Excessive serine from the bone marrow microenvironment impairs megakaryopoiesis and thrombopoiesis in Multiple Myeloma.

Thrombocytopenia is a major complication in a subset of patients with multiple myeloma (MM). However, little is known about its development and significance during MM. Here, we show thrombocytopenia is linked to poor prognosis in MM. In addition, we identify serine, which is released from MM cells into the bone marrow microenvironment, as a key metabolic factor that suppresses megakaryopoiesis and thrombopoiesis. The impact of excessive serine on thrombocytopenia is mainly mediated through the suppression of megakaryocyte (MK) differentiation. Extrinsic serine is transported into MKs through SLC38A1 and downregulates SVIL via SAM-mediated tri-methylation of H3K9, ultimately leading to the impairment of megakaryopoiesis. Inhibition of serine utilization or treatment with TPO enhances megakaryopoiesis and thrombopoiesis and suppresses MM progression. Together, we identify serine as a key metabolic regulator of thrombocytopenia, unveil molecular mechanisms governing MM progression, and provide potential therapeutic strategies for treating MM patients by targeting thrombocytopenia.

Analysis of synthesis, accumulation and physicochemical properties of Tartary buckwheat starches affected by nitrogen fertilizer.

Nitrogen fertilizer is a crucial factor affecting the growth and grain quality of Tartary buckwheat. This study was to investigate the synthesis, accumulation, and physicochemical properties of Tartary buckwheat starches under four nitrogen levels (0, 90, 180, 270 kg N ha-1). The results showed that activities of four key enzymes, starch contents all first increased and then decreased with increasing nitrogen levels, and peaked at 180 kg N ha-1. All the starches showed typical A-type, while higher nitrogen levels significantly increased the relative crystallinity. The viscosities significantly decreased, onset, peak, and conclusion first decreased and then increased, while pasting temperature and gelatinization enthalpy increased with increasing nitrogen levels. Nitrogen fertilizer and year had significant effects on the synthesis, accumulation and physicochemical properties of Tartary buckwheat starch, and the nitrogen level of 180 kg N ha-1 was more suitable for planting in the northern area of the Loess Plateau.

Characterization of Cellular Heterogeneity and an Immune Subpopulation of Human Megakaryocytes.

Megakaryocytes (MKs) and their progeny platelets function in a variety of biological processes including coagulation, hemostasis, inflammation, angiogenesis, and innate immunity. However, the divergent developmental and cellular landscape of adult MKs remains mysterious. Here, by deriving the single-cell transcriptomic profiling of MKs from human adult bone marrow (BM), cellular heterogeneity within MKs is unveiled and an MK subpopulation with high enrichment of immune-associated genes is identified. By performing the dynamic single-cell transcriptomic landscape of human megakaryopoiesis in vitro, it is found that the immune signatures of MKs can be traced back to the progenitor stage. Furthermore, two surface markers, CD148 and CD48, are identified for mature MKs with immune characteristics. At the functional level, these CD148+ CD48+ MKs can respond rapidly to immune stimuli both in vitro and in vivo, exhibit high-level expression of immune receptors and mediators, and may function as immune-surveillance cells. The findings uncover the cellular heterogeneity and a novel immune subset of human adult MKs and should greatly facilitate the understanding of the divergent functions of MKs under physiological and pathological conditions.

Diverse effects of nitrogen fertilizer on the structural, pasting, and thermal properties of common buckwheat starch.

At present, the yield of common buckwheat, which is mainly grown in northern Shaanxi of China, is low and the grain quality is poor. Nitrogen is an important nutrient for the growth of common buckwheat, and appropriate nitrogen application can improve the grain quality. Nitrogen fertilizer could alter the starch granule morphology shapes and the granule size distribution. With increasing nitrogen levels, branch number, flower clusters number, grain number per plant, contents of protein and fat, size distribution of "C" granules, and percentages of light transmittance significantly increased, whereas amylose content and retrogradation decreased. All the samples displayed typical A-type X-ray diffraction patterns. Starch showed higher pasting temperature and gelatinization enthalpy but lower trough and final viscosities under high nitrogen levels. These results suggested N2 treatment was more suitable for common buckwheat growth, principal components and correlation analysis revealed that nitrogen fertilizer significantly affected the physicochemical properties of common buckwheat starches.

Decoding Human Megakaryocyte Development.

Despite our growing understanding of embryonic immune development, rare early megakaryocytes (MKs) remain relatively understudied. Here we used single-cell RNA sequencing of human MKs from embryonic yolk sac (YS) and fetal liver (FL) to characterize the transcriptome, cellular heterogeneity, and developmental trajectories of early megakaryopoiesis. In the YS and FL, we found heterogeneous MK subpopulations with distinct developmental routes and patterns of gene expression that could reflect early functional specialization. Intriguingly, we identified a subpopulation of CD42b+CD14+ MKs in vivo that exhibit high expression of genes associated with immune responses and can also be derived from human embryonic stem cells (hESCs) in vitro. Furthermore, we identified THBS1 as an early marker for MK-biased embryonic endothelial cells. Overall, we provide important insights and invaluable resources for dissection of the molecular and cellular programs underlying early human megakaryopoiesis.

LGR4, Not LGR5, Enhances hPSC Hematopoiesis by Facilitating Mesoderm Induction via TGF-Beta Signaling Activation.

Attempts to generate functional blood cells from human pluripotent stem cells (hPSCs) remain largely unsuccessful, mainly due to the lack of understanding of the regulatory network of human hematopoiesis. In this study, we identified leucine-rich-repeat-containing G-protein-coupled receptor 4 (LGR4) as an essential regulator of early hematopoietic differentiation of hPSCs. The deletion of LGR4 severely impairs mesoderm development, thereby limiting hematopoietic differentiation both in vitro and in vivo. In contrast, LGR5 is dispensable for hPSC hematopoiesis. The four R-spondin proteins show differential activities and dependencies on LGR4 in hematopoietic differentiation. The deletion of LGR4 almost entirely abolishes the enhancement induced by R-spondin1 and R-spondin3, but not R-spondin2. In addition, ZNRF3 is required for the response of R-spondin1-R-spondin3. At the mechanistic level, LGR4 regulates transforming growth factor beta (TGF-beta) signaling to control hematopoietic differentiation. Together, our results reveal vital roles of LGR4 in hematopoietic development and uncover distinct functions and underlying mechanisms for R-spondins.

Loss of Tet2 affects platelet function but not coagulation in mice.

Ten-eleven translocation 2 (TET2) functions as a methylcytosine dioxygenase that catalyzes the iterative oxidation of 5-methylcytosine to 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine. TET2 has been shown to be crucial for the maintenance and differentiation of hematopoietic stem cells, and its deletion and/or mutations results in the expansion of HSPCs, and leads to hematological malignancies. TET2 mutations were found in a variety of hematological disorders such as CMML (60%), MDS (30%), MPN (13%) and AML (20%). Interestingly, it was shown that CMML patients with TET2 mutation exhibited fewer platelets than CMML patients without TET2 mutation. However, the role and function of TET2 in platelet hemostasis and thrombogenesis is not well defined. Here in this study, using a genetically engineered Tet2 deletion mouse model, we found that the absence of Tet2 caused a decrease in the proportion of MEP cells and hyperploid megakaryocytes. Additionally, Tet2-deficient mice displayed impaired platelet activation and aggregation under stimulation of ADP and low concentrations of thrombin, although the modestly compromised platelet function and MEP differentiation in Tet2-deficient mice could be compensated without affecting blood coagulation function. Our study indicate that Tet2 deficiency leads to mild impairment of platelet function and thrombopoiesis in mice.

Common buckwheat-resistant starch as a suitable raw material for food production: A structural and physicochemical investigation.

Heat-moisture treatment (HMT) of starch is defined as a physical method to change its properties. Compared with maize and potato, starches from common buckwheat (Xinong9976 and Pingqiao2) were isolated and its morphology and physicochemical properties investigated by scanning electron microscope (SEM), X-ray diffraction (XRD), ATR-FTIR analysis, rapid viscosity analyzer (RVA) and differential scanning calorimeter (DSC) were studied before and after HMT. The experimental results showed that there were obvious differences between native starch (NS) and resistant starch (RS) of common buckwheat. HMT altered the A-type crystalline pattern and the degree of short-range order of common buckwheat starches and significantly decreased water solubility, swelling power (70-90 °C), freeze-thaw stability and pasting properties and increased oil and water absorption capacities, light transmittance as well as thermal stability. This study shows that the NS and RS of common buckwheat can be used as the suitable raw materials in food processing.

Effects of phosphate fertiliser on the physicochemical properties of Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) starch.

Phosphate fertilisation affects the growth, development and quality of Tartary buckwheat. In this study, the effect of different phosphorus levels, including 0, 15, 75, and 135 kg/ha (non-, low-, medium-, and high-phosphorus levels, respectively), on the characteristics of starch from Tartary buckwheat were investigated in 2015 and 2017. With increased phosphorus level, the median diameter of starch granules and the apparent amylose content initially decreased and then increased. All starch samples showed the features of A-type X-ray diffraction patterns. Starches under medium-phosphorus treatment showed higher relative crystallinity than those under non-phosphorus treatment, as well as the highest solubility, gelatinisation enthalpy and transmittance among all starches. Starches under low-phosphorus treatment exhibited higher pasting properties than those under non-phosphorus treatment. This research revealed that phosphorus treatments and year significantly affected the physicochemical properties of Tartary buckwheat starch, and can provide information for the applications of starch in the food and non-food industries.

Effects of nitrogen level on the physicochemical properties of Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) starch.

Nitrogen is an essential nutrient for Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) because this substance affects the yield and quality of Tartary buckwheat. The physicochemical characteristics of starch represent the important parameters of Tartary buckwheat quality. The effects of different levels of nitrogen application at different levels (0, 45, 135 and 225kg/ha in 2015 and 0, 90, 180 and 270kg/ha in 2017) on the physicochemical characteristics of Tartary buckwheat starch were studied. The amylose content, particle size and retrogradation of Tartary buckwheat starch with nitrogen were lower than those of without nitrogen. The structure complexity, pasting temperature, gelatinization enthalpy, relative crystallinity, light transmittance and solubility of the former were higher than those of the latter. Nitrogen application did not change the 'A'-type crystalline pattern of Tartary buckwheat starch. This study indicated that nitrogen level and years and the interactions among nitrogen fertilizer levels and years significantly affected the physicochemical properties of Tartary buckwheat starch. These integrated results also provided information about the management of fertilization conditions to obtain starches with special properties for applications in food or nonfood industries.

Enhanced biosynthesis of chiral phenyllactic acid from L-phenylalanine through a new whole-cell biocatalyst.

Phenyllactic acid (PLA) is a high-value compound, which was usually produced by lactic acid bacteria (LAB) as biocatalysts and glucose or phenylpyruvic acid (PPA) as starting materials for PLA synthesis in previous studies. However, the PLA produced using LAB is a racemic mixture. Besides, both glucose and PPA were unsatisfactory substrates, as the former could not produce high concentrations of PLA while the latter is not a renewable and green substrate. To overcome these drawbacks, in this study, a new biotransformation process was developed for chiral PLA production from L-phenylalanine via the intermediate PPA using recombinant Escherichia coli co-expressing L-amino acid deaminase, NAD-dependent L-lactate dehydrogenase or NAD-dependent D-lactate dehydrogenase, and formate dehydrogenase. After optimization, the recombinant E. coli produced L- and D-PLA at concentrations of 59.9 and 60.3 mM in 6 h, respectively. Hence, this process provides an effective and promising alternative method for chiral PLA production.

Rational Design of Bacillus coagulans NL01 l-Arabinose Isomerase and Use of Its F279I Variant in d-Tagatose Production.

d-Tagatose is a prospective functional sweetener that can be produced by l-arabinose isomerase (AI) from d-galactose. To improve the activity of AI toward d-galactose, the AI of Bacillus coagulans was rationally designed on the basis of molecular modeling and docking. After alanine scanning and site-saturation mutagenesis, variant F279I that exhibited improved activity toward d-galactose was obtained. The optimal temperature and pH of F279I were determined to be 50 °C and 8.0, respectively. This variant possessed 1.4-fold catalytic efficiency compared with the wild-type (WT) enzyme. The recombinant Escherichia coli overexpressing F279I also showed obvious advantages over the WT in biotransformation. Under optimal conditions, 67.5 and 88.4 g L-1 d-tagatose could be produced from 150 and 250 g L-1 d-galactose, respectively, in 15 h. The biocatalyst constructed in this study presents a promising alternative for large-scale d-tagatose production.

A new eremophilane glycoside from the fruits of Physalis pubescens and its cytotoxic activity.

Physalis pubescens L. has been used as a traditional folk medicine in China. Comprehensive studies aimed at identifying the bioactive constituents are still lacking. In the course of searching bioactive secondary metabolites from P. pubescens L., phytochemical investigation of the fruits of P. pubescens L. led to the isolation of one new eremophilane glycoside, 1ß,3α-dihydroxy-7α-eremophila-911-dien-3-O-[α-L-arabinopyranosyl-(l→6)]-ß-D-glucopyranoside (1), and six known glycosides (2-7). Their structures were established via extensive spectroscopic data including 1D and 2D NMR and HRESIMS. Cytotoxic effects against six human cancer cell lines (C4-2B, A375, A375-S2, 786-O, Caki-2, and ACHN) and one human normal hepatic cell line (L02) were evaluated and compounds 1 and 2 exhibited moderate cytotoxicity against C4-2B and A375 cell lines with IC50 values in the range of 6.78-28.3 µM. The study indicates that the fruits of P. pubescens L. have the potential to be used for drug discovery.

Combined oridonin with cetuximab treatment shows synergistic anticancer effects on laryngeal squamous cell carcinoma: involvement of inhibition of EGFR and activation of reactive oxygen species-mediated JNK pathway.

Epidermal growth factor receptor (EGFR), a transmembrane glycoprotein, is expressed at high levels in a large proportion of laryngeal squamous cell carcinoma (LSCC). Cetuximab (Cet), an anti-EGFR monoclonal antibody, has limited clinical outcome for patients with head and neck squamous cell carcinoma. Our previous studies showed that oridonin (ORI), a natural and safe kaurene diterpenoid isolated from Rabdosia rubescens, inhibited cell growth in HEp-2 cells through inhibition of EGFR phosphorylation. The aim of the present study was to determine whether ORI could improve the anticancer efficacy of Cet on LSCC. We observed that the combination with Cet and ORI synergistically inhibited cell growth associated with Fas-mediated apoptosis and G2/M phase arrest in two LSCC cell lines (HEp-2 and Tu212 cells). Moreover, combination treatment caused cell death associated with suppression of p-EGFR and activation of reactive oxygen species (ROS)-mediated JNK pathway. In nude mice bearing HEp-2 xenografts, ORI plus Cet caused a significant tumor regression through induction of apoptosis and inhibition of proliferation with no side-effect. Together, our findings suggest that the combination of ORI and Cet has the potential to enhance tumor responses and may significantly improve therapeutic outcomes in LSCC.