The association between sleep duration, bedtime, and cognitive ability in Chinese adults: Evidence from the China family panel studies.

Introduction: Dementia is marked by a steady decline or worsening in cognitive abilities, affecting memory, logic, and social competencies. While many studies suggest a potential link between the amount of sleep and dementia risk, the outcomes are not yet consistent. This research delved into the relationship between sleep length and bedtime on cognitive abilities using an extensive dataset from the China Family Panel Studies (CFPS) from 2014 to 2020. Methods: Data from 175,702 observations were collected, including cognitive function test data from 22,848 participants. Various cognitive tests were used to assess cognitive function. Restricted cubic spline (RCS) models were used for data analysis. Results: The optimal sleep duration for cognitive function was found to be 6-7 h, and the optimal bedtime was generally between 22:00-23:00. Longitudinal analysis revealed that sleep duration four years prior had a significant impact on current cognitive function. After accounting for various factors, those who slept for 7-8 h and over 8 h displayed lower cognitive function scores. Conversely, individuals sleeping less than 6 h had higher scores on the Vocabulary Test. Bedtime before 22:00 was associated with lower scores on the Vocabulary Test and Mathematical Test. Subgroup analyses based on age, gender, and urban residence showed variations in optimal sleep duration for different populations. Propensity Score Matching (PSM) analysis supported the findings. Conclusions: Maintaining a sleep duration of 6-7 h and a regular bedtime between 22:00-23:00 is important for optimizing cognitive performance.

Engineering the ß-galactosidase from Aspergillus oryzae for making lactose-free and no-sugar-added yogurt.

Yogurt usually contains 5-7% sugar and 3-5% lactose. As ß-galactosidases can hydrolyze lactose and improve sweetness, they have the potential to produce lactose-free (LF) and no-sugar-added (NSA) yogurt. In this study, ß-galactosidase AoBgal35A from Aspergillus oryzae was engineered by site-saturation mutagenesis. Results of 19 variants of T955 residue showed that lactose hydrolysis rate of T955R-AoBgal35A was up to 90.7%, much higher than 78.5% of the wild type. Moreover, the optimal pH of T955R-AoBgal35A was shifted from pH 4.5 to pH 5.5 and the optimal temperature decreased from 60°C to 50°C. The mutant T955R-AoBgal35A was successfully expressed in Komagatella pastoris, which produced extracellularly 4528 U/mL of ß-galactosidase activity. The mutant T955R-AoBgal35A was used to produce LF yogurt. Streptococcus thermophilus counts of LF yogurt increased from 7.9 to 9.5 lg cfu/g, significantly higher than that of the control group (8.9 lg cfu/g). Residual lactose content of LF yogurt was 0.13%, meeting the requirement of "lactose-free" label (<0.5%, GB 28050-2011, China). Furthermore, sugar in yogurt was replaced by whey powder to produce LF-NSA yogurt. The optimal addition content of whey powder was 7.5%. The texture, WHC and titratable acidity of LF and LF-NSA yogurt achieved good stability during the shelf life. Therefore, this study provides an insight for technological implications of ß-galactosidases in the dairy industry.

Protein Engineering of a Novel ß-Galactosidase from Thermus scotoductus for Efficient Synthesis of Lacto-N-Neotetraose from Chitin Powder.

A novel ß-galactosidase (TsGal48) from Thermus scotoductus was cloned, and the enzyme was biochemically characterized. TsGal48 catalyzed the synthesis of lacto-N-neotetraose (LNnT) from lactose via the transglycosylation reaction with a maximal yield of 20%, which is the highest yield for the synthesis of LNnT so far. To further improve the yield of LNnT, TsGal48 was successfully engineered by directed evolution and site-saturation mutagenesis. A mutated ß-galactosidase (mTsGal48) was selected and characterized. mTsGal48 produced LNnT with a yield of 27.7 g/L, which is 1.4-fold higher than that of TsGal48 (19.7 g/L). Then, a developed strategy for LNnT synthesis from chitin powder was provided in a 30 L bioreactor. The reaction process included chitin powder hydrolysis, lacto-N-triose II (LNT2) synthesis, and LNnT synthesis. The reaction time was reduced from 44 to 17 h in chitin powder hydrolysis and LNT2 synthesis. The content of LNnT was up to 25 g/L in the multienzyme system. The green and efficient route may be suitable for large-scale production of LNnT from chitin powder.

Efficient Biosynthesis of Difucosyllactose via De Novo GDP-l-Fucose Pathway in Metabolically Engineered Escherichia coli.

Difucosyllactose (DFL) is an important component of human milk oligosaccharides (HMOs) and has significant benefits for the growth and development of infants. So far, a few microbial cell factories have been constructed for the production of DFL, which still have problems of low production and high cost. Herein, a high-level de novo pathway DFL-producing strain was constructed by multistep optimization strategies in Escherichia coli BL21star(DE3). We first efficiently synthesized the intermediate 2'-fucosyllactose (2'-FL) in E. coli BL21star(DE3) by the advisable stepwise strategy. The truncated α-1,3/4-fucosyltransferase (Hp3/4FT) was then introduced into the engineered strain to achieve de novo biosynthesis of DFL. ATP-dependent protease (Lon) and GDP-mannose hydrolase (NudK) were deleted, and mannose-6-phosphate isomerase (ManA) was overexpressed to improve GDP-l-fucose accumulation. The regulator RcsA was overexpressed to fine-tune the expression level of pathway genes, thereby increasing the synthesis of DFL. The final strain produced 6.19 g/L of DFL in the shake flask and 33.45 g/L of DFL in the 5 L fermenter, which were the highest reported titers so far. This study provides a more economical, sustainable, and effective strategy to produce the fucosylated human milk oligosaccharides (HMOs).

Structural and functional analysis of SpGlu64A: a novel glycoside hydrolase family 64 laminaripentaose-producing ß-1,3-glucanase from Streptomyces pratensis.

Laminaripentaose (L5)-producing ß-1,3-glucanases can preferentially cleave the triple-helix curdlan into ß-1,3-glucooligosaccharides, especially L5. In this study, a newly identified member of the glycoside hydrolase family 64, ß-1,3-glucanase from Streptomyces pratensis (SpGlu64A), was functionally and structurally characterized. SpGlu64A shared highest identity (30%) with a ß-1,3-glucanase from Streptomyces matensis. The purified SpGlu64A showed maximal activity at pH 7.5 and 50 °C, and exhibited strict substrate specificity toward curdlan (83.1 U·mg-1). It efficiently hydrolyzed curdlan to produce L5 as the end product. The overall structure of SpGlu64A consisted of a barrel domain and a mixed (α/ß) domain, which formed an unusually wide groove with a crescent-like structure. In the two complex structures (SpGlu64A-L3 and SpGlu64A-L4), two oligosaccharide chains were captured and the triple-helical structure was relatively compatible with the wide groove, which suggested the possibility of binding to the triple-helical ß-1,3-glucan. A catalytic framework (ß6-ß9-ß10) and the steric hindrance formed by the side chains of residues Y161, N163, and H393 in the catalytic groove were predicted to complete the exotype-like cleavage manner. On the basis of the structure, a fusion protein with the CBM56 domain (SpGlu64A-CBM) and a mutant (Y161F; by site-directed mutation) were obtained, with 1.2- and 1.7-fold increases in specific activity, respectively. Moreover, the combined expression of SpGlu64A-CBM and -Y161F improved the enzyme activity by 2.63-fold. The study will not only be helpful in understanding the reaction mechanism of ß-1,3-glucanases but will also provide a basis for further enzyme engineering.

Construction of an engineered Escherichia coli for effective synthesis of 2'-fucosyllactose via the salvage pathway.

2'-Fucosyllactose (2'-FL) is one of the important functional oligosaccharides in breast milk. So far, few attempts on biosynthesis of 2'-FL by the salvage pathway have been reported. Herein, the salvage pathway enzyme genes were introduced into the E. coli BL21star(DE3) for synthesis of 2'-FL. The 2'-FL titer increased from 1.56 to 2.13 g/L by deleting several endogenous genes on competitive pathways. The α-1,2-fucosyltransferase (WbgL) was selected, and improved the 2'-FL titer to 2.88 g/L. Additionally, the expression level of pathway enzyme genes was tuned through optimizing the plasmid copy number. Furthermore, the spatial distribution of WbgL was enhanced by fusing with the MinD C-tag. After optimizing the fermentation conditions, the 2'-FL titer reached to 7.13 g/L. The final strain produced 59.22 g/L of 2'-FL with 95% molar conversion rate of lactose and 92% molar conversion rate of fucose in a 5 L fermenter. These findings will contribute to construct a highly efficient microbial cell factory to produce 2'-FL or other HMOs.

Antimicrobial efficacy of aloe-emodin mediated photodynamic therapy against antibiotic-resistant Pseudomonas aeruginosa in vitro.

Multidrug-resistant Pseudomonas aeruginosa is a common pathogen that causes topical infections following burn injuries. Antimicrobial photodynamic therapy (aPDT) has emerged as a promising approach for treating antibiotic-resistant bacterial infections. The objective of this study was to evaluate the aPDT efficacy of aloe-emodin (AE), which is a photosensitizer extracted from traditional Chinese herbs, on antibiotic-sensitive and antibiotic-resistant P. aeruginosa in vitro. In this study, we confirmed the effectiveness of AE-mediated aPDT against both standard and MDR P. aeruginosa, explored the effects of irradiation time and AE concentration on bacterial survival in AE-mediated aPDT, and observed the structural damage of P. aeruginosa by using transmission electron microscope. Our results showed that neither AE nor light irradiation alone caused cytotoxic effects on P. aeruginosa. However, AE-mediated aPDT effectively inactivated both antibiotic-sensitive and antibiotic-resistant P. aeruginosa. The transmission electron microscope investigation showed that aPDT mediated by AE primarily caused damage to the cytoplasm and cell membrane. Our findings suggest that AE is a photosensitizer in the aPDT of MDR P. aeruginosa-caused topical infections following burn injuries. Future investigations will concentrate on the safety and efficacy of AE-mediated aPDT in animal models and clinical trials.

Manno-oligosaccharides from Cassia Seed Gum Attenuate Atherosclerosis through Inflammation Modulation and Intestinal Barrier Integrity Improvement in ApoE-/- Mice.

SCOPE: Manno-oligosaccharides from cassia seed gum (CMOS) have demonstrated anti-inflammatory and regulatory effects on cholesterol metabolism. However, their protective effects against the progression of atherosclerosis (AS) and underlying molecular mechanisms have not been investigated. This study investigates the anti-atherosclerotic effects of CMOS on ApoE-/- mice. METHODS AND RESULTS: CMOS are supplemented in atherosclerotic male ApoE-/- mice fed with a high-fat-high-cholesterol diet (HFHCD). After the 12-week intervention, CMOS at 1200 mg kg-1 ·bw d-1 significantly decrease the atherosclerotic lesion area by 0.63-fold and the aortic arch lesion size by 0.63-fold when compared to the HFHCD group. Moreover, inflammation in atherosclerotic lesions is reduced by CMOS intervention, and the levels of serum lipids and inflammatory cytokines are decreased. The number of goblet cells and the expression of intestinal epithelial tight junction proteins in the H-CMOS group increase, thus indicating that CMOS can restore intestinal barrier integrity in atherosclerotic mice. Furthermore, CMOS reshape the unbalanced gut microbiota in ApoE-/- mice caused by HFHCD, and reduce the relative abundance of Desulfovibrio and Faecalibaculum that exhibits positive relationships with inflammation. CONCLUSION: CMOS inhibit inflammation, alter intestinal barrier integrity, and regulate gut microbiota to attenuate AS in ApoE-/- mice.

Development of a two-enzyme system in Aspergillus niger for efficient production of N-acetyl-ß-D-glucosamine from powdery chitin.

A chitinase (PbChi70) from Paenibacillus barengoltzii was engineered by directed evolution to enhance its hydrolysis efficiency towards powder chitin. Through two rounds of screening, a mutant (mPbChi70) with a maximum specific activity of 73.21 U/mg was obtained, which is by far the highest value ever reported. The mutant gene was further transformed into Aspergillus niger FBL-B (ΔglaA) which could secrete high level of endogenously ß-N-acetylglucosaminidase (GlcNAcase), thus a two-enzyme expression system was constructed. The highest chitinase activity of 61.33 U/mL with GlcNAcase activity of 353.1 U/mL was obtained in a 5-L fermentor by high-cell density fermentation. The chitin-degrading enzyme cocktail was used for the bioconversion of GlcNAc from powder chitin directly, and the highest conversion ratio reached high up to 71.9 % (w/w) with GlcNAc purity ≥95 % (w/w). This study may provide an excellent chitinase as well as a double enzyme cocktail system for efficient biological conversion of chitin materials.

Role of Contrast-Enhanced Ultrasound With the Enhancement Pattern and Qualitative Analysis for Differentiating Hypovascular Solid Renal Lesions.

OBJECTIVE: The aim of the work described here was to explore the clinical value of contrast-enhanced ultrasound (CEUS) with the enhancement pattern and qualitative analysis in distinguishing different types of hypovascular solid renal lesions. METHODS: A total of 140 patients with 140 renal tumors (all diagnosed by pathology), which manifested hypo-enhancement on CEUS, were included in this study. We compared conventional ultrasound (US) and CEUS features in five common hypovascular renal tumors, including renal angiomyolipoma (RAML), clear cell renal cell carcinoma (ccRCC), renal pelvic urothelial carcinoma (RPUC), papillary renal cell carcinoma (pRCC) and chromophobe renal cell carcinoma (chRCC). The diagnostic value of conventional US and qualitative parameters of CEUS for differentiating hypovascular solid renal lesions were evaluated. RESULTS: The mean age of patients with a benign renal lesion was younger than that of patients with a malignant renal lesion (p < 0.05). Echogenicity and qualitative parameters such as wash-out, perfusion defects and perilesional rim-like enhancement (PRE) in the two groups differed significantly (all p values <0.05). Benign renal lesions exhibited mainly slow wash-out, whereas malignant renal lesions exhibited predominantly fast wash-out on CEUS (p < 0.05). There were significant differences in echogenicity, such as between RAML and ccRCC, between RAML and RPUC and between RAML and pRCC (all p values <0.05). The rates of appearance of perfusion defect in ccRCC (48%, 13/27) and pRCC (53%, 10/19) were significantly higher than the rate in RAML (14%, 6/43) (p < 0.05). The rates of appearance of PRE in ccRCC (15%, 4/27), pRCC (26%, 5/19) and chRCC (24%,4/17) were significantly higher than the rate in RAML (9%, 4/43) (p < 0.05). CONCLUSION: CEUS with the enhancement pattern and qualitative analysis may be helpful in distinguishing malignant from benign hypovascular renal lesions.

High-level production of a novel alginate lyase (FsAly7) from Flammeovirga sp. for efficient production of low viscosity soluble dietary fiber from sodium alginate.

Sodium alginate is one of the most abundant sustainable gum source for dietary fiber production. However, the preparation efficiencies of low viscosity soluble dietary fiber from sodium alginate remain low. Here, a novel alginate lyase gene (FsAly7) from Flammeovirga sp. was identified and high-level expressed in Pichia pastoris for low viscosity soluble dietary fiber production. The highest enzyme production of 3050 U mL-1 was achieved, which is by far the highest yield ever reported. FsAly7 was used for low viscosity soluble dietary fiber production from sodium alginate, and the highest degradation rate of 85.5 % was achieved under a high substrate content of 20 % (w/v). The molecular weight of obtained soluble dietary fiber converged to 10.75 kDa. FsAly7 catalyzed the cleavage of glycosidic bonds in alginate chains with formation of unsaturated non-reducing ends simultaneously in the degradation process, thus altered the chemical structures of hydrolysates. The soluble dietary fiber exhibited excellent properties, including low viscosity, high oil adsorption capacity activity (2.20 ± 0.03 g g-1) and high emulsifying activity (60.05 ± 2.96 mL/100 mL). This investigation may provide a novel alginate lyase catalyst as well as a solution for the efficient production of low viscosity soluble dietary fiber from sodium alginate.

Progress of near-infrared-II fluorescence in precision diagnosis and treatment of colorectal cancer.

Colorectal cancer is a malignant tumour with high incidence and mortality worldwide; therefore, improving the early diagnosis of colorectal cancer and implementing a targeted "individualized treatment" strategy is of great concern. NIR-II fluorescence imaging is a large-depth, high-resolution optical bioimaging tool. Around the NIR-II window, researchers have developed a variety of luminescent probes, imaging systems, and treatment methods with colorectal cancer targeting capabilities, which can be visualized and image-guided in clinical surgery. This article aims to overcome the difficulties in diagnosing and treating colorectal cancer. The present review summarizes the latest results on using NIR-II fluorescence for targeted colorectal cancer imaging, expounds on the application prospects of NIR-II optical imaging for colorectal cancer, and discusses the imaging-guided multifunctional diagnosis and treatment platforms.

Pathogenicity and functional analysis of CFAP410 mutations causing cone-rod dystrophy with macular staphyloma.

Background: Cone-rod dystrophy (CORD) caused by pathogenic variants in CFAP410 is a very rare disease. The mechanisms by which the variants caused the disease remained largely unknown. CFAP410 pathogenic variants were identified in a cone-rod dystrophy with macular staphyloma patient. We explored the pathogenicity and performed functional analysis of two compound heterozygous mutations. Methods: A 6-year-old boy complained decreased vision for 1 year, underwent ocular examinations together with systemic X-ray check. Blood sample was taken for targeted next generation sequencing (Tg-NGS). Pathogenicity of identified variants was determined by ACMG guideline. Mutated plasmids were constructed and transferred to HEK293T cells. Cell cycle, protein stability, and protein ubiquitination level was measured. Results: The best-corrected visual acuity of proband was 0.20 bilaterally. Fundus showed macular staphyloma and uneven granular pigment disorder in the periphery of the retina. SS-OCT showed thinning and atrophy of the outer retina, residual ellipsoid zone (EZ) in the fovea. Scotopic and photopic ERG responses severe reduced. Two heterozygous missense pathogenic variants, c.319 T > C (p.Tyr107His) and c.347 C > T (p.Pro116Leu) in exon 4 of the CFAP410, were found and were pathogenic by the ACMG guideline. In vitro, pathogenic variants affect cell cycle. Immunofluorescence and western blotting showed that the mutant proteins decreased expression levels protein stability. Meanwhile, co-IP data suggested that ubiquitination level was altered in cells transferred with the mutated plasmids. Conclusion: Compound heterozygous pathogenic variants c.319 T > C and c.347 C > T in CFAP410 caused CORD with macular staphyloma. The pathogenic mechanisms may be associated with alternations of protein stability and degradation through the ubiquitin-proteasome pathway.

Biochemical characterization of a novel ß-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose.

Lacto-N-tetraose (LNT) is one of the most important components of human milk oligosaccharides, which has various beneficial health effects. ß-Galactosidase is an important enzyme used in dairy processing. The transglycosylation activity of ß-galactosidases offers an attractive approach for LNT synthesis. In this study, we reported for the first time the biochemical characterization of a novel ß-galactosidase (LzBgal35A) from Lacticaseibacillus zeae. LzBgal35A belongs to glycoside hydrolases (GH) family 35 and shared the highest identity of 59.9% with other reported GH 35 members. The enzyme was expressed as soluble protein in Escherichia coli. The purified LzBgal35A displayed optimal activity at pH 4.5 and 55°C. It was stable within the pH range of 3.5 to 7.0 and up to 60°C. Moreover, LzBgal35A could catalyze the synthesis of LNT via transferring the galactose residue from o-nitrophenyl-ß-galactopyranoside to lacto-N-triose II. Under optimal conditions, the conversion rate of LNT reached 45.4% (6.4 g/L) within 2 h, which was by far the highest yield of LNT synthesized through a ß-galactosidase-mediated transglycosylation reaction. This study demonstrated that LzBgal35A has great potential application in LNT synthesis.

Releasing Bioactive Compounds from Brown Seaweed with Novel Cold-Adapted Alginate Lyase and Alcalase.

Seaweeds are considered to be third-generation renewable biomasses, the comprehensive utilization of which has drawn increasing attention in recent years. A novel cold-active alginate lyase (VfAly7) was identified from Vibrio fortis and biochemically characterized for brown seaweed utilization. The alginate lyase gene was high-level expressed in Pichia pastoris, with an enzyme yield of 560 U/mL and a protein content of 9.8 mg/mL by high-cell density fermentation. The recombinant enzyme was most active at 30 °C and pH 7.5, respectively. VfAly7 was a bifunctional alginate lyase with both poly-guluronate and poly-mannuronate hydrolysis activities. On the basis of VfAly7, a bioconversion strategy for the utilization of brown seaweed (Undaria pinnatifida) was developed. The obtained AOSs showed stronger prebiotic activity towards tested probiotics when compared to that of commercial fructooligosaccharides (FOSs), while the obtained protein hydrolysates displayed strong xanthine oxidase inhibitory activity with IC50 of 3.3 mg/mL. This study provided a novel alginate lyase tool as well as a biotransformation route for the utilization of seaweeds.

Dietary neoagarotetraose extends lifespan and impedes brain aging in mice via regulation of microbiota-gut-brain axis.

INTRODUCTION: Dietary oligosaccharides can impact the gut microbiota and confer tremendous health benefits. OBJECTIVES: The aim of this study was to determine the impact of a novel functional oligosaccharide, neoagarotetraose (NAT), on aging in mice. METHODS: 8-month-old C57BL/6J mice as the natural aging mice model were orally administered with NAT for 12 months. The preventive effect of NAT in Alzheimer's disease (AD) mice was further evaluated. Aging related indicators, neuropathology, gut microbiota and short-chain fatty acids (SCFAs) in cecal contents were analyzed. RESULTS: NAT treatment extended the lifespan of these mice by up to 33.3 %. Furthermore, these mice showed the improved aging characteristics and decreased injuries in cerebral neurons. Dietary NAT significantly delayed DNA damage in the brain, and inhibited reduction of tight junction protein in the colon. A significant increase at gut bacterial genus level (such as Lactobacillus, Butyricimonas, and Akkermansia) accompanied by increasing concentrations of SCFAs in cecal contents was observed after NAT treatment. Functional profiling of gut microbiota composition indicated that NAT treatment regulated the glucolipid and bile acid-related metabolic pathways. Interestingly, NAT treatment ameliorated cognitive impairment, attenuated amyloid-ß (Aß) and Tau pathology, and regulated the gut microbiota composition and SCFAs receptor-related pathway of Alzheimer's disease (AD) mice. CONCLUSION: NAT mitigated age-associated cerebral injury in mice through gut-brain axis. The findings provide novel evidence for the effect of NAT on anti-aging, and highlight the potential application of NAT as an effective intervention against age-related diseases.

Synthesis of 2'-fucosyllactose from apple pomace-derived xyloglucan oligosaccharides by an α-L-fucosidase from Pedobacter sp. CAU209.

2'-Fucosyllactose (2'-FL) is known for its ability to provide various health benefits to infants, such as gut maturation, pathogen resistance, improved immunity, and nervous system development. However, the production of 2'-FL using α-L-fucosidases is hindered by the lack of low-cost natural fucosyl donors and high-efficiency α-L-fucosidases. In this work, a recombinant xyloglucanase from Rhizomucor miehei (RmXEG12A) was applied to produce xyloglucan-oligosaccharide (XyG-oligos) from apple pomace. Then, an α-L-fucosidase gene (PbFucB) was screened from the genomic DNA of Pedobacter sp. CAU209 and expressed in Escherichia coli. The capability of purified PbFucB to catalyze XyG-oligos and lactose to synthesize 2'-FL was further evaluated. The deduced amino acid sequence of PbFucB shared the highest identity (38.4%) with that of other reported α-L-fucosidases. PbFucB showed the highest activity at pH 5.5 and 35 °C. It catalyzed the hydrolysis of 4-nitrophenyl-α-L-fucopyranoside (pNP-Fuc, 20.3 U mg-1), 2'-FL (8.06 U mg-1), and XyG-oligos (0.43 U mg-1). Furthermore, PbFucB demonstrated a high enzymatic conversion rate in 2'-FL synthesis with pNP-Fuc or apple pomace-derived XyG-oligos as donors and lactose as acceptor. Under the optimized conditions, PbFucB converted 50% of pNP-Fuc or 31% of the L-fucosyl residue in XyG-oligos into 2'-FL. This work elucidated an α-L-fucosidase that mediates the fucosylation of lactose and provided an efficient enzymatic strategy to synthesize 2'-FL either from artificial pNP-Fuc or natural apple pomace-derived XyG-oligos. KEY POINTS: • Xyloglucan-oligosaccharide (XyG-oligos) was produced from apple pomace by a xyloglucanase from Rhizomucor miehei. • An α-L-fucosidase (PbFucB) from Pedobacter sp. CAU209 shared the highest identity (38.4%) with reported α-L-fucosidases. •PbFucB synthesized 2'-FL using apple pomace-derived XyG-oligos and lactose with a conversion ratio of 31%.

Nondigestible Functional Oligosaccharides: Enzymatic Production and Food Applications for Intestinal Health.

Nondigestible functional oligosaccharides are of particular interest in recent years because of their unique prebiotic activities, technological characteristics, and physiological effects. Among different types of strategies for the production of nondigestible functional oligosaccharides, enzymatic methods are preferred owing to the predictability and controllability of the structure and composition of the reaction products. Nondigestible functional oligosaccharides have been proved to show excellent prebiotic effects as well as other benefits to intestinal health. They have exhibited great application potential as functional food ingredients for various food products with improved quality and physicochemical characteristics. This article reviews the research progress on the enzymatic production of several typical nondigestible functional oligosaccharides in the food industry, including galacto-oligosaccharides, xylo-oligosaccharides, manno-oligosaccharides, chito-oligosaccharides, and human milk oligosaccharides. Moreover, their physicochemical properties and prebiotic activities are discussed as well as their contributions to intestinal health and applications in foods.

Hyperechoic mass of the right ventricle in adults: A rare presentation of an intracardiac teratoma.

Cardiac teratomas are very rare primary tumors; most are intrapericardial, while a few are intracardiac. Furthermore, most reported intracardiac teratomas are in the pediatric population, with few cases of secondary metastases from testicular teratomas reportedly manifesting in adults. Here, we report a rare case of a mature cystic teratoma in the right ventricle complicated by a bicuspid aortic valve (BAV) in an adult. Echocardiography and enhanced computed tomography (CT) were performed, and the mass was surgically excised. A pathological examination confirmed the diagnosis of a mature cystic teratoma. Meanwhile, mechanical valve replacement of the aortic valve was performed. No tumor recurrence or symptoms occurred in the 2-year follow-up. This is the first report of an adult primary intracardiac teratoma with solid hyperechoic findings on echocardiography and a BAV.

Directed evolution of a ß-N-acetylhexosaminidase from Haloferula sp. for lacto-N-triose II and lacto-N-neotetraose synthesis from chitin.

In our previous study, a ß-N-acetylhexosaminidase (HaHex74) from Haloferula sp. showing high human milk oligosaccharides (HMOs) synthesis ability was identified and characterized. In this study, HaHex74 was further engineered by directed evolution and site-saturation mutagenesis to improve its transglycosylation activity for HMOs synthesis. A mutant (mHaHex74) with improved transglycosylation activity (HaHex74-Asn401Ile/His394Leu) was obtained and characterized. mHaHex74 exhibited maximal activity at pH 5.5 and 35 °C, respectively, which were distinct from that of HaHex74 (pH 6.5 and 45 °C). Moreover, mHaHex74 showed the highest LNT2 conversion ratio of 28.2% from N,N'-diacetyl chitobiose (GlcNAc2), which is 2.2 folds higher than that of HaHex74. A three-enzyme cascade reaction for the synthesis of LNT2 and LNnT from chitin was performed in a 5-L reactor, and the contents of LNT2 and LNnT reached up to 15.0 g L1 and 4.9 g L1, respectively. Therefore, mHaHex74 maybe a good candidate for enzymatic synthesis of HMOs.