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Sulfated polysaccharides from sea cucumber Stichopus japonicus (SCSPsj) and Lactobacillus gasseri synergistically ameliorate the overweight in gut microbiota-altered mice through improving gut microbiota dysbiosis and metabolic disorders.

Sea cucumber sulfated polysaccharides (SCSPsj) enhance the relative abundance of Lactobacillus by promoting biofilm formation and mucus binding, and further regulate the metabolism in a humanized microbiota mice model.

It has been proposed that the proportions of the human face are crucial for facial aesthetics. If this is the case, we should describe the relationship among proportions of face components quantitatively. This study aims to develop a mathematical model of facial proportions to provide a quantitative description of facial attractiveness. Furthermore, we expect that plastic surgeons can use models in clinical work to enhance communication efficiency between doctors and patients. Face alignment technique was used to analyse 5500 frontal faces with diverse properties (male/female, Asian/Caucasian, ages) to obtain the ratios among the nose length ($ {N}_{L} $), the nasal base width ($ N $), and the inner canthus width ($ {E}_{I} $). A mathematical model ($ {N}_{L}^{2} = a{E}_{I}\mathrm{*}{N}_{L}+b{E}_{I}\mathrm{*}N+cN\mathrm{*}{N}_{L} $) was developed to describe the relationship among these proportions. To validate the effectiveness of this approach, we simulated the post-operative photos using Adobe Photoshop. Our findings show that the ratio of nose length to nose width, the ratio of inner canthus width to nose length and the ratio of inner canthus to nose width play a significant role in determining facial attractiveness. These results provide a possible strategy to quantitatively describe the relationship among human face proportions.

Background The kidney function change after Transcatheter Aortic Valve Replacement (TAVR) in diabetes and/or hypertension patients is currently unknown. Methods A total of 242 severe Aortic Stenosis patients with diabetes mellitus and/or hypertension who underwent TAVR were analyzed. Patients were categorized into 3 groups based on the percent change [(discharge eGFR – baseline eGFR) x 100/baseline eGFR] in eGFR: improved ≥ 10%, no change, and declined ≥ 10%. Results Of these patients, 93 (38.4%) had an improvement in eGFR ≥ 10%, 117 (48.3%) had no changes, and 32 (13.2%) had a decline in eGFR of more than 10%. Patients with unchanged kidney function may be with lower STS scores (5.09 [3.58–8.34] % vs. 7.04 [4.52–9.91] % vs. 7.41 [3.52–10.97] %, p = 0.003), and lower perivascular disease (14.5% vs. 26.9% vs. 40.6%, p = 0.003) when compared with improved and declined kidney function patients, respectively. Patients with improved kidney function had lower eGFR (45.0 [33.9–60.3] mL/min/1.73 m2 vs. 57.4 [43.4–70.7] mL/min/1.73 m2 vs. 56.6 [44.2–76.8] mL/min/1.73 m2, p = 0.000), and lower LVEF (55.1 [40.9–62.0] % vs. 60.4 [53.4–66.1] % vs. 59.9 [52.9–67.8] %, p = 0.002) than the unchanged or declined group, respectively. Moreover, patients with worsening kidney function had higher rate of in-hospital (12.5% vs. 1.7% vs. 0%, p = 0.002) and 30-day (15.6% vs. 1.7% vs. 0%, p = 0.000) mortality. Independent predictors of eGFR changes were STS scores, LVEF, baseline eGFR, and unplanned cardiopulmonary bypass. Conclusions Nearly 40% of diabetic and/or hypertensive patients experience an intermediate improvement after TAVR. Patients with declining renal function suffer more terrible outcomes in mortality.

Saccharomyces cerevisiae is commonly used as a starter culture for alcoholic beverages but is less applied in meat products. In this study, the effects of S. cerevisiae LXPSC1 on sour meat during fermentation were investigated. It was found that samples inoculated with S. cerevisiae LXPSC1 (Sc group) had better sensory characteristics and higher levels of pH, ethanol, free amino acids (FAAs), volatile organic compounds (VOCs). The bacterial communities of both groups were dominated by Lactobacillus sakei, Weissella hellenica, and Lactobacillus plantarum, which might play a role in reducing pH and a

Membrane fouling and wetting limit the applications of membrane distillation (MD) for wastewater treatment, especially when treating the wastewater with a high concentration of low surface tension substances such as oil and surfactants. In this paper, virgin polyvinylidene fluoride (PVDF) membrane was modified by polydimethylsiloxane (PDMS) to enhance anti-wetting ability. Then a thin polydopamine (PDA) layer was coated as a reaction platform for further modification. Polyethyleneimine (PEI) was cross-linked with PDA to form a uniform and stable layer, through hydrogen bonds and electrostatic interaction to immobilize hydrophilic MXene, which formed a Janus MXene-PVDF membrane. The MXene layer was the key for superoleophobicity and high liquid entry pressure (LEP) of membrane, capable of mitigating membrane fouling and wetting when dealing with low surface tension wastewater (LSTW). From the experiments results, pristine PVDF membrane showed severe fouling and wetting with flux decline and salt leakage during treatment of LSTW (surfactants containing water, oil-in-water emulsion and sodium dodecyl sulfate stabilized oil-in-water emulsion). However, under the same conditions, the Janus MXene-PVDF membrane exhibited remarkably stable flux (9.3 kg m

To target a carotenoid biosynthetic gene in the oleaginous yeast Rhodosporidium toruloides by using the Agrobacterium-mediated transformation (AMT) method.The RHTO_04602 locus of R. toruloides NP11, previously assigned to code the carotenoid biosynthetic gene CRTI, was amplified from genomic DNA and cloned into the binary plasmid pZPK-mcs, resulting in pZPK-CRT. A HYG-expression cassette was inserted into the CRTI sequence of pZPK-CRT by utilizing the restriction-free clone strategy. The resulted plasmid was used to transform R. toruloides cells according to the AMT method, leading to a few white transformants. Sequencing analysis of those transformants confirmed homologous recombination and insertional inactivation of CRTI. When the white variants were transformed with a CRTI-expression cassette, cells became red and produced carotenoids as did the wild-type strain NP11.Successful homologous targeting of the CrtI locus confirmed the function of RHTO_04602 in carotenoids biosynthesis in R. toruloides. It provided valuable information for metabolic engineering of this non-model yeast species.

Rhodosporidium toruloides has been reported as a potential biotechnological microorganism to produce carotenoids. The most commonly used molecular and genetic manipulation methods based on Agrobacterium-mediated transformation (ATMT). However, this method was of relatively lower transformation efficiency. In this study, we optimized the ATMT method for R. toruloides on account of the promoter on T-DNA, the ratio of A. tumefaciens to R. toruloides NP11, acetosyringone concentration, cocultivation temperature and time, and a transformation efficiency of 2,369 cells per 105 recipient cells was obtained and was 24 times as that of the previous report. With this optimized method, four redder mutants and four yellower mutants were selected out with torularhodin and β-carotene production preference, respectively. The highest torularhodin production was 1,638.15 µg/g dry cell weight in A1-13. The yellower mutants were found to divert the metabolic flux from torularhodin and torulene to γ-carotene and β-carotene, and the proportion of γ-carotene and β-carotene were all over 92%. TAIL-PCR was carried out to found T-DNA insertion in these mutants, and insertion hotspot was found. RT-qPCR results showed that CTA1 genes in these mutants were closely related to the synthesis of total carotenoids, especially torularhodin, and was a potenial metabolic engineering site in the future.

Monodisperse metal clusters provide a unique platform for investigating magnetic exchange within molecular magnets. Herein, the core-shell structure of the monodisperse molecule magnet of [Gd52 Ni56 (IDA)48 (OH)154 (H2 O)38 ]@SiO2 (1 a@SiO2 ) was prepared by encapsulating one high-nuclearity lanthanide-transition-metal compound of [Gd52 Ni56 (IDA)48 (OH)154 (H2 O)38 ]⋅(NO3 )18 ⋅164 H2 O (1) (IDA=iminodiacetate) into one silica nanosphere through a facile one-pot microemulsion method. 1 a@SiO2 was characterized using transmission electron microscopy, N2 adsorption-desorption isotherms, and inductively coupled plasma-atomic emission spectrometry. Magnetic investigation of 1 and 1 a revealed J1 =0.25 cm(-1) , J2 =-0.060 cm(-1) , J3 =-0.22 cm(-1) , J4 =-8.63 cm(-1) , g=1.95, and z J=-2.0×10(-3) cm(-1) for 1, and J1 =0.26 cm(-1) , J2 =-0.065 cm(-1) , J3 =-0.23 cm(-1) , J4 =-8.40 cm(-1) g=1.99, and z J=0.000 cm(-1) for 1 a@SiO2 . The z J=0 in 1 a@SiO2 suggests that weak antiferromagnetic coupling between the compounds is shielded by silica nanospheres.

Flavourzyme is known to promote protein decomposition, resulting in more peptides and amino acids which can improve the quality of fermented foods. In this study, the effects of flavourzyme addition on the fermentation of Suanzhayu fish were investigated. The results showed that the addition of 50 U/g flavourzyme reduced the water activity (aw) of products and promoted the release of trichloroacetic acid (TCA)-soluble peptides and free amino acids (FAAs). Thus, the stability of the product was improved and its nutritional value was increased. In addition, with the addition of flavourzyme, Lactobacillus and Saccharomyces more quickly became the dominant genera in the fermentation. Furthermore, the formation of alcohols, aldehydes, and esters was promoted in flavourzyme addition group. Redundant analysis (RDA) indicated that Lactobacillus and Lactococcus play important roles in the formation of flavors, especially for the characteristic flavors of Suanzhayu. Flavourzyme addition may be a novel method to greatly improve the properties of Suanzhayu and shorten the fermentation time.

Lipid droplets (LDs) are ubiquitous organelles that serve as a neutral lipid reservoir and a hub for lipid metabolism. Manipulating LD formation, evolution, and mobilization in oleaginous species may lead to the production of fatty acid-derived biofuels and chemicals. However, key factors regulating LD dynamics remain poorly characterized. Here we purified the LDs and identified LD-associated proteins from cells of the lipid-producing yeast Rhodosporidium toruloides cultured under nutrient-rich, nitrogen-limited, and phosphorus-limited conditions. The LD proteome consisted of 226 proteins, many of which are involved in lipid metabolism and LD formation and evolution. Further analysis of our previous comparative transcriptome and proteome data sets indicated that the transcription level of 85 genes and protein abundance of 77 proteins changed under nutrient-limited conditions. Such changes were highly relevant to lipid accumulation and partially confirmed by reverse transcription-quantitative PCR. We demonstrated that the major LD structure protein Ldp1 is an LD marker protein being upregulated in lipid-rich cells. When overexpressed in Saccharomyces cerevisiae , Ldp1 localized on the LD surface and facilitated giant LD formation, suggesting that Ldp1 plays an important role in controlling LD dynamics. Our results significantly advance the understanding of the molecular basis of lipid overproduction and storage in oleaginous yeasts and will be valuable for the development of superior lipid producers.

Directed evolution-based protein engineering usually generates large library contained insoluble mutants because of structural disturbance by mutation. To reduce the workload and costs, it is crucial to identify and eliminate those insoluble variants prior to dedicated analysis. Here, we demonstrate a method to visualize soluble protein mutants by using monomeric red fluorescent protein (mRFP) as a fusion tag. A plasmid was devised to express nicotinic acid mononucleotide adenylyltransferase (NadD) fused with a GGGS-linked mRFP tag at the C-terminus. The plasmid was subjected to site saturation mutagenesis within the nadD gene, used to transform Escherichia coli DH10B competent cells, leading to colonies with different red intensities. It was found that the fluorescence intensity of the cell culture correlated positively with the content of NadD-mRFP mutant in the supernatant. Mutation at position 132 led to a library of which most colonies lost the red phenotype, indicating that the position had a key role for proper protein folding. Similarly, mRFP enabled identification of soluble mutants of other enzymes including 1-deoxy-D-xylulose-5-phosphate reductoisomerase and phosphite dehydrogenase. These data suggested that mRFP can serve as a fusion reporter for visualizing soluble protein mutants to facilitate more efficient library screening in directed evolution.

Limiting nitrogen supply has been routinely used as the master regulator to direct lipid biosynthesis. However, this strategy does not work with nitrogen-rich substrates, such as Jerusalem artichoke (JA), a fructose-based biomass, while it is difficult to obtain a high carbon-to-nitrogen (C/N) molar ratio. In this study, an alternative strategy to promote lipid accumulation by the oleaginous yeast Trichosporon fermentans CICC 1368 was developed by limiting phosphorous supply, and this strategy was implemented with JA hydrolysate as substrate. We showed that lipid accumulation was directly correlated with the C/P ratio of the culture media for T. fermentans. The time course of cell growth and lipid production was analyzed in a media with an initial C/P ratio of 6342, and the cellular lipid content could reach up to 48.5% of dry biomass. Moreover, JA hydrolysates were used as substrate for microbial lipid accumulation, under high C/P molar ratio condition, lipid yield, lipid content, and lipid coefficient increased by 10, 30, and 34%, respectively. It showed that by limiting phosphorus, the conversion of sugar into lipids can be improved effectively. Limiting phosphorus provides a promising solution to the problem of microbial lipid production with nitrogen-rich natural materials.

Oleaginous yeast Lipomyces starkeyi, a promising strain of great biotechnical importance, is able to accumulate over 60% of its cell biomass as triacylglycerols (TAGs). It is promising to directly produce the derivatives of TAGs, such as long-chain fatty acid methyl esters and alkanes, in L. starkeyi. However, techniques for genetic modification of this oleaginous yeast are lacking, thus, further research is needed to develop genetic tools and functional elements. Here, we used two exogenous promoters (pGPD and pPGK) from oleaginous yeast Rhodosporidium toruloides to establish a simpler Agrobacterium-mediated transformation (AMT) method for L. starkeyi. Hygromycin-resistant transformants were obtained on antibiotic-contained plate. Mitotic stability test, genotype verification by PCR, and protein expression confirmation all demonstrated the success of this method. Furthermore, the strength of these two promoters was evaluated at the phenotypic level on a hygromycin-gradient plate and at the transcriptional level by real-time quantitative PCR. The PGK promoter strength was 2.2-fold as that of GPD promoter to initiate the expression of the hygromycin-resistance gene. This study provided an easy and efficient genetic manipulation method and elements of the oleaginous yeast L. starkeyi for constructing superior strains to produce advanced biofuels.

This study was designed to explore the temperature effects on bacterial communities and metabolites, as well as their relationships during the fermentation of sour meat, a traditional fermented meat product in the ethnic minority regions of China. Results showed that reduction of pH and increase of lactic acid and free amino acid contents occurred (p 0.05) as the fermentation temperature and time increased, and the tendency was more apparent at higher temperature. During the fermentation, Lactobacillus gradually replaced other genera, and higher the temperature, more rapid was the process. Both the number and amount of volatile organic compounds increased at higher temperatures. Hexanal, benzaldehyde, nonanal, (E,E)-2,4-decadienal, 1-octen-3-ol and octanal were identified as the key volatile organic compounds produced by Lactobacillus in sour meat as main contributors to odor as confirmed by variable importance in the projection analysis. Redundancy analysis and Pearson correlation showed positive correlation between Lactobacillus and desired product characteristics, such as higher content of lactic acid, free amino acids, volatile organic compounds, and lower pH and water activity values, which may represent a better quality and longer shelf life after fermentation at higher temperature. Therefore fermentation at 20 °C and 25 °C are proposed as optimum temperatures for sour meat production.

Early biochemical studies have demonstrated that lipid accumulation by oleaginous yeasts is linked to the activity of the NAD(+)-dependent isocitrate dehydrogenase (Idh). However, molecular study of Idh of oleaginous microorganisms remains limited. Here, we present the cloning of a mitochondrial NAD(+)-specific Idh from Rhodosporidium toruloides (RtIdh), an excellent microbial lipid producer that uses carbohydrates as the carbon source. The evolutionary relationship analyses among RtIdhs and other yeast Idhs revealed that RtIdh had a closer relationship with the Idhs of Ustilago maydis and Schizophyllum commune. We expressed the RtIDH gene in the yeast Saccharomyces cerevisiae idhΔ mutant. Under the nitrogen-limited condition, the intracellular lipid content and extracellular citrate concentration of the culture of the S. cerevisiae idhΔ carrying the RtIDH gene increased as the carbon/nitrogen molar ratio of the media increased, while the wild-type S. cerevisiae strain showed no correlation. Our data provided valuable information for elucidating the molecular mechanism of microbial oleaginicity and for engineering microorganisms to produce metabolites of fatty acid pathway.

The basidiomycetous yeast Rhodosporidium toruloides represents an excellent producer for microbial lipids and carotenoids. However, further rational engineering of this unconventional yeast remains challenging partially because of the absence of efficient and reliable transformation method. In this study, we developed an Agrobacterium-mediated transformation (ATMT) protocol for effective gene integration into the R. toruloides genome. Both haploid and diploid strains were successfully modified, and the integration was confirmed by colony PCR, Western blot analysis and genome walking. We further demonstrated that multiple genes could be integrated by consecutive ATMT, leading to engineered strains simultaneously resistant to multiple antibiotics. Our results provided a practical method for functional integration and expression of exogenous genes in R. toruloides, which should facilitate the development of genetic tools and the construction of superior strains to produce biofuel molecules and biochemicals.

Recent studies have suggested that synovium-derived mesenchymal stem cells (SMSCs) may be promising candidates for tissue engineering and play an important role in cartilage regeneration. However, the mechanisms of SMSC chondrogenesis remain to be identified and characterized. The aim of this study was to evaluate the activation of the RhoA/Rho kinase (ROCK) pathway, as well as the manner by which it may contribute to chondrogenesis and the actin cytoskeletal organization of rat temporomandibular SMSCs in response to transforming growth factor-β1 (TGF-β1). Primary isolated SMSCs were treated with TGF-β1, and their actin organization was examined by fluorescein isothiocyanate-phalloidin staining. The specific biochemical inhibitors, C3 transferase, Y27632 and SB431542, were employed to evaluate the function of RhoA/ROCK and Smads. The effect of C3 transferase and Y27632 on the gene expression of chondrocyte-specific markers was evaluated by quantitative real-time polymerase chain reaction. To examine the effect of Y27632 on Smad2/3 phosphorylation induced by TGF-β1, western blot analysis was also performed. The stimulation of TGF-β1 in SMSCs resulted in the activation of the RhoA/ROCK pathway and concomitantly induced cytoskeletal reorganization, which was specifically blocked by C3 transferase and Y27632. The TGF-β-induced gene expression of Sox9, type I collagen, type II collagen and aggrecan was also inhibited by both C3 transferase and Y27632, at different levels. Y27632 treatment reduced the phosphorylation of Smad2/3 in a concentration-dependent manner. These results demonstrate the RhoA/ROCK activation regulates chondrocyte-specific gene transcription and cytoskeletal organization induced by TGF-β1 by interacting with the Smad pathway. This may have significant implications for the successful utilization of SMSCs as a cell source for articular cartilage tissue engineering.

CdBiO2Cl is synthesized by solid state reaction of BiOCl and CdO (alumina crucible, 700 °C, 24 h) and characterized by powder XRD, UV/VIS diffuse reflectance spectroscopy, and TB-LMTO band structure calculations.

A novel visible-light-driven layered photocatalyst of Bi-based PbBiO2Cl is prepared by solid-state reaction. The optical band gap of PbBiO2Cl is determined to be 2.45 eV by UV–vis diffuse reflectance spectroscopy. Generally, the photocatalytic activity for degrading methyl orange (MO) over PbBiO2Cl is higher than that over anatase-type TiO2 under UV light and visible light illumination. The RuO2 loading over PbBiO2Cl leads to an obvious increase in photocatalytic performance. The internal electric fields between [PbBiO2] and [Cl] slabs are considered to be useful for the efficient separation of electron–hole pairs upon photoexcitation.

A Bi-based oxychloride Na 0.5 Bi 1.5 O 2 Cl with a layered structure as a novel efficient photocatalyst was studied in the present paper. The powder was synthesized by a solid state reaction method. It was characterized by X-ray diffraction, scanning electron microscope and UV–vis diffuse reflectance spectrum. Degradation of methyl orange was used to evaluate the photocatalytic activity. The as-synthesized Na 0.5 Bi 1.5 O 2 Cl has a smaller optical band gap of 3.04 eV than BiOCl ( E g = 3.44 eV). It possesses a fair visible-light-response ability. The UV-induced photocatalytic activity follows the decreasing order of BiOCl > Na 0.5 Bi 1.5 O 2 Cl > TiO 2 , different from the order of Na 0.5 Bi 1.5 O 2 Cl > TiO 2 > BiOCl under visible light irradiation. The dispersion of Pt over Na 0.5 Bi 1.5 O 2 Cl leads to an obvious increase in the photocatalytic performance. The internal electric fields between [Na 0.5 Bi 1.5 O 2 ] and [Cl] slabs favor the efficient separation of photostimulated electron–hole pairs.

A Bi-based oxychloride Bi(3)O(4)Cl with a layered structure as a novel efficient photocatalyst was studied in the present paper. The compound synthesized by a solid-state reaction method has a band gap of 2.79 eV. The material possesses a fair visible-light-induced photocatalytic activity. Generally, the photocatalytic efficiency of Bi(3)O(4)Cl for degrading methyl orange (MO) is higher than that of anatase TiO(2) under UV light illumination. The dispersion of Ag over Bi(3)O(4)Cl leads to an obvious increase in the photocatalytic performance. The MO decolorization over Bi(3)O(4)Cl is mainly initiated by a photocatalytic process. The photocatalytic activity is discussed in close connection with the crystal structure and the electronic structure in details.