Biological evaluation of novel derivatives of the orange pigments from Monascus sp. as inhibitors of melanogenesis.

Nitrogenous derivatives of the two orange pigments from Monascus sp. with anti-melanogenic activities were prepared using fermentation and chemical synthesis. The pigments were produced in a 5 l jar fermentor. A total of 33 derivatives were synthesized via incorporation of L-amino acids and amines into the pigments. Two derivatives with high inhibitory melanin-synthesizing activities and low cell toxicities were selected based on testing using B16F10 cells. Glutamic acid and (S)-(+)-1-amino-2-propanol derivatives showed high inhibitory activities against melanogenesis. Both the reaction and expression of tyrosinase, an important enzyme in the melanin-synthesizing pathway, were inhibited by the glutamic acid derivative in a dose-dependent manner. The (S)-(+)-1-amino-2-propanol derivative inhibited expression of tyrosinase in cells, but not the tyrosinase reaction. TRP1 and TRP2, other important proteins in melanin-synthesis, were not affected by the two derivatives.

Preparation of eutectic substrate mixtures for enzymatic conversion of ATC to L-cysteine at high concentration levels.

High concentration eutectic substrate solutions for the enzymatic production of L-cysteine were prepared. Eutectic melting of binary mixtures consisting of D,L-2-amino-Δ(2)-thiazoline-4-carboxylic acid (ATC) as a substrate and malonic acid occurred at 39 °C with an ATC mole fraction of 0.5. Formation of eutectic mixtures was confirmed using SEM, SEM-EDS, and XPS surface analyses. Sorbitol, MnSO4, and NaOH were used as supplements for the enzymatic reactions. Strategies for sequential addition of five compounds, including a binary ATC mixture and supplements, during preparation of eutectic substrate solutions were established. Eutectic substrate solutions were stable for 24 h. After 6 h of enzymatic reactions, a 550 mM L-cysteine yield was obtained from a 670 mM eutectic ATC solution.

Enhancing the dispersibility of Gelidium amansii-derived microfibrillated cellulose through centrifugal fractionation.

Hydrothermal pretreatment is useful for microfibrillated cellulose (MFC) preparation due to its safety, but the remaining hemicellulose might affect MFC properties. This study aimed to investigate the effect of centrifugation time on hemicellulose removal and the physicochemical properties of MFC obtained after hydrothermal pretreatment and micro-fibrillation. In this study, centrifugation was applied to the MFC suspension at varying duration times. Composition analysis and Fourier transform infrared spectra indicated that fractionated MFC has no hemicellulose content after 10, 20, and 30 min centrifugation. It also showed an approximately 5 times higher than 0.5 % g/g of initial solid concentration, indicated by a lower gel concentration point, than unfractionated MFC. Scanning electron microscope images of the fractionated MFC for 30 min (MFC2C) presented thin, long cellulose fibrils of 517 nm in average diameter and 635-10,000 nm in length that induced a slower sedimentation rate. MFC2C dispersion was also improved by autoclave sterilization by regulating cellulose structure, rheology, and crystallinity. As a result, MFC dispersibility can be enhanced by removing hemicellulose through simple centrifugation.

The role of Bifidobacterium in longevity and the future of probiotics.

This review explores the role and health impacts of probiotics, focusing specifically on Bifidobacterium spp. It highlights the functionalities that Bifidobacteria can provide, underscored by the historical evolution of definitions and technological advancements related to probiotics. By examining the association between Bifidobacteria and longevity, this review suggests new avenues for health enhancement. Highlighting case studies of centenarians, it presents examples related to human aging, illuminating the potential links to longevity through research on Bifidobacterium strains found in centenarians. This review not only emphasizes the importance of current research but also advocates for further investigation into the health benefits of Bifidobacteria, underlining the necessity for continuous study in the nutraceutical field.

Mucoadhesive chitosan microcapsules for controlled gastrointestinal delivery and oral bioavailability enhancement of low molecular weight peptides.

A bioactive compound, collagen peptide (CP), is widely used for biological activities such as anti-photoaging and antioxidant effects, with increased oral bioavailability because of its low molecular weight and high hydrophilicity. However, controlling release time and increasing retention time in the digestive tract for a more convenient oral administration is still a challenge. We developed CP-loaded chitosan (CS) microcapsules via strong and rapid ionic gelation using a highly negative phytic acid (PA) crosslinker. The platform enhanced the oral bioavailability of CP with controlled gastrointestinal delivery by utilizing the mucoadhesiveness and tight junction-opening properties of CS. CS and CP concentrations varied from 1.5 to 3.5% and 0-30%, respectively, for optimal and stable microcapsule synthesis. The physicochemical properties, in vitro release profile with intestinal permeability, in vivo oral bioavailability, in vivo biodistribution, anti-photoaging effect, and antioxidant effect of optimized CS microcapsules were analyzed to investigate the impact of controlling parameters. The structure of CS microcapsules was tuned by PA diffused gradient ionic cross-linking degree, resulting in a controlled CP release region in the gastrointestinal tract. The optimized microcapsules increased Cmax, AUC, and tmax by 1.5-, 3.4-, and 8.0-fold, respectively. Furthermore, CP in microcapsules showed anti-photoaging effects by downregulating matrix metalloproteinases-1 via antioxidant effects. According to our knowledge, this is the first study to microencapsulate CP for oral bioavailability enhancement. The peptide delivery method employed is simple, economical, and can be applied to customize bioactive compound administration.

Hyaluronic acid hydrolysis using vacuum ultraviolet TiO2 photocatalysis combined with an oxygen nanobubble system.

Advanced technologies for producing high-quality low molecular weight hyaluronic acid (LMW-HA) are required from the perspective of cost-efficiency and biosafety. Here, we report a new LMW-HA production system from high molecular weight HA (HMW-HA) using vacuum ultraviolet TiO2 photocatalysis with an oxygen nanobubble system (VUV-TP-NB). The VUV-TP-NB treatment for 3 h resulted in a satisfactory LMW-HA (approximately 50 kDa measured by GPC) yield with a low endotoxin level. Further, there were no inherent structural changes in the LMW-HA during the oxidative degradation process. Compared with conventional acid and enzyme hydrolysis methods, VUV-TP-NB showed similar degradation degree with viscosity though reduced process time by at least 8-fold. In terms of endotoxin and antioxidant effects, degradation using VUV-TP-NB demonstrated the lowest endotoxin level (0.21 EU/mL) and highest radical scavenging activity. This nanobubble-based photocatalysis system can thus be used to produce biosafe LMW-HA cost-effectively for food, medical, and cosmetics applications.

Thermoresponsive semi-interpenetrating gelatin-alginate networks for encapsulation and controlled release of scent molecules.

Plant-based meats, which are nutritious foods from non-animal sources, provide clues for addressing the negative externalities associated with conventional meat production. Interest in plant-based meat has increased and is driving the rapid growth of its market. Plant-based meat should be equipped with a temperature-dependent scent release system similar to the scent release mechanism of conventional meat, to deliver a desirable meat-like flavor to consumers and obtain higher market acceptance. In this study, we prepared thermoresponsive gelatin-alginate hybrid hydrogels to control the release of scent molecules. The polymer network of gelatin-alginate hydrogels was reinforced by a semi-interpenetrating network (sIPN). sIPN formation conferred resistance to external stimuli, such as shear force, swelling, and temperature, resulting in a sustained release of the meat scent. In addition, controlled size microcapsules fabricated from the same composition via an electrostatic extrusion process showed a sustained release pattern of the loaded scent at 70 °C, and the scent release rate was precisely controlled within an approximately 2-fold range by adjusting the alginate concentration. These observations suggest the potential use of edible biological macromolecules as food additives that can control the release of scent molecules from the plant-based meat during cooking.

High-Fat Diet-Induced Obese Effects of Adipocyte-Specific CXCR2 Conditional Knockout in the Peritoneal Tumor Microenvironment of Ovarian Cancer.

Obesity contributes to ovarian cancer (OC) progression via tumorigenic chemokines. Adipocytes and OC cells highly express CXCR2, and its ligands CXCL1/8, respectively, indicating that the CXCL1/8-CXCR2 axis is a molecular link between obesity and OC. Here, we investigated how the adipocyte-specific CXCR2 conditional knockout (cKO) affected the peritoneal tumor microenvironment of OC in a high-fat diet (HFD)-induced obese mouse model. We first generated adipocyte-specific CXCR2 cKO in mice: adipose tissues were not different in crown-like structures and adipocyte size between the wild-type (WT) and cKO mice but expressed lower levels of CCL2/6 compared to the obese WT mice. HFD-induced obese mice had a shorter survival time than lean mice. Particularly, obese WT and cKO mice developed higher tumors and ascites burdens, respectively. The ascites from the obese cKO mice showed increased vacuole clumps but decreased the floating tumor burden, tumor-attached macrophages, triglyceride, free fatty acid, CCL2, and TNF levels compared to obese WT mice. A tumor analysis revealed that obese cKO mice attenuated inflammatory areas, PCNA, and F4/80 compared to obese WT mice, indicating a reduced tumor burden, and there were positive relationships between the ascites and tumor parameters. Taken together, the adipocyte-specific CXCR2 cKO was associated with obesity-induced ascites despite a reduced tumor burden, likely altering the peritoneal tumor microenvironment of OC.

In vivo evaluation of the anti-obesity effects of combinations of Monascus pigment derivatives.

The prevention and treatment of obesity using naturally derived compounds is desirable in terms of marketing and safety in the nutraceutical and functional food markets. One of the noticeable effects of Monascus pigment derivatives is the inhibition/deactivation of lipid metabolism. Our earlier studies reported that threonine (Thr), tryptophan (Trp), and 2-(p-tolyl)-ethylamine (TEA) derivatives of Monascus pigment showed cholesterol-lowering, lipase-inhibitory, and adipogenic differentiation-inhibitory activities, respectively. In this work, we investigated the in vivo anti-obesity effects of a combination of Thr, Trp and TEA derivatives. C57BL/6 mice were fed a high-fat diet (HFD) and simultaneously administered one of three 1 : 1 mixtures of Thr, Trp, and TEA derivatives. After 10 weeks of feeding, the weight gains of mice fed with three combined derivatives decreased by 20.3-37.9%, compared to mice fed the HFD. The epididymal adipose tissue (EAT) weights of mice fed with the combined derivatives decreased by 42.3-60.5% compared to the HFD group, and their EAT size decreased. Transverse micro-CT imaging revealed reduction of the subcutaneous and visceral fat layers of test mice. Our results confirm that Monascus-fermented pigment derivatives have in vivo anti-obesity effects and their combinations provide a higher efficacy in the reduction of body weight and EAT weights as well as lipid accumulation in mice. The key to accomplishing high anti-obesity effect was combining Thr and Trp derivatives, which provide higher effectiveness than other combined derivatives. These observations offer a potential use of Monascus pigment derivatives as a therapeutic approach to prevention and/or treatment of obesity.

Microbial biocatalysis of quercetin-3-glucoside and isorhamnetin-3-glucoside in Salicornia herbacea and their contribution to improved anti-inflammatory activity.

Salicornia herbacea (glasswort) is a traditional Asian medicinal plant which exhibits multiple nutraceutical and pharmaceutical properties. Quercetin-3-glucoside and isorhamnetin-3-glucoside are the major flavonoid glycosides found in S. herbacea. Multiple researchers have shown that flavonoid glycosides can be structurally transformed into minor aglycone molecules, which play a significant role in exerting physiological responses in vivo. However, minor aglycone molecule levels in S. herbacea are very low. In this study, Bifidobacterium animalis subsp. lactis AD011, isolated from infant feces, catalyzed >85% of quercetin-3-glucoside and isorhamnetin-3-glucoside into quercetin and isorhamnetin, respectively, in 2 h, without breaking down flavonoid backbones. Functionality analysis demonstrated that the quercetin and isorhamnetin produced showed improved anti-inflammatory activity vs. the original source molecules against lipopolysaccharide induced RAW 264.7 macrophages. Our report highlights a novel protocol for rapid quercetin and isorhamnetin production from S. herbacea flavonoids and the applicability of quercetin and isorhamnetin as nutraceutical molecules with enhanced anti-inflammatory properties.

Production and Characterization of Anti-Inflammatory Monascus Pigment Derivatives.

The prevention and treatment of chronic inflammation using food-derived compounds are desirable from the perspectives of marketing and safety. Monascus pigments, widely used as food additives, can be used as a chronic inflammation treatment. Orange Monascus pigments were produced by submerged fermentation in a 5 L bioreactor, and multiple orange Monascus pigment derivatives with anti-inflammatory activities were synthesized using aminophilic reaction. A total of 41 types of pigment derivatives were produced by incorporating amines and amino acids into the orange pigments. One derivative candidate that inhibited nitric oxide (NO) production in Raw 264.7 cells and exhibited low cell cytotoxicity was identified via in vitro assay. The 2-amino-4 picoline derivative inhibited NO production of 48.4%, and exhibited cell viability of 90.6%. Expression of inducible NO synthase, an important enzyme in the NO synthesis pathway, was suppressed by such a derivative in a dose-dependent manner. Therefore, this derivative has potential as a functional food colorant with anti-inflammatory effects.

Cationic cellulose nanocrystals complexed with polymeric siRNA for efficient anticancer drug delivery.

Surface-modified cellulose nanocrystals (CNCs) were developed for efficient delivery of polymeric siRNA in cancer cells. Cationic CNCs were synthesized using the sequential process of hydrothermal desulfation and chemical modification following which, polymeric siRNA obtained using from a two-step process of rolling circle transcription and Mg2+ chelation was complexed with the modified CNCs by electrostatic interaction. The complexation efficiency was optimized for high drug loading and release in the cytoplasmic environment. The resultant nanocomplex showed significantly enhanced enzymatic stability, gene knockdown efficacy, and apoptosis-induced in vitro therapeutic effect. Our results suggest CNCs as a promising carbohydrate-based delivery platform which could be utilized for RNAi-mediated cancer therapeutics.

Synthesis of high-strength microcrystalline cellulose hydrogel by viscosity adjustment.

Developing hydrogels with enhanced mechanical strength is desirable for bio-related applications. For such applications, cellulose is a notable biopolymer for hydrogel synthesis due to its inherent strength and stiffness. Here, we report the viscosity-adjusted synthesis of a high-strength hydrogel through the physical entanglement of microcrystalline cellulose (MCC) in a solvent mixture of tetrabutylammonium fluoride/dimethyl sulfoxide (TBAF/DMSO). MCC was strategically dissolved with TBAF in DMSO at a controlled ratio to induce the formation of a liquid crystalline phase (LCP), which was closely related to the viscosity of the cellulose solution. The highest viscosity was obtained at 2.5% MCC and 3.5% TBAF, leading to the strongest high-strength MCC hydrogel (strongest HS-MCC hydrogel). The resulting hydrogel exhibited a high compressive strength of 0.38MPa and a densely packed structure. Consequently, a positive linear correlation was determined between the viscosity of the cellulose solution and the mechanical strength of the HS-MCC hydrogel.

Nanoparticle-Patterned Multicompartmental Chitosan Capsules for Oral Delivery of Oligonucleotides.

Orally administered antisense therapy has been introduced as an effective approach for treating cancer in the gastrointestinal tract. However, its practical application has been limited by the instability of oligonucleotides and their inefficient delivery. To overcome these problems, we synthesized size-dependent, oligonucleotide nanoparticle-patterned chitosan/phytic acid (ODN/CS/PA) capsules with protective shields via a three-step process of self-assembly, nanoparticle encapsulation, and shell formation. The multicompartmental capsule size and oligonucleotide nanoparticle-loading pattern were controlled by applying different potentials during the electrostatic extrusion process used for nanoparticle encapsulation. Over 95% of encapsulated oligonucleotides were protected from nuclease digestion (DNase I) and, depending on their size, showed 40-75% protection against simulated gastric fluid. Their controlled release from capsules correlated with the cellular delivery of released nanoparticles and the inhibition of protein expression in cancer cells. Specifically, large capsules showed approximately 32-fold greater delivery to cancer cells than nonencapsulated nanoparticles. We also confirmed delivery of oligonucleotide nanoparticles to the small intestine and colon of rats following oral administration. These findings demonstrate that the multicompartmental ODN/CS/PA capsules can facilitate efficient oral delivery of oligonucleotides for cancer treatment.

Clinical and Biological Evaluations of Biodegradable Collagen Matrices for Glaucoma Drainage Device Implantation.

Purpose: To characterize the clinical and biological properties of biodegradable collagen matrices (BCMs) for possible glaucoma drainage device implantation. Methods: A total of 68 refractory glaucoma eyes, followed up postoperatively for at least 6 months, were consecutively enrolled after retrospective chart review. The BCM-augmented Ahmed valve implantations (BAAVI) using our Ologen-6 and Ologen-7 valves were performed and compared with a conventional method. Complete surgical success was defined as an IOP of ≤21 mm Hg (IOP 1) or ≤17 mm Hg (IOP 2) without antiglaucoma medications. Qualified success was defined as an IOP ≤21 mm Hg with or without antiglaucoma medications. The biological properties of each BCM were assessed by enzymatic degradation rates via collagenase under ocular physiological conditions. Results: The mean ages and preoperative IOPs were similar for the groups. In the conventional, BAAVI with Ologen-6, and BAAVI with Ologen-7 groups, complete success rates with target IOP 1 were 29.2%, 40.0%, and 66.7%; those with target IOP 2 were 12.5%, 30.0%, and 45.8%; qualified success rates were 45.8%, 55.0%, and 75.0%, respectively. The enzymatic degradation rate of Ologen-7 was significantly slower than that of Ologen-6 (12.5 × 10-3 vs. 28.8 × 10-3). Conclusions: The surgical success rate was highest in the Ologen-7 BAAVI group, with the lowest dependency on postoperative antiglaucoma medication use compared with the conventional and Ologen-6 BAAVI groups. The clinical results correlated with the different biological and physicochemical properties based on the degree of enzymatic degradation and on the structural morphology.

Novel derivatives of monascus pigment having a high CETP inhibitory activity.

The cholesteryl ester transfer protein (CETP), inhibition of which assists in maintaining a high level of high-density lipoprotein cholesterol in the blood, is a target for anti-atherosclerosis treatments. Orange monascus pigment was produced by a Monascus species in a 5 L jar fermenter and various derivative compounds were synthesised by incorporating 19 different L-amino acids into the orange pigment. Among them, the L-Thr and L-Tyr derivatives exhibited high inhibitory activities against the CETP reaction. The inhibitory activities of the L-Thr and L-Tyr derivatives increased in a dose-dependent manner, resulting in IC50 values of 1.0 and 2.3 µM, respectively. When CETP reactions in the presence of the derivatives were performed, the inhibition modes of the L-Thr and L-Tyr derivatives were non-competitive with inhibition constant (Ki) values of 2.7 and 4.3 µM, respectively.