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Pregnancy alters many physiological systems, including the maternal gut microbiota. Diet is a key regulator of this system and can alter the host immune system to promote inflammation. Multiple perinatal disorders have been associated with inflammation, maternal metabolic alterations, and gut microbial dysbiosis, including gestational diabetes mellitus, pre-eclampsia, preterm birth, and mood disorders. However, the effects of high-inflammatory diets on the gut microbiota during pregnancy have yet to be fully explored. We aimed to address this gap using a system-based approach to characterize associations among dietary inflammatory potential, a measure of diet quality, and the gut microbiome during pregnancy. Forty-seven pregnant persons were recruited prior to 16 weeks of gestation. Participants completed a food frequency questionnaire (FFQ) and provided fecal samples. Dietary inflammatory potential was assessed using the Dietary Inflammatory Index (DII) from the FFQ data. Fecal samples were analyzed using 16S rRNA amplicon sequencing. Differential taxon abundances with respect to the DII score were identified, and the microbial metabolic potential was predicted using PICRUSt2. Inflammatory diets were associated with decreased vitamin and mineral intake and a dysbiotic gut microbiota structure and predicted metabolism. Gut microbial compositional differences revealed a decrease in short-chain fatty acid producers such as Faecalibacterium, and an increase in predicted vitamin B12 synthesis, methylglyoxal detoxification, galactose metabolism, and multidrug efflux systems in pregnant individuals with increased DII scores. Dietary inflammatory potential was associated with a reduction in the consumption of vitamins and minerals and predicted gut microbiota metabolic dysregulation.

The heterotrimeric flavin adenine dinucleotide (FAD) dependent glucose dehydrogenase derived from Burkholderia cepacia (BcGDH) has many exceptional features for its use in glucose sensing—including that this enzyme is capable of direct electron transfer with an electrode in its heterotrimeric configuration. However, this enzyme's high catalytic activity towards not only glucose but also galactose presents an engineering challenge. To increase the substrate specificity of this enzyme, it must be engineered to reduce its activity towards galactose while maintaining its activity towards glucose. To aid in these mutagenesis studies, the crystal structure composed of BcGDH's small subunit and catalytic subunit (BcGDHγα), in complex with D -glucono-1,5-lactone was elucidated and used to construct the three-dimensional model for targeted, site-directed mutagenesis. BcGDHγα was then mutated at three different residues, glycine 322, asparagine 474 and asparagine 475. The single mutations that showed the greatest glucose selectivity were combined to create the resulting mutant, α-G322Q-N474S-N475S. The α-G322Q-N474S-N475S mutant and BcGDHγα wild type were then characterized with dye-mediated dehydrogenase activity assays to determine their kinetic parameters. The α-G322Q-N474S-N475S mutant showed more than a 2-fold increase in V max towards glucose and this mutant showed a lower activity towards galactose in the physiological range (5 mM) of 4.19 U mg−1, as compared to the wild type, 86.6 U mg−1. This resulting increase in specificity lead to an 81.7 gal/glc % activity for the wild type while the α-G322Q-N474S-N475S mutant had just 10.9 gal/glc % activity at 5 mM. While the BcGDHγα wild type has high specificity towards galactose, our engineering α-G322Q-N474S-N475S mutant showed concentration dependent response to glucose and was not affected by galactose. • X-ray crystal structure of catalytic subunit of DET-type GDH in complex with 1,5-Glucono-δ-lactone was elucidated. • Three residues, G322, N474 and N475, were solicited to be responsible for enzyme substrate specificity. • G322Q/N474S/N475S mutant improved specificity toward glucose by decreasing activity toward galactose. • Structural model provided the impact of mutations, confirming the narrowed substrate binding pocket in the mutant. [ABSTRACT FROM AUTHOR]

α-Galactosidase (α-GAL) is a class of hydrolase that releases galactose from galacto-oligosaccharides and synthetic substrates such as pNPG. In this study, the production of α-GAL by Actinoplanes utahensis B1 in submerged fermentation was enhanced by using statistical methods. The effects of temperature, pH, and inoculum percentage on enzyme secretion were optimized using BBD of RSM. The optimized process was scaled up from the shake flask to the laboratory scale (5 L) and to pilot scale (30 L) using KLa based scale-up strategy. By using BBD, a maximum yield of 62.5 U/mL was obtained at a temperature of 28 °C, a pH of 6.9, and an inoculum of 6.4%. Scale-up was performed successfully and achieved a yield of 74.4 U/mL and 76.8 U/mL in laboratory scale and pilot scale fermenters. The TOST was performed to validate the scale-up strategy and the results showed a confidence level of 95% for both scales indicating the perfect execution of scale-up procedure. Through the implementation of BBD and scale-up strategy, the overall enzyme yield has been significantly increased to 76%. This is the first article to explore the scale-up of α-GAL from the A. utahensis B1 strain and provide valuable insights for industrial applications. [ABSTRACT FROM AUTHOR]

In classic galactosemia (CG) patients, aldose reductase (AR) converts galactose to galactitol. In a phase 1/2, placebo‐controlled study (NCT04117711), safety, pharmacokinetics (PK), and pharmacodynamics (PD) of govorestat were evaluated after single and multiple ascending doses (0.5‐40 mg/kg) in healthy adults (n = 81) and CG patients (n = 14). Levels of govorestat in plasma and cerebrospinal fluid (CSF) and blood levels of galactitol, galactose, and galactose‐1‐phosphate (Gal‐1p) were measured for population PK and PK/PD analyses. Govorestat was well tolerated. Adverse event frequency was comparable between placebo and govorestat. Govorestat PK displayed a 2‐compartment model with sequential zero‐ and first‐order absorption, and no effect of demographic factors. Multiple‐dose PK of govorestat was linear in the 0.5‐40 mg/kg range, and CSF levels increased dose dependently. Elimination half‐life was ∼10 h. PK/PD modeling supported once‐daily dosing. Change from baseline in galactitol was −15% ± 9% with placebo and −19% ± 10%, −46% ± 4%, and −51% ± 5% with govorestat 5, 20, and 40 mg/kg, respectively, thus was similar for 20 and 40 mg/kg. Govorestat did not affect galactose or Gal‐1p levels. In conclusion, govorestat displayed a favorable safety, PK, and PD profile in humans, and reduced galactitol levels in the same magnitude (∼50%) as in a rat model of CG that demonstrated an efficacy benefit on neurological, behavioral, and ocular outcomes. [ABSTRACT FROM AUTHOR]

Inadequate levels of monounsaturated (MUFA) and polyunsaturated (PUFA) fatty acids have been reported as a risk factor for neurodegenerative diseases, including Alzheimer's disease. Avocado oil (Persea americana Mill., Laucaceae) represents a source of bioactive compounds with a relative abundance of omega-3, omega-6 polyunsaturated, and omega-9 monounsaturated fatty acids. The present study investigated the effects of extra-virgin avocado oil on the D-galactose-induced Alzheimer's disease model in ovariectomized Wistar rats. The cognitive dysfunctions were induced by D-galactose administration (150 mg.kg−1 i.p) and/or ovariectomy in 54 female rats for 70 days. The rats were concomitantly treated with extra-virgin avocado oil (0.25, 0.5, and 1 mL.kg−1, p.o), donepezil (1 mg.kg−1, p.o), and estradiol valerate (1 mg.kg−1, p.o). Memory disorders were evaluated using the Object Recognition, Y-Maze, and Morris water maze tests. Some biochemical and histological parameters regarding memory function were evaluated on hippocampus homogenate and tissue. D-galactose administration and ovariectomy significantly induced learning and memory impairments, decreased relative hippocampal weight (p < 0.001), the levels of acetylcholine (p < 0.001), glutamate (p < 0.001), reduced glutathione (p < 0.001), catalase (p < 0.05), and superoxide dismutase (p < 0.001) activities, and an increase (p < 0.001) in the levels of methylglyoxal, malondialdehyde, and nitrites. The treatment with extra-virgin avocado oil at all tested doses reversed or prevented the negative effects induced by ovariectomy and/or by D-galactose. Taken together, these results suggest that avocado oil possesses neuroprotective properties and can be consumed or supplemented to prevent the onset of Alzheimer's disease. [ABSTRACT FROM AUTHOR]

AbstractPrangos uechtritzii Boiss & Hausskn stands out for its rich bioactive constituents including prantschimgin (PRA), imperatorin (IMP), suberosin (SUB), adicardin (ADI), and oxypeucedanin hydrate (OPH) in the Apiaceae family. Although these molecules contribute to several biological activities, their mitochondrial toxicity were not illuminated in depth with the appropriate in vitro and in silico models. Cell viability studies investigated the cytotoxic activities of molecules in HepG2 cells by replacing glucose with galactose due to Warburg effects. Mitochondrial toxicity (mitotoxicity) parameters such as cellular adenosine triphosphate (ATP) and mitochondrial membrane potential (MMP) levels were assessed with cytotoxic concentrations of selected molecules. Molecular docking and dynamics studies were also conducted against mitochondrial electron transport chain (ETC) complexes (I-V) with selected compounds. In vitro results showed that PRA, SUB, and IMP reduced cell viability more in galactose media compared to high glucose media in a dose-dependent manner. PRA, IMP, and SUB decreased ATP levels and MMP, especially in the galactose medium. The in silico study revealed that PRA, IMP, and SUB might bind to complexes I-V at different levels. The docking study demonstrated that PRA had the highest binding potential with the complexes, higher than the standard ligands in some cases. The molecular dynamics (MD) simulation study showed that PRA formed stable complexes with complexes II, III, and IV. In addition, PRA was anticipated to remain inside the binding site of complex II most stably during the 230 ns simulation period. Our study suggests that PRA, IMP, and SUB exhibit mitotoxicity. [ABSTRACT FROM AUTHOR]

Background: Exanthematous drug eruption and infectious mononucleosis (IM) are both exanthematous diseases. Current research on exanthematous drug eruption and IM mainly targets identifying these disorders, the resulting differences at the metabolism level have not yet been systematically analyzed. Materials and methods: A total of 30 cases of exanthematous drug eruption and IM, 10 patients without exanthema and 10 healthy volunteers were enrolled, 3 mL of fasting venous blood was collected, the serum metabolite content was detected by gas chromatography‐mass spectrometry metabolomics. Results: A total of 165 metabolites were identified, exhibiting significant differences in plasma metabolic trends between exanthematous drug eruption and IM, and pinpointed 28 potential biomarkers. Notable changes were seen in the metabolic activities of the pentose phosphate pathway (PPP), tricarboxylic acid cycle (TCA‐cycle), and galactose metabolism, characterized by increased levels of gluconate, gluconolactone, glucose, galactaric acid, and mannose, along with decreased amounts of pyruvic acid, succinic acid, malic acid, and glycerol, indicating an impairment in the exanthematous drug eruption group's capacity to endure oxidative stress and regulate energy metabolism. In contrast to its medication without rash counterpart, the exanthematous drug eruption group's plasma displayed distinct metabolic routes, predominantly in the processing of arginine and proline, along with the TCA. This resulted in a marked reduction in urea levels and a rise in pyruvate, citrate, and ornithine, indicating hypoxic stress as the primary cause of these rashes. In contrast to the healthy control group, the IM group showed 26 potential biomarkers, marked by increased levels of ketoglutaric acid, malic acid, pyruvic acid, and oxoglutaric acid, and reduced amounts of glutamine, galacturonic acid, arachidonic acid, trimethylphosphonic acid ester, gluconolactone, and indole acetic acid. Mainly, the metabolic pathways included the TCA, breaking down alanine, aspartate and glutamate metabolism, and the processing of D‐glutamine and D‐glutamate metabolism, underscoring the body's crucial role in generating energy and inflammatory agents through the citric acid cycle. Conclusions: The comparison of serum metabolomic features of exanthematous drug eruptions and IM outlines a unique pattern closely related to the differences in the pathogenesis of these two exanthematous diseases. [ABSTRACT FROM AUTHOR]

Cataracts are a disease that reduces vision due to opacity formation of the lens. Diabetic cataracts occur at young age and progress relatively quickly, so the development of effective treatment has been awaited. Several studies have shown that pyruvate inhibits oxidative stress and glycation of lens proteins, which contribute to onset of diabetic cataracts. However, detailed molecular mechanisms have not been revealed. In this study, we attempted to reduce galactose‐induced opacity by pyruvate with rat ex vivo model. Rat lenses were extracted and cultured in galactose‐containing medium to induce lens opacity. After opacity had developed, continued culturing with pyruvate in the medium resulted in a reduction of lens opacity. Subsequently, we conducted microarray analysis to investigate the genes that contribute to the therapeutic effect. We performed quantitative expression measurements using RT‐qPCR for extracted genes that were upregulated in cataract‐induced lenses and downregulated in pyruvate‐treated lenses, resulting in the identification of 34 candidate genes. Functional analysis using the STRING database suggests that metallothionein‐related factors (Mt1a, Mt1m, and Mt2A) and epithelial‐mesenchymal transition‐related factors (Acta2, Anxa1, Cd81, Mki67, Timp1, and Tyms) contribute to the therapeutic effect of cataracts. [ABSTRACT FROM AUTHOR]

To understand the adaptation mechanism of vetiver under copper (Cu) stress, the effects of 5 concentrations of Cu (0, 40, 60, 120, and 150 mg/L CuSO4) on the morphological and physiological responses of vetiver roots were studied. The differential protein expression in vetiver roots treated with Cu-free (CK) and 60 mg/L CuSO4 was analyzed by 4D label-free quantitative proteomics technology. The results showed that the root length, root volume, and root tip number of vetiver initially increased and then decreased with increasing Cu concentration. The maximum values of root length and root volume appeared at 60 mg/L, which increased by 13.91% and 85.16%, respectively, compared with CK; copper ions (Cu2+) and malondialdehyde (MDA) contents increased with increasing Cu concentration and increased by 1626.40% ~ 2197.46% and 3.61% ~ 76.07% compared with CK, respectively; superoxide dismutase (SOD) increased first and then decreased and increased by 126.52% at 60 mg/L compared with CK; soluble protein reduced by 11.76% at 60 mg/L compared with CK (P < 0.05). A total of 1676 differentially expressed proteins were identified in vetiver roots after Cu stress, of which 765 were upregulated and 911 were downregulated. The differentially expressed proteins mainly occur in subcellular structures such as the cytoplasm, nucleus, and tonoplast; are enriched in amino sugar and nucleotide sugar metabolism, phenylpropanoid biosynthesis, cyanoamino acid metabolism, galactose metabolism, steroid biosynthesis, and other pathways; are involved in hydrolase, internal transferases, pyruvate carboxykinase, etc.; and identified the c-terminal domains of cupin, serine carboxypeptidase, peroxidase, and α-l-arabinofuranosidase. The key proteins, including USP, FRK2, HXK3, RFS6, EXPB, OMT2, AS, SODA3, Pod, ZB8, and PRX7, were significantly expressed and are involved in vetiver's regulation of energy metabolism, root morphological growth, and oxidative stress to increase the tolerance of plants to Cu stress. This study further elaborated on the adaptive mechanisms of vetiver under Cu stress and provided a theoretical basis for the ecological restoration of tailings vegetation. [ABSTRACT FROM AUTHOR]

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Postharvest litchi is susceptible to browning that limits the development of litchi industry. Hydrogen sulfide (H2S) is an important bioactive molecule that can regulate many physiological processes. This study examined the effects of exogenous H2S on pericarp browning and related physiological mechanisms in postharvest litchi. The results exhibited that exogenous H2S treatment delayed the browning of litchi pericarp and reduced the damage to cell membrane integrity during storage. This treatment inhibited the energy losses of litchi fruit by increasing the activities of H+-ATPase, Ca2+- ATPase, cytochrome C oxidase (CCO) and succinate dehydrogenase (SDH) and regulating the expression of energy metabolism-related genes, including LcAtpB, LcSnRK2, LcAAC1, LcAOX1 and LcUCP1. In addition, H2S treatment increased the levels of fructose, glucose, sucrose, inositol, galactose and sorbose in litchi fruit, and promoted sucrose synthesis by regulating the activities of sucrose phosphate synthase (SPS), sucrose synthase (SS), acid invertase (AI) and neutral invertase (NI). Based on the current findings, we suggest that exogenous H2S enhances the energy supply and antioxidant activity of litchi by modulating energy and sugar metabolism, thereby inhibiting fruit browning and senescence. These results indicated that H2S treatment is an effective approach to maintaining the quality of litchi fruit and extending its shelf life. [ABSTRACT FROM AUTHOR]

Background: Premature ovarian insufficiency (POI) is a condition characterized by a substantial decline or loss of ovarian function in women before the age of 40. However, the pathogenesis of POI remains to be further elucidated, and specific targeted drugs which could delay or reverse ovarian reserve decline are urgently needed. Abnormal DNA damage repair (DDR) and cell senescence in granulosa cells are pathogenic mechanisms of POI. Ubiquitin-specific protease 14 (USP14) is a key enzyme that regulates the deubiquitylation of DDR-related proteins, but whether USP14 participates in the pathogenesis of POI remains unclear. Methods: We measured USP14 mRNA expression in granulosa cells from biochemical POI (bPOI) patients. In KGN cells, we used IU1 and siRNA-USP14 to specifically inhibit USP14 and constructed a cell line stably overexpressing USP14 to examine its effects on DDR function and cellular senescence in granulosa cells. Next, we explored the therapeutic potential of IU1 in POI mouse models induced by D-galactose. Results: USP14 expression in the granulosa cells of bPOI patients was significantly upregulated. In KGN cells, IU1 treatment and siUSP14 transfection decreased etoposide-induced DNA damage levels, promoted DDR function, and inhibited cell senescence. USP14 overexpression increased DNA damage, impaired DDR function, and promoted cell senescence. Moreover, IU1 treatment and siUSP14 transfection increased nonhomologous end joining (NHEJ), upregulated RNF168, Ku70, and DDB1, and increased ubiquitinated DDB1 levels in KGN cells. Conversely, USP14 overexpression had the opposite effects. Intraperitoneal IU1 injection alleviated etoposide-induced DNA damage in granulosa cells, ameliorated the D-galactose-induced POI phenotype, promoted DDR, and inhibited cell senescence in ovarian granulosa cells in vivo. Conclusions: Upregulated USP14 in ovarian granulosa cells may play a role in POI pathogenesis, and targeting USP14 may be a potential POI treatment strategy. Our study provides new insights into the pathogenesis of POI and a novel POI treatment strategy. [ABSTRACT FROM AUTHOR]

Advances in modern medicine have extended human life expectancy, leading to a world with a gradually aging society. Aging refers to a natural decline in the physiological functions of a species over time, such as reduced pain sensitivity and reaction speed. Healthy-level physiological pain serves as a warning signal to the body, helping to avoid noxious stimuli. Physiological pain sensitivity gradually decreases in the elderly, increasing the risk of injury. Therefore, geriatric health care receives growing attention, potentially improving the health status and life quality of the elderly, further reducing medical burden. Health food is a geriatric healthcare choice for the elderly with Ganoderma tsuage (GT), a Reishi type, as the main product in the market. GT contains polysaccharides, triterpenoids, adenosine, immunoregulatory proteins, and other components, including anticancer, blood sugar regulating, antioxidation, antibacterial, antivirus, and liver and stomach damage protective agents. However, its pain perception-related effects remain elusive. This study thus aimed at addressing whether GT could prevent pain sensitivity reduction in the elderly. We used a galactose-induced animal model for aging to evaluate whether GT could maintain pain sensitivity in aging mice undergoing formalin pain test, hot water test, and tail flexes. Our results demonstrated that GT significantly improved the sensitivity and reaction speed to pain in the hot water, hot plate, and formalin tests compared with the control. Therefore, our animal study positions GT as a promising compound for pain sensitivity maintenance during aging. [ABSTRACT FROM AUTHOR]

Sarcopenia is a significant geriatric syndrome that involves the loss of skeletal muscle mass and strength. Due to its substantial endocrine role, the metabolic microenvironment of skeletal muscle undergoes changes with age. Examining the pathogenesis of sarcopenia through focusing on metabolic dysregulation could offer insights for developing more effective intervention strategies. In this study, we analyzed the transcriptomics data to identify specific genes involved in the regulation of metabolism in skeletal muscle during the development of sarcopenia. Three machine learning algorithms were employed to screen key target genes exhibiting strong correlations with metabolism, which were further validated using RNA‐sequencing data and publicly accessible datasets. Among them, the metabolic enzyme nicotinamide N‐methyltransferase (NNMT) was elevated in sarcopenia, and predicted sarcopenia with an area under the curve exceeding 0.7, suggesting it as a potential therapeutic target for sarcopenia. As expected, inhibition of NNMT improved the grip strength in aging mice and alleviated age‐related decline in the mass index of the quadriceps femoris muscles and whole‐body lean mass index. Additionally, the NNMTi treatment increased the levels of nicotinamide adenine dinucleotide (NAD+) content, as well as PGC1α and p‐AMPK expression in the muscles of both the D‐galactose‐treated mouse model and naturally aging mouse model. Overall, this work demonstrates NNMT as a promising target for preventing age‐related decline in muscle mass and strength. [ABSTRACT FROM AUTHOR]

Although PDO Pecorino Sardo is one of the oldest traditional cheeses of Sardinia, Italy, data on its nutritional properties and food safety are lacking. In particular, significant amounts of lactose and galactose may be a health concern for consumers. The primary objective of this study is to quantify, using a validated GC-FID method, the residual lactose and galactose content in "maturo" (i.e., ripened for at least two months) Protected Denomination of Origin (PDO) Pecorino Sardo cheese. A statistically representative sampling from seven dairies distributed throughout Sardinia has been selected for this aim. In addition to lactose and galactose, two of their metabolites (i.e., glucose and tagatose, respectively) and a bioactive polyol like myo-inositol were quantified. The concentration of lactose (mean 26 mg kg−1, range 4–90 mg kg−1) was below the strictest limit set in the European Union (i.e., 100 mg kg−1), while the galactose content was found to be in an amount (mean: 76 mg kg−1, range: 10–200 mg kg−1) that even patients afflicted with severe galactosemia, albeit with some circumspection, could consume this cheese. Ripening (two to four months) had no significant effect on the amount of all analytes, while a slight decrease in galactose levels was observed during the manufacturing season. Finally, the amounts of glucose, tagatose, and myo-inositol are constant in the range of a few tens of mg kg−1. [ABSTRACT FROM AUTHOR]

The current study was designed to compare in vivo efficacy between beeswax alcohol (BWA) and coenzyme Q10 (CoQ10) to treat fatty liver changes, oxidative stress, and damages in major organs of zebrafish by 12 weeks with high-cholesterol (HC) and galactose (Gal) supplementation. At week 12, the HC control and HC+Gal control groups showed 96% and 92% survivability, respectively, while co-supplementation of the 0.5% BWA and 1.0% BWA groups exhibited 96% and 100% survivability. However, co-supplementation of the 0.5% CoQ10 and 1.0% CoQ10 groups revealed the lowest survivability, around 92% and 89%, respectively. The 0.5% BWA and 1.0% BWA groups showed 21% (p < 0.001) and 41% (p < 0.001), respectively, lower total cholesterol (TC) than the HC+Gal control, while the 1.0% CoQ10 group showed only 15% lower TC than the control. Interestingly, the 0.5% BWA and 1.0% BWA groups showed 22% (p < 0.001) and 38% (p < 0.001), respectively, lower triglyceride (TG) than the HC+Gal control. However, both the 0.5% CoQ10 and 1.0% CoQ10 groups showed similar TG levels as the control, suggesting that CoQ10 supplementation had no effect on lowering serum TG. The 1.0% BWA group showed the highest plasma HDL-C and HDL-C/TC (%) up to 3.2-fold and 5.5-fold, respectively, higher than those of the HC+Gal control, while the 1.0% CoQ10 group showed 2.4-fold and 2.8-fold higher plasma HDL-C and HDL-C/TC (%), respectively, than the control. The plasma aspartate transaminase (AST) and alanine transaminase (ALT) levels were lowest in the 1.0% BWA group, 51% and 72%, respectively, lower than HC+Gal control, suggesting the lowest extent of hepatic damage. In hepatic tissue, neutrophil infiltration and interleukin (IL)-6 production were the lowest in the 1.0% BWA group, around 67% and 85%, respectively, lower than the HC+Gal control. Fatty liver change, cellular apoptosis, and cell senescence in hepatic tissue were remarkably lowered in the 1.0% BWA group, while the CoQ10 group showed much less effect than the BWA group. In kidney, ovary, and testis tissue, the 1.0% BWA group showed the lowest production of reactive oxygen species, the extent of cellular senescence, and cellular apoptosis with the healthiest cell morphology. In conclusion, supplementation of BWA remarkably protected the liver, kidney, ovary, and testis from oxidative damage by cholesterol and galactose consumption, with the least serum AST and ALT levels, inflammatory parameters, and senescence markers. [ABSTRACT FROM AUTHOR]

Steroidal alkaloids are FDA-approved drugs (e.g. , Zytiga) and promising drug candidates/leads (e.g. , cyclopamine); yet many of the ≥697 known steroidal alkaloid natural products remain underutilized as drugs because it can be challenging to scale their biosynthesis in their producing organisms. Cyclopamine is a steroidal alkaloid produced by corn lily (Veratrum spp.) plants, and it is an inhibitor of the Hedgehog (Hh) signaling pathway. Therefore, cyclopamine is an important drug candidate/lead to treat human diseases that are associated with dysregulated Hh signaling, such as basal cell carcinoma and acute myeloid leukemia. Cyclopamine and its semi-synthetic derivatives have been studied in (pre)clinical trials as Hh inhibitor-based drugs. However, challenges in scaling the production of cyclopamine have slowed efforts to improve its efficacy and safety profile through (bio)synthetic derivatization, often limiting drug development to synthetic analogs of cyclopamine such as the FDA-approved drugs Odomzo, Daurismo, and Erivedge. If a platform for the scalable and sustainable production of cyclopamine were established, then its (bio)synthetic derivatization, clinical development, and, ultimately, widespread distribution could be accelerated. Ongoing efforts to achieve this goal include the biosynthesis of cyclopamine in Veratrum plant cell culture and the semi-/total chemical synthesis of cyclopamine. Herein, this work advances efforts towards a promising future approach: the biosynthesis of cyclopamine in engineered microorganisms. We completed the heterologous microbial production of verazine (biosynthetic precursor to cyclopamine) from simple sugars (i.e. , glucose and galactose) in engineered Saccharomyces cerevisiae (S. cerevisiae) through the inducible upregulation of the native yeast mevalonate and lanosterol biosynthetic pathways, diversion of biosynthetic flux from ergosterol (i.e. , native sterol in S. cerevisiae) to cholesterol (i.e. , biosynthetic precursor to verazine), and expression of a refactored five-step verazine biosynthetic pathway. The engineered S. cerevisiae strain that produced verazine contains eight heterologous enzymes sourced from seven different species. Importantly, S. cerevisiae -produced verazine was indistinguishable via liquid chromatography-mass spectrometry from both a commercial standard (Veratrum spp. plant-produced) and Nicotiana benthamiana -produced verazine. To the best of our knowledge, this is the first report describing the heterologous production of a steroidal alkaloid in an engineered yeast. Verazine production was ultimately increased through design-build-test-learn cycles to a final titer of 83 ± 3 μg/L (4.1 ± 0.1 μg/g DCW). Together, this research lays the groundwork for future microbial biosynthesis of cyclopamine, (bio)synthetic derivatives of cyclopamine, and other steroidal alkaloid natural products. [Display omitted] • Biosynthesis of verazine (83 μg/L) from glucose and galactose in S. cerevisiae. • First heterologous steroidal alkaloid produced in an engineered yeast. • Biosynthesis of verazine via transient expression of its pathway in N. benthamiana. • S. cerevisiae , N. benthamiana , and commercial verazine were indistinguishable. [ABSTRACT FROM AUTHOR]

Objective(s): To study the anti-aging effect of (-)-α-bisabolol ((-)-α-bis) on the skin and preliminarily clarify its mechanism. Materials and Methods: Human skin fibroblasts (HSF) were induced senescence by D-Galactose. Senescence β-galactosidase staining was utilized to evaluate the senescence of HSF. TNF-α, IL6, IL-8, IL-1β, CCL-2, CCL-5, and MMP-9 in senescence-as-sociated secretory phenotype (SASP) were detected by RT-qPCR. Meanwhile, aged BALB/c mice were applied topically with 0.5% and 2%(-)-α-bis gel for 30 days continuously to evaluate anti-aging parameters on the skin such as surface measurement, the Trans Epidermal Water Loss (TEWL), and skin barrier index of dorsal skin. Then, HE staining, Masson staining, and IHC were applied to measure epidermal thickness, collagen fiber content in the dermis, and content of dermal collagen I, respectively. Last, SOD, MDA, and HYP contents of the back skin tissue of mice were also detected. Results: (-)-α-Bis reduced the expression of senescence-associated β-galactosidase (SA-β-gal) and expression levels of SASP in HSF cells stimulated by D-Gal (P<0.05). Mice aged 9 months were applied locally with (-)-α-bis gel to improve skin aging, the TEWL and skin barrier index of dorsal skin, and ameliorate the epidermal thickness and contents of dermal collagen fibers and collagen I (P<0.05). Furthermore, (-)-α-bis up-regulated the mRNA expression levels of elastin and collagen III effectively (P<0.05). Conclusion: (-)-α-Bis can delay the senescence of HSF cells by reducing the expression of SA-β-gal and SASP factors in vitro. Improved skin barrier function as well as SASP is responsible for the delay of skin aging in vivo. [ABSTRACT FROM AUTHOR]

As a protein extracted from soybeans, soy protein isolate (SPI) may undergo the Maillard reaction (MR) with co-existing saccharides during the processing of soy-containing foods, potentially altering its structural and functional properties. This work aimed to investigate the effect of mono- and polysaccharides on the structure and functional properties of SPI during MR. The study found that compared to oat β-glucan, the reaction rate between SPI and D-galactose was faster, leading to a higher degree of glycosylation in the SPI–galactose conjugate. D-galactose and oat β-glucan showed different influences on the secondary structure of SPI and the microenvironment of its hydrophobic amino acids. These structural variations subsequently impact a variety of the properties of the SPI conjugates. The SPI–galactose conjugate exhibited superior solubility, surface hydrophobicity, and viscosity. Meanwhile, the SPI–galactose conjugate possessed better emulsifying stability, capability to produce foam, and stability of foam than the SPI–β-glucan conjugate. Interestingly, the SPI–β-glucan conjugate, despite its lower viscosity, showed stronger hypoglycemic activity, potentially due to the inherent activity of oat β-glucan. The SPI–galactose conjugate exhibited superior antioxidant properties due to its higher content of hydroxyl groups on its molecules. These results showed that the type of saccharides had significant influences on the SPI during MR. [ABSTRACT FROM AUTHOR]

Aalok Basu,1 Arunsajee Sae-be,1 Thanaphon Namporn,1 Orasa Suriyaphan,2 Pongtip Sithisarn,3 Jiraporn Leanpolchareanchai,1 Piyaporn Plommaithong,1 Apichat Chatsukit,1 Khanit Sa-ngiamsuntorn,4 Parichart Naruphontjirakul,5 Pakatip Ruenraroengsak1,6 1Department of Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand; 2Department of Food Chemistry, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand; 3Department of Pharmacognosy, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand; 4Department of Biochemistry, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand; 5Biological Engineering Program, Faculty of Engineering, King Mongkut’s University of Technology Thonburi, Bangkok, Thailand; 6Centre of Molecular Targeting and Integrated Drug Development, Faculty of Pharmacy, Mahidol University, Bangkok, ThailandCorrespondence: Pakatip Ruenraroengsak, Division of Pharmaceutical Technology, Department of Pharmacy, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Road, Rajathevi, Bangkok, 10400, Thailand, Email pakatip.rue@mahidol.ac.thBackground: Sorafenib-resistant (SR) hepatocellular carcinoma (HCC) is a current serious problem in liver cancer treatment. Numerous phytochemicals derived from plants exhibit anticancer activity but have never been tested against drug-resistant cells.Methods: Avocado seed extract (APE) isolated by maceration was analysed for its phytochemical composition and anticancer activity. Novel design charge-switchable pH-responsive nanocarriers of aminated mesoporous silica nanoparticles with conjugated galactose (GMSN) were synthesised for delivering APE and their physicochemical properties were characterized. The drug loading efficiency (%LE) and entrapment efficiency (%EE) were evaluated. Anticancer activity of APE loaded GMSN was measured against HCC (HepG2, Huh-7) and SR-HCC (SR-HepG2).Results: Anticancer activity of APE against non-resistant HepG2 (IC50 50.9 ± 0.83 μg mL− 1), Huh-7 (IC50 42.41 ± 1.88 μg mL− 1), and SR-HepG2 (IC50 62.58 ± 2.29 μg mL− 1) cells was confirmed. The APE loaded GMSN had a diameter of 131.41 ± 14.41 nm with 41.08 ± 2.09%LE and 44.96 ± 2.26%EE. Galactose functionalization (55%) did not perturb the original mesoporous structure. The GMSN imparted positive surface charges, 10.3 ± 0.61mV at acidic medium pH 5.5 along with rapid release of APE 45% in 2 h. The GMSN boosted cellular uptake by HepG2 and SR-HepG2 cells, whereas the amine functionalized facilitated their endosomal escape. Their anticancer activity was demonstrated in non-resistant HCC and SR-HCC cells with IC50 values at 30.73 ± 3.14 (HepG2), 21.86 ± 0.83 (Huh-7), 35.64 ± 1.34 (SR-HepG2) μg mL− 1, respectively, in comparison to the control and non-encapsulated APE.Conclusion: APE loaded GMSN is highly effective against both non-resistant HCC and SR-HCC and warrants further in vivo investigation. Keywords: liver cancer, hepatocellular carcinoma, galactose, sorafenib drug resistance, avocado, mesoporous silica nanoparticles

Sodium-dependent glucose transporters (SGLTs) couple a downhill Na+ ion gradient to actively transport sugars. Here, we investigate the impact of the membrane potential on vSGLT structure and function using sugar uptake assays, double electron-electron resonance (DEER), electrostatic calculations, and kinetic modeling. Negative membrane potentials, as present in all cell types, shift the conformational equilibrium of vSGLT towards an outward-facing conformation, leading to increased sugar transport rates. Electrostatic calculations identify gating charge residues responsible for this conformational shift that when mutated reduce galactose transport and eliminate the response of vSGLT to potential. Based on these findings, we propose a comprehensive framework for sugar transport via vSGLT, where the cellular membrane potential facilitates resetting of the transporter after cargo release. This framework holds significance not only for SGLTs but also for other transporters and channels.

• 4-NP induced reproduction, development, and locomotion toxicity in C. elegans. • 4-NP significantly altered amino acids and energy metabolism of C. elegans. • Estrogen signaling pathway contributed to the toxicity of 4-NP. • Brood size and body length was mediated by glycerol-3P and glucose-1,6P2, respectively. 4-Nitrophenol (4-NP), as a toxic and refractory pollutant, has generated significant concern due to its adverse effects. However, the potential toxic effects and mechanism remained unclear. In this study, the reproduction, development, locomotion and reactive oxygen species (ROS) production of Caenorhabditis elegans were investigated to evaluate the 4-NP toxicity. We used metabolomics to assess the potential damage mechanisms. The role of metabolites in mediating the relationship between 4-NP and phenotypes was examined by correlation and mediation analysis. 4-NP (8 ng/L and 8 µg/L) caused significant reduction of brood size, ovulation rate, total germ cells numbers, head thrashes and body bends, and an increase in ROS. However, the oosperm numbers in uterus, body length and body width were decreased in 8 µg/L. Moreover, 36 differential metabolites were enriched in the significant metabolic pathways, including lysine biosynthesis, β-alanine metabolism, tryptophan metabolism, pentose phosphate pathway, pentose and glucuronate interconversions, amino sugar and nucleotide sugar metabolism, starch and sucrose metabolism, galactose metabolism, propanoate metabolism, glycerolipid metabolism, and estrogen signaling pathway. The mechanism of 4-NP toxicity was that oxidative stress caused by the perturbation of amino acid, which had effects on energy metabolism through disturbing carbohydrate and lipid metabolism, and finally affected the estrogen signaling pathway to exert toxic effects. Moreover, correlation and mediation analysis showed glycerol-3P, glucosamine-6P, glucosamine-1P, UDP-galactose, L-aspartic acid, and uracil were potential markers for the reproduction and glucose-1,6P2 for developmental toxicity. The results provided insight into the pathways involved in the toxic effects caused by 4-NP and developed potential biomarkers to evaluate 4-NP toxicity. [Display omitted] [ABSTRACT FROM AUTHOR]

The compounds 2′,3′,4′,6′-tetra-O-acetyl-β-D-glucopyranosyl N′-cyano-N-phenylcarbamimidothioate (C22H25N3O9S, 5a), 2′,3′,4′,6′-tetra-O-acetyl-β-D-galactopyranosyl N′-cyano-N-phenylcarbamimidothioate, (C22H25N3O9S, 5b), 2′,3′,4′,6′-tetra-O-acetyl-β-D-galactopyranosyl N′-cyano-N-methylcarbamimidothioate (C17H23N3O9S, 5c), and 2′,3′,4′,6′-tetra-O-acetyl-β-D-galactopyranosyl N′-cyano-N-p-tolylcarbamimidothioate (C23H27N3O9S, 5d) all crystallize in P212121 with Z = 4. For all four structures, the configuration across the central (formal) C=N(CN) double bond of the carbamimidothioate group is Z. The torsion angles C5—O1—C1—S (standard sugar numbering) are all close to 180°, confirming the β position of the substituent. Compound 5b involves an intramolecular hydrogen bond N—H...O1; in 5c this contact is the weaker branch of a three-centre interaction, whereas in 5a and 5d the H...O distances are much longer and do not represent significant interactions. The C—N bond lengths at the central carbon atom of the carbamimidothioate group are almost equal. All C—O—C=O torsion angles of the acetyl groups correspond to a synperiplanar geometry, but otherwise all four molecules display a high degree of conformational flexibility, with many widely differing torsion angles for equivalent groups. In the crystal packing, 5a, 5c and 5d form layer structures involving the classical hydrogen bond N—H...Ncyano and a variety of ‘weak’ hydrogen bonds C—H...O or C—H...S. The packing of 5b is almost featureless and involves a large number of borderline ‘weak’ hydrogen bonds. In an appendix, a potted history of wavelength preferences for structure determination is presented and it is recommended that, even for small organic crystals in non-centrosymmetric space groups, the use of Mo radiation should be considered.

Abstract Background The development of alternative carbon sources is important for reducing the cost of heterotrophic microalgae cultivation. Among cheap feedstocks, galactose is one of the most abundant sugars and can be easily obtained from many natural biomasses. However, it is generally difficult to be utilized by microalgae. In addition, the mechanism of its low utilization efficiency in heterotrophic cultivation is still unknown. Results Among seven tested carbon sources, only glucose and acetate could be efficiently utilized by C. sorokiniana in heterotrophic cultivation while there were no apparent signs of utilization of other carbohydrates, including galactose, in regular heterotrophic cultivation. However, galactose could be utilized in cultures with high inoculation sizes. This confirmed that C. sorokiniana has a complete pathway for transporting and assimilating galactose under dark conditions, but the rate of galactose utilization is quite low. In addition, the galactose utilization was greatly enhanced in mixotrophic cultures, which indicated that galactose utilization could be enhanced by additional pathways that can enhance cell growth. Based on above results, a mixed carbon source culture strategy was proposed to improve the utilization rate of galactose, and a significant synergistic effect on cell growth was achieved in cultures using a mixture of galactose and acetate. Conclusions This study indicated that the galactose metabolism pathway may not be inherently deficient in Chlorophyta. However, its utilization rate was too low to be detected in regular heterotrophic cultivation. Mixed carbon source culture strategy was confirmed effective to improve the utilization rate of galactose. This study contributes to a deeper understanding of the utilization ability of difficultly utilized substrates in the heterotrophic cultivation of microalgae, which is of great significance for reducing the cost of heterotrophic cultivation of microalgae.

Objective To investigate the effect and mechanism of galactose on the expression of sialic-acid-binding immunoglobulin-like lectin 9 (Siglec-9) glycan ligand on human primarily cultured pulmonary artery endothelial cells. Methods The expression of Siglec-9 glycan ligand on the endothelial cell surface and the type of glycosidic bonds at the end of this ligand were identified by flow cytometry. Endothelial cells were treated with L-glucose, glucose, N-acetylglucose, mannose, N-acetylmannose, galactose, sialic acid and sucrose for 48 h, and the expression of Siglec-9 glycan ligand in endothelial cells was detected by flow cytometry. The endothelial cells were divided into the control group and the galactose group (n=3). Western blotting, flow cytometry, and immunofluorescence assay were employed to investigate the impact of galactose on the Siglec-9 glycan ligand in endothelial cells. After α2-3, 6, 8 sialidase treatment for endothelial cells, Western blotting was used to detected the effect of galactose on the recovery time of Siglec-9 glycan ligand in endothelial cells. Endothelial cells were treated with galactose, Western blotting was used to detect and analyze the expression levels of intracellular sialic acid synthetases (GNE, NANS, NANP, CMAS, NPL and ST3Gals), and the mRNA expression levels of the relevant proteins were verified by RT-qPCR. Western blotting was used to detect the changes of Siglec-9 glycan ligand level in endothelial cells after siRNA knockdown in NANP, CMAS and ST3Gal-Ⅲ. The effect of the increase of Siglec-9 ligand on apoptosis and phagocytosis of macrophages was analyzed by macrophage co-culture experiments. Results Endothelial cells showed the expression of Siglec-9 ligand and the ligand was a sialic acid glycoprotein linked to α 2-3 sialic acid at the end. Compared with the control group, the expression level of this ligand on endothelial cells in the galactose group was increased significantly (P < 0.01), but the addition of galactose had no effect on the self-recovery time of the ligand. Compared with the control group, the expression levels of NANP, CMAS and ST3Gal-Ⅲ in endothelial cells treated with galactose were increased (P < 0.05, P < 0.01), respectively. The results of RT-qPCR verification were consistent with those results. After the expression of NANP, CMAS or ST3Gal-Ⅲ was inhibited, the level of Siglec-9 glycan ligand in endothelial cells was decreased (P < 0.01). In the co-culture experiment, compared with the untreated group, galactose-treated endothelial cells promoted the apoptosis of macrophages (P < 0.01) and reduced their phagocytosis (P < 0.05). Conclusion Galactose up-regulates the level of Siglec-9 glycan ligand on endothelial cells through the NANP-CMAS-ST3Gal-Ⅲ pathway, thereby inhibiting the activity of macrophages.

Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Recent studies provide increasing evidence for a relevant role of lifestyle factors including diet in the pathogenesis of neuroinflammatory diseases such as multiple sclerosis (MS). While the intake of saturated fatty acids and elevated salt worsen the disease outcome in the experimental model of MS by enhanced inflammatory but diminished regulatory immunological processes, sugars as additional prominent components in our daily diet have only scarcely been investigated so far. Apart from glucose and fructose, galactose is a common sugar in the so-called Western diet. Methods: We investigated the effect of a galactose-rich diet during neuroinflammation using myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis (MOG-EAE) as a model disease. We investigated peripheral immune reactions and inflammatory infiltration by ex vivo flow cytometry analysis and performed histological staining of the spinal cord to analyze effects of galactose in the central nervous system (CNS). We analyzed the formation of advanced glycation end products (AGEs) by fluorescence measurements and investigated galactose as well as galactoseinduced AGEs in oligodendroglial cell cultures and induced pluripotent stem cellderived primary neurons (iPNs). Results: Young mice fed a galactose-rich diet displayed exacerbated disease symptoms in the acute phase of EAE as well as impaired recovery in the chronic phase. Galactose did not affect peripheral immune reactions or inflammatory infiltration into the CNS, but resulted in increased demyelination, oligodendrocyte loss and enhanced neuro-axonal damage. Ex vivo analysis revealed an increased apoptosis of oligodendrocytes isolated from mice adapted on a galactose-rich diet. In vitro, treatment of cells with galactose neither impaired the maturation nor survival of oligodendroglial cells or iPNs. However, incubation of proteins with galactose in vitro led to the formation AGEs, that were increased in the spinal cord of EAE-diseased mice fed a galactose-rich diet. In oligodendroglial and neuronal cultures, treatment with galactose-induced AGEs promoted enhanced cell death compared to control treatment. Conclusion: These results imply that galactose-induced oligodendrocyte and myelin damage during neuroinflammation may be mediated by AGEs, thereby identifying galactose and its reactive products as potential dietary risk factors for neuroinflammatory diseases such as MS. [ABSTRACT FROM AUTHOR]

Ageing is the most prominent risk for osteoarthritis (OA) development. This study aimed to investigate the role of phosphoinositide‐specific phospholipase Cγ (PLCγ) 1, previously linked to OA progression, in regulating age‐related changes in articular cartilage and subchondral bone. d‐galactose (d‐Gal) was employed to treat chondrocytes from rats and mice or injected intraperitoneally into C57BL/6 mice. RTCA, qPCR, Western blot and immunohistochemistry assays were used to evaluate cell proliferation, matrix synthesis, senescence genes and senescence‐associated secretory phenotype, along with PLCγ1 expression. Subchondral bone morphology was assessed through micro‐CT. In mice with chondrocyte‐specific Plcg1 deficiency (Plcg1flox/flox; Col2a1‐CreERT), articular cartilage and subchondral bone were examined over different survival periods. Our results showed that d‐Gal induced chondrocyte senescence, expedited articular cartilage ageing and caused subchondral bone abnormalities. In d‐Gal‐induced chondrocytes, diminished PLCγ1 expression was observed, and its further inhibition by U73122 exacerbated chondrocyte senescence. Plcg1flox/flox; Col2a1‐CreERT mice exhibited more pronounced age‐related changes in articular cartilage and subchondral bone compared to Plcg1flox/flox mice. Therefore, not only does d‐Gal induce senescence in chondrocytes and age‐related changes in articular cartilage and subchondral bone, as well as diminished PLCγ1 expression, but PLCγ1 deficiency in chondrocytes may also accelerate age‐related changes in articular cartilage and subchondral bone. PLCγ1 may be a promising therapeutic target for mitigating age‐related changes in joint tissue. [ABSTRACT FROM AUTHOR]

The compounds 2′,3′,4′,6′-tetra-O-acetyl-β-D-gluco­pyranosyl N′-cyano-N-phenyl­carbamimido­thio­ate (C22H25N3O9S, 5a), 2′,3′,4′,6′-tetra-O-acetyl-β-D-galacto­pyranosyl N′-cyano-N-phenyl­carbamimido­thio­ate, (C22H25N3O9S, 5b), 2′,3′,4′,6′-tetra-O-acetyl-β-D-galacto­pyranosyl N′-cyano-N-methyl­carbamimido­thio­ate (C17H23N3O9S, 5c), and 2′,3′,4′,6′-tetra-O-acetyl-β-D-galacto­pyranosyl N′-cyano-N-p-tolyl­carbamimido­thio­ate (C23H27N3O9S, 5d) all crystallize in P212121 with Z = 4. For all four structures, the configuration across the central (formal) C=N(CN) double bond of the carbamimido­thio­ate group is Z. The torsion angles C5—O1—C1—S (standard sugar numbering) are all close to 180°, confirming the β position of the substituent. Compound 5b involves an intra­molecular hydrogen bond N—H⋯O1; in 5c this contact is the weaker branch of a three-centre inter­action, whereas in 5a and 5d the H⋯O distances are much longer and do not represent significant inter­actions. The C—N bond lengths at the central carbon atom of the carbamimido­thio­ate group are almost equal. All C—O—C=O torsion angles of the acetyl groups correspond to a synperiplanar geometry, but otherwise all four mol­ecules display a high degree of conformational flexibility, with many widely differing torsion angles for equivalent groups. In the crystal packing, 5a, 5c and 5d form layer structures involving the classical hydrogen bond N—H⋯Ncyano and a variety of ‘weak’ hydrogen bonds C—H⋯O or C—H⋯S. The packing of 5b is almost featureless and involves a large number of borderline ‘weak’ hydrogen bonds. In an appendix, a potted history of wavelength preferences for structure determination is presented and it is recommended that, even for small organic crystals in non-centrosymmetric space groups, the use of Mo radiation should be considered. [ABSTRACT FROM AUTHOR]

Objective • Exploring the clinical efficacy of neomycin and sakubactria valsartan in the treatment of patients with chronic heart failure (CHF) and atrial fibrillation. This study investigates the potential benefits of combining neomycin with sakubactria valsartan, a medication with a background of demonstrated efficacy in cardiovascular conditions, to address the complex challenges presented by chronic heart failure and atrial fibrillation. Methods • Using a single-center clinical randomized trial, 111 patients with CHF complicated with atrial fibrillation who were treated in the cardiovascular department of Xingtai Third Hospital from June 2019 to March 2021 were randomly divided into two groups. In the control group, 56 patients received treatment with Western Medicine Foundation + Shakubatra valsartan. In the experimental group, consisting of 55 patients, the treatment was identical to the control group, with the additional administration of neomycin.. After 12 weeks of continuous treatment, the echocardiograms, electrocardiogram parameters, and Differences in changes in serum soluble growth stimulating gene 2 protein (sST2) and galactose agglutinin 3 (Gal-3), clinical efficacy, and incidence of adverse reactions. Results • Before treatment, no significant differences existed in LVEF, LVEDV, FS, and SV between the experimental and control groups (P > .05). Post-treatment, both groups exhibited significant improvements in these parameters, with the experimental group showing statistically higher values (P < .05).Similarly, pretreatment comparisons of Pd, sST2, Gal-3, and NT-proBNP revealed no significant differences between the groups (P > .05). After treatment, both groups showed significant reductions, with the experimental group demonstrating lower values (P < .05). Clinical efficacy assessment post-treatment showed significant differences. The experimental group had a basic cure rate of 45.45%, a significant effective rate of 43.64%, and an effective rate of 10.91%, while the control group had rates of 28.57%, 48.21%, and 23.21%, respectively (P < .05).Adverse reactions occurred in 9 and 4 patients in the experimental and control groups, respectively. The severity was not significant, and treatment was uninterrupted (P > 0.05).The treatment improved heart function and reduced atrial fibrillation occurrences, holding clinical significance by potentially enhancing patients’ quality of life and decreasing cardiovascular events. These results highlight the clinical significance of this treatment, which may help improve patients’ quality of life and reduce the occurrence of cardiovascular events. Conclusion • The treatment of patients with CHF combined with atrial fibrillation using neomycin and sakubactria valsartan can more effectively improve their cardiac function and alleviate the condition of atrial fibrillation, which is worthy of clinical promotion and application. In actual clinical practice, physicians and healthcare providers may consider incorporating this treatment into their treatment regimens, especially for patients who need to improve heart function and reduce the risk of atrial fibrillation. Additionally, further research and clinical trials can further validate these findings to ensure their effectiveness and safety. These insights will help the medical community better understand how to apply this treatment to real patients and maximize its clinical effectiveness. [ABSTRACT FROM AUTHOR]

Sialic acid is a unique sugar moiety that resides in the distal and most accessible position of the glycans on mammalian cell surface and extracellular glycoproteins and glycolipids. The potential for sialic acid to obscure underlying structures has long been postulated, but the means by which such structural changes directly affect biological processes continues to be elucidated. Here, we appraise the growing body of literature detailing the importance of sialic acid for the generation, differentiation, function and death of haematopoietic cells. We conclude that sialylation is a critical post‐translational modification utilized in haematopoiesis to meet the dynamic needs of the organism by enforcing rapid changes in availability of lineage‐specific cell types. Though long thought to be generated only cell‐autonomously within the intracellular ER‐Golgi secretory apparatus, emerging data also demonstrate previously unexpected diversity in the mechanisms of sialylation. Emphasis is afforded to the mechanism of extrinsic sialylation, whereby extracellular enzymes remodel cell surface and extracellular glycans, supported by charged sugar donor molecules from activated platelets. [ABSTRACT FROM AUTHOR]

Copyright of Modern Food Science & Technology is the property of Editorial Office of Modern Food Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The study aims to investigate the anti-aging effect of Silybum marianum extract (SME) on skin aging mice induced by D-galactose (D-gal). The mice were divided into 6 groups. The mouse model of skin aging was induced by subcutaneous injection of D-gal (1 g/ (kg·d)) on the back for 6 weeks. Body weight, skin water content, skin oxidative stress index, HYP and HA levels of mice in each group were measured. HE staining was used to evaluate the skin injury. The relative mRNA expressions of collagen I, collagen III, MMP-1 and MMP-3 in skin were detected by RT-qPCR. The relative expressions of Nrf2 (nucleus), Keap1 and HO-1 in skin were detected by Western blotting. The results show that percutaneous application of all three doses of SME increase the skin water content, alleviate the skin injury, increase the levels of SOD, CAT and GSH-Px, decrease the level of MDA, increased the levels of HYP and HA, increase the relative expression of collagen I and collagen III mRNA, decrease the relative expression of MMP-1 and MMP-3 mRNA, increase the relative expression of Nrf2 (nucleus) and HO-1 protein, and decrease the relative expression of Keap1 protein in D-gal-induced aging mice (P<0.05). ML385 inhibits the activation of Nrf2 pathway and weakens the anti-aging effect of SME (P<0.05). This study shows that percutaneous application of SME has a good anti-aging effect on skin aging mice induced by D-gal, and Nrf2 pathway mediates the anti-aging effect of SME. SME may be a kind of anti-aging daily skin care material with high development value. [ABSTRACT FROM AUTHOR]

Increased production of advanced glycation end products (AGEs) among reducing sugars (glucose, fructose, galactose, or ribose) and amino acids/proteins via non-enzymatic Maillard reaction can be found in lifestyle-related disease (LSRD), metabolic syndrome (MetS), and obesity and immune-related diseases. Increased serum levels of AGEs may induce aging, diabetic complications, cardiovascular diseases (CVD), neurodegenerative diseases (NDD), cancer, and inflamm-aging (inflammation with immunosenescence). The Maillard reaction can also occur among reducing sugars and lipoproteins or DNAs to alter their structure and induce immunogenicity/genotoxicity for carcinogenesis. AGEs, as danger-associated molecular pattern molecules (DAMPs), operate via binding to receptor for AGE (RAGE) or other scavenger receptors on cell surface to activate PI3K-Akt-, P38-MAPK-, ERK1/2-JNK-, and MyD88-induced NF-κB signaling pathways to mediate various pathological effects. Recently, the concept of "inflamm-aging" became more defined, and we have unveiled some interesting findings in relation to it. The purpose of the present review is to dissect the potential molecular basis of inflamm-aging in patients with diabetes and immune-mediated diseases caused by different AGEs. [ABSTRACT FROM AUTHOR]

The Neurospora crassa genome has a gene cluster for the synthesis of galactosaminogalactan (GAG). The gene cluster includes the following: (1) UDP-glucose-4-epimerase to convert UDP-glucose and UDP-N-acetylglucosamine to UDP-galactose and UDP-N-acetylgalactosamine (NCU05133), (2) GAG synthase for the synthesis of an acetylated GAG (NCU05132), (3) GAG deacetylase (/NCW-1/NCU05137), (4) GH135-1, a GAG hydrolase with specificity for N-acetylgalactosamine-containing GAG (NCU05135), and (5) GH114-1, a galactosaminidase with specificity for galactosamine-containing GAG (NCU05136). The deacetylase was previously shown to be a major cell wall glycoprotein and given the name of NCW-1 (non-GPI anchored cell wall protein-1). Characterization of the polysaccharides found in the growth medium from the wild type and the GAG synthase mutant demonstrates that there is a major reduction in the levels of polysaccharides containing galactosamine and N-acetylgalactosamine in the mutant growth medium, providing evidence that the synthase is responsible for the production of a GAG. The analysis also indicates that there are other galactose-containing polysaccharides produced by the fungus. Phenotypic characterization of wild-type and mutant isolates showed that deacetylated GAG from the wild type can function as an adhesin to a glass surface and provides the fungal mat with tensile strength, demonstrating that the deacetylated GAG functions as an intercellular adhesive. The acetylated GAG produced by the deacetylase mutant was found to function as an adhesive for chitin, alumina, celite (diatomaceous earth), activated charcoal, and wheat leaf particulates. [ABSTRACT FROM AUTHOR]

Fever is one of the most common clinical conditions and is characterized by pyrogenic infection, malignancy, inflammation, and tissue damage, among others. Ellagic acid (EA) can inhibit the expression of related proteins on the pathway by blocking the nuclear factor kappa-B(NF-κB) signaling pathway, inhibit the levels of pro-inflammatory factors interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α), increase the level of anti-inflammatory factor IL-10, and effectively alleviate inflammatory symptoms. In addition, EA can also reduce the levels of malondialdehyde(MDA) and nitric oxide(NO) in the body, increase the activities of superoxide dismutase (SOD), glutathione (GSH), and catalase(CAT), scavenge oxidative free radicals, inhibit lipid oxidation, and achieve antipyretic and anti-inflammatory effects. The purpose of this study was to establish the relationship between EA and various inflammatory markers, such as TNF-α, IL-6, IL-1β, prostaglandin E2(PGE2), and cyclic adenosine monophosphate(cAMP), and clarify the mechanism of the cyclooxidase-2(COX-2)/NF-κB signaling pathway. Combined with the metabolomics analysis, our study revealed the effects of EA on multiple endogenous biomarkers, reflecting the characteristics of a multi-component, multi-target, and multi-pathway mechanism. Compared to lipopolysaccharide (LPS)- treated animals, subsequent administration of EA significantly lowered the LPS-induced rectal temperature increase (p < 0.05 or p < 0.01), significantly increased serum SOD and GSH levels (p < 0.05 or p < 0.01), and significantly decreased serum MDA, IL-1β, IL-6, and TNF-α levels (p < 0.05 or p < 0.01). In addition, compared to LPS-treated animals, subsequent administration of EA significantly decreased cerebrospinal fluid cAMP and PGE2 levels (p < 0.05 or p < 0.01), significantly decreased cAMP, significantly increased 5-HT levels (p < 0.05 or p < 0.01), and significantly down-regulated p-NF-κB p65 and COX-2 protein levels in the hypothalamus. Subsequent gas chromatography mass spectrometry(GC-MS) metabolite analysis indicated that 12 differential metabolites were detected in serum isolated 4 h after LPS treatment, and 10 differential metabolites were detected in serum collected 7 h after LPS treatment. Next, Pearson correlation analysis was used to systematically characterize the relationship between the identified metabolites and TNF-α, IL-6, MDA, SOD, PGE2, and cAMP. The levels of propionic acid, pyridine, and L-valine were up-regulated by EA, which inhibited the expression of MDA, IL-1β, and TNF-α and increased the activity of GSH. The levels of inositol, urea, and 2-monopalmitin were down-regulated by EA, which inhibited the expression of MDA, IL-1β, and TNF-α, increased the activity of SOD and GSH, reduced the inflammatory response, and alleviated the oxidative stress state. Combined with the results of the metabolic pathway analysis, we suggest that the pathways of the galactose metabolism, synthesis and degradation of ketone bodies, as well as ascorbic acid and aldehyde acid metabolism are closely related to the antipyretic and anti-inflammatory effects of EA. Our study established the relationship between EA and various inflammatory markers, such as TNF-α, IL-6, IL-1β, PGE2, and cAMP, and clarified the mechanism of the COX-2/NF-κB signaling pathway. Combined with the metabolomics analysis, our study revealed the effects of EA on multiple endogenous biomarkers, reflecting the characteristics of a multi-component, multi-target, and multi-pathway mechanism. [ABSTRACT FROM AUTHOR]

Obtaining information about cellular interactions is fundamental to the elucidation of physiological and pathological processes. Proximity labeling technologies have been widely used to report cellular interactions in situ; however, the reliance on addition of tag molecules typically restricts their application to regions where tags can readily diffuse, while the application in, for example, solid tissues, is susceptible. Here, we propose an "in‐situ‐tag‐generation mechanism" and develop the GalTag technology based on galactose oxidase (GAO) for recording cellular interactions within three‐dimensional biological solid regions. GAO mounted on bait cells can in situ generate bio‐orthogonal aldehyde tags as interaction reporters on prey cells. Using GalTag, we monitored the dynamics of cellular interactions and assessed the targeting ability of engineered cells. In particular, we recorded, for the first time, the footprints of Bacillus Calmette‐Guérin (BCG) invasion into the bladder tissue of living mice, providing a valuable perspective to elucidate the anti‐tumor mechanism of BCG. [ABSTRACT FROM AUTHOR]

• Age-related cognitive decline was detected through rodent temporal order recognition. • Poor temporal order memory is rescued by D 1 dopamine agonists. • Optimal effective doses vary markedly among individual rats of different age groups. • Population effectiveness may be influenced by agonist functional selectivity. Dopamine D 1 receptor agonists improve spatial working memory, but their effects on temporal order memory, particularly prone to the effects of aging, have not been studied. Two D 1 agonists, PF6256142 (PF) and 2-methyldihydrexidine (2MDHX), were examined for their effects in a rodent temporal order recognition task. Our results are consistent with the hypothesis that there is an age-related decline in rodent temporal order memory. The data also show that either agonist rescues the poor memory performance with a large effective size. Interestingly, the optimal effective dose varied among individual rats of different age groups. PF showed greater potency for older rats, whereas 2MDHX showed better overall population effectiveness. Both PF and 2MDHX have high intrinsic activity at rodent D 1 -mediated cAMP synthesis. Conversely, at D 1 -mediated β-arrestin recruitment, PF has essentially no intrinsic activity, whereas 2MDHX is a super-agonist. These findings suggest that D 1 agonists have potential to treat age-related cognitive decline, and the pattern of functional selectivity may be useful for developing drugs with an improved therapeutic index. [ABSTRACT FROM AUTHOR]

The global demand for plant oil has reached unprecedented levels and is relevant in all industrial sectors. Driven by the growing awareness for environmental issues of traditional plant oils and the need for eco-friendly alternatives, microbial oil emerges as a promising product with significant potential. Harnessing the capabilities of oleaginous microorganisms is an innovative approach for achieving sustainable oil production. To increase economic feasibility, it is crucial to explore feedstocks such as agricultural waste streams as renewable resource for microbial bioprocesses. The fungal model Ustilago maydis is one promising organism in the field of microbial triglyceride production. It has the ability to metabolize a wide variety of carbon sources for cell growth and accumulates high amounts of triglycerides intracellularly. In this study we asked whether this large variety of usable carbon sources can also be utilized for triglyceride production, using corn stover saccharides as a showcase. Our experiments revealed metabolization of the major saccharide building blocks present in corn stover, demonstrating the remarkable potential of U. maydis. The microorganism exhibited the capacity to synthesize triglycerides using the saccharides glucose, fructose, sucrose, xylose, arabinose, and galactose as carbon source. Notably, while galactose has been formerly considered as toxic to U. maydis, we found that the fungus can metabolize this saccharide, albeit with an extended lag phase of around 100 hours. We identified two distinct methods to significantly reduce or even prevent this lag phase, challenging previous assumptions and expanding the understanding of U. maydis metabolism. Our findings suggest that the two tested methods can prevent long lag phases on feedstocks with high galactose content and that U. maydis can produce microbial triglycerides very efficiently on many different carbon sources. Looking forward, exploring the metabolic capabilities of U. maydis on additional polymeric components of corn stover and beyond holds promise for innovative applications, marking a significant step toward environmentally sustainable bioprocessing technologies. [ABSTRACT FROM AUTHOR]

Wolfberry, esteemed as a traditional Chinese medicinal material and functional food, is replete with nutrients and boasts a diverse array of health benefits, including hypoglycemic, antitumor, antioxidant, anti-inflammatory, and immune-enhancing properties. Notably, inflammation is a pivotal factor in the onset and progression of numerous diseases. Despite this, there is a paucity of research on the comprehensive evaluation of the components found in different wolfberries, and the exploration of their primary active components is limited. To address this issue, we conducted a comprehensive targeted metabolomics analysis, employing statistical methods such as principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), KEGG pathway analysis, and volcano plots to delineate the compositional differences among red, black, and yellow wolfberries. Furthermore, we investigated the antiinflammatory effects of their primary components through in vitro experiments. Our analysis revealed a total of 1,104 chemical compositions in the three wolfberries, with alkaloids, phenolic acids, flavonoids, and lipids being the predominant nutritional components. KEGG enrichment analysis indicated that these compositions were primarily involved in the biosynthesis of secondary metabolites, ABC transport, and galactose metabolism pathway. Moreover, our study demonstrated that quercetin exhibited dose-dependent anti-inflammatory activity in LPS-stimulated HUVECs. It effectively inhibited the production of inflammatory factors such as TNF-α, MCP-1, and IL-1β, while also downregulating the gene and protein expression levels of ICAM-1 and VCAM-1. In conclusion, our findings indicate that there are variations in compositions among the three wolfberries, with flavonoids being the most abundant, and in vitro studies also confirmed the anti-inflammatory potential of quercetin. It is worth noting that Lycium ruthenicum contains higher levels of antioxidant components and possesses greater nutritional value, providing valuable insights for the future development and utilization of the three wolfberries. [ABSTRACT FROM AUTHOR]

2-Keto-3-deoxy-galactonate (KDGal) serves as a pivotal metabolic intermediate within both the fungal d-galacturonate pathway, which is integral to pectin catabolism, and the bacterial DeLey-Doudoroff pathway for d-galactose catabolism. The presence of KDGal enantiomers, l-KDGal and d-KDGal, varies across these pathways. Fungal pathways generate l-KDGal through the reduction and dehydration of d-galacturonate, whereas bacterial pathways produce d-KDGal through the oxidation and dehydration of d-galactose. Two distinct catabolic routes further metabolize KDGal: a nonphosphorolytic pathway that employs aldolase and a phosphorolytic pathway involving kinase and aldolase. Recent findings have revealed that l-KDGal, identified in the bacterial catabolism of 3,6-anhydro-l-galactose, a major component of red seaweeds, is also catabolized by Escherichia coli, which is traditionally known to be catabolized by specific fungal species, such as Trichoderma reesei. Furthermore, the potential industrial applications of KDGal and its derivatives, such as pyruvate and d- and l-glyceraldehyde, are underscored by their significant biological functions. This review comprehensively outlines the catabolism of l-KDGal and d-KDGal across different biological systems, highlights stereospecific methods for discriminating between enantiomers, and explores industrial application prospects for producing KDGal enantiomers. Key points: • KDGal is a metabolic intermediate in fungal and bacterial pathways • Stereospecific enzymes can be used to identify the enantiomeric nature of KDGal • KDGal can be used to induce pectin catabolism or produce functional materials [ABSTRACT FROM AUTHOR]

The research utilized network pharmacology to explore how Yixin Fumai granules mitigate sinoatrial node fibrosis caused by D-galactose in older mice by altering inflammatory reactions. Effective ingredients and targets of Yixin Fumai granules (including ginsenoside, ophiopogonin, schisandrin, astragaloside, danshensu, and ligustilide) were screened from traditional Chinese medicine systems pharmacology and BATMANtraditional Chinese medicine databases. Targeted diseases were pinpointed using GeneCards, Online Mendelian Inheritance in Man, and Disgenet databases. By employing Cytoscape 3.7.2 software, a network targeting drug-effective ingredients-targets was created, accompanied by a protein-protein interaction network formed via the Search Tool for the Retrieval of Interacting Genes databases. To forecast the fundamental processes, analyses of Gene Ontology functional enrichment and Kyoto Encyclopedia of Genes and Genomes pathway enrichment were conducted utilizing the R language. The expression levels of the Nucleotide oligomerization domain-like receptor protein 3, caspase-1, gasdermin D, interleukin-1 beta, and interleukin-18 genes were assessed through reverse transcription polymerase chain reaction. A total of 127 components were identified, with major components including quercetin, beta-sitosterol, kaempferol, naringenin, ginsenoside Rb1, and beta-eudesmol. There were 129 therapeutic targets identified, including key targets such as tumor necrosis factor, protein kinase B serine/threonine kinase 1, interleukin-6, tumor protein P53, interleukin-1 beta, and vascular endothelial growth factor A. The experimental findings revealed that the Yixin Fumai granules group mice, in contrast to the model group, showed enhanced mental health, a standard diet, notably better coat color, and zero fatalities. The echocardiographic results showed a rise in ejection fraction and fractional shortening, a faster heart rate, a reduction in left ventricular internal diameter end systole and left ventricular internal diameter end diastole, and an increase in left ventricular posterior wall end systole and left ventricular posterior wall end diastole in the Yixin Fumai granules group relative to the model group. This study preliminarily elucidated the active ingredients, targets, and pathways through which Yixin Fumai granules treat sick sinus syndrome using network pharmacology and further validated its mechanism through experimental verification. These findings provide new evidence and research directions for treating sick sinus syndrome induced by aging. [ABSTRACT FROM AUTHOR]

Background: Hippeastrum reticulatum (HR) belongs to the Amaryllidaceae, a well-known ornamental family. Objectives: This study was designed to reveal in vivo anti-acetylcholinesterase (AChE) and antioxidant activities and evaluate the acute toxicity of the alkaloid fraction (AF). Methods: The D-galactose-induced aging model and the scopolamine-induced cognitive-deficit model were used for the in vivo tests. The median lethal dose (LD50) of the AF was determined using the Litchfield and Wilcoxon method. Results: On the aging model, the AF at 10 and 15 mg/kg significantly improved the antioxidant markers including superoxide dismutase, glutathione peroxidase, and malonyl dialdehyde. Upon the scopolamine-injected mice, the AF at 10 and 15 mg/kg notably downregulated cerebral AChE in mice. The LD50 of the HR AF is 90.2 mg/kg. Conclusion: These findings provide strong evidence for the safety and advantages of utilizing HR in treating Alzheimer's disease. [ABSTRACT FROM AUTHOR]

Acute pancreatitis (AP) is a life-threatening inflammatory disease with no specific therapy. Excessive cytoplasmic Ca2+ elevation and intracellular ATP depletion are responsible for the initiation of AP. Inhibition of Ca2+ release–activated Ca2+ (CRAC) channels has been proposed as a potential treatment, and currently, a novel selective CRAC channel inhibitor CM4620 (Auxora, CalciMedica) is in Phase 2b human trials. While CM4620 is on track to become the first effective treatment for AP, it does not produce complete protection in animal models. Recently, an alternative approach has suggested reducing ATP depletion with a natural carbohydrate galactose. Here, we have investigated the possibility of using the smallest effective concentration of CM4620 in combination with galactose. Protective effects of CM4620, in the range of 1-100 n m, have been studied against necrosis induced by bile acids, palmitoleic acid, or l-asparaginase. CM4620 markedly protected against necrosis induced by bile acids or asparaginase starting from 50 n m and palmitoleic acid starting from 1 n m. Combining CM4620 and galactose (1 m m) significantly reduced the extent of necrosis to near-control levels. In the palmitoleic acid-alcohol–induced experimental mouse model of AP, CM4620 at a concentration of 0.1 mg/kg alone significantly reduced edema, necrosis, inflammation, and the total histopathological score. A combination of 0.1 mg/kg CM4620 with galactose (100 m m) significantly reduced further necrosis, inflammation, and histopathological score. Our data show that CM4620 can be used at much lower concentrations than reported previously, reducing potential side effects. The novel combination of CM4620 with galactose synergistically targets complementary pathological mechanisms of AP. Graphical Abstract [ABSTRACT FROM AUTHOR]

Chamuangone is an exotic medicinal compound isolated from Garcinia cowa leaves. It shows great potential against inflammation, neurodegeneration, and oxidation. Through green extraction method, chamuangone-rich rice bran oil (CRBO) containing 1.97 mg/mL of chamuangone was obtained. This research study investigates the therapeutic effects of CRBO on D-galactose- and aluminum chloride (AlCl3)-induced Alzheimer's disease in rat model. In all, 30 rats were divided into six groups and treated for 42 days as follows: control group (vehicle), disease control group (AlCl3 150 mg/kg + D-galactose 150 mg/kg p.o.), standard group (rivastigmine 3 mg/kg p.o.), and CRBO treatment groups with doses of 2.5, 5.0, and 7.5 mg/kg p.o. Neurobehavioral studies were performed on completion of 42 days. Furthermore, biochemical, neurochemical, and histopathological analyses were performed. CRBO treatment mitigated impairment in neuromuscular coordination, mentation and cognition in behavioral tasks. It potentially recovered (p < 0.001) the declined level of catalase, superoxide dismutase, reduced glutathione and lipid peroxidation. Neurochemical analysis revealed the restoration of serotonin, noradrenaline, and dopamine concentration in the brain after CRBO treatment. Histopathological examination of vital organs revealed recovery in CRBO-treated rats in a dose-dependent manner. Owing to neurotoxic effects of inducing agents, the level of acetylcholinesterase decreased in disease control group; however, CRBO treatment recovered the declined activity of this enzyme. Therefore, it is concluded that CRBO significantly recovered the neurodegenerative effects through multiple pathways, such as amelioration of oxidative stress and neuronal plasticity. Therefore, this innovative green remedy can be considered as a natural alternative for treating neurodegenerative disorders. [ABSTRACT FROM AUTHOR]

The capture and reduction of atmospheric dinitrogen gas to ammonium can be accomplished through the enzyme nitrogenase in a process known as biological nitrogen fixation (BNF), by a class of microbes known as diazotrophs. The diazotroph Azotobacter vinelandii is a model organism for the study of aerobic nitrogen fixation, and in recent years has been promoted as a potential producer of biofertilizers. Prior reports have demonstrated the potential to partially deregulate BNF in A. vinelandii, resulting in accumulation and extracellular release of ammonium. In many cases, deregulation requires the introduction of transgenic genes or elements to yield the desired phenotype, and the long‐term stability of these strains has been reported to be somewhat problematic. In this work, we constructed two strains of A. vinelandii where regulation can be precisely controlled without the addition of any foreign genes or genetic markers. Regulation is maintained through native promoters found in A. vinelandii that can be induced through the addition of extraneous galactose. These strains result in varied degrees of regulation of BNF, and as a result, the release of extracellular ammonium is controlled in a precise, and galactose concentration‐dependent manner. In addition, these strains yield high biomass levels, similar to the wild‐type A. vinelandii strain and are further able to produce high percentages of the bioplastic polyhydroxybutyrate. [ABSTRACT FROM AUTHOR]

Estimating feed efficiency (FE) in dairy sheep is challenging due to the high cost of systems that measure individual feed intake. Identifying proxies that can serve as effective predictors of FE could make it possible to introduce FE into breeding programs. Here, 39 Assaf ewes in first lactation were evaluated regarding their FE by 2 metrics, residual feed intake (RFI) and feed conversion ratio (FCR). The ewes were classified into high, medium and low groups for each metric. Milk samples of the 39 ewes were subjected to untargeted metabolomics analysis. The complete milk metabolomic signature was used to discriminate the FE groups using partial least squares discriminant analysis. A total of 41 and 26 features were selected as the most relevant features for the discrimination of RFI and FCR groups, respectively. The predictive ability when utilizing the complete milk metabolomic signature and the reduced data sets were investigated using 4 machine learning (ML) algorithms and a multivariate regression method. The orthogonal partial least squares algorithm outperformed other ML algorithms for FCR prediction in the scenarios using the complete milk metabolite signature (R2 = 0.62 ± 0.06) and the 26 selected features (R2 = 0.62 ± 0.15). Regarding RFI predictions, the scenarios using the 41 selected features outperformed the scenario with the complete milk metabolite signature, where the multilayer feedforward artificial neural network (R2 = 0.18 ± 0.14) and extreme gradient boosting (R2 = 0.17 ± 0.15) outperformed other algorithms. The functionality of the selected metabolites implied that the metabolism of glucose, galactose, fructose, sphingolipids, amino acids, insulin, and thyroid hormones was at play. Compared with the use of traditional methods, practical applications of these biomarkers might simplify and reduce costs in selecting feed-efficient ewes. [ABSTRACT FROM AUTHOR]

Saccharomyces cerevisiae is a genetically tractable, affordable, and extensively documented eukaryotic single-cell model organism. This budding yeast is amenable for the development of genetic and biochemical experiments and is frequently used to investigate the function, activity, and mechanism of mammalian proteins. However, yeast contains a cell wall that hinders select assays including organelle isolation. Lytic enzymes, with Zymolyase as the most effective and frequently used tool, are utilized to weaken the yeast cell wall resulting in yeast spheroplasts. Spheroplasts are easily lysed by, for example, osmotic-shock conditions to isolate yeast nuclei or mitochondria. However, during our studies of the DNA repair enzyme tyrosyl-DNA phosphodiesterase I (Tdp1), we encountered a negative effect of Zymolyase. We observed that Zymolyase treatment affected the steady-state protein levels of Tdp1. This was revealed by inconsistencies in technical and biological replicate lysates of plasmid-born galactose-induced expression of Tdp1. This off-target effect of Zymolyase is rarely discussed in articles and affects a select number of intracellular proteins, including transcription factors and assays such as chromatin immunoprecipitations. Following extensive troubleshooting, we concluded that the culprit is the Ser-protease, Zymolyase B, component of the Zymolyase enzyme mixture that causes the degradation of Tdp1. In this study, we report the protocols we have used, and our final protocol with an easy, affordable adaptation to any assay/protocol involving Zymolyase. [ABSTRACT FROM AUTHOR]

Cold‐active enzymes support life at low temperatures due to their ability to maintain high activity in the cold and can be useful in several biotechnological applications. Although information on the mechanisms of enzyme cold adaptation is still too limited to devise general rules, it appears that very diverse structural and functional changes are exploited in different protein families and within the same family. In this context, we studied the cold adaptation mechanism and the functional properties of a member of the glycoside hydrolase family 1 (GH1) from the Antarctic bacterium Marinomonas sp. ef1. This enzyme exhibits all typical functional hallmarks of cold adaptation, including high catalytic activity at 5 °C, broad substrate specificity, low thermal stability, and higher lability of the active site compared to the overall structure. Analysis of the here‐reported crystal structure (1.8 Å resolution) and molecular dynamics simulations suggest that cold activity and thermolability may be due to a flexible region around the active site (residues 298–331), whereas the dynamic behavior of loops flanking the active site (residues 47–61 and 407–413) may favor enzyme‐substrate interactions at the optimal temperature of catalysis (Topt) by tethering together protein regions lining the active site. Stapling of the N‐terminus onto the surface of the β‐barrel is suggested to partly counterbalance protein flexibility, thus providing a stabilizing effect. The tolerance of the enzyme to glucose and galactose is accounted for by the presence of a "gatekeeping" hydrophobic residue (Leu178), located at the entrance of the active site. [ABSTRACT FROM AUTHOR]

Under specific conditions of oxygen availability and lactose hydrolysis, Brettanomyces claussenii OYL-201 can convert glucose in dairy coproducts into ethanol, leaving the galactose almost untouched. This approach could be applied specifically to whey permeate (WP) to develop foods with low-glycemic loads and other potential health benefits. The aims of this research are to optimize the fermentation of WP by B. claussenii, to maximize the production of ethanol and galactose, and to characterize various products obtained with this approach. For this purpose, five fermentation factors were studied to determine their impacts on ethanol and galactose: temperature (20–40 °C), substrate concentration (5–15%TS), lactase enzyme/substrate ratio (0–40 IU/ g lactose), inoculation level (6–8 log cfu/mL), and time (6–30 days). Linear models, containing quadratic and interaction effects, were built for the optimization of both responses. Optimal levels were predicted for the maximum outputs of ethanol and galactose simultaneously, which utilized the following parameters: 15%TS, 37 IU / g lactose, 28 °C, 7.5 log cfu/mL, and 30 days, which together were predicted to produce 4.0%v/v ethanol and 51 g/L galactose in the final product. These parameters were applied to 18-L fermentations, and the fermentates were processed via distillation and freeze-drying. As a result, four product streams were obtained: a fermented product with 3.4%v/v ethanol and 56 g/L galactose, a 45%v/v ethanol distillate, a galactose-rich drink base (63 g/L), and a galactose-rich powder (55%w/w). These results demonstrate that it is possible to maximize the production of ethanol and galactose from WP and to develop novel, potentially functional bioproducts from this stream. [ABSTRACT FROM AUTHOR]