Phylogenomics and biogeographical diversification of Collabieae (Orchidaceae) and its implication in the reconstruction of the dynamic history of Asian evergreen broadleaved forests.

The tribe Collabieae (Epidendroideae, Orchidaceae) comprises approximately 500 species. Generic delimitation within Collabieae are confusing and phylogenetic interrelationships within the Collabieae have not been well resolved. Plastid genomes and nuclear internal transcribed spacer (ITS) sequences were used to estimate the phylogenetic relationships, ancestral ranges, and diversification rates of Collabieae. The results showed that Collabieae was subdivided into nine clades with high support. We proposed to combine Ancistrochilus and Pachystoma into Spathoglottis, merge Collabium and Chrysoglossum into Diglyphosa, and separate Pilophyllum and Hancockia as distinctive genera. The diversification of the nine clades of Collabieae might be associated with the uplift of the Himalayas during the Late Oligocene/Early Miocene. The enhanced East Asian summer monsoon in the Late Miocene may have promoted the rapid diversification of Collabieae at a sustained high diversification rate. The increased size of terrestrial pseudobulbs may be one of the drivers of Collabieae diversification. Our results suggest that the establishment and development of evergreen broadleaved forests facilitated the diversification of Collabieae.

Genome sequence of a Sobemovirus from the feces of Northern Pintail (Anas acuta).

Northern pintail (Anas acuta) is a migratory waterfowl that can transmit various viruses. The genome sequence of a Sobemovirus was determined using metagenomic sequencing from the feces of northern pintail (Anas acuta) in Xinjiang, northwest China. The virus possesses a linear RNA molecule of 4177 bp and is most closely related to isolates SoMV-WA (GenBank accession no. HM163159.1) and ATCC PV-109 (GenBank accession no. GQ845002.2), with a nucleotide identity of 86.7%. The virus encodes four open reading frames (ORF) coding for four proteins, and phylogenetic analysis of capsid protein and RNA-dependent RNA polymerase (RdRp) showed that the strain was clustered into the species Sowbane Mosaic Virus (SoMV). The amino acid sequence identity of capsid protein was 89.6-90.9% to other isolates of SoMV, but 17.6-31.4% similar to other strains in the genus Sobemovirus, indicating a strain of Sowbane Mosaic Virus. This is the first report of SoMV in the feces of wild birds and in China, and it suggested that northern pintail likely plays an alternative role in the transmission of SoMV.

Diversification in Qinghai-Tibet Plateau: Orchidinae (Orchidaceae) clades exhibiting pre-adaptations play critical role.

We explore the origins of the extraordinary plant diversity in the Qinghai-Tibetan Plateau (QTP) using Orchidinae (Orchidaceae) as a model. Our results indicate that six major clades in Orchidinae exhibited substantial variation in the temporal and spatial sequence of diversification. Our time-calibrated phylogenetic model suggests that the species-richness of Orchidinae arose through a combination of in situ diversification, colonisation, and local recruitment. There are multiple origins of species-richness of Orchidinae in the QTP, and pre-adaptations in clades from North Temperate and alpine regions were crucial for in situ diversification. The geographic analysis identified 29 dispersals from Asia, Africa and Europe into the QTP and 15 dispersals out. Most endemic species of Orchidinae evolved within the past six million years.

Isolation and Assessment of a Highly-Active Anti-Inflammatory Exopolysaccharide from Mycelial Fermentation of a Medicinal Fungus Cs-HK1.

The purpose of this work was to fractionate the complex exopolysaccharide (EPS) from a medicinal fungus Ophiocordyceps sinensis Cs-HK1 based on the molecular weight (MW) range and to assess the in vitro anti-inflammatory activity of different EPS fractions in THP-1 cell culture. The lower MW fraction (EPS-LM-1) showed a much higher anti-inflammatory activity. EPS-LM-1 was identified as a heteropolysaccharide consisting of mannose, glucose, and galactose residues with an average MW of 360 kDa. EPS-LM-1 significantly inhibited the lipopolysaccharide-induced inflammatory responses with the effective concentrations for 50% inhibition below 5 µg/mL on a few major proinflammatory markers. With such a notable in vitro anti-inflammatory activity, EPS-LM-1 is a promising candidate for the development of a new anti-inflammation therapy.

A Comparative Study of Associated Microbiota Between Pig Farm and Pig Slaughterhouse in Guangdong, China.

The goal of this study was to compare the microbiota in different pig-present settings in China. Bioaerosol samples from pig farms and slaughterhouses and nasal samples from pig farmers and slaughterhouse workers were collected in Guangdong, southern China. The bacterial genomic DNA was isolated and subjected to 16S sequencing. The data were analyzed using QIIME2 with the DADA2 pipeline. A total of 14,923,551 clean reads and 2785 operational taxonomic units (OTUs) were obtained, which were mostly grouped into 4 phyla (Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria) and 220 families. The microbiota richness of nasal samples in pig-present workers was higher than that of bioaerosols collected in the vicinity of the pig enclosures. There were 31.7% (620/1954) shared OTUs between pig farm bioaerosols and pig farmers which was higher than that between pig slaughterhouses and slaughterhouse workers (23.4%, 364/1553) (p < 0.001). Acinetobacter and Pseudomonas were the most abundant in pig-present bioaerosols, and Staphylococcus, Pseudomonas, and Corynebacterium were dominant bacterial genus in pig farmers. The bacterial patterns are also specific to the location of sample collected. The results suggest that bioaerosol microbiota interact with human nasal microbes in the vicinity of the pig farm enclosures, providing the basis for further analysis of microbial transmission across hosts in pig-present settings.

Transcutaneous electrical acupoint stimulation for post-traumatic stress disorder: Assessor-blinded, randomized controlled study.

AIM: Transcutaneous electrical acupoint stimulation (TEAS) has the potential to alleviate post-traumatic stress disorder (PTSD). The purpose of this study was to determine whether adding TEAS to sertraline or cognitive behavioral therapy (CBT) could improve the anti-PTSD efficacy. METHODS: In this randomized controlled trial, 240 PTSD patients (60 in each group) were assigned to receive simulated TEAS combined with sertraline (group A) or with CBT (group B), active TEAS combined with CBT (group C), or active TEAS combined with CBT plus sertraline (group D) for 12 weeks. The outcomes were measured using the Clinician-Administered PTSD Scale, PTSD Check List-Civilian Version, and 17-item Hamilton Rating Scale for Depression. RESULTS: While PTSD symptoms reduced over time in all patients, groups C and D had markedly greater improvement in both PTSD and depressive measures than groups A and B in all post-baseline measurement points, with moderate to very large effect sizes of 0.484-2.244. Groups C and D also had a significantly higher rate than groups A and B on clinical response (85.0% and 95.0% vs 63.3% and 60.0%, P < 0.001) and on remission (15.0% and 25.0% vs 3.3% and 1.7%, P < 0.001). The incidence of adverse events was similar between groups A and D and between groups B and C. CONCLUSIONS: Additional TEAS augments the anti-PTSD and antidepressant efficacy of antidepressants or CBT, without increasing the incidence of adverse effects. TEAS could serve as an effective intervention for PTSD and comorbid depression. This trial was registered with www.chictr.org (no.: ChiCTR1800017255).

Three-phase partitioning as an elegant and versatile platform applied to nonchromatographic bioseparation processes.

In recent years, a big trend has been the development of rapid, green, efficient, economical, and scalable approaches for the separation and purification of bioactive molecules from natural sources, which can be used in food, cosmetics, and medicine. As a new nonchromatographic bioseparation technology, three-phase partitioning (TPP) is attracting the attention of a growing number of scientists and engineers. Although a number of studies have been published in the last 40 years regarding the extraction, separation, and purification of numerous bioactive molecules using TPP systems, a background review on TPP partitioning fundamentals and its applications is much needed. Therefore, the present review focuses in detail on the TPP separation process, including the definition of TPP, partitioning mechanisms, parameters for establishing the suitable condition to form precipitate such as concentration of ammonium sulfate, content of tert-butanol, pH and temperature, and the application for separation and purification of protein, enzyme, plant oil, polysaccharide, and other small molecule organic compounds. In addition, the possible directions of future developments in TPP technology are discussed. The review presents a good opportunity, as well as a challenge for scientists, to understand the detailed partitioning rule and to take better use of TPP for the production and separation of various bioactive molecules, which have been intensively applied in the food and medical fields.

Improving the water solubility of Monascus pigments under acidic conditions with gum arabic.

BACKGROUND: Monascus pigments (Mps) are natural food colorants and their stability in acidic solutions is important for application in the food industry. This study aimed to evaluate the use of gum arabic (GA) as a stabilizer for maintaining the solubility of Mps in an acidic aqueous solution exposed to a high temperature, and to analyze the molecular interactions between GA and Mps. RESULTS: Mps dispersed (0.2 g kg-1 ) in deionized water at pH 3.0-4.0 without GA formed precipitates but remained in a stable solution in the presence of GA (1 g kg-1 ). The significant improvement of Mps water solubility under acidic conditions was attributed to the formation of Mps-GA complexes, as indicated by a sharp increase in the fluorescence intensity. The results on particle size, zeta potential, and transmission electron microscopy further suggested that molecular binding of Mps to GA, electrostatic repulsion, and steric hindrance of GA were contributing factors to preventing the aggregation of Mps in acidic solutions. A mechanistic model was presented for GA-Mps interactions and complex structures. CONCLUSION: GA was proven to be an effective stabilizer of natural food colorants in acidic solutions. © 2016 Society of Chemical Industry.

Synthesis, antitumor evaluation and molecular docking studies of [1,2,4]triazolo[4,3-b][1,2,4,5]tetrazine derivatives.

A series of [1,2,4]triazolo[4,3-b][1,2,4,5]tetrazine derivatives have been synthesized and evaluated for their antitumor activities. These compounds exhibited potent antiproliferative activities against MCF-7, Bewo and HL-60 cells and c-Met kinase inhibitory activities. Three compounds were highly effective against MCF-7, Bewo and HL-60 cells with IC50 values in 1.09-2.24µM. Molecular docking was further performed to study the inhibitor-c-Met kinase interactions, and the results show that compound 4j was potently bound to the c-Met kinase with three hydrogen bonds. The further research on acute toxicity and in vivo antitumor activity of compound 4j to ICR (Institute of Cancer Research) mice were carried out, and found 4j with a certain toxicity but good efficacy in vivo. Based on the preliminary results, it is deduced that compound 4j with potent c-Met kinase inhibitory activity may be a potential anticancer agent.

Formation and Physiochemical Properties of Silver Nanoparticles with Various Exopolysaccharides of a Medicinal Fungus in Aqueous Solution.

Natural polysaccharides are the most widely used biopolymers for green synthesis of eco-friendly silver nanoparticles (AgNPs). In a previous study, a high molecular weight (MW) fraction of exopolysaccharides (EPS) produced by a medicinal fungus Cs-HK1 has been shown useful for green and facile synthesis of AgNPs in water. This study was to further evaluate the effects of molecular properties of EPS on the formation, stability and properties of AgNPs with different EPS fractions at various pH conditions. Three EPS fractions (P0.5, P2.0 and P5.0: MW high to low and protein content low to high) were reacted with silver nitrate at various pH 3.0-8.0 in water. The most favorable pH range was 5.5-8.0 for the formation and stable dispersion of AgNPs. At a given pH, the maximum amount of AgNPs was produced with P5.0, and the minimum with P0.5. The shape, size and physiochemical properties of AgNPs were strongly affected by the molecular characteristics of EPS (MW and conformation). The results may be helpful for understanding the factors and mechanisms for formation of stable AgNPs with natural polysaccharides and the interactions between AgNPs and the polysaccharide hydrocolloids in water.

[Pharmacokinetics Study on Curcumin Hydroxypropyl-ß-Cyclodextrin Phospholipid Complex in Rats].

OBJECTIVE: To compare the in vivo pharmacokinetics of curcumin hydroxypropyl-ß-cyclodextrin phospholipid complex, curcumin hydroxypropyl-ß-cyclodextrin and curcumin phospholipid complex, and to discuss the advantage of hydroxypropyl-ß-cyclodextrin phospholipid complex as carrier. METHODS: Drawing blood after SD rats were oral administered with the above preparations and free drug at 50 mg/kg( corresponding to curcumin) , and the blood concentration were determined by HPLC. RESULTS: The AUC0-∞ of curcumin hydroxypropyl-ß-cyclodextrin phospholipid complex was(1 126. 20 ± 323. 24) g/(L . h), which was 5. 89, 1. 49 and 1. 17 times as curcumin (191. 08 ± 43. 27) µg/( L . h), curcumin phospholipid complex(754. 93 ± 55. 33) µg/(L . h), curcumin hydroxypropyl-ß- cyclodextrin(961. 21 ± 253. 65) µg/(L . h). CONCLUSION: The curcumin hydroxypropyl-ß-cyclodextrin phospholipid complex has a better absorption property than curcumin phospholipid complex and curcumin hydroxypropyl-ß-cyclodextrin, which is more beneficial to improve the bioavailability.

Optimization of exopolysaccharide production in submerged culture of Daedalea dickinsii and its antioxidant activity.

The major nutrients, pH and temperature were evaluated for the exopolysaccharide (EPS) production by Daedalea dickinsii in submerged culture to derive an optimal medium composition and conditions as follows: 50 g/L maltose, 5 g/L soy peptone, 5 mM CaCl(2), at pH 6.0 and 28 °C. A purified EPS fraction was attained from gel filtration chromatography and its major molecular characteristics were determined. FT-IR spectral analysis revealed the prominent characteristic groups of polyhydric alcohols. GC analysis and NMR spectrum showed its major molecular composition of glucose and galactose. Furthermore, thermogravimetric analysis indicated its degradation temperature (T(d)) of 189 °C. The antioxidant activity of the EPS fraction showed a correlation with the molecular properties. It might be attributed to the functional groups in the EPS fraction, which can donate electrons to reduce the radicals to a more stable form or react with the free radicals to terminate the radical chain reaction.

Isolation and structure characterization of an antioxidative glycopeptide from mycelial culture broth of a medicinal fungus.

A novel glycopeptide (Cs-GP1) with an average molecular weight (Mw) of 6.0 kDa was isolated and purified by column chromatography from the lower Mw fraction of exopolysaccharide (EPS) produced by a medicinal fungus Cordyceps sinensis Cs-HK1. Its carbohydrate moiety was mainly composed of glucose and mannose at 3.2:1.0 mole ratio, indicating an O-linked glycopeptide. The peptide chain contained relatively high mole ratios of aspartic acid, glutamic acid and glycine (3.3-3.5 relative to arginine) but relatively low ratios of tyrosine and histidine. The peptide chain sequence analyzed after trypsin digestion by LC-MS was KNGIFQFGEDCAAGSISHELGGFREFREFLKQAGLE. Cs-GP1 exhibited remarkable antioxidant capacity with a Trolox equivalent antioxidant capacity of 1183.8 µmol/g and a ferric reducing ability of 611.1 µmol Fe(II)/g, and significant protective effect against H2O2-induced PC12 cell injury at a minimum dose of 10 µg/mL. This is the first report on the structure and bioactivity of an extracellular glycopeptide from the Cordyceps species.

[Amplicon density-weighted algorithms for analyzing dissimilarity and dynamic alterations of RAPD polymorphisms of Cordyceps sinensis].

OBJECTIVE: To examine the dynamic maturational alterations of random amplified polymorphic DNA (RAPD) molecular marker polymorphism resulted from differential expressions of multiple fungi in the caterpillar body, stroma and ascocarp portion of Cordyceps sinensis (Cs). METHODS: Used the fuzzy, integral RAPD molecular marker polymorphism method with 20 random primers; used density-weighted cluster algorithms and ZUNIX similarity equations; compared RAPD polymorphisms of the caterpillar body, stroma and ascocarp of Cs during maturation; and compared RAPD polymorphisms of Cs and Hirsutella sinensis (Hs). RESULTS: Density-unweighted algorithms neglected the differences in density of the DNA amplicons. Use of the density-weighted ZUNIX similarity equations and the clustering method integrated components of the amplicon density differences in similarity computations and clustering construction and prevented from the loss of the information of fungal genomes. An overall similarity 0.42 (< the overall dissimilarity 0.58) was observed for all compartments of Cs at different maturation stages. The similarities for the stromata or caterpillar bodies of Cs at 3 maturational stages were 0.57 or 0.50, respectively. During Cs maturation, there were dynamic Low→High→Low alterations of the RAPD polymorphisms between stromata and caterpillar bodies dissected from the same pieces of Cs. The polymorphic similarity was the highest (0.87) between the ascocarp and mature stroma, forming a clustering clade, while the premature stroma and caterpillar body formed another clade. These 2 clades merged into one cluster. Another clade containing the maturing stroma and caterpillar body merged with mature caterpillar body, forming another cluster. The RAPD polymorphic similarities between Hs and Cs samples were 0.55-0.69. Hs were separated from Cs clusters by the out-group control Paecilomyces militaris. CONCLUSION: The wealthy RAPD polymorphisms change dynamically in the Cs compartments with maturation. The different RAPD polymorphism for Hs from those for Cs supports the hypothesis of integrated micro-ecosystem Cs with multiple fungi, but does not support the "single fungal species" hypothesis for Cs and the anamorph-teleomorph connection between Hs and Cs.

Pathogen Discovery in the Post-COVID Era.

Pathogen discovery plays a crucial role in the fields of infectious diseases, clinical microbiology, and public health. During the past four years, the global response to the COVID-19 pandemic highlighted the importance of early and accurate identification of novel pathogens for effective management and prevention of outbreaks. The post-COVID era has ushered in a new phase of infectious disease research, marked by accelerated advancements in pathogen discovery. This review encapsulates the recent innovations and paradigm shifts that have reshaped the landscape of pathogen discovery in response to the COVID-19 pandemic. Primarily, we summarize the latest technology innovations, applications, and causation proving strategies that enable rapid and accurate pathogen discovery for both acute and historical infections. We also explored the significance and the latest trends and approaches being employed for effective implementation of pathogen discovery from various clinical and environmental samples. Furthermore, we emphasize the collaborative nature of the pandemic response, which has led to the establishment of global networks for pathogen discovery.

Anti-bacterial effects, and metabolites derived from bifidobacterial fermentation of an exopolysaccharide of Cs-HK1 medicinal fungus.

This study was to investigate the antibacterial effects and metabolites derived from bifidobacterial fermentation of an exopolysaccharide EPS-LM produced by a medicinal fungus Cordyceps sinensis, Cs-HK1. EPS-LM was a partially purified polysaccharide fraction which was mainly composed of Man, Glc and Gal at 7.31:12.95:1.00 mol ratio with a maximum molecular weight of 360 kDa. After fermentation of EPS-LM in two bifidobacterial cultures, B. breve and B. longum, the culture digesta showed significant antibacterial activities, inhibiting the proliferation and biofilm formation of Escherichia coli. Based on untargeted metabolomic profiling of the digesta, the levels of short chain fatty acids, carboxylic acids, benzenoids and their derivatives were all increased significantly (p < 0.01), which probably contributed to the enhanced antibacterial activity by EPS-LM. Since EPS-LM was only slightly consumed for the bifidobacterial growth, it mainly stimulated the biosynthesis of bioactive metabolites in the bifidobacterial cells. The results also suggested that EPS-LM polysaccharide may have a regulatory function on the bifidobacterial metabolism leading to production of antibacterial metabolites, which may be of significance for further exploration.

Enhancement of chitosan-based film physicochemical and storage properties by interaction with proanthocyanidin and natural deep eutectic solvent.

Deep eutectic solvent (DES) has been recognized as a promising plasticizer for the preparation of biodegradable food packaging films. In addition, DES-plasticized chitosan (CS) films could also serve as a favorable carrier for loading active components. In this work, a ternary composite film was fabricated by plasticizing chitosan with DES and the active ingredient proanthocyanidin (PA) was used as a cross-linking agent. The incorporation of PAs significantly enhanced the toughness, elasticity, and hydrophobicity of the ternary CS-DES-PA composite films. It achieved antioxidant and bacteriostatic functions. In particular, the ternary CS-DES-PA composite films had a thickness of 0.16 ± 0.01 µm, a tensile strength of 2.63 ± 0.48 MPa, and an elongation about 73.22 %. They also have improved water resistance, UV blocking, with a high-water contact angle of 88.4° and a low water swelling of 5 % on the surface of the film. Meanwhile, the PAs in the film could slow down the browning of litchi fruits. This ternary blended film (CS-DES-PA) achieves better compatibility of the active ingredient in the film-forming substrate. It also provides a green and biodegradable packaging material for food packaging.

Xantolisweimingii (Sapotaceae), a new species from the Yuanjiang River basin, Yunnan, southwest China.

Xantolisweimingii sp. nov. (Sapotaceae) is described and illustrated from Yunnan, southwest China. The new species is morphologically most similar to X.tomentosa (Roxb.) Raf., but differs from the latter in the ovate or obovate leaves, entirely glabrous corollas, lanceolate, ca. 5 mm long staminodes, fringed at the base. We provided a distribution map and a preliminary conservation assessment for the new species. Additionally, an updated dichotomous key to all known species of Xantolis is presented.

Fractionation, characterization, and assessment of nutritional and immunostimulatory protein-rich polysaccharide-protein complexes isolated from Lentinula edodes mushroom.

This study aimed to fractionate and characterize the protein-rich polysaccharide-protein (PSP) complexes from a well-known edible mushroom, Lentinula edodes, and assess their nutritional and immunostimulatory properties. Crude PSP isolated from the mushroom water extract was purified by anion exchange chromatography, yielding fractions PSP-F1 and PSP-F2 containing 66.1 % and 74.0 % protein, respectively. Both fractions exhibited primarily ß-sheet and random-coil protein structures, though the crude PSP fraction exhibited an additional α-helix structure. On SDS-PAGE, PSP-F1 showed two molecular weight bands, one below 10 kDa and another at 34 kDa, and PSP-F2 showed several bands, one below 10 kDa and others between 34 and 95 kDa. The nutritional value of essential and non-essential amino acid profiles was in the order of PSP-F2 > PSP-F1 > crude PSP; the amino acid ratio coefficient values of the crude PSP, PSP-F1, and PSP-F2 were 63 %, 67 %, and 72 %, respectively. The combination of PS and PSP fractions exhibited stronger immunoactivity than PSP-F1 or PSP-F2 alone. PSP-F2 showed a higher immunostimulatory activity than PSP-F1 in RAW264.7 cell culture. PSP-F2 was also more abundant of easily absorbed high-quality proteins. The results provide useful references for dietary and medicinal uses of PSP fractions in L. edodes and other edible mushrooms.