Search

The utilization of hydrolytic enzymes in various industrial processes worldwide has gained more attention than chemical catalysts due to the high selectivity of enzymes, their ease of control, and their negligible environmental impact, as they produce very small amounts of byproducts. Xylanase is one such enzyme that catalyzes the breakdown of the β-1,4 linkage of xylan, the second most abundant renewable heteropolysaccharide and hemicellulosic constituent of the plant cell wall. Naturally, xylanase can be obtained from various sources such as mollusks, insects, plants, animals, and various microorganisms (bacteria, fungi, yeast, and algae). The utilization of xylanase could greatly improve the overall economics of processing lignocellulosic materials for the generation of monosaccharides, liquid fuels, and chemicals. Microbial xylanase is suitable for applications in food and feed, paper and pulp, textile, pharmaceutical, and biorefining industries. It has gained global attention due to its substrate specificities, biochemical properties, and various biotechnological applications. This review focuses on xylanase production, sources, fermentation processes, modes of action, purification methods, and applications in various industries. [ABSTRACT FROM AUTHOR]

The modes of formation and release of secretion are complex processes that occur in secretory ducts and their description has great divergence in some species. The use of modern techniques to detect hydrolytic enzymes, cytoskeleton arrangement and indicators of programmed cell death may help clarify the processes involved during the ontogeny of that gland. The goal of our study was to analyze subcellular changes during schizogenous formation and secretion production and release into the lumen in resin ducts of Kielmeyera appariciana. Our results demonstrate the participation of pectinase through the loosening of the central cells of the rosette, which subsequently split from each other through polarized growth mediated by a rearrangement of the microtubules. The resin is mainly synthesized in plastids and endoplasmic reticulum and is observed inside vesicles and small vacuoles. The secretion release is holocrine and occurs through programmed cell death related to the release of reactive oxygen species, causing cytoplasm darkening, chromatin condensation, vacuole rupture and plastid and mitochondria degeneration. Cellulase activity was identified prior to the rupture of the cell wall, causing the release of secretion into the lumen of the duct. The participation of the cytoskeleton was observed for the first time during schizogeny of ducts as well as programmed cell death as part of the process of the release of holocrine secretion. This type of secretion release may be a key innovation in Kielmeyera since it has not been observed in ducts of any other plant thus far. [ABSTRACT FROM AUTHOR]

Objectives Antibiotic resistance can arise as a mutation to adapt to stress or be mediated by horizontal gene transfer. This study aimed at identifying the resistance determinants present in the mobile genetic elements of prophages and plasmids within multidrug-resistant (MDR) extraintestinal Escherichia coli. Materials and Methods Thirty-five anonymized MDR E. coli isolates of nonintestinal infections were confirmed for their antimicrobial resistance to six categories of antimicrobials by the disk diffusion test. Genes coding for beta-lactamases and carbapenemases in bacterial genome, plasmid, and prophage fractions were separately determined by polymerase chain reaction. Transducing ability of prophages carrying resistance genes was determined. Results Twenty-six isolates were positive for the gene blaCTX-M , nine for blaTEM , one each for blaKPC and blaVIM , thirteen for blaNDM, and seven for blaOXA. A majority of these isolates carried these determinants in plasmids and prophage fractions. Twenty-one percent of the prophage fractions (4 of 19) were able to successfully transfer resistance to sensitive isolates. Conclusion This study indicates blaCTX-M , blaTEM, and blaNDM genes that are reported most frequently in MDR isolates are more frequent in the plasmid and prophage fractions thus supporting for increased mobility. [ABSTRACT FROM AUTHOR]

Enzymatic degradation of polyethylene terephthalic acid (PET) has been gaining increasing importance. This has resulted in a significant increase in the search for newer enzymes and the development of more efficient enzyme-based systems. Due to the lack of a standard screening process, screening new enzymes has relied on other assays to determine the presence of esterase activity. This, in turn, has led to various nomenclatures and methods used to describe them and measure their activity. Since all PET-hydrolyzing enzymes are α/β hydrolases, they catalyze a serine nucleophilic attack and cleave an ester bond. They are lipases, esterases, cutinases and hydrolases. This has been used interchangeably, leading to difficulties while comparing results and evaluating progress. This review discusses the varied enzyme nomenclature being adapted, the different assays and analysis methods reported, and the strategies used to increase PET-hydrolyzing enzyme efficiency. A section on the various ways to quantify PET hydrolysis is also covered. [ABSTRACT FROM AUTHOR]

Background/Aim: Hematotoxicity is a lifethreatening condition that has become the major cause of drug discontinuation in patients with acute lymphoblastic leukemia (ALL). The nudix hydrolase 15 (NUDT15) gene polymorphism (c.415C>T) is reported to have an association with the hematotoxicity of 6-mercaptopurine (6-MP) as maintenance therapy in patients with ALL. However, the prevalence of this genetic polymorphism in the Indonesian population is unknown. This study aimed to assess the frequency of NUDT15 polymorphism among Indonesian pediatric patients with ALL and its association with the hematotoxicity of 6-MP. Patients and Methods: A total of 101 stored DNA samples from pediatric patients with ALL receiving 6-MP treatment were used for genetic testing. Direct sequencing was conducted to determine the NUDT15 c.415C>T genotype. Chi-square or Fisher's exact test were employed to examine the association between the NUDT15 c.415C>T genotype and hematotoxicity. Results: All (100%) of the DNA samples from patients with ALL treated with 6-MP exhibited a homozygous variant of the NUDT15 c.415C>T genotype, 70.3% of which showed hematotoxicity to some extent. We found no significant differences in NUDT15 gene polymorphism among patients with ALL with different states of hematotoxicity. Conclusion: The observed high frequency of NUDT15 c.415C>T in our study population might explain the elevated prevalence of 6-MPassociated hematotoxicity in pediatric patients with ALL within the Indonesian population. Our study provides new insight regarding the NUDT15 gene polymorphism and its relation to hematotoxicity. Further studies are required to determine the necessity of adjusting the initial dose of 6-MP for Indonesian pediatric patients with ALL. [ABSTRACT FROM AUTHOR]

Biotechnological processes are essential for producing climate-friendly high-value chemicals or pharmaceutical compounds, which can include steps catalyzed by enzymes. Therefore, establishing new, robust, and cheap enzyme production processes is desirable. One possible way to enhance processes is through the use of the spore display method. Spore display can present heterologous proteins on the surface of bacterial spores, offering numerous advantages in a range of biotechnological applications. This study demonstrates the implementation of the spore display method in Paenibacillus polymyxa, achieved by modifying the spore surface, incorporating an anchoring protein, and attaching green fluorescent protein to it, allowing the visualization of fluorescent spores. Following the initial experiment, a native lipase (Lip3), a heterologous lipase (LipA) from Bacillus subtilis, a native esterase (PnbA) from P. polymyxa, and a lipoyl synthase were expressed during sporulation and displayed on the spore surface. The activity profiles were determined in the temperature range from 4 °C to 70 °C. The PnbA reached its optimum at 4 °C, whereas the LipA from B. subtilis showed 4.4-fold higher activity at 42 °C compared to the control. Furthermore, we explored a possible new technique for the purification of enzymes with the TEV cleavage site between the anchor and the protein of interest. Finally, we showed a not-yet-described side activity of the lipoyl synthase over a wide temperature range. [ABSTRACT FROM AUTHOR]

As genomic databases expand and artificial intelligence tools advance, there is a growing demand for efficient characterization of large numbers of proteins. To this end, here we describe a generalizable pipeline for high-throughput protein purification using small-scale expression in E. coli and an affordable liquid-handling robot. This low-cost platform enables the purification of 96 proteins in parallel with minimal waste and is scalable for processing hundreds of proteins weekly per user. We demonstrate the performance of this method with the expression and purification of the leading poly(ethylene terephthalate) hydrolases reported in the literature. Replicate experiments demonstrated reproducibility and enzyme purity and yields (up to 400 µg) sufficient for comprehensive analyses of both thermostability and activity, generating a standardized benchmark dataset for comparing these plastic-degrading enzymes. The cost-effectiveness and ease of implementation of this platform render it broadly applicable to diverse protein characterization challenges in the biological sciences. [ABSTRACT FROM AUTHOR]

Synthesis and in vitro testing of a prodrug designed for the controlled delivery of the anticancer drug camptothecin within pancreatic cancer cells are reported. Our study reveals a non-conventional pharmacokinetic release characterized by an exponential pattern before reaching the half-life (t1/2) and a linear pattern thereafter. The release mechanism was triggered either by hydrolytic enzymes and/or by the acid microenvironment of cancer cells. [ABSTRACT FROM AUTHOR]

Peptidoglycan (PG) sacculi surround the cytoplasmic membrane, maintaining cell integrity by withstanding internal turgor pressure. During cell growth, PG endopeptidases cleave the crosslinks of the fully closed sacculi, allowing for the incorporation of new glycan strands and expansion of the peptidoglycan mesh. Outer-membrane-anchored NlpI associates with hydrolases and synthases near PG synthesis complexes, facilitating spatially close PG hydrolysis. Here, we present the structure of adaptor NlpI in complex with the endopeptidase MepS, revealing atomic details of how NlpI recruits multiple MepS molecules and subsequently influences PG expansion. NlpI binding elicits a disorder-to-order transition in the intrinsically disordered N-terminal of MepS, concomitantly promoting the dimerization of monomeric MepS. This results in the alignment of two asymmetric MepS dimers respectively located on the two opposite sides of the dimerization interface of NlpI, thus enhancing MepS activity in PG hydrolysis. Notably, the protein level of MepS is primarily modulated by the tail-specific protease Prc, which is known to interact with NlpI. The structure of the Prc-NlpI-MepS complex demonstrates that NlpI brings together MepS and Prc, leading to the efficient MepS degradation by Prc. Collectively, our results provide structural insights into the NlpI-enabled avidity effect of cellular endopeptidases and NlpI-directed MepS degradation by Prc. The outer-membrane-anchored protein NlpI recruits multiple MepS endopeptidases to enhance peptidoglycan breakdown. It also connects MepS to the protease Prc for efficient degradation, providing insights into how NlpI regulates cell wall remodeling. [ABSTRACT FROM AUTHOR]

In the last decades, biocatalysis has offered new perspectives for the synthesis of (chiral) amines, which are essential building blocks for pharmaceuticals, fine and bulk chemicals. In this regard, amidases have been employed due to their broad substrate scope and their independence from expensive cofactors. To expand the repertoire of amidases, tools for their rapid identification and characterization are greatly demanded. In this work an ultra-high throughput growth selection assay based on the production of the folate precursor p-aminobenzoic acid (PABA) is introduced to identify amidase activity. PABA-derived amides structurally mimic the broad class of commonly used chromogenic substrates derived from p-nitroaniline. This suggests that the assay should be broadly applicable for the identification of amidases. Unlike conventional growth selection assays that rely on substrates as nitrogen or carbon source, our approach requires PABA in sub-nanomolar concentrations, making it exceptionally sensitive and ideal for engineering campaigns that aim at enhancing amidase activities from minimally active starting points, for example. The presented assay offers flexibility in the adjustment of sensitivity to suit project-specific needs using different expression systems and fine-tuning with the antimetabolite sulfathiazole. Application of this PABA-based assay facilitates the screening of millions of enzyme variants on a single agar plate within two days, without the need for laborious sample preparation or expensive instruments, with transformation efficiency being the only limiting factor. Key points: • Ultra-high throughput assay (tens of millions on one agar plate) for amidase screening • High sensitivity by coupling selection to folate instead of carbon or nitrogen source • Highly adjustable in terms of sensitivity and expression of the engineering target [ABSTRACT FROM AUTHOR]

In recent years, there has been increasing interest in studying gut microbiomederived hydrolases in relation to oral drug metabolism, particularly focusing on natural product drugs. Despite the significance of natural product drugs in the field of oral medications, there is a lack of research on the regulatory interplay between gut microbiome-derived hydrolases and these drugs. This review delves into the interaction between intestinal microbiome-derived hydrolases and natural product drugs metabolism from three key perspectives. Firstly, it examines the impact of glycoside hydrolases, amide hydrolases, carboxylesterase, bile salt hydrolases, and epoxide hydrolase on the structure of natural products. Secondly, it explores how natural product drugs influence microbiome-derived hydrolases. Lastly, it analyzes the impact of interactions between hydrolases and natural products on disease development and the challenges in developing microbial-derived enzymes. The overarching goal of this review is to lay a solid theoretical foundation for the advancement of research and development in new natural product drugs and personalized treatment. [ABSTRACT FROM AUTHOR]

The sulfolipid sulfoquinovosyl diacylglycerol (SQDG), produced by plants, algae, and cyanobacteria, constitutes a major sulfur reserve in the biosphere. Microbial breakdown of SQDG is critical for the biological utilization of its sulfur. This commences through release of the parent sugar, sulfoquinovose (SQ), catalyzed by sulfoquinovosidases (SQases). These vanguard enzymes are encoded in gene clusters that code for diverse SQ catabolic pathways. To identify, visualize and isolate glycoside hydrolase CAZY‐family 31 (GH31) SQases in complex biological environments, we introduce SQ cyclophellitol‐aziridine activity‐based probes (ABPs). These ABPs label the active site nucleophile of this enzyme family, consistent with specific recognition of the SQ cyclophellitol‐aziridine in the active site, as evidenced in the 3D structure of Bacillus megaterium SQase. A fluorescent Cy5‐probe enables visualization of SQases in crude cell lysates from bacteria harbouring different SQ breakdown pathways, whilst a biotin‐probe enables SQase capture and identification by proteomics. The Cy5‐probe facilitates monitoring of active SQase levels during different stages of bacterial growth which show great contrast to more traditional mRNA analysis obtained by RT‐qPCR. Given the importance of SQases in global sulfur cycling and in human microbiota, these SQase ABPs provide a new tool with which to study SQase occurrence, activity and stability. [ABSTRACT FROM AUTHOR]

Silyl ethers fulfil a fundamental role in synthetic organic chemistry as protecting groups and their selective cleavage is an important factor in their application. We present here for the first time two enzymes, SilE−R and SilE−S, which are able to hydrolyse silyl ethers. They belong to the stress‐response dimeric A/B barrel domain (DABB) family and are able to cleave the Si−O bond with opposite enantiopreference. Silyl ethers containing aromatic, cyclic or aliphatic alcohols and, depending on the alcohol moiety, silyl functions as large as TBDMS are accepted. The X‐ray crystal structure of SilE−R, determined to a resolution of 1.98 Å, in combination with mutational studies, revealed an active site featuring two histidine residues, H8 and H79, which likely act synergistically as nucleophile and Brønsted base in the hydrolytic mechanism, which has not previously been described for enzymes. Although the natural function of SilE−R and SilE−S is unknown, we propose that these 'silyl etherases' may have significant potential for synthetic applications. [ABSTRACT FROM AUTHOR]

Biomass from various types of organic waste was tested for possible use in hydrogen production. The composition consisted of lignified samples, green waste, and kitchen scraps such as fruit and vegetable peels and leftover food. For this purpose, the enzymatic pretreatment of organic waste with a combination of five different hydrolytic enzymes (cellulase, amylase, glucoamylase, pectinase and xylase) was investigated to determine its ability to produce hydrogen (H2) with the hydrolyzate produced here. In course, the anaerobic rod-shaped bacterium T. neapolitana was used for H2 production. First, the enzymes were investigated using different substrates in preliminary experiments. Subsequently, hydrolyses were carried out using different types of organic waste. In the hydrolysis carried out here for 48 h, an increase in glucose concentration of 481% was measured for waste loads containing starch, corresponding to a glucose concentration at the end of hydrolysis of 7.5 g·L−1. In the subsequent set fermentation in serum bottles, a H2 yield of 1.26 mmol H2 was obtained in the overhead space when Terrific Broth Medium with glucose and yeast extract (TBGY medium) was used. When hydrolyzed organic waste was used, even a H2 yield of 1.37 mmol could be achieved in the overhead space. In addition, a dedicated reactor system for the anaerobic fermentation of T. neapolitana to produce H2 was developed. The bioreactor developed here can ferment anaerobically with a very low loss of produced gas. Here, after 24 h, a hydrogen concentration of 83% could be measured in the overhead space. [ABSTRACT FROM AUTHOR]

Background and Aims: Oral bacteria play an important role in oral diseases, due to their high adaptability to different environmental areas of the mouth. In this article, an attempt was made to describe the molecular mechanisms involved in the physiological relationships of oral and dental environment bacteria and their pathogenic significance with molecular approaches. Materials and Methods: The present systematic review was written based on the advanced and standard search of keywords including Oral bacteria, Biofilm, and Dental diseases in PubMed, Springer, Scopus, Medline, Google Scholar, Science Direct, and Web of Science databases. For this purpose, an advanced and systematic search of articles published from 1993 to 2023 was conducted to compile the present article. Results: Bacteria in the oral cavity have nutritional adaptations that are important for living in pathogen-host relationships, including adapting to proteolytic living conditions, using the host's glycome as a nutritional interface. This includes the use of host-derived sialic acid and other glycosidases in oral bacteria. Some of these bacteria adhere to surfaces such as salivary, epithelial proteins, and glycans, which ultimately lead to biofilm formation. Bacteria living in the oral environment are constantly exposed to a wide range of stress-causing factors and oxidative stress in the biofilm. Conclusion: Dental caries, pulp, periapical, and periodontitis diseases (including gingivitis) are among the most common bacterial diseases. Among them, tooth decay caused by the presence of Streptococcus mutans is the most common dental disease due to the production of acids from carbohydrate fermentation which is characterized by the demineralization of tooth structure. [ABSTRACT FROM AUTHOR]

As a crucial hydrolytic enzyme, urease plays a vital role in anaerobic biological treatment. It is well-known that manganese ions are abundant in landfill leachate, but their concentration fluctuates significantly. However, few studies have investigated the effect and mechanism of different concentrations of Mn2+ on urease activity during anaerobic biological treatment of landfill leachate. This paper aimed to investigate the effects and mechanisms of different concentrations of Mn2+ on urease activity. The results showed that an appropriate amount of Mn2+ could significantly enhance urease activity, while a high concentration of Mn2+ could inhibit it. Insight into the mechanisms behind this phenomenon, various methods such as Zeta potential, particle size, ultraviolet spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy, and statistical analysis were employed in our study. Research suggested that, on one hand, Mn2+ may form hydrogen bonds with the side chain amino or carboxyl groups of urease amino acid residues, affecting the structure of urease through hydrogen bonding. Additionally, Mn2+ also binds to urease through hydrophobic interactions. On the other hand, the C–OH and C-N functional groups in urease have a strong affinity for Mn2+, and changes in these functional groups can greatly enhance the activity of urease. Furthermore, under the action of high concentrations of Mn2+, while the structure of urease becomes more stable, there is also a steric hindrance phenomenon that affects the substrate from entering the catalytic center. Therefore, studying the mechanism of Mn2+ affecting urease activity has significant biological significance and provides a new perspective for exploring the impact of metals on anaerobic bioprocessing of landfill leachate. [ABSTRACT FROM AUTHOR]

Poultry's digestive tract lacks hydrolytic phytase enzymes, which results in chelation of dietary minerals, vital amino acids, proteins, and carbohydrates, phytate‐phosphate unavailability, and contamination of the environment due to phosphorus. Therefore, it is necessary to use exogenous microbial phytases as feed additive to chicken feed to catalyze the hydrolysis of dietary phytate. Potential sources of microbial isolates that produce desired phytases for chicken feed supplementation have been isolated from agricultural croplands. It is achievable to isolate phytase‐producing bacteria isolates using both broth and agar phytase screening media. Potential substrates for submerged fermentation (SmF) for bacterial phytase production and solid‐state fermentation (SSF) for fungal phytase production include rice and wheat bran. Following fermentation, saturated ammonium sulphate precipitation is typically used to partially purify microbial culture filtrate. The precipitate is then desalted. Measurements of the pH optimum and stability, temperature optimum and stability, metal ions stability, specificity and affinity to target substrate, proteolysis resistance, storage stability, and in vitro feed dephosphorylation are used to perform an enzymatic evaluation of phytase as an additive for poultry feed. The growth of the feed phytase market is primarily due to the expansion of chicken farms to meet the demand for meat and eggs from humans. The Food and Drug Administration in the USA and the European Food and Safety Authority are primarily in charge of putting rules pertaining to feed phytase use in chicken feed into effect. Conclusively, important components of the production of phytase additives for poultry feed include identifying a reliable source for potential microbe isolation, selecting an economical method of phytase production, thoroughly characterizing the biochemical properties of phytase, and comprehending the size and regulation of the current feed phytase market. [ABSTRACT FROM AUTHOR]

Background: The high rate of stoma placement during emergency laparotomy for secondary peritonitis is a paradigm in need of change in the current fast-track surgical setting. Despite growing evidence for the feasibility of primary bowel reconstruction in a peritonitic environment, little data substantiate a surgeons' choice between a stoma and an anastomosis. The aim of this retrospective analysis is to identify pre- and intraoperative parameters that predict the leakage risk for enteric sutures placed during source control surgery (SCS) for secondary peritonitis. Methods: Between January 2014 and December 2020, 497 patients underwent SCS for secondary peritonitis, of whom 187 received a primary reconstruction of the lower gastro-intestinal tract without a diverting stoma. In 47 (25.1%) patients postoperative leakage of the enteric sutures was directly confirmed during revision surgery or by computed tomography. Quantifiable predictors of intestinal suture outcome were detected by multivariate analysis. Results: Length of intensive care, in-hospital mortality and failure of release to the initial home environment were significantly higher in patients with enteric suture leakage following SCS compared to patients with intact anastomoses (p < 0.0001, p = 0.0026 and p =0.0009, respectively). Reduced serum choline esterase (sCHE) levels and a high extent of peritonitis were identified as independent risk factors for insufficiency of enteric sutures placed during emergency laparotomy. Conclusions: A preoperative sCHE < 4.5 kU/L and generalized fecal peritonitis associate with a significantly higher incidence of enteric suture insufficiency after primary reconstruction of the lower gastro-intestinal tract in a peritonitic abdomen. These parameters may guide surgeons when choosing the optimal surgical procedure in the emergency setting. [ABSTRACT FROM AUTHOR]

In the current study, we compared the production of extracellular lignocellulose degrading enzymes and bioethanol from the spent mushroom substrate (SMS) of Calocybe indica and Volvariella volvacea. From SMS at different stages of the mushroom development cycle, ligninolytic and hydrolytic enzymes were analysed. The activities of lignin-degrading enzymes, including lignin peroxidase (LiP), laccase, and manganese peroxidase (MnP) were maximal in the spawn run and primordial stages, while hydrolytic enzymes including xylanase, cellobiohydrolase (CBH), and carboxymethyl cellulase (CMCase) showed higher activity during fruiting bodies development and at the end of the mushroom growth cycle. SMS of V. volvacea showed relatively lower ligninase activity than the SMS of C. indica, but had the maximum activity of hydrolytic enzymes. The enzyme was precipitated with acetone and further purified with the DEAE cellulose column. The maximum yield of reducing sugars was obtained after hydrolysis of NaOH (0.5 M) pretreated SMS with a cocktail of partially purified enzymes (50% v/v). After enzymatic hydrolysis, the total reducing sugars were 18.68 ± 0.34 g/l (SMS of C. indica) and 20.02 ± 0.87 g/l (SMS of V. volvacea). We observed the highest fermentation efficiency and ethanol productivity (54.25%, 0.12 g/l h) obtained from SMS hydrolysate of V. volvacea after 48 h at 30 ± 2 °C, using co-culture of Saccharomyces cerevisiae MTCC 11,815 and Pachysolen tannophilus MTCC 1077. [ABSTRACT FROM AUTHOR]

The recently discovered Type 9 Secretion System (T9SS) is present in bacteria of the Fibrobacteres–Bacteroidetes–Chlorobi superphylum, which are key constituents of diverse microbiomes. T9SS is instrumental in the extracellular secretion of over 270,000 proteins, including peptidases, sugar hydrolases, metal ion‐binding proteins, and metalloenzymes. These proteins are essential for the interaction of bacteria with their environment. This mini‐review explores the extensive array of proteins secreted by the T9SS. It highlights the diverse functions of these proteins, emphasizing their roles in pathogenesis, bacterial interactions, host colonization, and the overall health of the ecosystems inhabited by T9SS‐containing bacteria. [ABSTRACT FROM AUTHOR]

In this review article, a perspective on the immobilization of various hydrolytic enzymes onto magnetic nanoparticles for synthetic organic chemistry applications is presented. After a first part giving short overview on nanomagnetism and highlighting advantages and disadvantages of immobilizing enzymes on magnetic nanoparticles (MNPs), the most important hydrolytic enzymes and their applications were summarized. A section reviewing the immobilization techniques with a particular focus on supporting enzymes on MNPs introduces the reader to the final chapter describing synthetic organic chemistry applications of small molecules (flavour esters) and polymers (polyesters and polyamides). Finally, the conclusion and perspective section gives the author's personal view on further research discussing the new idea of a synergistic rational design of the magnetic and biocatalytic component to produce novel magnetic nano‐architectures. [ABSTRACT FROM AUTHOR]

Penicillium italicum, a major postharvest pathogen, causes blue mold rot in citrus fruits through the deployment of various virulence factors. Recent studies highlight the role of the epigenetic reader, SntB, in modulating the pathogenicity of phytopathogenic fungi. Our research revealed that the deletion of the SntB gene in P. italicum led to significant phenotypic alterations, including delayed mycelial growth, reduced spore production, and decreased utilization of sucrose. Additionally, the mutant strain exhibited increased sensitivity to pH fluctuations and elevated iron and calcium ion stress, culminating in reduced virulence on Gannan Novel oranges. Ultrastructural analyses disclosed notable disruptions in cell membrane integrity, disorganization within the cellular matrix, and signs of autophagy. Transcriptomic data further indicated a pronounced upregulation of hydrolytic enzymes, oxidoreductases, and transport proteins, suggesting a heightened energy demand. The observed phenomena were consistent with a carbon starvation response potentially triggering apoptotic pathways, including iron-dependent cell death. These findings collectively underscored the pivotal role of SntB in maintaining the pathogenic traits of P. italicum, proposing that targeting PiSntB could offer a new avenue for controlling citrus fungal infections and subsequent fruit decay. [ABSTRACT FROM AUTHOR]

Purpose: Polysorbates are the most commonly used surfactants in formulations to stabilize therapeutic proteins against interfacial stresses. Polysorbates can undergo oxidative or enzyme-mediated hydrolytic degradation to produce free fatty acids (FFAs) and subvisible particles in formulations. To determine which product related variables contribute to PS20 degradation, we investigated the effects of storage temperature, formulation, pH, presence of hydrolytic enzymes, and specific fatty acid composition on different grades of PS20 in relation to their PS20 degradation profile and consequently the quality of protein drug products. Methods: Bevacizumab and T-DM1 were reformulated in the freshly prepared therapeutic protein formulations containing either compendial PS20 or non-compendial PS20 with high % lauric acid and spiked with exogenous esterase or lipase. The release of FFAs and formation of particles were monitored at 4°C and 37°C. Protein quality was assessed for secondary structures, purity, and biological activity. Results: Hydrolytic release of FFAs and formation of subvisible particles were found to be dependent on grades of PS20, types of enzymes used, incubation temperature, and pH. Esterase- or lipase-mediated degradation of PS20 and formation of subvisible particles in drug formulation showed no significant impact on the biological activity and stability of therapeutic proteins against degradation or aggregation. Conclusions: Our study suggests that degradation of PS20 and formation of FFA particles depend on the fatty acid composition of PS20, types of hydrolytic enzymes, pH, and temperature. The presence of FFA subvisible particles showed no significant impact on the purity and biological activity of the therapeutic proteins under the tested conditions. [ABSTRACT FROM AUTHOR]

Biofilms, that are the community of microorganisms attached on biotic or abiotic surfaces coated with a self-produced matrix, composed by exopolymeric substances, dominate in all habitats on the surface of the Earth, except in the oceans, accounting for ~80% of bacterial and archaeal cells. Hydrolytic enzymes have a number of industrial applications and have been indicated as an alternative to the traditional chemical methods that are used to eradicate microbial biofilms. In this study, we evaluated the ability of enzymatic extracts produced by Bacillus subtilis 170 to remove multispecies biofilms, formed by the interaction of Bacillus subtilis and Escherichia coli strains. After culture in liquid medium, containing inorganic nitrogen sources, the bacterial hydrolytic extracts showed protease (250 U/mL) activity. Cell-free supernatants of B. subtilis 170 strain with proteolytic activity were the most effective, and promoted the complete removal of Bacillus subtilis and Escherichia coli dual-species biofilms. Of the treatments using cell-free supernatants of B.subtilis 170 with proteolytic enzyme activities with 250 U/mL, total biofilm degradation was observed for both dual-species biofilms in this study. Thus, the hydrolases produced by Bacillus subtilis 170 strains evaluated here are highlighted as an interesting tool in the fight against microbial biofilms. [ABSTRACT FROM AUTHOR]

Hydrolases are the most popular enzymes, and among the most valuable in biotechnological applications. Some hydrolases, such as lipases, esterases, proteases, cellulases and amylases, are used in the food industry and the production of biopharmaceuticals, biofuels, biopolymers and detergents. Of special interest are those obtained from thermophilic microorganisms. Although there is great microbial diversity in extreme environments, the investigations aimed at detecting and isolating enzymes with potential for polyester degradation such as polyethylene terephthalate (PET) are limited. In this work, we explored the metagenomic library of an oil-enriched soil sample from the "Los Humeros" geothermal field by means of in silico probes in search for enzymes potentially able to degrade polyesters. Using conserved motifs and activity-relevant sites of reported polyester hydrolases, we designed probes that allowed us to identify 6 potential polyester hydrolases in the metagenome. Three-dimensional structure prediction revealed a canonical α/β fold and a cap covering the active site of the enzymes. The catalytic triads were composed of Ser, His and Asp. Structural comparison, substrate binding site analysis and molecular docking suggested their potential as polyester hydrolases, particularly cutinases and PETases. An enzyme, REC98271, was cloned, expressed and characterized, showing thermophilic properties and preference for short-chain substrates. These findings contribute to our understanding of enzyme diversity in "Los Humeros" metagenome and their potential applications in biodegradation and recycling processes. [ABSTRACT FROM AUTHOR]

The west coast of India (WCI) is signified by increased primary productivity during the southwest monsoon season. In this study, the bacterial counts and the cultivable communities were enumerated from water depths and sediment samples at stations, off Goa, Mangalore and Trivandrum along the WCI during the monsoon season. The results reveal that the bacterial morphotypes isolated from this study included representatives from Firmicutes (70%), Proteobacteria (20%) and Actinobacteria (5%) phylum. Phylum Firmicutes had a major representation at Trivandrum and Mangalore locations compared to those from Goa. However, the Proteobacteria phylum was more prevalent in the Goa location compared to the other two locations. The morphotypes at the genera level also showed distinct spatial variations and taxa specific to each of the three locations could also be identified through Canonical correspondence analysis (CCA). Spatial variations in hydrolytic enzyme activities were also observed, with bacterial morphotypes isolated from water samples of Trivandrum exhibiting comparatively higher activities than the Goa and Mangalore sediment samples. These findings enhance our understanding of bacterial diversity and their functional roles along the WCI, emphasizing their significance in biogeochemical cycling processes. [ABSTRACT FROM AUTHOR]

Background: We previously reported that binge ethanol induces atrophy of the spleen, a key immune organ, in adolescent male F344 rats. Because there are significant sex effects in immune function, we investigated whether binge ethanol exerts sex‐dependent effects on the spleen, including producing splenic atrophy. Methods: We gave F344 rats ethanol (4.8 g/kg/day; 52% w/v; i.g.) on postnatal days [PND] 36 ~ 38 and sacrificed them on PND 39 for spleen collection. We performed immunophenotyping analysis of splenic cells and examined the expression of 158 genes related to alcohol metabolism, epigenetic modification, and immune regulation in the spleens of adolescent (PND 39) male and female rats. Results: Following a 3‐day ethanol exposure, a loss of body weight, and absolute and relative spleen weight, was seen only in male adolescent rats. Ethanol altered the relative proportions of lymphocyte subtypes in both sexes with different patterns. We also found that 3‐day ethanol exposure induced sex‐dependent gene expression changes in spleen. Among the 158 genes studied, the expression of only three genes was significantly increased in female rats. However, the expression of 30 genes was significantly increased/decreased in male rats. Female rats had greater expression of alcohol metabolizing enzyme genes in the spleen under physiological conditions and when stimulated by binge ethanol. The genes are involved in epigenetic modification were differentially expressed in a sex‐dependent manner. Conclusion: We found that male adolescent rats were more sensitive to binge ethanol than female rats. Differential expression of the genes related to alcohol metabolism and epigenetic modification (of DNA methyltransferase and histone deacetylases) between the sexes could account for the observed sex‐dependent responses to binge ethanol in adolescent rats. [ABSTRACT FROM AUTHOR]

Ganoderma boninense is a white-rot fungus that causes basal stem rot (BSR) disease in the oil palm. Potential natural inhibitors, such as gallic acid, thymol, propolis, and carvacrol, were assessed for their antagonistic effects against G. boninense. These naturally occurring phenolic compounds have also been utilised to inhibit hydrolytic and ligninolytic enzymes produced by the pathogen. Mycelial inhibition was dose-dependent in the presence of different concentrations of phenolic compounds, including, for example, in cellulase enzyme inhibition (GA mg/mL = 94%, THY 0.25 mg/mL = 90%, PRO 3.5 mg/mL = 92.5%, and CARV 0.15 mg/mL = 90.3%). A significant difference was observed revealing that gallic acid had the greatest inhibitory effect on the secretion of hydrolytic and ligninolytic enzymes, especially at 40 mM GA (cellulase = 0.337 U/mL, amylase = 0.3314 U/mL, xylanase = 0.211 U/mL, laccase = 0.4885 U/mL, lignin peroxidase = 0.218 U/mL, and manganese peroxidase = 0.386 U/mL). The growth and secretion of enzymes (inhibitory action) are inversely proportional to the concentration of phenolic compounds. Phenolic compounds have a greater potential as inhibitory agents and suppress the production of hydrolytic and ligninolytic enzymes. The selected phenolic compounds were evaluated for their ability to alter the morphology and integrity of G. boninense mycelia. The reduction in cell viability of G. boninense has been explained by research on morphological disruption, such as branching patterns, hyphal length, and rigidity of fungal cells, which eventually interrupt the secretion of enzymes. These studies highlight the efficacy of phenolic compounds in treating Ganoderma. In addition, these findings proved that naturally occurring phenolic compounds could be a substitute for chemical controls and other synthetic fungicides to eradicate the occurrence of BSR in oil palms, thus avoiding a situation that is difficult to overcome. [ABSTRACT FROM AUTHOR]

Atopic Dermatitis is the most common eczematous inflammatory skin condition, presenting with lesions that typically appear as poorly demarcated erythematous and scaly papules and plaques. The lesions most commonly occur on flexural surfaces of the knees, elbows, and wrists and are associated with moderate to severe itching. This article focuses on the clinical presentation of atopic dermatitis and treatment options. Other related topics include epidemiology, pathogenesis, risk factors, triggers, and differential diagnoses. [ABSTRACT FROM AUTHOR]

The maintenance of the root stem cell niche identity in Arabidopsis relies on the delicate balance of reactive oxygen species (ROS) levels in root tips; however, the intricate molecular mechanisms governing ROS homeostasis within the root stem cell niche remain unclear. In this study, we unveil the role of ATP hydrolase superfamily protein 1 (ASP1) in orchestrating root stem cell niche maintenance through its interaction with the redox regulator cystathionine β-synthase domain-containing protein 3 (CBSX3). ASP1 is exclusively expressed in the quiescent center (QC) cells and governs the integrity of the root stem cell niche. Loss of ASP1 function leads to enhanced QC cell division and distal stem cell differentiation, attributable to reduced ROS levels and diminished expression of SCARECROW and SHORT ROOT in root tips. Our findings illuminate the pivotal role of ASP1 in regulating ROS signaling to maintain root stem cell niche homeostasis, achieved through direct interaction with CBSX3. [ABSTRACT FROM AUTHOR]

The study aims to investigate plant growth promotion (PGP) activities of thermophilic bacteria isolated from the Jakrem hot spring in Meghalaya, North-East India, and determine their effect on Brassica juncea's growth. The bacteria were isolated by a culture-dependent approach following a serial dilution method in a nutrient agar medium. All the isolates were determined for PGP attributes such as indole acetic acid, phosphate solubilization, hydrolytic enzymes, and siderophore production. The potent bacterial isolates were characterized by 16S rDNA sequencing and phylogenetic analysis. Altogether, 53 bacterial isolates were obtained, most belonging to the genus Bacillus. Of the total isolates, 37.7% exhibited both PGP and hydrolytic enzyme activities. Three isolates, namely JAB1, JAB8, and JAB100, showed promising PGP and were identified as Bacillus velezensis, B. proteolyticus, and Bacillus sp., respectively. The PGP attributes of these isolates were determined in vivo on B. juncea, and their effects were measured in terms of shoot and root length biomass and biochemical contents. It was observed that combined inoculation of all three isolates significantly enhanced the growth and development of B. juncea, evident by increased shoot and root length, fresh and dry weight, and higher levels of protein, phenol, flavonoid, and chlorophyll content compared to the control. In conclusion, the study highlights the potential application of thermophilic Bacillus spp. from hot springs as bioinoculants to enhance crop productivity in sustainable agricultural practices. [ABSTRACT FROM AUTHOR]

Microorganisms with multiple ecological functions can be a useful biotechnological resource in integrated pest- and disease-management programs. This work aimed to investigate the potential endophytic and virulent effects of a strain of Purpureocillium lilacinum on organic cultivation in Brazil. Specifically, the strain's ability to establish itself as an endophyte in common bean, soybean, and sunflower plants when inoculated via seed was evaluated. Furthermore, its antifungal activity against phytopathogens and its pathogenicity and virulence against insects of the order Lepidoptera, Coleoptera, and Hemiptera were evaluated. Furthermore, the strain was evaluated for its biochemical and physiological characteristics. For virulence bioassays, the experiments were conducted under a factorial scheme (2 × 3), with the following factors: (a) fungal inoculation and control without inoculum and (b) types of inocula (blastospores, aerial conidia, and metabolites). The treatments were sprayed on insect species at different stages of development. In summary, it was found that the SBF054 strain endophytically colonized the common bean, with partial recovery from the root tissues of soybean and sunflower plants, 30 days after inoculation; suppressed 86% of Rhizoctonia solani mycelial growth in an in vitro assay; and controlled eggs, nymphs, and Euschistus heros adults. These multifunctional abilities are mainly attributed to the strain's mechanisms of producing metabolites, such as organic acids, soluble nutrients, and hydrolytic enzymes. [ABSTRACT FROM AUTHOR]

Abstract In this work, we studied 46 bacterial isolates to determine their potential for polyethylene terephthalate (PET) degradation. Only eight lipase-positive isolates were pre-selected to continue analysis. The isolates belonged to the genera Micrococcus, Acinetobacter, Pseudomonas, Bacillus, and Pseudochrobactrum. In addition to lipase, other enzymatic activities such as protease, carboxymethyl cellulase, xylanase, and chitinase were detected. PET sheets were pretreated with temperature, UV light, HNO3, and Stenotrophomonas pavanii. All isolates were tested against pretreated PET for 60 days. All isolates displayed lipase activity, and some hydrolase activities. The chemical changes of PET sheets were determined using Fourier-transform infrared spectroscopy. Peaks observed at 3000-3500 cm-1 corresponding to hydroxyl group suggested degradation. The physical changes of PET observed with the naked eye and with SEM consisted of erosion, bubbles, scales, bumps, pits, and a shiny surface. Pseudomonas soli was selected as the best PET degrader, according to the notable changes in the infrared spectrum and macroscopic changes compared to the other genera and the control. In addition, this isolate did not show xylanase activity and it lacks the chitinase gene too. It is possible that not only lipase activity could be associated with the PET degradation, but also another enzymatic activity not detected in this study. Although the genus Pseudomonas has been reported as a plastic degrader, the P. soli species is novel in the degradation of PET, as well as the other genera mentioned here. Finally, these findings contribute to the knowledge of bacterial species for PET degradation.

Congenital disorders of glycosylation (CDG) and Niemann-Pick type C (NPC) disease are inborn errors of metabolism that can both present with infantile-onset severe liver disease and other multisystemic manifestations. Plasma bile acid and N-palmitoyl-O-phosphocholineserine (PPCS) are screening biomarkers with proposed improved sensitivity and specificity for NPC. We report an infant with ATP6AP1-CDG who presented with cholestatic liver failure and elevated plasma oxysterols and bile acid, mimicking NPC clinically and biochemically. On further investigation, PPCS, but not the bile acid derivative N-(3β,5α,6β-trihydroxy-cholan-24-oyl) glycine (TCG), were elevated in plasma samples from individuals with ATP6AP1-, ALG1-, ALG8-, and PMM2-CDG. These findings highlight the importance of keeping CDG within the diagnostic differential when evaluating children with early onset severe liver disease and elevated bile acid or PPCS to prevent delayed diagnosis and treatment.

Enterocytes modulate the extent of postprandial lipemia by storing dietary fats in cytoplasmic lipid droplets (cLDs). We have previously shown that the integrin ligand MFGE8 links absorption of dietary fats with activation of triglyceride (TG) hydrolases that catabolize cLDs for chylomicron production. Here, we identify CES1D as the key hydrolase downstream of the MFGE8-αvβ5 integrin pathway that regulates catabolism of diet-derived cLDs. Mfge8 knockout (KO) enterocytes have reduced CES1D transcript and protein levels and reduced protein levels of the transcription factor HNF4γ. Both Ces1d and Hnf4γ KO mice have decreased enterocyte TG hydrolase activity coupled with retention of TG in cLDs. Mechanistically, MFGE8-dependent fatty acid uptake through CD36 stabilizes HNF4γ protein level; HNF4γ then increases Ces1d transcription. Our work identifies a regulatory network that regulates the severity of postprandial lipemia by linking dietary fat absorption with protein stabilization of a transcription factor that increases expression of hydrolases responsible for catabolizing diet-derived cLDs.

BackgroundBiocatalysis offers a promising path for plastic waste management and valorization, especially for hydrolysable plastics such as polyethylene terephthalate (PET). Microbial whole-cell biocatalysts for simultaneous PET degradation and growth on PET monomers would offer a one-step solution toward PET recycling or upcycling. We set out to engineer the industry-proven bacterium Pseudomonas putida for (i) metabolism of PET monomers as sole carbon sources, and (ii) efficient extracellular expression of PET hydrolases. We pursued this approach for both PET and the related polyester polybutylene adipate co-terephthalate (PBAT), aiming to learn about the determinants and potential applications of bacterial polyester-degrading biocatalysts.ResultsP. putida was engineered to metabolize the PET and PBAT monomer terephthalic acid (TA) through genomic integration of four tphII operon genes from Comamonas sp. E6. Efficient cellular TA uptake was enabled by a point mutation in the native P. putida membrane transporter MhpT. Metabolism of the PET and PBAT monomers ethylene glycol and 1,4-butanediol was achieved through adaptive laboratory evolution. We then used fast design-build-test-learn cycles to engineer extracellular PET hydrolase expression, including tests of (i) the three PET hydrolases LCC, HiC, and IsPETase; (ii) genomic versus plasmid-based expression, using expression plasmids with high, medium, and low cellular copy number; (iii) three different promoter systems; (iv) three membrane anchor proteins for PET hydrolase cell surface display; and (v) a 30-mer signal peptide library for PET hydrolase secretion. PET hydrolase surface display and secretion was successfully engineered but often resulted in host cell fitness costs, which could be mitigated by promoter choice and altering construct copy number. Plastic biodegradation assays with the best PET hydrolase expression constructs genomically integrated into our monomer-metabolizing P. putida strains resulted in various degrees of plastic depolymerization, although self-sustaining bacterial growth remained elusive.ConclusionOur results show that balancing extracellular PET hydrolase expression with cellular fitness under nutrient-limiting conditions is a challenge. The precise knowledge of such bottlenecks, together with the vast array of PET hydrolase expression tools generated and tested here, may serve as a baseline for future efforts to engineer P. putida or other bacterial hosts towards becoming efficient whole-cell polyester-degrading biocatalysts.

Water-filled sinkholes known locally as cenotes, found on the Yucatán Peninsula, have remarkable biodiversity. The primary objective of this study was to explore the biotechnological potential of Gram-positive cultivable bacteria obtained from sediment samples collected at the coastal cenote Pol-Ac in Yucatán, Mexico. Specifically, the investigation aimed to assess production of hydrolytic enzymes and antimicrobial compounds. 16 S rRNA gene sequencing led to the identification of 49 Gram-positive bacterial isolates belonging to the phyla Bacillota (n = 29) and Actinomycetota (n = 20) divided into the common genera Bacillus and Streptomyces, as well as the genera Virgibacillus, Halobacillus, Metabacillus, Solibacillus, Neobacillus, Rossellomorea, Nocardiopsis and Corynebacterium. With growth at 55ºC, 21 of the 49 strains were classified as moderately thermotolerant. All strains were classified as halotolerant and 24 were dependent on marine water for growth. Screening for six extracellular hydrolytic enzymes revealed gelatinase, amylase, lipase, cellulase, protease and chitinase activities in 93.9%, 67.3%, 63.3%, 59.2%, 59.2% and 38.8%, of isolated strains, respectively. The genes for polyketide synthases type I, were detected in 24 of the strains. Of 18 strains that achieved > 25% inhibition of growth in the bacterial pathogen Staphylococcus aureus ATCC 6538, 4 also inhibited growth in Escherichia coli ATCC 35,218. Isolates Streptomyces sp. NCA_378 and Bacillus sp. NCA_374 demonstrated 50–75% growth inhibition against at least one of the two pathogens tested, along with significant enzymatic activity across all six extracellular enzymes. This is the first comprehensive report on the biotechnological potential of Gram-positive bacteria isolated from sediments in the cenotes of the Yucatán Peninsula. [ABSTRACT FROM AUTHOR]

Chickpea pod borer (CPB) (Helicoverpa armigera) is one of the major pests, causing significant yield losses. The objectives were to screen chickpea mutants for pod borer resistance/tolerance under field conditions and identification of biochemical markers of tolerance. Chickpea mutant CM216-A/15 had highest leaf (25 trichomes/mm²) and stem trichome density (17 trichomes/mm²) with least pod damage at Kallur Kot and highest pod weight per plant (22.8 ± 2.6g) at AZRI. Higher total phenolic contents (TPCs) and antioxidant capacity were detected in tolerant mutants, i.e., CM216-A/15 and CM664/15. TPC was positively associated with pod yield and had negative correlation with pod damage. Mutants CM216-A/15, CM664/15, and CM766/15 depicted the highest resilience to CPB, owing to higher hairiness, better antioxidant defense response, and lower levels of hydrolytic enzymes and sugars. Identified biochemical markers like TPC, total oxidant status, superoxide dismutase, and pigments can be used for screening of CPB-tolerant/resistant mutants. [ABSTRACT FROM AUTHOR]

Low production costs and a potential feedstock supply make lignocellulosic ethanol (bioethanol) an important source of advanced biofuels. The physical and chemical preparation of this kind of lignocellulosic feedstock led to a high ethanol yield. In order to increase the yield of fermentable sugars, pretreatment is an essential process step that alters the lignocellulosic structure and improves its accessibility for the expensive hydrolytic enzymes. In this context, the chemical composition of sugarcane trash (dry leaves, green leaves, and tops) and jatropha (shell and seed cake) was determined to be mainly cellulose, hemicellulose, and lignin. Hydrogen peroxide and sodium hydroxide were applied in an attempt to facilitate the solubilization of lignin and hemicelluloses in five agrowastes. The extraction of hydrogen peroxide was much better than that of sodium hydroxide. A comparative study was done using SEM, EDXA, and FTIR to evaluate the difference between the two methods. The pretreated wastes were subjected to saccharification by commercial cellulases (30 IU/g substrate). The obtained glucose was fortified with nutrients and fermented statically by Saccharomyces cerevisiae F-307 for bioethanol production. The results revealed the bioethanol yields were 325.4, 310.8, 282.9, 302.4 and 264.0 mg ethanol/g treated agrowastes from green leaves of sugarcane, jatropha deolied seed cake, tops sugarcane, dry leaves of sugarcane, and jatropha shell, respectively. This study emphasizes the value of lignocellulosic agricultural waste as a resource for the production of biofuels as well as the significance of the extraction process. • The chemical composition of sugar cane trash was determined to be mainly cellulose, hemicellulose, and lignin. • Hydrogen peroxide was the most suitable for the pretreatment of different cellulose types. • The pretreatment wastes were subjected to a fermentation procedure after being saccharified by cellulase. • All pretreated wastes recorded high ethanol yields to different degrees. [ABSTRACT FROM AUTHOR]

Background: Sirtuins have an important role in the regulation of metabolic and biological processess. Thus, we hypothesized that foods that could activate sirtuins, known as "sirtfood", may improve health status. So, this study was aimed at investigating the association between the amount of sirtfood intake and the risk of major adverse cardiovascular events (MACE). Methods: In this cohort study, 2918 adults who had no history of MACE at the start of the study (2006–2008) participated and were followed up on until 2018. The amount of sirtfoods intake (servings per week) was computed using a validated food frequency questionnaire. Each patient's medical records were evaluated to detect MACE. The Cox proportional hazards model was applied to assess the association between the amount of sirtfood intake and the risk of MACE. Results: The median duration of the study was 10.6 years. The hazard ratio (HR) for the risk of MACE was 0.70 for the second (95% CI: 0.50, 0.98) and 0.60 (95% CI: 0.42, 0.86) for the third tertile of sirtfoods intake compared with the first tertile. This association was nonlinear, and sirtfoods consumption of more than five servings per week did not result in a lower risk of MACE. In addition, there was a significant interaction between age (P-interaction < 0.001) and sirtfoods intake in relation to MACE occurrence. When assessing sirtfood components, compared with the lowest intake, the highest amount of soy (HR: 0.74, 95% CI: 0.56, 0.99) and parsley (HR: 0.62, 95% CI: 0.47, 0.83) intake was related to a lower risk of MACE. Conclusion: Our results indicated an inverse association between a higher amount of sirtfood intake and a lower risk of MACE incidents. This association was nonlinear, and having more than five servings of sirtfood per week did not reduce the risk of MACE any further. [ABSTRACT FROM AUTHOR]

Background: Lipoid Proteinosis (LP), also termed Urbach-Wiethe disease, is an enigmatic genodermatosis marked by the systemic deposition of hyaline material. With its etiology rooted in ECM1 mutations, LP's neuropathological spectrum has been hypothesized to involve an array of neurodegenerative biomarkers, underscoring a potential for substantial neurobiological implications. This study endeavored to elucidate the serum concentrations of neurodegenerative biomarkers--phosphorylated Tau (pMAPT), S100B, Neuron-Specific Enolase (NSE), Neurofilament Light Chain (NEFL), and Glial Fibrillary Acidic Protein (GFAP)--in LP patients, seeking to establish their diagnostic utility for the condition. Materials and Methods: Fifteen LP patients and 15 matched healthy controls were enrolled. Serum levels of the biomarkers were quantified using ELISA, and their predictive power was assessed through binary logistic regression and Receiver Operating Characteristic (ROC) analysis. Results: Elevated serum levels of NSE, NEFL, and GFAP were observed in LP subjects relative to healthy counterparts, reaching statistical significance (p<0.05). In contrast, pMAPT and S100B levels did not differ appreciably. GFAP is considered a predictive marker for LP with an area under the curve (AUC) value of 0.813 and a 95% confidence interval (CI) of 0.658-0.968 (p=0.003). Conclusions: The study underscores a distinctive neurodegenerative profile in LP, with NSE, NEFL, and GFAP concentrations significantly amplified. These biomarkers, particularly GFAP, may represent novel indicators for LP, offering prospective biomarker-based diagnostic strategies. The insights garnered herein pave the way for advanced understanding and clinical management of LP, delineating a novel avenue for future high-impact research. [ABSTRACT FROM AUTHOR]

This study presents an innovative method for synthesizing β‐amino carbonylated compounds, specifically 2‐[phenyl(phenylamino)methyl] cyclohexanone, achieving high conversions and diastereomeric ratios. Using trypsin or α‐chymotrypsin in both free and immobilized forms on titanate nanotubes (NtsTi), synthesized through alkaline hydrothermal methods, successful immobilization yields were attained. Notably, α‐chymotrypsin, when free, displayed a diastereoselective synthesis of the anti‐isomer with 97 % conversion and 16 : 84 (syn : anti) diastereomeric ratio, which slightly decreased upon immobilization on NtsTi. Trypsin, in its free form, exhibited diastereoselective recognition of the syn‐isomer, while immobilization on NtsTi (trypsin/NtsTi) led to an inversion of diastereomeric ratio. Both trypsin/NtsTi and α‐chymotrypsin/NtsTi demonstrated significant catalytic efficiency over five cycles. In conclusion, NtsTi serves as an effective support for trypsin and α‐chymotrypsin immobilization, presenting promising prospects for diastereoselective synthesis and potential industrial applications. Furthermore, it offers promising prospects for the diastereoselective synthesis of 2‐[phenyl(phenylamino)methyl] cyclohexanone through multicomponent Mannich reaction and future industrial application. [ABSTRACT FROM AUTHOR]

The main objectives of the present investigation was to characterize and recognize potential indigenous Rhizobium spp. associated with red clover nodules, as well as to evaluate the influence of prospective rhizobia inoculation (individual and consortia) on common bean plant growth. Total of 8 rhizobia like bacteria were isolated and were investigated for their specific PGP traits, production of hydrolytic enzymes. Results revealed, four isolates synthesize IAA (22.3 ± 1.45 to 88.6 ± 1.45 μg mL−1), three isolates solubilized phosphate (40.69 ± 1.25 to 216.3 ± 1.31 μg mL−1), three isolates solubilized potassium, produced siderophore, and showed chitinase production, two isolates possess ACC deaminase activity, one isolate was able to solubilize zinc, 5 isolates produced β- 1, 3-glucanase (0.38 ± 0.005 to 2.82 ± 0.011 % units) and four isolates produced cellulase. Two bacterial isolates found to exhibit most of the PGP activities, were selected and identified on the basis of phenotypic, biochemical tests and 16S rRNA sequencing, as Rhizobium tropici IHTF-1 and Rhizobium mayense IHTF-2. Interaction of these potential rhizobium strains individually and consortia with common bean plants under greenhouse conditions boosted root and shoot length, fresh and dry biomass compared to un-inoculated control plants. SEM study revealed that both the strains, IHTF-1 and IHTF-2 colonized common bean roots. These results imply that Rhizobium spp., identified in this work, can be used as bio-inoculants to increase common bean plant production in a sustainable way. [Display omitted] [ABSTRACT FROM AUTHOR]