that CDs-MPs could be embedded into the cortex of SBS and enter the plant with cell division and elongation, leading to an increase in pore size on the cell wall surface. After entering the root system, CDs-MPs will pass through the Casparian strip and migrate in the vessels. Then, CDs-MPs enter the leaves through vascular bundles, and the distribution and size of epicuticular wax on leaves have changed. Furthermore, SBS showed resistant growth and increased levels of oxidative response when exposed to MPs/CDs-MPs. It is the first study to demonstrate the application of leaf-derived CDs in the prevention of MPs pollution by revealing the migration behavior of submicrometre plastics in SBS.

Evaluating the effectiveness of coconut oil (Cocus nucifera) anti-bacterial benefits in caries prevention: A literature review

Background: Caries is the damage to the hard tooth tissue, which is enamel and dentine due to bacteria, with one of them being Streptococcus mutans. Caries is the most widespread non-infectious disease, with 2 billion people in the world suffering from permanent tooth caries and 514 million children suffering from primary tooth caries. The abundance of natural resources in Indonesia can be used as a way to prevent caries, such as using coconut oil which is known to have antibacterial benefits. Method: Analysis of several scientific findings and research with publication limitation over the last 5 years, recorded after January 1st, 2018. Results: Based on the results of analysis from several studies, coconut oil is proven to contain lauric acid which can be converted into monolaurin which is an active antibacterial agent. Monolaurin will cover the surface of the bacterial cell wall (peptidoglycan) and penetrate so that metabolism and cell permeability are disrupted and ultimately the cell lysis and cell death occurs. Studies show the effect of coconut oil on the incubation of bacteria that successfully inhibit colonization. Conclusion: It is proven that the content of coconut oil can effectively prevent the growth of the bacteria causing caries. Coconut oil has the potential to be an important step in preventing caries in the future.

Sweet specificities of the root extracellular trap of perennial ryegrass (Lolium perenne), a fructan accumulating plant

Perennial ryegrass (Lolium perenne) is a fructan-accumulating plant constituting one of the most important grassland species with high herbage production, nutritive value and digestibility for grazing cattle. Although fructans were reported to be involved in plant defense acting as antioxidants or stress signals, their contribution in root protection is still to be explored. In roots, atypical defense is provided by the “Root Extracellular Trap” or “RET” at the root-soil interface. The molecular composition and structural organization of the RET are essential to provide root defense against pathogen attacks and abiotic stresses. The RET was reported to be mainly composed of polysaccharides (homogalacturonan, xylogalacturonan, xyloglucan) and proteoglycans such as arabinogalactan proteins (AGPs). Our aim is to characterize the RET composition of L. perenne using cell imaging techniques and a wide range of monoclonal antibodies directed against epitopes from cell wall glycomolecules and to investigate the potential presence of fructans. Interestingly, we found that both mucilage and cell wall surface of border cells were enriched in AGP epitopes. An increased amount of the AGP-containing mucilage was produced by L. perenne root tip in response to both elicitor and osmotic stress. Fructan epitopes were also detected in root cap cells and appeared to be released in the RET under stress conditions. Taken together our findings suggest that AGPs together with fructans are involved in root response of L. perenne to environmental stresses.

Biosynthesis of UDP-α-N-Acetyl-d-mannosaminuronic Acid and CMP-β-N-Acetyl-d-neuraminic Acid for the Capsular Polysaccharides of Campylobacter jejuni.

Campylobacter jejuni is a human pathogen and a leading cause of food poisoning in North America and Europe. The exterior surface of the bacterial cell wall is attached to a polymeric coat of sugar molecules known as the capsular polysaccharide (CPS) that helps protect the organism from the host immune response. The CPS is composed of a repeating sequence of common and unusual sugar residues. In the HS:11 serotype of C. jejuni, we identified two enzymes in the gene cluster for CPS formation that are utilized for the biosynthesis of UDP-α-N-acetyl-d-mannosaminuronic acid (UDP-ManNAcA). In the first step, UDP-α-N-acetyl-d-glucosamine (UDP-GlcNAc) is epimerized at C2 to form UDP-α-N-acetyl-d-mannosamine (UDP-ManNAc). This product is then oxidized by a NAD+-dependent C6-dehydrogenase to form UDP-ManNAcA. In the HS:6 serotype (C. jejuni strain 81116), we identified three enzymes that are required for the biosynthesis of CMP-β-N-acetyl-d-neuraminic acid (CMP-Neu5Ac). In the first step, UDP-GlcNAc is epimerized at C2 and subsequently hydrolyzed to form N-acetyl-d-mannosamine (ManNAc) with the release of UDP. This product is then condensed with PEP by N-acetyl-d-neuraminate synthase to form N-acetyl-d-neuraminic acid (Neu5Ac). In the final step, CMP-N-acetyl-d-neuraminic acid synthase utilizes CTP to convert this product into CMP-Neu5Ac. A bioinformatic analysis of these five enzymes from C. jejuni serotypes HS:11 and HS:6 identified other bacterial species that can produce UDP-ManNAcA or CMP-Neu5Ac for CPS formation.

Horizontally Arranged Zn Platelet Deposition Regulated by Bi2O3/Bi toward High-Rate and Dendrite-Free 3D Zn Composite Anode.

Aqueous Zn-metal battery is considered as a promising energy-storage system. However, uncontrolled zinc dendrite growth is the main cause of short-circuit failure in aqueous Zn-based batteries. One of the most efficient and convenient strategies to alleviate this issue is to introduce appropriate zincophilic nucleation sites to guide zinc metal deposition and regulate crystal growth. Herein, this work proposes Bi2O3/Bi nanosheets anchored on the cell wall surface of the 3D porous conductive host as the Zn deposition sites to modulate Zn deposition behavior and hence inhibit the zinc dendrite growth. Density functional theory and experimental results demonstrate that Bi2O3 has a super zinc binding energy and strong adsorption energy with zinc (002) plane, as a super-zincophilic nucleation site, which results in the deposition of zinc preferentially along the horizontal direction of (002) crystal plane, fundamentally avoids the formation of Zn dendrites. Benefiting from the synergistic effect Bi2O3/Bi zincophilic sites and 3D porous structure in the B-BOGC host, the electrochemical performance of the constructed Zn-based battery is significantly improved. As a result, the Zn anode cycles for 1500cycles at 50mAcm-2 and 1.0 mAhcm-2. Meanwhile, the Zn@B-BOGC//MnO2 full cell can operate stably for 2000 cycles at 2.0Ag-1.

Anti-Candida Antibodies of Patients with Invasive Candidiasis Inhibit Growth, Alter Cell Wall Structure, and Kill Candida albicans In Vitro

Invasive candidiasis (IC), caused by Candida yeasts, particularly Candida albicans, poses a significant threat with high mortality rates. Diagnosis is challenging due to Candida's common presence in human microbiota. To address this, our research group developed an immunofluorescence assay detecting Candida albicans Germ Tube Antibodies (CAGTA) in IC patients. CAGTA, indicative of invasive processes, is associated with a lower mortality rate in ICU patients. Based on this premise, this study aims to provide results regarding the lack of knowledge about the potential activity of CAGTA against invasive infections in humans caused by the fungus Candida albicans. Therefore, in order to characterize the activity of CAGTA produced by patients with IC, we used sera from 29 patients with IC caused by either C. albicans or non-albicans Candida species. Whole serum IgG antibodies were fractionated into anti-blastospores, CAGTA-enriched, and purified CAGTA and the assessments included XTT colorimetric assays for metabolic activity, CFU counts for viability, and microscopy for growth, viability, and morphological analysis. The CAGTA-enriched IgG fraction significantly reduced the metabolic activity and viability of C. albicans compared to anti-blastospores. Purified CAGTA altered germ tube cell wall surfaces, as revealed by electron microscopy, and exhibited fungicidal properties by DiBAC fluorescent staining. In conclusion, antibodies in response to invasive candidiasis have antifungal activity against Candida albicans, influencing metabolic activity, viability, and cell wall structure, leading to cell death. These findings suggest the potential utility of CAGTA as diagnostic markers and support the possibility of developing immunization protocols against Candida infections.