Converting Enzyme Research Articles

Взаимосвязь между течением COVID-19 и уровнем половых гормонов у мужчин.

Introduction. The new coronavirus infection remains a pressing health problem today. The new mutagenic strain (omicron) and its variants (stealth-omicron, «centaur»), like its predecessors (beta, delta), also causes multisystem damage to the body. A pressing issue in the study of the pathogenic properties of the pathogen is the effect of the virus on changes in the level of sex hormones in men. Purpose of the work: to analyze modern ideas about the course of COVID-19 and the level of sex hormones in men. Materials and methods. A search for publications was carried out in the international MEDLINE database on the PubMed platform, in the scientific electronic library eLibrary.ru and in the GoogleScholar. Search terms included the following combinations of words: «COVID-19», «SARS-CoV-2», «male fertility», «hypogonadism», «testis», «angiotensin converting enzyme 2», «reproductive health», «testosterone», «estradiol», «hypothalamic-pituitary-gonadal axis». A total of 130 publications were analyzed, of which 43 publications were selected for this review. Conference abstracts, short communications, and duplicate publications were excluded. Results. According to a review of scientific publications, the level of male sex hormones and the course of the new coronavirus infection are interrelated. The novel coronavirus infection has a negative impact on male sex hormone levels via ACE-2 as well as cellular transmembrane serine protease 2 (TMPRSS2), due to increased levels of pro-inflammatory cytokines that increase metabolic clearance and suppress the gonadal axis. Conclusions. COVID-19 and sex hormones have a relationship. There is still debate and there is no consensus on how the level of sex hormones affects the course of COVID-19. Therefore, we believe that this issue remains relevant to this day, given that outbreaks of new strains of SARS-CoV-2 arise periodically.

Antioxidant capacity and peptidomic analysis of in vitro digested Camelina sativa L. Crantz and Cynara cardunculus co-products

In recent decades, the food system has been faced with the significant problem of increasing food waste. Therefore, the feed industry, supported by scientific research, is attempting to valorise the use of discarded biomass as co-products for the livestock sector, in line with EU objectives. In parallel, the search for functional products that can ensure animal health and performances is a common fundamental goal for both animal husbandry and feeding. In this context, camelina cake (CAMC), cardoon cake (CC) and cardoon meal (CM), due valuable nutritional profile, represent prospective alternatives. Therefore, the aim of this work was to investigate the antioxidant activity of CAMC, CC and CM following in vitro digestion using 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), Ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays. Total phenolic content (TPC) and angiotensin converting enzyme (ACE) inhibitory activity, actively involved in modulating antioxidant properties, were also studied. Further, a peptidomic analysis was adopted to substantiate the presence of bioactive peptides after in vitro digestion. The results obtained confirmed an interesting nutritional profile of CAMC, CC and CM and relevant antioxidant and ACE inhibitory activities. In particular, considering antioxidant profile, CM and CC revealed a significantly higher (10969.80 ± 18.93 mg TE/100 g and 10451.40 ± 149.17 mg TE/100 g, respectively; p < 0.05) ABTS value than CAMC (9511.18 ± 315.29 mg TE/100 g); a trend also confirmed with the FRAP assay (306.74 ± 5.68 mg FeSO4/100 g; 272.84 ± 11.02 mg FeSO4/100 g; 103.84 ± 3.27 mg FeSO4/100 g, for CC, CM and CAMC, respectively). Similar results were obtained for TPC, demonstrating the involvement of phenols in modulating antioxidant activity. Finally, CAMC was found to have a higher ACE inhibitory activity (40.34 ± 10.11%) than the other matrices. Furthermore, potentially bioactive peptides associated with ACE inhibitory, anti-hypertensive, anti-cancer, antimicrobial, antiviral, antithrombotic, DPP-IV inhibitory and PEP-inhibitory activities were identified in CAMC. This profile was broader than that of CC and CM. The presence of such peptides corroborates the antioxidant and ACE profile of the sample. Although the data obtained report the important antioxidant profile of CAMC, CC, and CM and support their possible use, future investigations, particularly in vivo trials will be critical to evaluate and further investigate their effects on the health and performance of farm animals.

Impact of Angiotensin Converting Enzyme Inhibitors on Pathologic Complete Response With Neoadjuvant Chemotherapy for Muscle Invasive Bladder Cancer

IntroductionThe renin-angiotensin system (RAS) has been demonstrated to modulate cell proliferation, desmoplasia, angiogenesis and immunosuppression. We examined the association of RAS inhibitors (RASi)—namely angiotensin converting enzyme inhibitors (ACEi) and angiotensin receptor blockers (ARB)—with neoadjuvant chemotherapy (NAC) for muscle-invasive bladder cancer (MIBC) preceding radical cystectomy (RC). Patients and MethodsWe retrospectively investigated concurrent RASi use with NAC prior to RC in 302 patients with MIBC from 3 academic institutions. Outcomes included pathologic complete response (pCR) and overall survival (OS). Pathologic features, performance status (PS), clinical stage, type/number of cycles of NAC, and toxicities were collected. ResultsOverall pCR rate was 26.2% and 5-year OS was 62%. Concurrent ACEi intake with NAC approached significance for association with pCR (odds ratio [OR] = 1.71; 95% CI, 0.94-3.11; P = .077). Patients with cT3/4N0-N1 disease receiving ACEi had higher pCR rates (30.8% vs. 17.7%, P = .056) than those not on ACEi. Female sex had a statistically significant favorable interaction for pCR with ACEi intake (P = .044). ACEi intake was not associated with OS, while pCR, PS and lower clinical stage were significantly associated with improved OS. ConclusionACEi intake is potentially associated with increased pCR in patients with MIBC receiving NAC prior to RC, and this association is more pronounced in patients with higher clinical stage of disease at the initiation of therapy and female sex. Our data suggest the potential relevance of the RAS as a therapeutic target in aggressive MIBC.

Enzymatic conversion of camellia seed oil into glycerol esters: Synthesis and characterization

AbstractThe conversion of triacylglycerols in edible oils into diacylglycerols (DAGs) is of great significance for obtaining products with health benefits. Camellia seed oil (C‐oil), which is rich in oleic acid and linoleic acid, is an excellent raw material for the production of DAGs. In this study, single factor optimization experiments were carried out for hydrolysis and esterification respectively. Using Lipozyme® RM IM as catalyst, the maximum percent of C‐oil hydrolysis reached 87.14% at the reaction temperature of 60°C, reaction time of 24 h, water content of 30% and enzyme addition amount of 4%. The maximum content of camellia seed oil diacylglycerol (C‐DAG) reached 62.49% under the conditions of Lipozyme® RM IM as catalyst, vacuum system, 3% enzyme addition, 2% water addition, reaction temperature of 50°C and substrate molar ratio of free fatty acid to glycerol of 1:1. The high content of DAG was obtained by a coupled method, which eliminated the purification steps and reduced production costs. C‐oil and C‐DAG have been characterized by GC, TG, DSC, and GC‐IMS. Our results showed that the enzymatic coupling method did not affect the structural of the substances, but did affect the crystallization and melting properties of the oils. Moreover, the taste of C‐DAG was more delicate than C‐oil. Finally, the reaction mechanism was analyzed using FTIR spectroscopy, revealing that C‐oil was primarily hydrolyzed into free fatty acids. C‐DAG exhibited ester C‐O stretching vibrations in the range 1280–1030 cm−1, indicating successful esterification reaction between camellia seed oil free fatty acids (C‐FFAs) and glycerol catalyzed by lipases.

Human angiotensin-converting enzyme 2–specific antisense oligonucleotides reduce infection with SARS-CoV-2 variants

BackgroundThe Spike protein mutation of SARS-CoV-2 led to decreased protective effect of various vaccines and monoclonal antibodies, suggesting that blocking SARS-CoV-2 infection by targeting host factors would make the therapy more resilient against virus mutations. Angiotensin converting enzyme 2 (ACE2) is the host receptor of SARS-CoV-2 and its variants, as well as many other coronaviruses. Down-regulation of ACE2 expression in the respiratory tract may prevent viral infection. Antisense oligonucleotides (ASOs) can be rationally designed based on sequence data, require no delivery system, and can be administered locally. ObjectiveWe sought to design ASOs that can block SARS-CoV-2 by down-regulating ACE2 in human airway. MethodsACE2-targeting ASOs were designed using a bioinformatic method and screened in cell lines. Human primary nasal epithelial cells cultured at the air-liquid interface and humanized ACE2 mice were used to detect the ACE2 reduction levels and the safety of ASOs. ASOs pretreated nasal epithelial cells and mice were infected and then used to detect the viral infection levels. ResultsASOs reduced ACE2 expression on mRNA and protein level in cell lines and in human nasal epithelial cells. Furthermore they efficiently suppressed virus replication of three different SARS-CoV-2 variants in human nasal epithelial cells. In vivo, ASOs also down-regulated human ACE2 in humanized ACE2 mice and thereby reduced viral load, histopathological changes in lungs, and they increased survival of mice. ConclusionACE2-targeting ASOs can effectively block SARS-COV-2 infection. Our study provides a new approach for blocking SARS-CoV-2 and other ACE2-targeting virus in high-risk populations.

Effects of Evodiamine on Behavior and Hippocampal Neurons through Inhibition of Angiotensin-Converting Enzyme and Modulation of the Renin Angiotensin Pathway in a Mouse Model of Post-Traumatic Stress Disorder.

Post-traumatic stress disorder (PTSD) is a persistent psychiatric condition that arises following exposure to traumatic events such as warfare, natural disasters, or other catastrophic incidents, typically characterized by heightened anxiety, depressive symptoms, and cognitive dysfunction. In this study, animals subjected to single prolonged stress (SPS) were administered evodiamine (EVO) and compared to a positive control group receiving sertraline. The animals were then assessed for alterations in anxiety, depression, and cognitive function. Histological analysis was conducted to examine neuronal changes in the hippocampus. In order to predict the core targets and related mechanisms of evodiamine intervention in PTSD, network pharmacology was used. The metabolic markers pre- and post-drug administration were identified using nontargeted serum metabolomics techniques, and the intersecting Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were screened. Finally, the core targets were validated through molecular docking, enzyme-linked immunosorbent assays, and immunofluorescence staining to confirm the anti-PTSD effects and mechanisms of these targets. As well as improving cognitive impairment, evodiamine reversed anxiety- and depression-like behaviors. It also inhibited the reduction in the number of hippocampal neuronal cells and Nissl bodies in SPS mice inhibited angiotensin converting enzyme (ACE) levels in the hippocampus of SPS mice, and modulated the renin angiotensin pathway and its associated serum metabolites in brain tissue. Evodiamine shows promise as a potential candidate for alleviating the symptoms of post-traumatic stress disorder.

The lipopolysaccharide-TLR4 axis regulates hepatic glutaminase 1 expression promoting liver ammonia build-up as steatotic liver disease progresses to steatohepatitis

IntroductionAmmonia is a pathogenic factor implicated in the progression of metabolic-associated steatotic liver disease (MASLD). The contribution of the glutaminase 1 (GLS) isoform, an enzyme converting glutamine to glutamate and ammonia, to hepatic ammonia build-up and the mechanisms underlying its upregulation in metabolic-associated steatohepatitis (MASH) remain elusive. MethodsMultiplex transcriptomics and targeted metabolomics analysis of liver biopsies in dietary mouse models representing the whole spectra of MASLD were carried out to characterize the relevance of hepatic GLS during disease pathological progression. In addition, the acute effect of liver-specific GLS inhibition in hepatic ammonia content was evaluated in cultured hepatocytes and in in vivo mouse models of diet-induced MASLD. Finally, the regulatory mechanisms of hepatic GLS overexpression related to the lipopolysaccharide (LPS)/Toll-like receptor 4 (TLR4) axis were explored in the context of MASH. ResultsIn mouse models of diet-induced MASLD, we found that augmented liver GLS expression is closely associated with the build-up of hepatic ammonia as the disease progresses from steatosis to steatohepatitis. Importantly, the acute silencing/pharmacological inhibition of GLS diminishes the ammonia burden in cultured primary mouse hepatocytes undergoing dedifferentiation, in steatotic hepatocytes, and in a mouse model of diet-induced steatohepatitis, irrespective of changes in ureagenesis and gut permeability. Under these conditions, GLS upregulation in the liver correlates positively with the hepatic expression of TLR4 that recognizes LPS. In agreement, the pharmacological inhibition of TLR4 reduces GLS and hepatic ammonia content in LPS-stimulated mouse hepatocytes and hyperammonemia animal models of endotoxemia. ConclusionsOverall, our results suggest that the LPS/TLR4 axis regulates hepatic GLS expression promoting liver ammonia build-up as steatotic liver disease progresses to steatohepatitis.

PATH-04. OPTIMIZING CELL-FREE DNA ASSAYS FOR LOW INPUT LIQUID BIOPSY SAMPLES FROM PEDIATRIC BRAIN TUMOR PATIENTS

Abstract BACKGROUND Genetic and epigenetic profiling have transformed biological understanding, diagnostics, and therapeutic approaches for pediatric central nervous system (CNS) tumors. Obtaining tumor tissue may be associated with neurosurgical risk, may not be feasible in some cases, and is not routinely performed during follow-up or at relapse. Liquid biopsies (LBs) have emerged as a minimally invasive, serially collectable source of tumor-derived DNA. Here, we collected cerebrospinal fluid (CSF) and serum LBs from a large, unselected, cross-entity pediatric CNS tumor cohort (n=55 patients) for method optimization. METHODS Cell-free DNA (cfDNA) isolated from CSF and serum samples was analyzed 1) by low-coverage whole genome sequencing for copy number variation (CNV)-based tumor detection and 2) by enzymatic conversion (EC)-based DNA methylation profiling using the EPIC DNA methylation array and the Heidelberg Brain Tumor Classifier. All library protocols were optimized for LB samples with low cfDNA amounts in the picogram range. Matched tumor samples were used as reference. RESULTS Our lcWGS-based cfDNA profiling protocol had a high technical success rate across the whole cohort (&amp;gt;95%), despite very low cfDNA amounts for most CSF samples. The majority of CSF LBs captured tumor-derived CNVs and displayed a high fraction of tumor-derived cfDNA (&amp;gt;5%, range 0-78%). For serum LBs, only few samples showed detectable CNVs. Proof-of-concept methylation analyses leveraging a new EC-based workflow allowed molecular classification using CSF, particularly for embryonal brain tumors (11/15 samples). Clinical workup of selected cases illustrates the potential of LBs to aid as diagnostic tool, monitor disease, and characterize tumor evolution. CONCLUSIONS Tackling a major roadblock of clinical translation of LBs, this study delineates new methods tailored to low input cfDNA samples. Results highlight the clinical utility of LBs for the management of brain tumor patients, providing a strong rationale for prospective validation within pediatric neuro-oncology trials.

Missed opportunity: a clinical data linkage study of guideline‐directed medical therapy and clinical outcomes of patients discharged with acute coronary syndrome who attended cardiac rehabilitation programs

AbstractBackgroundAlthough guidelines recommend guideline‐directed medical therapy (GDMT) for patients with acute coronary syndrome (ACS), implementation is limited in clinical practice.AimTo assess the level of GDMT in ACS patients after discharge who attended cardiac rehabilitation (CR) programs and association with clinical outcomes.MethodA cross‐sectional study was conducted in 13 rural and 10 metropolitan CR programs via all modes of delivery (face‐to‐face, telephone, or general practice‐hybrid) operating in South Australia, Australia. ACS patients were included if they were ≥18 years of age and were referred and attended CR programs with medication details recorded in their hospital discharge summary. GDMT was assessed according to the Australian clinical guidelines for the management of acute coronary syndromes 2016. Prescription of all the four recommended medication classes was considered optimal. Logistic regression and χ2 test were used for association.ResultsOf the 1229 patients included, 74.6% were male and 41.1% had acute myocardial infarction. Only 39.7% of patients received optimal prescription. Prescription of any three or two medication class combinations occurred for 78.3% and 94.1% of patients, respectively. Optimal GDMT was associated with fewer hospital admissions (odds ratio = 0.647, 95% confidence interval 0.424–0.987, p = 0.043) with no significant gender association. Women were less likely to be prescribed angiotensin converting enzyme inhibitors (p = 0.003), angiotensin receptor blockers (p = 0.007), statins (p = 0.005), and any two (p &lt; 0.001) and three combinations (p = 0.023) of medication classes.ConclusionGDMT prescription was suboptimal in patients with ACS before attendance at CR. Primary care and CR clinicians have missed an opportunity to implement best practice guideline recommendations, particularly for women.

Assay for okadaic acid O-acyl transferase using HPLC-FLD.

Dinophysistoxin 1 (DTX1, 1) and okadaic acid (OA, 2), produced by the dinoflagellates Dinophysis spp. and Prorocentrum spp., are primary diarrhetic shellfish toxins (DSTs), which may cause gastric illness in people consuming such as bivalves. Both compounds convert to dinophysistoxin 3 (DTX3, 3; generic name for 1 and 2 with fatty acids conjugated at 7-OH) in bivalves. The enzyme okadaic acid O-acyl transferase (OOAT) is a membrane protein found in the microsomes of the digestive glands of bivalves. In this study, we established an in vitro enzymatic conversion reaction using 4-nitro-2,1,3-benzoxadiazole (NBD)-OA (4), an OA derivative conjugated with (R)-(-)-4-nitro-7-(3-aminopyrrolidin-1-yl)-2,1,3-benzoxadiazole (NBD-APy) on 1-CO2H, as a substrate. We detected the enzymatically produced 3, NBD-7-O-palmitoyl-OA (NBD-Pal-OA), using high-performance liquid chromatography-fluorescence detection. We believe that an OOAT assay using 4 will facilitate the fractionation and isolation of OOAT in the future.

Bidirectional effects of the tryptophan metabolite indole-3-acetaldehyde on colorectal cancer.

Colorectal cancer (CRC) has a high incidence and mortality. Recent studies have shown that indole derivatives involved in gut microbiota metabolism can impact the tumorigenesis, progression, and metastasis of CRC. To investigate the effect of indole-3-acetaldehyde (IAAD) on CRC. The effect of IAAD was evaluated in a syngeneic mouse model of CRC and CRC cell lines (HCT116 and DLD-1). Cell proliferation was assessed by Ki-67 fluorescence staining and cytotoxicity tests. Cell apoptosis was analysed by flow cytometry after staining with Annexin V-fluorescein isothiocyanate and propidium iodide. Invasiveness was investigated using the transwell assay. Western blotting and real-time fluorescence quantitative polymerase chain reaction were performed to evaluate the expression of epithelial-mesenchymal transition related genes and aryl hydrocarbon receptor (AhR) downstream genes. The PharmMapper, SEA, and SWISS databases were used to screen for potential target proteins of IAAD, and the core proteins were identified through the String database. IAAD reduced tumorigenesis in a syngeneic mouse model. In CRC cell lines HCT116 and DLD1, IAAD exhibited cytotoxicity starting at 24 h of treatment, while it reduced Ki67 expression in the nucleus. The results of flow cytometry showed that IAAD induced apoptosis in HCT116 cells but had no effect on DLD1 cells, which may be related to the activation of AhR. IAAD can also increase the invasiveness and epithelial-mesenchymal transition of HCT116 and DLD1 cells. At low concentrations (< 12.5 μmol/L), IAAD only exhibited cytotoxic effects without promoting cell invasion. In addition, predictions based on online databases, protein-protein interaction analysis, and molecular docking showed that IAAD can bind to matrix metalloproteinase-9 (MMP9), angiotensin converting enzyme (ACE), poly(ADP-ribose) polymerase-1 (PARP1), matrix metalloproteinase-2 (MMP2), and myeloperoxidase (MPO). Indole-3-aldehyde can induce cell apoptosis and inhibit cell proliferation to prevent the occurrence of CRC; however, at high concentrations (≥ 25 μmol/L), it can also promote epithelial-mesenchymal transition and invasion in CRC cells. IAAD activates AhR and directly binds MMP9, ACE, PARP1, MMP2, and MPO, which partly reveals why it has a bidirectional effect.

The Effects of Exogenous Benzoic Acid on the Physicochemical Properties, Enzyme Activities and Microbial Community Structures of Perilla frutescens Inter-Root Soil.

This study analyzed the effects of benzoic acid (BA) on the physicochemical properties and microbial community structure of perilla rhizosphere soil. The analysis was based on high-throughput sequencing technology and physiological and biochemical detection. The results showed that with the increase in BA concentration, soil pH significantly decreased, while the contents of total nitrogen (TN), alkaline nitrogen (AN), available phosphorus (AP), and available potassium (AK) significantly increased. The activities of soil conversion enzymes urease and phosphatase significantly increased, but the activities of catalase and peroxidase significantly decreased. This indicates that BA can increase soil enzyme activity and improve nutrient conversion; the addition of BA significantly altered the composition and diversity of soil bacterial and fungal communities. The relative abundance of beneficial bacteria such as Gemmatimonas, Pseudolabrys, and Bradyrhizobium decreased significantly, while the relative abundance of harmful fungi such as Pseudogymnoascus, Pseudoeurotium, and Talaromyces increased significantly. Correlation analysis shows that AP, AN, and TN are the main physicochemical factors affecting the structure of soil microbial communities. This study elucidates the effects of BA on the physicochemical properties and microbial community structure of perilla soil, and preliminarily reveals the mechanism of its allelopathic effect on the growth of perilla.

Alteration of circulating ACE2-network related microRNAs in patients with COVID-19

Angiotensin converting enzyme 2 (ACE2) serves as the primary receptor for the SARS-CoV-2 virus and has implications for the functioning of the cardiovascular system. Based on our previously published bioinformatic analysis, in this study we aimed to analyze the diagnostic and predictive utility of miRNAs (miR-10b-5p, miR-124-3p, miR-200b-3p, miR-26b-5p, miR-302c-5p) identified as top regulators of ACE2 network with potential to affect cardiomyocytes and cardiovascular system in patients with COVID-19. The expression of miRNAs was determined through qRT-PCR in a cohort of 79 hospitalized COVID-19 patients as well as 32 healthy volunteers. Blood samples and clinical data of COVID-19 patients were collected at admission, 7-days and 21-days after admission. We also performed SHAP analysis of clinical data and miRNAs target predictions and advanced enrichment analyses. Low expression of miR-200b-3p at the seventh day of admission is indicative of predictive value in determining the length of hospital stay and/or the likelihood of mortality, as shown in ROC curve analysis with an AUC of 0.730 and a p-value of 0.002. MiR-26b-5p expression levels in COVID-19 patients were lower at the baseline, 7 and 21-days of admission compared to the healthy controls (P < 0.0001). Similarly, miR-10b-5p expression levels were lower at the baseline and 21-days post admission (P = 0.001). The opposite situation was observed in miR-124-3p and miR-302c-5p. Enrichment analysis showed influence of analyzed miRNAs on IL-2 signaling pathway and multiple cardiovascular diseases through COVID-19-related targets. Moreover, the COVID-19-related genes regulated by miR-200b-3p were linked to T cell protein tyrosine phosphatase and the HIF-1 transcriptional activity in hypoxia. Analysis focused on COVID-19 associated genes showed that all analyzed miRNAs are strongly affecting disease pathways related to CVDs which could be explained by their strong interaction with the ACE2 network.

Investigation of Cyrene organosolv fractionation of softwood biomass and alkaline post-incubation

Cyrene organosolv fractionation effectively extracted 78 % of lignin from recalcitrant softwood biomass pine at a mild temperature of 120 °C. However, the enzymatic conversion of the fractionated cellulose-rich solid did not improve significantly. Alkali post-incubation of the fractionated cellulose fraction notably enhanced the glucan conversion. This phenomenon has also been observed in other organosolv processes, but how this approach transforms pretreated biomass has not yet been comprehensively investigated. In this study, small-angle X-ray scattering (SAXS) was employed to understand the structural changes in biomass during fractionation and post-incubation at the nanometer scale. No lignin aggregation was found on the microfibrils, whereas the distance between the microfibrils increased after pretreatment and decreased after alkaline post-incubation. These results suggests that the Cyrene molecules remained between the microfibrils and were removed by post-incubation. In addition, the pine lignin recovered after pretreatment was characterized by NMR to understand the impact of Cyrene pretreatment on the lignin structure.

Angiotensin-I converting enzyme (ACE) inhibitory bioaccessible peptides obtained from protein extracts of Macrocystis pyrifera and Chondracanthus chamissoi seaweeds

Hypertension is a paramount chronic health concern, underscoring the critical need for preventive measures in public health. In recent years, research has discovered natural anti-hypertensive agents, including those derived from macroalgae. This study focuses on enzymatically hydrolyzing protein extracts from Macrocystis pyrifera and Chondracanthus chamissoi seaweeds to yield angiotensin I-converting enzyme (ACE) inhibitory peptides, potentially applicable in both food and bioactive products. ACE inhibitory activity was evaluated for enzymatic hydrolysates generated using bromelain, papain, and alcalase. Notably, hydrolysates produced using alcalase demonstrated the highest effect, yielding 74 % ACE inhibition for M. pyrifera and 58 % for C. chamissoi protein extracts. Optimization efforts focused on hydrolysis conditions, varying hydrolysis time and substrate-to-enzyme ratio, resulting in peak ACE inhibition of 79.9 % for M. pyrifera (5.3 h, 12.7 S:E ratio) and 70.3 % for C. chamissoi (6 h, 10 S:E ratio).Further analysis explored the relationship between molecular size separation and ACE inhibitory activity in algae hydrolysates. Fractionation via ultrafiltration revealed that fractions below 3 kDa exhibited the highest ACE inhibitory potency. Additionally, in vitro digestion studies employing pepsin and pancreatin treatments demonstrated the bioaccessibility of the seaweed-derived bioactive peptides, with maintained or enhanced bioactivity post-digestion. These findings underscore the potential of protease hydrolysates from M. pyrifera and C. chamissoi as sources of ACE inhibitory activity. Consequently, they present an exciting natural source of functional ingredients or nutraceuticals for hypertension management, with the benefits of easy implementation, profitability, and sustainability in their production process.

DNA methylation modification in Idiopathic pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and irreversible interstitial lung disease with a prognosis worse than lung cancer. It is a fatal lung disease with largely unknown etiology and pathogenesis, and no effective therapeutic drugs render its treatment largely unsuccessful. With continuous in-depth research efforts, the epigenetic mechanisms in IPF pathogenesis have been further discovered and concerned. As a widely studied mechanism of epigenetic modification, DNA methylation is primarily facilitated by DNA methyltransferases (DNMTs), resulting in the addition of a methyl group to the fifth carbon position of the cytosine base, leading to the formation of 5-methylcytosine (5-mC). Dysregulation of DNA methylation is intricately associated with the advancement of respiratory disorders. Recently, the role of DNA methylation in IPF pathogenesis has also received considerable attention. DNA methylation patterns include methylation modification and demethylation modification and regulate a range of essential biological functions through gene expression regulation. The Ten-Eleven-Translocation (TET) family of DNA dioxygenases is crucial in facilitating active DNA demethylation through the enzymatic conversion of the modified genomic base 5-mC to 5-hydroxymethylcytosine (5-hmC). TET2, a member of TET proteins, is involved in lung inflammation, and its protein expression is downregulated in the lungs and alveolar epithelial type II cells of IPF patients. This review summarizes the current knowledge of pathologic features and DNA methylation mechanisms of pulmonary fibrosis, focusing on the critical roles of abnormal DNA methylation patterns, DNMTs, and TET proteins in impacting IPF pathogenesis. Researching DNA methylation will enchance comprehension of the fundamental mechanisms involved in IPF pathology and provide novel diagnostic biomarkers and therapeutic targets for pulmonary fibrosis based on the studies involving epigenetic mechanisms.

Effect of cellulose reducing ends and primary hydroxyl groups modifications on cellulose-cellulase interactions and cellulose hydrolysis.

Cellulose reducing ends are believed to play a vital role in the cellulose recalcitrance to enzymatic conversion. However, their role in insoluble cellulose accessibility and hydrolysis is not clear. Thus, in this study, reducing ends of insoluble cellulose derived from various sources were modified by applying reducing and/or oxidizing agents. The effects of cellulose reducing ends modification on cellulose reducing ends, cellulose structure, and cellulose accessibility to cellulase were evaluated along with the impact on cellulose hydrolysis with complete as well purified cellulase components. Sodium borohydride (NaBH4) reduction and sodium chlorite-acetic acid (SC/AA) oxidation were able to modify more than 90% and 60% of the reducing ends, respectively, while the bicinchoninic acid (BCA) reagent applied for various cycles oxidized cellulose reducing ends to various extents. X-ray diffractograms of the treated solids showed that these treatments did not change the cellulose crystalline structure and the change in crystallinity index was insignificant. Surprisingly, it was found that the cellulose reducing ends modification, either through selective NaBH4 reduction or BCA oxidation, had a negligible impact on cellulose accessibility as well on cellulose hydrolysis rates or final conversions with complete cellulase at loadings as low as 0.5 mg protein/g cellulose. In fact, in contrast to what is traditionally believed, modifications of cellulose reducing ends by these two methods had no apparent impact on cellulose conversion with purified cellulase components and their synergy. However, SC/AA oxidation resulted in significant drop in cellulose conversion (10%-50%) with complete as well purified cellulase components. Nonetheless, further research revealed that the cause for drop in cellulose conversion for the SC/AA oxidation case was due to primary hydroxyl groups (PHGs) oxidation and not the oxidation of reducing ends. Furthermore, it was found that the PHGs modification affects cellulose accessibility and slows the cellulase uptake as well resulting in significant drop in cellulose conversions.

A novel series of metazoan L/D peptide isomerases

The function of endogenous cell-cell signaling peptides rely on their interactions with cognate receptors, which in turn are influenced by the peptides' structures, necessitating a comprehensive understanding of the suite of post-translational modifications (PTMs) of the peptide. Herein, we report the initial characterization of putative peptide isomerase enzymes extracted from R. norvegicus, A. californica, and B. taurus tissues. These enzymes are both tissue and substrate-specific across all three organisms. Notably, the lungs of the mammalian species, and the central nervous system of the mollusk displayed the highest isomerase activity among the examined tissues. In-vitro enzymatic conversion was observed for several endogenous peptides, such as the tetrapeptide GFFD in A. californica, and mammalian neuropeptide FF in R. norvegicus and B. taurus. To understand their mode of action, we explored the effects of several inhibitors on these enzymes, which suggests common active site residues. While further characterization of these enzymes is required, the investigations emphasize a widespread and overlooked enzyme activity related to the creation of bioactive peptides.

The association of the ACE gene I/D polymorphism and in-game performance in elite Rugby Union players

Genetic variation has been proposed as a factor in human athletic performance. The angiotensin converting enzyme (ACE) gene has been studied extensively albeit with heterogeneous results. The insertion (I) and deletion (D) alleles have been associated with altered serum and tissue ACE, as well as endurance- and power-based performance respectively. It is suggested the distinction in rugby union performance can be partly attributed to variations in the ACE gene. The aim of this study was to identify associations between the ACE I/D genetic polymorphism and in-game performance in elite (≥ 5 games at top-flight) rugby union athletes. The athlete cohort consisted of 347 Caucasian men, divided into positional groups (backs and forwards) and positional subgroups (front five, back row, halves, centres, and back three). Ethical approval was provided by Manchester Metropolitan University with written informed consent obtained from all participants. Genotyping and genotype analysis was performed using a combination of the X9 High-Throughput Genomics System (Standard BioTools) and the StepOnePlus™ Real-Time PCR System (Applied Biosystems) and their respective corresponding software. IBM SPSS Statistics 28 (SPSS, Chicago, IL) software was used to conduct Pearson's Chi-square (χ2) test of independence comparing genotype and allelic frequencies and the Kruskal-Wallis H and Mann-Whitney U tests to investigate differences between genotypes. Genotypic and allelic frequencies did not differ at the population level, nor at the positional group/subgroup levels. Performance did not differ between genotypes nor alleles at the population and positional group levels. Back row II genotypes beat more defenders than IDs (P = 0.008) but completed fewer tackles than both IDs (P = 0.05) and DDs (P = 0.013). Halfback DDs missed fewer tackles than both IDs (P = 0.003) and IIs (P = 0.01). Centre DD genotypes made more carries than IIs (P = 0.006). Back three IIs scored more tries than both IDs (P = 0.005) and DDs (P = 0.007) and made more clean breaks than IDs (P = 0.004). Most of the existing literature has been concerned with identifying the ACE gene as a marker of elite performance. To our knowledge, this is the first study to elucidate how the inter-personal differences in rugby union performance are influenced by the ACE I/D polymorphism. The findings from this study may provide novel insight into the specialisation of training and rehabilitation modalities for athletes, based on their genetic makeup.

Sex-specific transgenerational effects on murine thyroid gland imposed by ancestral exposure to neonicotinoid thiacloprid

Neonicotinoids, a relatively new widely used class of insecticide is used in agriculture to control insect populations. We examined the capacity of ancestral exposure to the neonicotinoid thiacloprid (thia) to induce transgenerational effects on thyroid tissue. Pregnant outbred Swiss female mice were exposed to thia at embryonic days E6.5 to E15.5 using 0, 0.6, and 6 mg/kg/day doses. Thyroid paraffin sections were prepared for morphology analysis. We apply ELISA method to measure T4 and TSH levels, RT-qPCR for gene expression analysis, ChIP-qPCR techniques for sperm histone H3K4me3 analysis, and immunofluorescence microscopy and western blots for protein detection. We observed an alteration in the morphology of thyroids in both males and females in the F3 generation. We observed an increase in T4 hormone in F1 females and a significant T4 level decrease in F3 males. T4 changes in F1 females were associated with a TSH increase. We found that the amount of Iodothyronine Deiodinase 1 (DIO1) (an enzyme converting T4 to T3) was decreased in both F1 and F3 generations in female thyroids. GNAS protein which is important for thyroid function has increased in female thyroids. Gene expression analysis showed that the expression of genes encoding thyroid gland development, chromatin, biosynthesis and transport factors were affected in the thyroid gland in both sexes in F1 and F3. The analysis of sperm histone H3K4me3 showed that H3K4me3 occupancy at the Dio1 locus has decreased while Thyroglobulin (Tg) and Matrix Metallopeptidase 2 (Mmp2) genes have increased H3K4me3 occupancy in the sperm of F3 mice. Besides, DNA methylation analysis of our previously published datasets showed that, in the sperm of F1 and F3 thia-derived mice, several genes related to thyroid function show consistent alterations. Our data suggest that ancestral exposure to thiacloprid affects thyroid function not only in exposed but also in indirectly exposed F3 generation.