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A straightforward synthetic route towards DAB-1 scaffolded dimeric iminosugars is described here, starting from readily available bis-glycosylamines. The method allows the integration of a variety of linkages (aryl, alkyl, polyethyleneglycol chains) between both iminosugars through the choice of the bis-amine used in the first step. Moreover, an additional substituent (allyl, ethynyl) may be inserted into the structure via nucleophilic addition of an organometallic reagent to the starting bis-glycosylamine. A symmetrical ethynyl-iminosugar proved susceptible to intramolecular Glaser coupling, affording the corresponding macrocyclic structure. Dimeric iminosugars were tested towards a series of commercial glycosidases to uncover potencies and selectivities when compared to DAB-1, their monomeric counterpart. Whereas a significant drop in inhibition potencies was observed towards glucosidases, some compounds displayed unexpected potent inhibition of β-galactosidase. [ABSTRACT FROM AUTHOR]

Background: Keloid is a fibro-proliferative dermal benign growth affecting the Asian population. In India, ear keloids are common, often resulting from ear piercing, a prevalent cultural practice. The resultant ear keloids pose aesthetic concerns, leading to significant psychological distress, and necessitating effective treatment. While various treatment options are available, their outcomes and recurrence rates vary, highlighting the need for individualized and optimal management strategies. Our study aimed to observe the combined effect of intralesional triamcinolone with hyaluronidase on keloid regression. Methods: This prospective observational study was conducted at our tertiary care institute over 1 and a half years. Fifty patients who met the inclusion criteria were enrolled and received intralesional triamcinolone injection with hyaluronidase. Keloid regression was assessed using the Vancouver Scar Scale (VSS), while the visual analogue score (VAS) and patient satisfaction score (PSS) provided subjective evaluations of symptom relief. All patients underwent compression therapy. Patients were evaluated at every visit and then at 1 year for symptom relief, keloid regression, and complications if any. Results: Demographic data of all patients were recorded. Ear keloids were predominantly observed in females, with a male-to-female ratio of 18:32. The most common etiology was trauma following ear piercing, accounting for 46% of cases. All patients showed improvement in VSS, VAS, and PSS scores in the follow-up visits. A total of 98% of patients demonstrated a complete treatment response, with only a single instance of recurrence. Conclusion: Intralesional triamcinolone with hyaluronidase provides satisfactory symptomatic relief and has lower recurrence rates. All patients showed improvement in the Vancouver Scar Scale and had enhanced VAS and PSS scores. [ABSTRACT FROM AUTHOR]

Purpose: Myeloid sarcoma (MS) is a rare extramedullary tumor of myeloid origin, often underdiagnosed or misdiagnosed, particularly in children. This study aims to evaluate the clinicopathological features, immunophenotypes, therapeutic approaches, and prognosis to enhance patient management. Methods: This retrospective, single-center case series examined 14 patients diagnosed with myeloid sarcoma through pathological analysis. Results: The median age at diagnosis was 7 years, with a male-to-female ratio of 8:6. The median disease duration at diagnosis was 1 month, and all patients had unilateral eye involvement. The most common presentation was proptosis, followed by restricted eye movement and eyelid swelling. Immunohistochemical analysis revealed that MPO (myeloperoxidase) and lysozyme were the most frequently expressed markers, followed by CD56, α1-antitrypsin (AACT), and CD34. Eleven patients were treated with chemotherapy according to the acute myeloid leukemia (AML) regimen. The median follow-up time for these patients was 13 months. No relapses occurred; one patient died, and three were lost to follow-up. The Kaplan–Meier survival analysis estimated a 5-year overall survival (OS) and event-free survival (EFS) rate of 87.5%. Conclusion: Diagnosing myeloid sarcoma (MS) is often challenging. An adequate tumor biopsy and comprehensive immunohistochemical analysis are essential for an accurate diagnosis. Early and consistent systemic chemotherapy can result in long-term survival. [ABSTRACT FROM AUTHOR]

Simple Summary: Antibiotics were extensively used in the pigeon breeding industry previously, leading to the spread of antibiotic-resistant genes (ARGs) and their deposition in gut microbes, which has become a major public health concern. In this study, we found the distinct gut microbiota and functional compositions between young and older pigeons in Nanjing, Jiangsu Province, China, by using a metagenomic analysis. Streptococcus and Escherichia were the most abundant genera in young and older pigeons, respectively. The gene ontology (GO) analysis highlighted the significant enrichment of integral components of the membrane, ATP binding, and DNA binding in pigeon gut microbiota. The CAZy functional classification showed that glycoside hydrolases, glycosyl transferases, and carbohydrate esterases were the most abundant in pigeon gut microbes. Moreover, 142 ARGs conferring multidrug resistance, tetracycline, and aminoglycoside resistance were identified, representing more than 70% of the fecal resistomes; the most abundant gene in young pigeons was tetracycline-tetW, while in older pigeons, it was multidrug-acrB. Additionally, we found the highest abundance of resistance genes in older pigeons, indicating a high level of antibiotic resistance. Background: Antibiotics were extensively used in the pigeon breeding industry previously to promote growth and prevent disease, leading to the spread of antibiotic-resistant genes (ARGs) in gut microbes, which has become a major public health concern. Methods: A metagenomic analysis was performed to investigate the gut microbial communities and ARGs in young and older pigeons in Nanjing, Jiangsu Province, China. Results: There were obviously distinct gut microbiota and functional compositions between young and older pigeons. Both Pseudomonadota and Uroviricota were dominant in young and older pigeons. Although sharing 24 gut microbiota phyla between young and older pigeons, Bacillota and Pseudomonadota were the dominant microbial phyla in them, respectively. Besides the shared metabolic pathways and biosynthesis of secondary metabolites, biosynthesis of amino acids was the most abundant Kyoto Encyclopedia of Genes and Genomes (KEGG) function in young pigeons, while microbial metabolism in diverse environments was abundant in older pigeons. A total of 142 ARGs conferring multidrug resistance, tetracycline, and aminoglycoside resistance were identified; the most abundant gene in young pigeons was tetracycline-tetW, while in older pigeons, it was multidrug-acrB. Conclusions: Our findings revealed significant differences in the gut microbial communities and ARGs between young and older pigeons. This study enhances our understanding of pigeon gut microbiota and antibiotic resistomes, contributing to knowledge-based sustainable pigeon meat production. [ABSTRACT FROM AUTHOR]

Glycoside hydrolase family 65 (GH65) includes glycoside hydrolases active on various α-glucosides. We previously demonstrated that the GH65 enzyme from Flavobacterium johnsoniae (FjGH65A) is a kojibiose hydrolase and determined its 3-dimensional structure. In this study, the effects of glucosidase inhibitors on FjGH65A and their complex structures were analyzed to elucidate their inhibition mechanism. FjGH65A was competitively inhibited by 1-deoxynojirimycin (DNJ) and noncompetitively inhibited by castanospermine (CSP) with K i values of 2.95 and 3.69 µ m , respectively. The crystal structures of FjGH65A complexed with the inhibitors indicated that DNJ was bound to subsite −1 of FjGH65A, while CSP was bound to subsites −1 and +1 of FjGH65A. Compared with the glucose complex structure, the conformation of Tyr337 was changed in the CSP complex structure. These results provide new structural insights into the mechanism of inhibition against GH65 α-glucoside hydrolases. [ABSTRACT FROM AUTHOR]

A new fungal species, Penicillium psychrofluorescens sp. nov. is described as a member of section Torulomyces. The new species is sister to P. catalonicum, and was isolated from soil collected from Robinson Ridge, East Antarctica, following enrichment cultivation under oligotrophic conditions supplemented with excess hydrogen gas. Penicillium psychrofluorescens is named for its intense autofluorescence derived from a combination of compounds that may include NADPH, porphyrins, and secondary metabolites, such as polyketides. Comparative genomics with both Antarctic and mesophilic Penicillium spp. shows that Penicillium psychrofluorescens has a wide repertoire of glycoside hydrolases, but almost no polysaccharide lyases, has comparably large effector proteins, lacks the machinery to use nitrate as an N-source, but has the genes for the assimilation of phosphorus from phosphonates via oxidative pathway. The strain was found to have 30 biosynthetic gene clusters (BGCs), the majority of which were unrelated to known compound BGCs. Given the remarkable diversity of natural products already characterised from Penicillium spp. and the presence of >30 BGCs with low similarity to known genes, there is biotechnological potential within this novel species that is yet to be explored. [ABSTRACT FROM AUTHOR]

Background: Chitinases (CHIs) are glycosidases that degrade chitin, playing critical roles in plant responses to both abiotic and biotic stress. Despite their importance, the CHI family's systematic analysis and evolutionary pattern in F. tataricum (Tartary buckwheat) yet to be explored. Results: This study analyzed their phylogenetic relationships, conserved motifs, gene structures, syntenic relationships, physiological functions, and biochemical properties. This research identified 26 FtCHIs and examined their expression patterns under different salt stress conditions and across various tissues. Differential expression analysis revealed a significant upregulation of multiple FtCHIs in response to salt stress, which RT-qPCR further validated. Additionally, subcellular localization experiments demonstrated that Ft_chitinaseIV-2 is localized in vacuoles. The results of transient·transformation showed that·overexpression of Ft_chitinaseIV-2 could·enhance the salt tolerance of plants. Conclusions: The findings provide new insights into the role of CHIs in stress tolerance and lay the groundwork for future research on the functional characterization of FtCHIs. Understanding the molecular mechanisms of CHI-mediated stress responses could contribute to developing stress-resistant crops. [ABSTRACT FROM AUTHOR]

Simple Summary: Breast Cancer (BC) is an important disease causing death of many women worldwide. Here the relationship between gastroenteral microbionta and metabolism in the context of BC is investigated and described in detail. The interrelation between BC, metabolite abnormalities and reprogramming, and micronenvironment is described. All information about the various mechanisms by which these bacterial residents may influence disease initiation, progression, and treatment response is collected and summarized. We will see that gut-based biomarkers, and synergy between conventional therapies and microbiome interventions, together will soon be able to open-up new gates for breast cancer therapy. Breast cancer (BC), a major cause of death among women worldwide, has traditionally been linked to genetic and environmental factors. However, emerging research highlights the gut microbiome's significant role in shaping BC development, progression, and treatment outcomes. This review explores the intricate relationship between the gut microbiota and the breast tumor microenvironment, emphasizing how these microbes influence immune responses, inflammation, and metabolic pathways. Certain bacterial species in the gut either contribute to or hinder BC progression by producing metabolites that affect hormone metabolism, immune system pathways, and cellular signaling. An imbalance in gut bacteria, known as dysbiosis, has been associated with a heightened risk of BC, with metabolites like short-chain fatty acids (SCFAs) and enzymes such as β-glucuronidase playing key roles in this process. Additionally, the gut microbiota can impact the effectiveness of chemotherapy, as certain bacteria can degrade drugs like gemcitabine and irinotecan, leading to reduced treatment efficacy. Understanding the complex interactions between gut bacteria and BC may pave the way for innovative treatment approaches, including personalized microbiome-targeted therapies, such as probiotics and fecal microbiota transplants, offering new hope for more effective prevention, diagnosis, and treatment of BC. [ABSTRACT FROM AUTHOR]

Plant metabolites have a great potential for limiting the spread of harmful fungi. However, a better understanding of the mode-of-action of these molecules and the defense systems developed by fungi to resist them, is needed to assess the benefits/risks of using them as antifungal treatment. White-rot fungi are excellent models in this respect, as they have adapted to the hostile habitat that is wood. In fact, wood is a source of putative antifungal compounds that can be derived using extraction techniques. In this study, we demonstrated that esculin and esculetin, which are coumarins found in plants and wood, reduce the growth of the wood-rotting fungi Fomitiporia mediterranea, Phanerochaete chrysosporium , Pycnoporus cinnabarinus and Trametes versicolor. We have shown that extracellular strategies are developed by the fungi to deal with esculin, through the involvement of laccases, peroxidases and glycoside hydrolases, and intracellular strategies, mainly via upregulated protein translation. Comparative proteomic and metabolomic approaches revealed that, despite the fact that the species analysed are closely related (they all belong to the Agaricomycetes, and have the same trophic mode), their defense responses to esculin differ. [Display omitted] • Esculin is inhibitory to white-rot fungi. • White-rot fungi use various strategies to mitigate the activities of esculin. • Laccases, peroxidases and glycoside hydrolases are involved in esculin detoxification. • Regulation of the translation machinery is part of the cellular responses to esculin. • Esculin disturbs protein secretion in white-rot fungi. [ABSTRACT FROM AUTHOR]

Background: To investigate the long term effects of ionizing radiation (IR) on hematopoietic stem/progenitor cells (HSPCs), immune tissues and cells, and the effects of Siwu decoction (SWD) on immune senescence mice. Methods: C57BL/6 J mice were exposed to 6.0 Gy 60Co γ irradiation. After 8-weeks of IR, SWD (5, 10, 20 g/kg/d) was administered for 30 days. The changes of HSPCs in bone marrow (BM) and T, B type lymphocyte and natural killer (NK) cells in spleen were detected by flow cytometry. The changes of peripheral blood cells were also examined. Hematoxylin–eosin staining were used to detect the pathological lesions of hippocampus, spleen and thymus tissues. Absolute mouse telomere length quantification qPCR assay kit was used to measure the telomere length of BM cells. The expression of factors associated with inflammation and aging such as p16, β-galactosidase in spleen, thymus and BM was determined. Results: Administration of SWD could increase the proportion of LSK (Lin−, Sca-1 + , c-Kit−), multipotent progenitor cells and multipotent progenitor cells and decrease the proportion of common myeloid progenitors and granulocyte–macrophage progenitors in BM. The proportion of B cells and NK cells in spleen and the content of white blood cells, red blood cells, hemoglobin, lymphocytes and eosinophils in peripheral blood were increased, at the same time, the proportion of neutrophils and monocytes was reduced by SWD. The pathological lesions of hippocampus, spleen and thymus were improved. The expression of p16 and β-galactosidase in spleen, thymus and BM was reduced and shortening of the telomere of BM cells was inhibited after administration. In addition, SWD could reduce the content of Janus activated kinase (JAK) 1, JAK2 and signal transducer and activator of transcription 3 (STAT3) in BM and spleen. Conclusions: SWD could slow down IR-induced immune senescence by improving hematopoietic and immunologic injury. SWD might reduce the inflammation level of BM hematopoietic microenvironment by acting on JAK/STAT signaling pathway, while the immune damage of mice was improved by affecting the differentiation of HSPCs. The remission of hematopoietic and immunologic senescence was further demonstrated at the overall level. Highlights: Siwu decoction (SWD) could relieve IR-induced senescence in mice. SWD could improve the learning and memory ability and the pathological damage of hippocampus. SWD could ameliorate the lymphatic/myeloid differentiation bias of bone marrow (BM) hematopoietic stem/progenitor cells (HSPCs) and immune imbalance in peripheral blood and spleen. SWD could inhibit the shortening of telomere of BM cells. SWD could alleviate the inflammation level of hematopoietic microenvironment and spleen, and reduce the expression of senescence related molecules (p16, β-galactosidase, etc.) in BM, spleen and thymus. [ABSTRACT FROM AUTHOR]

Barnyard grass is one of the most serious rice weeds, often growing near paddy fields and therefore potentially serving as a bridging host for the rice blast fungus. In this study, we isolated three fungal strains from diseased barnyard grass leaves in a rice field. Using a pathogenicity assay, we confirmed that they were capable of causing blast symptoms on barnyard grass and rice leaves to various extents. Based on morphology characterization and genome sequence analyses, we confirmed that these three strains were Epicoccum sorghinum (SCAU-1), Pyricularia grisea (SCAU-2), and Exserohilum rostratum (SCAU-6). The established Avirulence (Avr) genes Avr-Pia, Avr-Pita2, and ACE1 were detected by PCR amplification in SCAU-2, but not in SCAU-1 or SCAU-6. Furthermore, the whole-genome sequence analysis helped to reveal the genetic variations and potential virulence factors relating to the host specificity of these three fungal pathogens. Based on the evolutionary analysis of single-copy orthologous proteins, we found that the genes encoding glycoside hydrolases, carbohydrate esterases, oxidoreductase, and multidrug transporters in SCAU-1 and SCAU-6 were expanded, while expansion in SCAU-2 was mainly related to carbohydrate esterases. In summary, our study provides clues to understand the pathogenic mechanisms of fungal isolates from barnyard grass with the potential to cause rice blast. [ABSTRACT FROM AUTHOR]

Introduction: For upper limb procedures using the supraclavicular approach to brachial plexus block, various adjuncts to Local Anaesthetics (LA) have been employed to improve the quality and duration of anaesthesia and postoperative analgesia without leading to any adverse side-effects or prolonging the period of motor block. Hyaluronidase is an enzyme used with other drugs to accelerate their dispersion and distribution. It catalyses the hydrolysis of a component of the extracellular matrix, hyaluronan, thereby lowering its viscosity and improving tissue permeability. Aim: To evaluate the sensory and motor blockade resulting from the addition of hyaluronidase as an adjuvant to LA in Peripheral Nerve Stimulator (PNS)-guided Supraclavicular Brachial Plexus Block (SBPB). Materials and Methods: This prospective, comparative, double-blinded, randomised controlled study was conducted in a Department of Anaesthesiology S.B.K Shah Medical Institute and Research Centre, Vadodara, Gujarat, India at a tertiary care hospital on 72 patients undergoing elective upper limb surgeries under PNS-guided SBPB over 18 months from November 2022 to August 2024. Group B (n=36) received inj. bupivacaine (0.5%) 13 mL, inj. lignocaine with adrenaline (2%) (1:200,000) 13 mL, and inj. normal saline (0.9%) 4 mL, for a total of 30 mL, and Group H (n=36) received inj. bupivacaine (0.5%) 10 mL, inj. lignocaine with adrenaline (2%) (1:200,000) 10 mL, inj. hyaluronidase (900 IU) 6 mL, and inj. normal saline (0.9%) 4 mL, for a total of 30 mL. Sensory and motor blockade and rescue analgesia were recorded at 0, 1, 3, 5, 7, and 10 hours. Categorical variables were analysed using the Chi-square test. To analyse continuous variables, the student's t-test was used. The p-value of less than 0.05 was deemed statistically significant. Results: The onset time of sensory and motor block was earlier with Group H (2.36±0.25 minutes and 4.2±0.29 minutes) than with Group B (4.54±0.25 minutes and 6.51±0.29 minutes), respectively (p<0.05). The sensory and motor block lasted longer in Group B (278.8±7 minutes and 266.8±6.78 minutes) than in Group H (263.2±7 minutes and 246.3±6.78 minutes) respectively, p<0.05. The duration of analgesia was longer with Group B (240.8±7.84 minutes) compared to Group H (220±7.84 minutes) p<0.05. No major adverse effects were observed. Conclusion: Addition of hyaluronidase to a lower volume of LA results in rapid onset of the block with minimal side-effects. [ABSTRACT FROM AUTHOR]

Background and Aims: Hyaluronidase‐facilitated subcutaneous immunoglobulin (fSCIG) consists of subcutaneous human immunoglobulin G (IgG) 10% with recombinant human hyaluronidase (rHuPH20) and can be administered at the same dose and interval as intravenous IgG (IVIG). fSCIG recently received US approval as maintenance therapy for adults with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) and European approval for adults and children with CIDP after stabilization with IVIG. Methods: ADVANCE‐CIDP 3 (NCT02955355) was an open‐label long‐term extension of the Phase 3 double‐blind randomized placebo‐controlled ADVANCE‐CIDP 1 study (NCT02549170) that examined fSCIG safety and efficacy as maintenance CIDP therapy. Primary outcomes were safety, tolerability, and immunogenicity. Efficacy was an exploratory outcome. Results: The study provided 220 patient‐years of follow‐up data from 85 patients. Median (range) exposure was 33 (0–77) months. Patients received fSCIG every 4 weeks (88.2%) or every 3 weeks (11.8%). Median (range) 4‐weekly IgG dose equivalent was 64.0 (28.0–200.0) g. Mean (standard deviation) infusion duration was 135.5 (62.8) minutes. Most adverse events (AEs) were mild or moderate and self‐limiting. Of the 1406 AEs, only 48 were severe and 30 were serious. fSCIG‐related AEs (n = 798) included infusion site reactions such as pain, redness, and pruritus. Three infusions (0.1%) were reduced in rate, interrupted, or stopped due to intolerability. Relapse occurred in 10 of 77 patients (13.0%); annual relapse rate was 4.5%. An anti‐rHuPH20 antibody titer ≥1:160 was detected in 14 of 84 patients (16.7%); patients who tested positive (≥1:160) had similar relapse rates versus those who tested negative (16.7% vs. 12.3%, respectively). Interpretation: ADVANCE‐CIDP 3 demonstrated favorable fSCIG long‐term safety and tolerability consistent with its established safety profile, and a low relapse rate, supporting use as maintenance CIDP treatment. [ABSTRACT FROM AUTHOR]

Endophytes, symbiotically with their host plants, are frequently found in the roots of Panax ginseng. To explore ginsenosides biotransformation by endophytic fungi, 26 such fungi were isolated from ginseng roots sourced from four distinct planting regions, and their glycosyl hydrolysis activities were measured. Of these endophytic, 20 isolates exhibited glycosidase enzymes capable of cleaving glycosidic bonds of glucose, arabinofuranose and arabinopyranose. Notably, the glycosidase BglNh gene was isolated and cloned from Nectria haematococca, one of the endophytic fungi. Enzymatic assessments unveiled that BglNh specifically catalyzes the hydrolysis of glucose residues located external to the C-3 position of protopanaxadiol ginsenosides under optimal reaction conditions (pH 6.0 and temperature 35 °C). Our findings demonstrate the efficient conversion of major ginsenosides Rb1, Rb2 and Rc to minor ginsenosides Gyp17, CO and CMc1, highlighting the potential of these fungi for ginsenoside biotransformation and bioactive compound production. [ABSTRACT FROM AUTHOR]

Zhejiang, China, has a tradition of wine-fermented foods, including Jinhua Dajiao steamed bread (JDSB). The traditional leavening agent, jiaotou, confers its properties and taste to JDSB. We examined microorganisms in jiaotou, their effects on processing, and the metabolome using metagenomic and metabolomic analyses. Metagenomics of microorganisms in jiaotou revealed a high average diversity, with Rhizopus sp. being the dominant bacteria. The enriched genes were most abundant in genes related to carbohydrate and amino acid metabolism, with a greater number of non-redundant genes for glycoside hydrolases and glycosyl transferases. The enriched genes may have a direct impact on the organoleptic quality of JDSB. Functional enrichment analysis of different metabolites revealed that pathways associated with linoleic acid, ABC transporter, alanine, aspartate, glutamate, and galactose metabolism were enriched during JDSB processing. We identified a strong correlation between metagenomic functional level and metabolomics pathway enrichment analyses.The results contribute to the improvement of JDSB processing. [ABSTRACT FROM AUTHOR]

The content of malvin (malvidin-3,5-diglucoside) was determined in a set of wines obtained from Cabernet Cortis and other three recently introduced grape-resistant hybrids (Cabernet Volos, Merlot Khantus and Merlot Khorus) produced in South Tyrol (Italy), to establish their conformity to legal requirements and suggest winemaking approaches for their production and use. Different skin maceration times and ageing periods were considered for the wines investigated. Malvin content exceeded the limit of 15 mg/L, imposed by the Italian government for the commercialization of wines, in 18 out of 21 samples and showed a significant decrease during wine ageing (by about 40–50% every 12-month period) with the scarce effect of maceration time and fermentation temperature. Single-cultivar wines became suitable for commercialization after ca. 7 years ageing, following traditional winemaking approaches. The use of two commercial enzymatic pectolytic preparations indicated for aroma enhancement and possessing β-glucosidase activity was also investigated at two different dosages, to increase the degradation rate of malvin in wines after winemaking. They are unsuitable for significant malvin content decrease within a 6-month time lapse. [ABSTRACT FROM AUTHOR]

Discovering more novel antimicrobial compounds has become a keen research problem. In this study, YA215 genome was sequenced by the Illumina HiSeq + PacBio sequencing platform. Genome assembly was performed by Unicycler software and the gene clusters responsible for secondary metabolite biosynthesis were predicted by antiSMASH. The genome comprised 3976514 bp and had a 46.56% G + C content. 3809 coding DNA sequences, 27 rRNAs, 86 tRNAs genes, and 79 sRNA were predicted. Strain YA215 was re-identified as Bacillus velezensis based on ANI and OrthoANI analysis. In the COG database, 23 functional groups from 3090 annotations were predicted. In the GO database, 2654 annotations were predicted. 2486 KEGG annotations linked 41 metabolic pathways. Glycosyl transferases, polysaccharide lyases, auxiliary activities, glycoside hydrolases, carbohydrate esterases, and carbohydrate-binding modules were predicted among the 127 annotations in the CAZy database. AntiSMASH analysis predicted that B. velezensis YA215 boasted 13 gene clusters involved in synthesis of antimicrobial secondary metabolites including surfactin, fengycin, macrolactin H, bacillaene, difficidin, bacillibactin, bacilysin, and plantazolicin. Three of the gene clusters (gene cluster 5, gene cluster 9, and gene cluster 10) have the potential to synthesize unknown compounds. The research underscore the considerable potential of secondary metabolites, identified in the genomic composition of B. velezensis YA215, as versatile antibacterial agents with a broad spectrum of activity against pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Background/Objectives: Inborn errors of immunity (IEI) encompass various congenital disorders, resulting in immunity defects and recurrent infections. Home-based subcutaneous immunoglobulin replacement therapy (scIgRT) is the best treatment option for those with primary antibody deficiency (PAD). However, the lack of standardized procedures in patient training remains a challenge. Our study investigates nurses' practice and perspectives, aiming to identify areas for improvement in at-home scIgRT practice. Methods: We prepared a structured survey regarding scIgRT, including needle choice experience and perception of adverse events, and distributed it among qualified nurses involved in patient training and scIgRT supervising. Results: We included 56 nurses with a median age of 50 years. Among them, 67.9% represented adult care providers, while 32.1% supervised IgRT in children. Most respondents (83.9%) used the classic or assisted with hyaluronidase scIgRT preparations. Single-channel needles were administered most commonly (85.7%). The needle length was mostly chosen solely by a nurse (57.1%) or in cooperation with the patient (23.2%). Next, 9 mm and 12 mm needles were used most often (92.9% and 78.6%, respectively). As expected, the 6 mm needle was more frequently applied for children compared to adults (n = 16, 88.9% vs. n = 11, 28.9%, p < 0.001), while 12 mm was primarily used in adults (n = 35, 92.1% vs. n = 9, 50.0%, p < 0.001). Visual skin fold assessment was the basis for the needle selection (58.9%), followed by the injection site rule (26.8%) or a choice between two available needle types for thinner or thicker patients (25.0%). Results of this survey indicate that, according to nurses' opinions presented in this survey, the needle length could be associated with local scIgRT adverse events, such as side needle leakage or local burning. Yet, it was likely unrelated to general adverse signs, such as headaches or dizziness. Most respondents (66.1%) indicated that, even if local adverse events occur, patients are reluctant to change scIgRT preparation or needle length. Most participants (69.6%) reported that the optimal administration technique needs to be discussed with the patient before and during scIgRT. Conclusions: This study sheds light on scIgRT practice in Poland, emphasizing deficiency in needle selection technique. Future research should focus on standardized training and advanced needle selection procedures on patient outcomes, investigating the correlation between needle strategies and adverse events, as well as the effectiveness of scIgRT. [ABSTRACT FROM AUTHOR]

This study leverages Shotgun metagenomics to assess the rumen microbial community and functionality in Angus bulls with differing residual feed intake-expected progeny difference (RFI-EPD) values, aiming to elucidate the microbial contributions to feed efficiency. Negative RFI-EPD bulls (NegRFI: n=10; RFI-EPD= -0.3883 kg/d) and positive RFI-EPD bulls (PosRFI: n=10; RFI-EPD=0.2935 kg/d) were selected from a group of 59 Angus bulls (average body weight (BW) = 428 ± 18.8 kg; 350 ± 13.4 d of age) fed a high-forage total mixed ration after a 60-d testing period. At the end of the 60-d period, rumen fluid samples were collected for bacterial DNA extraction and subsequent shotgun metagenomic sequencing. Results of the metagenome analysis revealed greater gene richness in NegRFI bulls, compared to PosRFI. Analysis of similarity revealed a small but noticeable difference (P =0.052; R-value = 0.097) in the rumen microbial community of NegRFI and PosRFI bulls. Linear Discriminant Analysis effect size (Lefse) was utilized to identify the differentially abundant taxa. The Lefse results showed that class Fibrobacteria (LDA = 5.1) and genus Fibrobacter (LDA = 4.8) were greater in NegRFI bulls, compared to PosRFI bulls. Relative abundance of the carbohydrate-active enzymes was also compared using Lefse. The results showed greater relative abundance of glycoside hydrolases and carbohydrate-binding modules such as GH5, CBM86, CBM35, GH43 , and CBM6 (LDA > 3.0) in NegRFI bulls whereas GH13 and GT2 were greater in PosRFI bulls. The distinct metabolic and microbial profiles observed in NegRFI, compared to PosRFI bulls, characterized by greater gene richness and specific taxa such as Fibrobacter , and variations in carbohydrate-active enzymes, underscore the potential genetic and functional differences in their rumen microbiome. These findings contribute to a deeper understanding of the interplay between rumen microbiota and feed efficiency in Angus bulls, opening avenues for targeted interventions and advancements in livestock management practices. [ABSTRACT FROM AUTHOR]

Green tides caused by Ulva prolifera occur annually in the Yellow Sea of China, and the massive amount of biomass decomposing during the demise stage of this green tide has deleterious ecological effects. Although microorganisms are considered key factors influencing algal bloom demise, an understanding of the microbial-algae interactions within the phycospheric microenvironment during this process is still lacking. Here, we focused on the variations in phycospheric microbial communities during the late stage of the green tide in three typically affected areas of the Yellow Sea via metagenomic sequencing analysis. In total, 16.9 million reads obtained from 18 metagenome samples were incorporated into the assembled contigs (13.4 Gbp). The phycosphere microbial community composition and diversity changed visibly during the demise of U. prolifera. The abundances of algae-lysing bacteria, Flavobacteriaceae at the family level and Alteromonas , Maribacter , and Vibrio at the genus level increased significantly in the phycosphere. In addition, the levels of glycoside hydrolases (GHs) and polysaccharide lyases (PLs) enzymes, which decompose U. prolifera polysaccharides in the phycosphere, were greater. Therefore, the degradation of algal polysaccharides can increase the efficiency of carbon metabolism pathways in the phycospheric microenvironment. Most of the genes detected in the phycosphere, especially norC, nrfA , and nasA , were associated with nitrogen metabolism pathways and showed dynamics related to the demise of the large amount of organic matter released by a green tide. Therefore, the demise of green tide algae may affect the potential carbon and nitrogen cycles of the phycospheric microenvironment by driving changes in the structure and diversity of microbial communities. Our research provides a novel perspective to better understand the ecological impact of U. prolifera during the green tide demise stage. [ABSTRACT FROM AUTHOR]

Background: Wood-feeding termites have been employed as sources of novel and highly efficient lignocellulolytic enzymes due to their ability to degrade lignocellulose efficiently. As a higher wood-feeding termite, Globitermes sulphureus (Blattaria: Termitidae) plays a crucial role as a decomposer in regions such as Vietnam, Singapore, Myanmar, and Yunnan, China. However, the diversity of its gut microbiome and carbohydrate-active enzymes (CAZymes) remains unexplored. Here, we analyzed the diversity of hindgut microbial communities and CAZymes in a higher wood-feeding termite, G. sulphureus, and a lower wood-feeding termite, Coptotermes formosanus (Blattaria: Rhinotermitidae). Results: 16S rRNA sequencing revealed that Spirochaetota, Firmicutes, and Fibrobacterota were the dominant microbiota in the hindgut of the two termite species. At the phylum level, the relative abundances of Proteobacteria and Bacteroidota were significantly greater in the hindgut of C. formosanus than in G. sulphureus. At the genus level, the relative abundances of Candidatus_Azobacteroides and Escherichia-Shigella were significantly lower in the hindgut of G. sulphureus than in C. formosanus. Metagenomic analysis revealed that glycoside hydrolases (GHs) with cellulases and hemicellulases functions were not significantly different between G. sulphureus and C. formosanus. Interestingly, the cellulases in G. sulphureus were mainly GH5_2, GH5_4, GH6, GH9, and GH45, while the hemicellulases were mainly GH11, GH8, GH10, GH11, GH26, and GH53. In C. formosanus, the cellulases were mainly GH6 and GH9, and the hemicellulases were mainly GH5_7, GH5_21, GH10, GH12, and GH53. In addition, β-glucosidase, exo-β-1,4-glucanase, and endo-β-1,4-glucanase activities did not differ significantly between the two termite species, while xylanase activity was higher in G. sulphureus than in C. formosanus. The bacteria encoding GHs in G. sulphureus were mainly Firmicutes, Fibrobacterota, and Proteobacteria, whereas Bacteroidota and Spirochaetota were the main bacteria encoding GHs in C. formosanus. Conclusions: Our findings characterized the microbial composition and differences in the hindgut microbiota of G. sulphureus and C. formosanus. Compared to C. formosanus, G. sulphureus is enriched in genes encoding for hemicellulase and debranching enzymes. It also highlights the rich diversity of GHs in the hindgut microbiota of G. sulphureus, including the GH5 subfamily, GH6, and GH48, with the GH6 and GH48 not previously reported in other higher termites. These results strengthen the understanding of the diversity of termite gut microbiota and CAZymes. [ABSTRACT FROM AUTHOR]

Background: Late-onset Pompe disease (LOPD), a rare autosomal recessive multisystemic disorder, substantially impacts patients' day-to-day activities, outcomes, and health-related quality of life (HRQoL). The PROPEL trial compared cipaglucosidase alfa plus miglustat (cipa+mig) with alglucosidase alfa plus placebo (alg+pbo) in adult patients with LOPD over 52 weeks and showed improved motor and respiratory function in patients switching treatment from standard-of-care enzyme replacement therapy (ERT) to cipa+mig at baseline. This study evaluated the impact of cipa+mig on patient-reported outcomes (PROs), including HRQoL in ERT-experienced patients, using data from PROPEL. Methods: PROs evaluated included the Subject's Global Impression of Change (SGIC), Patient-Reported Outcomes Measurement Information System (PROMIS) Physical Function Short Form 20a, PROMIS Fatigue Short Form 8a, Rasch-built Pompe-specific Activity (R-PAct), and European Quality of Life-5 Dimensions 5 Response Levels (EQ-5D-5L). The proportions of responders in the cipa+mig arm and the alg+pbo arm were compared via chi-squared or Fisher's exact test (patient-level responder analysis), and least squares (LS) mean differences were calculated for change from baseline at Week 52 of the PRO measures (group-level analysis). Results: At Week 52, patient-level SGIC responder and group-level SGIC analyses favored cipa+mig compared with alg+pbo across all SGIC domains (e.g. 90 vs. 59% responders in the cipa+mig vs. the alg+pbo group for SGIC ability to move around; P = 0.0005; and LS mean difference 0.385; P = 0.02). Similarly, PROMIS Physical Function and Fatigue domains numerically favored cipa+mig in both analyses (e.g. 50 vs. 40% responders in the cipa+mig vs. alg+pbo arm for PROMIS Physical Function; P = 0.37; and LS mean difference 3.1; P = 0.11). R-PAct for both treatment groups was similar in the patient-level responder analysis, but numerically favored alg+pbo in the group-level analysis (35% responders in both arms; P = 0.95; and LS mean difference −0.8; P = 0.48). Self-care, usual activities, and depression/anxiety domains of EQ-5D-5L numerically favored cipa+mig in both analyses (e.g. 20 vs. 12% responders in the cipa+mig vs. alg+pbo arm for EQ-5D-5L self-care; P = 0.54; and LS mean difference −0.108; P = 0.52). Conclusions: Overall, switching treatment from alglucosidase alfa to cipa+mig positively impacted PRO measurements during the double-blind period of PROPEL. Trial registration: NCT03729362; Registration date: November 1, 2018; https://clinicaltrials.gov/study/NCT03729362 Plain English summary: Late-onset Pompe disease (LOPD) is a rare, multisystemic inherited genetic disease that causes glycogen accumulation in muscles and other body organs, leading to muscle weakness and respiratory insufficiency. LOPD significantly impacts patients' day-to-day life. Enzyme replacement therapies (ERT) have greatly improved the lives of patients with LOPD. The first approved ERT for LOPD was alglucosidase alfa (alg). To evaluate the effects of a new treatment (cipaglucosidase alfa+miglustat [cipa+mig]) in adult patients with LOPD, two-thirds of patients were switched from alg to cipa+mig and the remaining patients continued receiving alg (alg+placebo [alg+pbo]). We used patient-reported outcome (PRO) questionnaires (asking patients how they feel) to assess changes in patient health. Groups were similar at baseline. Analyses showed that patients improved following cipa+mig treatment for all domains of the PROs Subject's Global Impression of Change (SGIC; overall physical well-being, effort of breathing, muscle strength, muscle function, ability to move around, activities of daily living, energy level, level of muscular pain) and the Patient-Reported Outcomes Measurement Information System (PROMIS; Physical Function, Fatigue) compared with when treated with alg+pbo. Rasch-built Pompe-specific Activity (R-PAct), a survey evaluating daily activities and social life of patients living with Pompe disease, showed that patients felt similar after cipa+mig and alg+pbo. European Quality of Life-5 Dimensions-5 Response Levels (EQ-5D-5L), a measure of health covering five dimensions, favored cipa+mig in the self-care, usual activities, pain/discomfort and depression/anxiety areas, and alg+pbo for mobility. Overall, changing treatment from alg to cipa+mig positively affects PROs and the patient's general well-being. [ABSTRACT FROM AUTHOR]

Background: Green synthesized nanoparticles have recently gained significant medicinal applications and oftentimes outperform their green sources. Selenium is of fundamental importance to human health, stemming from its distinctive physicochemical properties, such as antioxidant activity, inhibition of Lipid peroxidation, stabilization of membrane proteins, maintenance of membrane fluidity and modulation of cell signaling. Though reports have shown some therapeutic potential of Ackee plant parts such as antioxidant, anti-inflammatory, antimicrobial, neuroprotective, very few scientific proofs still exist in support of these effects. Methods: This study synthesized selenium nanoparticles (Se-NPs) from crude methanolic extracts of Ackee leaves (AKL) and Ackee arils (AKA), examined the photo-physical characteristics of the Se-NPs and determined the in-vitro antioxidant, antidiabetic, and anti-inflammatory potentials of AKL, AKA, and their Se-NPs using established protocols. Results: In both leaves and arils Se-NPs: UV spectroscopy revealed a qualitative absorbance at 310 nm; FTIR indicated multiple vibrations around 4000 cm−1- 400 cm−1; SEM images of 5 µm principally showed consistent size distribution of amorphous and granular shape at a magnification of 10,000X; while EDS spectra strongly confirm the presence of atomic Se compound at 30 kV. Various antioxidant activities assays carried out showed a range of approximately 4 to 60 times higher activities of the AKL, AKA, and Se-NPs than Ascorbic acid—the standard drug used. Furthermore, appreciable activities of more than 50% were obtained for alpha-amylase and alpha-glucosidase inhibitory activities, along with highly significant activities of haemoglobin glycosylation, glucose uptake, membrane stabilization, anti-arthritic, anti-haemolysis activities, when AKL, AKA, and Se-NPs were compared with standard drugs. Conclusion: Encouraging the development and utilization of AKL, AKA, and Se-NPs will provide tremendous therapeutic efficacy and bioavailability approaches towards the management of diabetes, inflammation, and other oxidative stress-related diseases. [ABSTRACT FROM AUTHOR]

Objectives: Purple blotch, caused by the necrotrophic pathogen Alternaria porri, is one of the most economically significant diseases of onion and allied crops. While the virulent nature of many Alternaria spp. has been identified, the pathogenic repertoire of A. porri is still unknown. The objective of this work was to sequence the genome of A. porri using the PacBio SMRT sequencing strategy and analyse the repertoire of CAZymes, secondary metabolites, secretome and effectors in A. porri. Our research group is working to identify onion germplasm with purple blotch resistance and to understand the genetics of the pathogen. The reported de-novo assembly will contribute to the analysis of potential variants and the gene repertoire contributing to the virulence and pathogenicity of the purple blotch pathogen. Data description: Long-read sequencing on a PacBio Sequel II system resulted in a 32.98 Mb (20 contigs) assembly with an N50 of 2, 657, 264 bp, the longest contig length of 5.05 Mb, and a GC content of 51.06%. The Benchmarking Universal Single-Copy Orthologs (BUSCO) analysis resulted in 99.7% genome completeness at the Dothideomycetes lineage, representing a high-quality genome assembly. AUGUSTUS ab initio analysis resulted in 9875 protein-coding genes. Of the 6776 pathogenicity-related genes, 537 genes with effector functions were identified. Likewise, the glycoside hydrolases (434) were the most dominant group of the total 837 predicted CAZymes. The assembled genome of A. porri showed distinctive similarities to the genomes of A. alternata and A. brassicicola, the causal agents of leaf blight of onion and leaf spot of Brassica crops, respectively. [ABSTRACT FROM AUTHOR]

Broadly neutralizing antibodies (bNAbs) isolated from HIV-1 infected donors are vaccine paradigms. These bNAbs recognize envelope glycoprotein trimers that carry 75–90 oligomannose and complex-type glycans. Although bNAbs and their precursors must navigate past glycans, they usually also make some glycan contacts. Glycan-modified vaccines may therefore be useful to initiate and guide bNAb development. Here, we describe two ways to modify Env glycans for possible vaccine use: 1) using a cocktail of glycosidases (termed "NGAF3" (N euraminidase, β- G alactosidase, N- A cetylglucosaminidase, endoglycosidase F3 (endo F3)) to deplete complex glycans to try to minimize bNAb-glycan clashes and 2) co-expressing β-1,4-galactosyltransferase 1 (B4G) and β-galactoside α-2,6 sialyltransferase 1 (ST6) during Env biosynthesis, creating bNAb-preferred glycan structures. Mass spectrometry revealed that NGAF3 removed glycan heads at 3/7 sites occupied by complex glycans. B4G overexpression resulted in hybrid glycan development whenever complex glycans were closely spaced. The glycan at position 611 in of Env's gp41 transmembrane subunit was uniquely isolated from the effects of both endo F3 and B4G. B4G and ST6 co-expression increased hybrid and sialylated glycan abundance, reducing glycan complexity. In rabbit vaccinations, B4G + ST6 virus-like particles (VLPs) induced less frequent, weaker titer NAbs, implying that ST6-mediated increased Env charge dampens vaccine antibodies. In some cases, vaccine sera preferentially neutralized B4G + ST6-modified pseudovirus. HIV-1+ donor plasma NAbs were generally more effective against B4G + ST6 modified pseudovirus, suggesting a preference for less complex and/or α-2,6 sialylated Env trimers. Collectively, our data suggest that B4G and ST6 Env modifications are best suited for intermediate or late vaccine shots. [ABSTRACT FROM AUTHOR]

Olea europaea L. (Oleaceae), is rich in phenolic content and has powerful antioxidant and antidiabetic activity. However, there are no medicinal products prepared due to this feature. Therefore, this study aims to characterize an O. europaea extract with strong antioxidant and antidiabetic properties and to prepare nanoformulations containing this extract. To determine the activities of the extracts prepared from the leaves of the plant, DPPH• and ABTS•+ scavenging, Fe+3 reducing activity, α-amylase, and α-glucosidase inhibition assays were performed. The oleuropein content of the absolute ethanol extract with the highest activity was analysed by HPLC. The characterized extract was loaded into liposomes and chitosan coated liposomes, and the long-term sustainability of their activity was investigated. The encapsulation efficiency was 65.2% for the liposome and 66.8% for the chitosan-coated liposome formulation. The amounts of the extracts released from the formulations were evaluated to exhibit antioxidant and antidiabetic activity. [ABSTRACT FROM AUTHOR]

Astragalus membranaceus is a traditional Chinese medicine with multiple pharmacological activities. Modern pharmacological research has found that Astragalus membranaceus extract has an inhibitory effect on α-glucosidase, however, which component can inhibit the activity of α-glucosidase and its degree of inhibition are unknown. To address this issue, this study used affinity ultrafiltration screening combined with UPLC-ESI-Orbitrap-MS technology to screen α-glucosidase inhibitors in Astragalus membranaceus. Using affinity ultrafiltration technology, we obtained the active components, and using UPLC-ESI-Orbitrap-MS technology, we quickly analyzed and identified them. As a result, a total of 8 ingredients were selected as α-glucosidase inhibitors. [ABSTRACT FROM AUTHOR]

The production of wheat is fundamentally interconnected with worldwide food security. The practice of deep tillage (DT) cultivation has shown advantages in terms of soil enhancement and the mitigation of diseases and weed abundance. Nevertheless, the specific mechanisms behind these advantages are unclear. Accordingly, we aimed to clarify the influence of DT on rhizosphere soil (RS) microbial communities and its possible contribution to the improvement of soil quality. Soil fertility was evaluated by analyzing several soil characteristics. High-throughput sequencing techniques were utilized to explore the structure and function of rhizosphere microbial communities. Despite lowered fertility levels in the 0–20 cm DT soil layer, significant variations were noted in the microbial composition of the DT wheat rhizosphere, with Acidobacteria and Proteobacteria being the most prominent. Furthermore, the abundance of Bradyrhizobacteria, a nitrogen-fixing bacteria within the Proteobacteria phylum, was significantly increased. A significant increase in glycoside hydrolases within the DT group was observed, in addition to higher abundances of amino acid and carbohydrate metabolism genes in the COG and KEGG databases. Moreover, DT can enhance soil quality and boost crop productivity by modulating soil microorganisms' carbon and nitrogen fixation capacities. [ABSTRACT FROM AUTHOR]

Glycosidase is an indispensable enzyme in the sugar industry. It is difficult to maintain the stability of natural glycosidase during long-term high-temperature sugar conversion, limiting its application in industrial production. Numerous studies have shown that protein engineering is an important means for enhancing the thermal stability of glycosidase. This article systematically reviews the latest advances in using protein engineering to improve the thermal stability of glycosidase, and dissects the directed evolution, rational design, and semi-rational design strategies used in protein engineering. It also evaluates the effectiveness of these strategies in enhancing the thermal stability of glycosidase and their potential industrial application value, and presents the advantages, limitations and challenges of each strategy. Finally, it suggests that different strategies should be jointly used to improve the thermal stability of glycosidase and gives an outlook on future research directions with a view to promoting the synergistic improvement of the thermal stability and activity of glycosidase.

This study aimed to comprehensively characterize the gut microbiota in diarrheal mink. We conducted Shotgun metagenomic sequencing on samples from five groups of diarrheal mink and five groups of healthy mink. The microbiota α-diversity and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology did not show significant differences between the groups. However, significant differences were observed in microbiota β-diversity and the function of carbohydrate-active enzymes (CAZymes) between diarrheal and healthy mink. Specifically, The relative abundance of Firmicutes was lower, whereas that of Bacteroidetes was higher in diarrheal mink. Fusobacteria were enriched as invasive bacteria in the gut of diarrheal mink compared with healthy mink. In addition, Escherichia albertii was identified as a new bacterium in diarrheal mink. Regarding functions, nicotinate and nicotinamide metabolism and glycoside hydrolases 2 (GH2) family were the enhanced KEGG orthology and CAZymes in diarrheal mink. Furthermore, the diversity and number of antibiotic-resistant genes were significantly higher in the diarrheal mink group than in the healthy group. These findings enhance our understanding of the gut microbiota of adult mink and may lead to new approaches to the diagnosis and treatment of mink diarrhea. [ABSTRACT FROM AUTHOR]

Guided by in silico analysis tools and biotransformation technology, new derivatives of natural compounds with heightened bioactivities can be explored and synthesized efficiently. In this study, in silico data mining and molecular docking analysis predicted that glucosides of skullcapflavone II (SKII) were new flavonoid compounds and had higher binding potential to oncogenic proteins than SKII. These benefits guided us to perform glycosylation of SKII by utilizing four glycoside hydrolases and five glycosyltransferases (GTs). Findings unveiled that exclusive glycosylation of SKII was achieved solely through the action of GTs, with Bacillus subtilis BsUGT489 exhibiting the highest catalytic glycosylation efficacy. Structure analysis determined the glycosylated product as a novel compound, skullcapflavone II‐6′‐O‐β‐glucoside (SKII‐G). Significantly, the aqueous solubility of SKII‐G exceeded its precursor, SKII, by 272‐fold. Furthermore, SKII‐G demonstrated noteworthy anti‐melanoma activity against human A2058 cells, exhibiting an IC50 value surpassing that of SKII by 1.4‐fold. Intriguingly, no substantial cytotoxic effects were observed in a murine macrophage cell line, RAW 264.7. This promising anti‐melanoma activity without adverse effects on macrophages suggests that SKII‐G could be a potential candidate for further preclinical and clinical studies. The in silico tool–guided synthesis of a new, highly soluble, and potent anti‐melanoma glucoside, SKII‐G, provides a rational design to facilitate the future discovery of new and bioactive compounds. [ABSTRACT FROM AUTHOR]

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Glycoside hydrolases have been implicated in a wide range of human conditions including lysosomal storage diseases. Consequently, many researchers have directed their efforts towards identifying new classes of glycoside hydrolase inhibitors, both synthetic and from natural sources. A large percentage of such inhibitors are reversible competitive inhibitors that bind in the active site often due to them possessing structural features, often a protonatable basic nitrogen atom, that mimic the enzymatic transition state. We report that mechanism-based small molecule galacto-like configured cyclohexenyl carbasugars form reversible covalent complexes with both α-galactosidase and β-galactosidase. In addition, we show that the β-galactosidase from Aspergillus oryzae reacts with three different carbasugar inhibitors, with three different second-order rate constants (kinact/Ki), to give the same enzyme–carbasugar covalent intermediate. The surprising observation that the α-galacto-configured inhibitor covalently labels the A. oryzae β-galactosidase highlights the catalytic versatility of glycoside hydrolases. We expect that cyclohexenyl covalent inhibitors will become an important class of compounds in the chemical biologist's tool box. [ABSTRACT FROM AUTHOR]

This study was designed to screen 6 lysosomal storage diseases (LSDs) in neonates using tandem mass spectrometry (MS/MS), and establish cutoff values for these LSDs with 3000 dried blood spots (DBS) samples. Cutoff values for α-L-iduronidase (IDUA), α-galactosidase (GLA), acid beta glucosidase (ABG), β-galactocerebrosidase (GALC), acid sphingomyelinase (ASM), and acid alpha glucosidase (GAA) were as follows: GLA, > 2.06 μmol/L·h; ABG, > 1.78 μmol/L·h; ASM, > 0.99 μmol/L·h; IDUA, > 1.33 μmol/L·h; GALC, > 0.84 μmol/L·h; and GAA, > 2.06 μmol/L·h. There were 30 positives in initial MS/MS screening test, and 15 samples were still positive with repeat testing. Their parents/guardians were recontacted and DBS samples were collected again for test. Only 1 child showed abnormal GAA enzyme activity after recontacting process, and was diagnosed with Pompe disease after genetic screening. Eventually, cutoff values of 6 specific enzyme activities were established and MS/MS is effective for early LSDs screening. [ABSTRACT FROM AUTHOR]

There has been recent interest in using the β-galactosidase of Bacillus circulans to produce prebiotic oligosaccharides, including galactooligosaccharides (GOS), lactulose and lactosucrose. The types and amounts of the transgalactosylation products produced depend on the selectivities of the enzyme for the various transgalactosylation and hydrolysis reactions that occur. To date, the selectivities of the β-galactosidase of B. circulans for these reactions have not been adequately characterized. In the current work, we undertake four case studies using literature data for systems in which different product mixtures are produced (i) GOS; (ii) GOS and disaccharides; (iii) GOS and lactulose; and (iv) GOS and lactosucrose. We analyze this data to obtain quantitative estimates of the relevant selectivities. We show that, in the production of GOS, the isoform β-Gal-A is significantly less selective against hydrolysis reactions than the other β-galactosidase isoforms. In the production of lactulose, the β-galactosidase of B. circulans gives high lactulose yields, despite having a 19- to 33-fold preference for producing GOS over lactulose. In the production of lactosucrose, the β-galactosidase of B. circulans also prefers to produce GOS. [Display omitted] • β-galactosidase of B. circulans characterized in four transglycosylation processes. • Isoform β-Gal-A less selective against 1° and 2° hydrolysis than other isoforms. • High lactulose yields, despite preference for producing galactooligosaccharides. • Enzyme prefers producing galactooligosaccharides over lactosucrose. • Method for estimating selectivities unaffected by inhibition or denaturation. [ABSTRACT FROM AUTHOR]

Aromase™ H2 is an enzymatic cocktail marketed to intensify the aroma, promote clarity and overall flavor of tea by releasing aromatic compounds and antioxidants that are naturally glycosylated. The hydrolysis of a wide range of glycoconjugates is the key to its action, which can arise from the substrate promiscuity of the main diglycosidase or other enzymes that are in the mixture. The characterization focusing on the activity toward flavonoids at different pH values allowed to increase the cocktail ratio of diglycosidase to α-rhamnosidase activity. In that way, the free disaccharide rutinose was selectively obtained in a laboratory scale reactor with 90 % conversion after 3.5 h reaction at pH 8. Purification was carried out by liquid-liquid extraction, recovering the aglycone hesperetin. Subsequent extraction with activated carbon allowed a recovery of 84 % of the rutinose. This work demonstrates the practical application of a commercial cocktail for the selective hydrolysis of rutinosylated compounds. [Display omitted] • Aromase™ H2 activities were measured using flavonoids and synthetic substrates. • Selectivity of the hydrolysis was increased by modifying the reaction conditions. • Hesperidin was hydrolyzed at the heterosidic bond rendering rutinose. • Rutinose and hesperetin were produced in a laboratory scale reactor and purified. [ABSTRACT FROM AUTHOR]

A comparative analysis of carbohydrate degradation proteins encoded in the currently available genomic sequences of bacteria of the family Sphaerochaetaceae, namely Sphaerochaeta associata GLS2T, S. globosa BuddyT, S. pleomorpha GrapesT, S. halotolerans 4-11T, S. halotolerans 585, Sphaerochaeta sp. S2, Sphaerochaeta sp. PS, and Parasphaerochaetacoccoides SPN1T was carried out. The genomes of Sphaerochaeta spp. encode a medium-sized and diverse set of proteins potentially involved in the degradation of different classes of carbohydrates, mainly oligosaccharides. All studied genomes encode glycoside hydrolases of the GH1, GH2, GH3, GH4, GH13, GH20, GH28, GH36, GH43, GH57, GH63, GH77 and GH105 families, as well as carbohydrate esterases of the CE8 and CE9 families. All studied bacteria, with the exception of P. coccoides SPN1T, have many genes of the GH31 family proteins. The studied members of Sphaerochaetaceae do not have genes coding for endo-β-acetylmuramidase (lysozyme) of the GH23 family, which participates in peptidoglycan turnover. However, the genomes of S. associata, S. globosa, Sphaerochaeta sp. PS, and S. pleomorpha contain exo-β-acetylmuramidase genes (GH171 family). A significant part of the genes encoding carbohydrate degradation enzymes have the closest homologues among representatives of the phyla Bacillota, Bacteroidota, and Pseudomonadota. The genomes of the studied bacteria encode proteins that could potentially be involved in the degradation of pectin. The ability of the Sphaerochaetaceae members to use pectin for growth, as well as the evolutionary origin of the genes encoding potential α-galacturonidase (GH4 family) and unsaturated glucuronyl/rhamnogalacturonyl hydrolase (GH105 family), which participate in the degradation of pectin components, were studied. [ABSTRACT FROM AUTHOR]

The interactions between hosts and the gut microbiota are intricate and can significantly affect the ecology and evolution of both parties. Various host traits, including taxonomy, diet, social behaviour, and external factors such as prey availability and the local environment, all play an important role in shaping composition and diversity of the gut microbiogta. In this study, we explored the impact of intestinal microorganisms on the host in adapting to their respective ecological niches in two species of snakes. We collected feces from Lycodon rufozonatus and Lycodon rosozonatus from different geographical locations and used 16S rRNA gene sequencing technology to sequence the v3‐v4 region. The results revealed that there was no significant difference in the alpha diversity of intestinal microorganisms between L. rufozonatus and L. rosozonatus. The gut microbiota of all individuals comprised four main phyla: Pseudomonadota, Bacteroidota, Bacillota, and Actinomycetota. At the genus level, the genus Salmonella dominated the enterobacterial microbiota in the samples from Hainan, while there was no obvious dominant genus in the enterobacterial microbiota of the samples from the other four localities. Comparative analysis of enzyme families annotated to the gut microbiota between L. rufozonatus and L. rosozonatus from the four sampling regions by CAZy carbohydrate annotation revealed that nine enzyme families differed significantly in terms of glycoside hydrolases (GHs). In addition, we compared the composition of gut microbial communities between L. rufozonatus and L. rosozonatus and investigated the impact of the differences on their functions. Our results will provide insights into the coevolution of host and gut microbes. [ABSTRACT FROM AUTHOR]

Objectives We investigated the potential of serum proteins for distinguishing clinical and molecular subtypes in patients with GCA. Methods Proximity extension assays were used to analyse 1463 proteins in serum samples from patients with new-onset GCA (n  = 16) and patients who have achieved remission (n  = 13). Unsupervised and supervised cluster analyses were performed. Results Unsupervised cluster analysis identified three distinct clusters based on the protein signature. Compared with cluster 2, patients of cluster 1 had fewer PMR symptoms, increased levels of macrophage migration inhibitory factor (MIF) and pronounced NF-κB, STAT5 and IL-1 signalling. The changes in serum proteins upon remission differed between cluster 1 and 2. Patients with cranial GCA were characterized by altered endothelial and Th17 signalling, whereas patients not responding to treatment within the GUSTO-trial showed increased Th1 and diminished B cell signalling. Patients with anterior ischaemic optic neuropathy displayed higher levels of CHI3L1 (YKL40) and MMP12, and reduced levels of TIMP3. Conclusion Protein profiling identified patient clusters in GCA with distinct proteomic features and therefore likely different pathophysiology. These unique proteomic footprints might lead to more targeted treatments in future. [ABSTRACT FROM AUTHOR]

The biochemical and structural characteristics of PtLam, a laminarinase from deep-sea Planctomycetota, have been extensively elucidated, unveiling the fundamental molecular mechanisms governing substrate recognition and enzymatic catalysis. PtLam functions as an exo-laminarinase with the ability to sequentially hydrolyze laminarin, cleaving glucose units individually. Notably, PtLam exhibits proficient transglycosylation capabilities, utilizing various sugar alcohols as acceptors, with lyxose, in particular, yielding exclusively transglycosylated products. Structural analysis of both apo-PtLam and its laminarin oligosaccharide-bound complex revealed significant conformational alterations in active residues upon substrate binding. Moreover, pivotal residues involved in substrate recognition were identified, with subsequent mutation assays indicating the contribution of positive subsites in modulating exo-hydrolysis and transglycosidic activities. These results enhance our comprehension of laminarin cycling mechanisms by marine Planctomycetota, while also providing essential enzyme components for laminarin hetero-oligosaccharide synthesis. IMPORTANCE The ubiquitous Planctomycetota, with distinctive physiological traits, exert a significant influence on global carbon and nitrogen fluxes. Their intimate association with algae suggests a propensity for efficient polysaccharide degradation; however, research on glycoside hydrolases derived from Planctomycetota remains scarce. Herein, we unveil the GH16 family laminarinase PtLam from deep-sea Planctomycetota, shedding light on its catalytic mechanisms underlying hydrolysis and transglycosylation. Our findings elucidate the enzymatic pathways governing the marine laminarin cycle orchestrated by Planctomycetota, thereby fostering the exploration of novel polysaccharide hydrolases with promising practical implications. [ABSTRACT FROM AUTHOR]

The upwelling phenomenon plays a vital role within marine ecosystems, transporting essential nutrients from the bottom to the surface and boosting biological productivity. However, the bacterial community structure in upwelling zones along the western coast of India (WCI) is understudied. This research systematically examines bacterial diversity across three seasons—pre-monsoon (PR), monsoon (MN), and post-monsoon (PM)—using next-generation sequencing. Our findings show distinct spatial patterns of bacterial communities in the Arabian Sea and demonstrate that ecological variations influence bacterial distribution in this dynamic environment. During MN, the bacterial community exhibited greater species diversity but lower overall abundance compared to PR and PM. Non-Metric MDS cluster analysis revealed a 78% similarity (at order level) between PR and PM, indicating that MN supports unique bacterial diversity. KEGG analysis showed significant seasonal variations in metabolic functions, with increased functional potential during MN. Additionally, Carbohydrate-Active enZymes (CAZymes) analysis revealed distinct seasonal profiles, among which the GH13 enzymes were the most prevalent glycoside hydrolases during MN, predominantly being sucrose phosphorylase and glucosidase, known for breaking down glucan deposits derived from phytoplankton. The CAZymes profiles supported taxonomic and KEGG pathway findings, reinforcing that microbial communities are seasonally distinct and functionally adapted to changing availability of nutrients. [ABSTRACT FROM AUTHOR]

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Glycoside Hydrolase family 65 (GH65) is a unique family of carbohydrate-active enzymes. It is the first protein family to bring together glycoside hydrolases, glycoside phosphorylases and glycosyltransferases, thereby spanning a broad range of reaction types. These enzymes catalyze the hydrolysis, reversible phosphorolysis or synthesis of various α-glucosides, typically α-glucobioses or their derivatives. In this review, we present a comprehensive overview of the diverse reaction types and substrate specificities found in family GH65. We describe the determinants that control this remarkable diversity, as well as the applications of GH65 enzymes for carbohydrate synthesis. Key points: • GH65 is the first CAZy family to contain hydrolases, phosphorylases and transferases • Distinct residues and loops are determinants of substrate specificity in family GH65 • GH65 enzymes hold strong potential for carbohydrate synthesis via coupled reactions [ABSTRACT FROM AUTHOR]

Tuberculosis has posed a serious threat to human health. It is imperative to investigate the geographic prevalence of tuberculosis and medication resistance, as this information is essential for informing strategies for its prevention and treatment. Drug resistance was identified using a proportion method. Drug-resistant genes and pathways were predicted using whole genome sequencing. The drug resistance range of bedaquiline was identified using the microporous plate two-fold dilution method, and drug resistance genes were studied using sequencing. The study revealed that 19.99% of the tuberculosis cases had multidrug resistance. The genes of M. tuberculosis are predominantly involved in the synthesis of ABC transporters, two-component systems, and bacterial secretion systems, as well as in energy production and conversion, and lipid transport and metabolism. The genes encode for 82.45% of carbohydrate-related enzymes such as glycoside hydrolases, glycosyl transferases, and carbohydrate esterases. The minimum inhibitory concentration (MIC) of bedaquiline against clinical strains was approximately 0.06 μg/mL, with identified mutations in drug-resistant genes Rv0678, atpE, and pepQ, specifically V152A, P62A, and T222N, respectively. The multidrug resistance tuberculosis development was attributed to the strong medication resistance exhibited. It was concluded that tuberculosis had presented a high level of drug resistance. Phenotypic resistance was related to genes, existing potential genetic resistance in M. tuberculosis. Bedaquiline was found to possess effective antibacterial properties against M. tuberculosis. [ABSTRACT FROM AUTHOR]

The valorization of lignocellulosic biomass, derived from various bio-waste materials, has received considerable attention as a sustainable approach to improve production chains while reducing environmental impact. Microbial enzymes have emerged as key players in the degradation of polysaccharides, offering versatile applications in biotechnology and industry. Among these enzymes, glycoside hydrolases (GHs) play a central role. Xylanases, in particular, are used in a wide range of applications and are essential for the production of xylose, which can be fermented into bioethanol or find use in many other industries. Currently, fungal secretomes dominate as the main reservoir of lignocellulolytic enzymes, but thermophilic microorganisms offer notable advantages in terms of enzyme stability and production efficiency. Here we present the genomic characterization of Geobacillus stearothermophilus GF16 to identify genes encoding putative enzymes involved in lignocellulose degradation. Thermostable GHs secreted by G. stearothermophilus GF16 were investigated and found to be active on different natural polysaccharides and synthetic substrates, revealing an array of inducible GH activities. In particular, the concentrated secretome possesses significant thermostable xylanase and β-xylosidase activities (5 ×103 U/L and 1.7 ×105 U/L, respectively), highlighting its potential for application in biomass valorization. We assessed the hemicellulose hydrolysis capabilities of various agri-food wastes using the concentrated secretome of the strain cultivated on xylan. An impressive 300-fold increase in xylose release compared to a commercially available cocktail was obtained with the secretome, underscoring the remarkable efficacy of this approach. [Display omitted] • Genome characterization of G. stearothermophilus GF16 reveals genes potentially involved in hemicellulose degradation. • The secretome of GF16 grown on a minimal medium enriched with 0.1 % xylan shows inducible xylanolytic properties. • G. stearothermophilus GF16 CAZymes are pivotal in converting agri-food wastes into value-added products, such as xylose. [ABSTRACT FROM AUTHOR]

Simple Summary: Immunoglobulin (Ig) replacement therapy (IgRT) consists of the administration of low-doses human polyclonal Igs for the treatment of primary and secondary hypogammaglobulinemia, characterized by low serum levels of immunoglobulins that is associated with recurrent infections and immune dysfunction. In this review, we focus on the application and efficacy of therapeutic Igs for the management of multiple myeloma (MM) patients affected by secondary hypogammaglobulinemia that is associated with poor patients' outcome. The use of IgRT restores physiological antibody levels and stimulates innate and adaptive immune responses as well. Therefore, in MM settings the IgRT has shown a significant positive impact on infection rates increasing the patients' overall health status that correlates to a decrease in long-term complications and hospitalization and to an improved therapeutic adherence and patients' quality of life. Immunoglobulin (Ig) replacement therapy (IgRT) consists of the administration of low-dose human polyclonal Igs for the treatment of primary and secondary hypogammaglobulinemia that are associated with recurrent infections and immune dysfunction. IgRT restores physiological antibody levels and induces an immunomodulatory effect by strengthening immune effector cells, thus reducing infections. Here, we describe the pharmacology of different Ig formulations with a particular focus on their mechanism of action as low-dose IgRT, including the direct anti-microbial effect and the immunomodulatory function. In addition, we describe the use of therapeutic Igs for the management of multiple myeloma (MM), a hematologic malignancy characterized by severe secondary hypogammaglobulinemia associated with poor patient outcome. In MM settings, IgRT prevents life-threatening and recurrent infections showing promising results regarding patient survival and quality of life. Nevertheless, the clinical benefits of IgRT are still controversial. A deeper understanding of the immune-mediated effects of low-dose IgRT will provide the basis for novel combined therapeutic options and personalized therapy in MM and other conditions characterized by hypogammaglobulinemia. [ABSTRACT FROM AUTHOR]

The rise of agro-industrial activities over recent decades has exponentially increased lignocellulose biomasses (LCB) production. LCB serves as a cost-effective source for fermentable sugars and other renewable chemicals. This study explores the use of microbial consortia, particularly thermophilic consortia, for LCB deconstruction. Thermophiles produce stable enzymes that retain activity under industrial conditions, presenting a promising approach for LCB conversion. This research focused on two microbial consortia (i.e., microbiomes) that were analyzed for enzyme production using a cheap medium, i.e., a mixture of spent mushroom substrate (SMS) and digestate. The secreted xylanolytic enzymes were characterized in terms of temperature and pH optima, thermal stability, and hydrolysis products from LCB-derived polysaccharides. These enzymes showed optimal activity aligning with common biorefinery conditions and outperformed a formulated enzyme mixture in thermostability tests in the digestate. Phylogenetic and genomic analyses highlighted the genetic diversity and metabolic potential of these microbiomes. Bacillus licheniformis was identified as a key species, with two distinct strains contributing to enzyme production. The presence of specific glycoside hydrolases involved in the cellulose and hemicellulose degradation underscores these consortia's capacity for efficient LCB conversion. These findings highlight the potential of thermophilic microbiomes, isolated from an industrial environment, as a robust source of robust enzymes, paving the way for more sustainable and cost-effective bioconversion processes in biofuel and biochemical production and other biotechnological applications. [ABSTRACT FROM AUTHOR]

AbstractAlpha glucosidases are multifunctional glycoside hydrolases with hydrolysis and transglycosylation ability. It can be utilized for the glycosidic bond synthesis or glycosylation of Ascorbic acid/Vitamin C to its stable analogue, Ascorbic acid 2 glucoside (AA2G), a compound with wide applications in cosmetics and pharma. The application of α-glucosidases for industrial scale transglycosylation is limited due to the low transglycosylation yield of free enzymes. Enzyme immobilization techniques could enable the development of efficient, reusable catalysts. Only a few glycoside hydrolases have been studied in immobilized form for transglycosylation reactions, and α-glucosidases are probably the least explored in this form. Transglycosylation activity of immobilized α-glucosidase from Aspergillus carbonarius BTCF 5 was studied for AA2G synthesis, where different immobilization techniques like calcium alginate encapsulation, adsorption on chitosan beads, covalent cross-linking on magnetic nanoparticles, and cross-linked enzyme aggregates (CLEA) were employed for the immobilization. The immobilization yield of calcium alginate encapsulated enzyme, enzyme immobilized on Fe-MNP support, enzyme immobilized on chitosan beads and as CLEA were 107%, 99%, 46% and 486%, respectively. CLEA was identified as the best immobilization technique for this bi-substrate reaction due to the high immobilization yield and activity retention (30% activity retained after 5 consecutive cycles). Enzyme immobilization increased the transglycosylation activity by 38%, yielding 118 mM AA2G against 72 mM by the free enzyme. This indicates the potential of immobilized α-glucosidase as a catalyst for synthesizing AA2G at an industrial scale. [ABSTRACT FROM AUTHOR]

Glycoside hydrolases (GHs) are important in metabolic processes involving diverse carbohydrate-based substances found inside plant tissues. Potatoes (Solanum tuberosum) are rich in starchy carbohydrates, suggesting the role of GHs in their metabolic pathways. In this study, we examine the GH superfamily in potato where 366 potential GHs were identified using a similarity search method. Genes were subjected to further characterisation to gain insights into their structural composition, functional properties and distribution patterns across tissue types. Several in silico methodologies were also employed to investigate the physicochemical features, conserved motifs, chromosomal mapping, duplication events, syntenic links with tomato (Solanum lycopersicum), subcellular localisations, secondary structures and phylogenetic relationships. Cis -elements in StGHs revealed that the promoters of StGHs contain cis -elements that are responsive to phytohormones that are involved in plant growth and development, and are associated with stress responses. RNA-seq data identified significant changes in expression levels of GH16, GH17 , GH18 , GH19 and GH28 members under stress conditions. Expression patterns of several GHs were confirmed using real time quantitative PCR in response to stress. StGH16.24 expression increased after 3 days of drought stress, whereas StGH16.30 continuously increased under salt stress. Potential interactions between potato miRNAs and StGH revealed 393 and 627 interactions under drought and salt stress, respectively. Our findings offer insights into specific functions of GHs in diverse developmental stages and stress-related challenges in potato and other plants. Glycoside hydrolases (GHs) are involved in plant defence, cell wall metabolism and stress response. A total of 366 GH superfamily genes were identified in potato (Solanum tuberosum). Differential expression analysis indicated the involvement of GHs in various tissues, abiotic stress and biotic stresses, with specific genes showing significant changes in expression levels under drought and salt stress. Real time qPCR analysis revealed that StGH16.24 expression increased significantly after 3 days of drought stress, validating the presence of ABRE, MYB and MYC cis -regulatory elements. This article belongs to the Collection Functional Genomics for Developing Climate Resilient Crops. [ABSTRACT FROM AUTHOR]

Glycoside hydrolases belonging to family 48 (GH48) are one of the most important cellulases responsible for the synergistic degradation of crystalline cellulose (CC) to glucose. Microbial DNA was extracted from soils and compost followed by PCR amplification to detect bacterial GH48 genes. Amplified fragments were cloned to develop DNA libraries to determine the diversity and community structure of bacterial genes responsible for cellulose degradation using GH48 genes. Soil bacterial communities were predominantly based upon unidentified GH48 genes (66%). When identification was possible, community composition analysis of soil samples showed 4 known bacterial phyla with Proteobacteria (23%), Actinobacteria (9%), Firmicutes (1%), and Bacteroidetes (1%). Phylogenetic analysis of unidentified clones showed 2% of clones clustered with a reference sequence from the Chloroflexota genus Herpetosiphon sp. Compost GH48 sequences were predominantly comprised of 3 different bacterial phyla belonging to Firmicutes (72%), Actinobacteria (16%), Myxococcota (5%) with 7% of sequences not matching any known bacteria. The most abundant microbial species in soil with GH48 genes was the Proteobacteria species Massilia violaceinigra (13%) while in compost the Firmicutes species Bacillus licheniformis showed the highest percentage (70%). Soil communities showed a higher genetic diversity than compost predominantly composed of unknown genetic sequences showing the potential to isolate and characterize new cellulases from soil. The contribution of different bacterial phyla, genera, and species to the synergistic CC degradation by GH48 genes demonstrated the importance of diverse bacterial communities to optimize carbon recycling in the environment. [ABSTRACT FROM AUTHOR]