Inhibitory role of bone marrow mesenchymal stem cells-derived exosome in non-small-cell lung cancer: microRNA-30b-5p,EZH2 and PI3K/AKT pathway

Exosomal microRNA (miRNA) exerts potential roles in non-small-cell lung cancer (NSCLC). The current study elucidated the role of miR-30b-5p shuttled by bone marrow mesenchymal stem cells (BMSCs)-derived exosomes in treating NSCLC. Bioinformatics analysis was performed with NSCLC-related miRNA microarray GSE169587 and mRNA data GSE74706 obtained for collection of the differentially expressed miRNAs and mRNAs. The relationship between miR-30b-5p and EZH2 was predicted and confirmed. Exosomes were isolated from BMSCs and identified. BMSCs-derived exosomes overexpressing miR-30b-5p were used to establish subcutaneous tumorigenesis models to study the effects of miR-30b-5p, EZH2 and PI3K/AKT signalling pathway on tumour growth. A total of 86 BMSC-exo-miRNAs were differentially expressed in NSCLC. Bioinfomatics analysis found that BMSC-exo-miR-30b-5p could regulate NSCLC progression by targeting EZH2, which was verified by in vitro cell experiments. Besides, the target genes of miR-30b-5p were enriched in PI3K/AKT signalling pathway. Animal experiments validated that BMSC-exo-miR-30b-5p promoted NSCLC cell apoptosis and prevented tumorigenesis in nude mice via EZH2/PI3K/AKT axis. Collectively, the inhibitory role of BMSC-derived exosomes-loaded miR-30b-5p in NSCLC was achieved through blocking the EZH2/PI3K/AKT axis.     

Genetic identification of exported proteins in Streptococcus pneumoniae

A strategy was developed to mutate and genetically identify exported proteins in Streptococcus pneumoniae. Vectors were created and used to screen pneumococcal DNA in Escherichia coli and S. pneumoniae for translational gene fusions to alkaline phosphatase (PhoA), Twenty five PhoA⁺ pneumococcal mutants were isolated and the loci from eight of these mutants showed similarity to known exported or membrane-associated proteins. Homologues were found to: (i) protein-dependent peptide permeases, (ii) penicillin-binding proteins, (iii) Cip proteases, (iv) two-component sensor regulators, (v) the phospho-enolpyruvate:carbohydrate phosphotransferase permeases, (vi) membrane-associated dehydrogenases, (vii) P-type (E₁E₂-type) cation transport ATPases, (viii) ABC transporters responsible for the translocation of the RTX class of bacterial toxins. Unexpectedly one PhoA⁺ mutant contained a fusion to a member of the DEAD protein family of ATP-dependent RNA helicases suggesting export of these proteins.     

-cyano-substituted analogoues of decarboxylated S-adenosylmethionine as enzyme activated, irreversible inhibitors of S-adenosylmethionine decarboxylase

A pair of -cyano analogues of decarboxylated S-adenosylmethionine (2a and 2b) were synthesized as potential enzyme activated, irreversible inhibitors of the[pyruvoyl enzyme S-adenosylmethionine decarboxylase (AdoMet-DC). Each of these analogues acts as an irreversible inactivator for ADoMet-DC from Escherichia coli (IC₅₀ values of 9 and 50 μM, respectively). These analogues also inactivate human AdoMet-DC, with K_I values of 246.6 and 7.2 μM, and k_inact values of 0.29 and 0.03 min⁻¹, respectively.     

Preparation of the pure diastereomeric forms of S- (5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2- cyclopentene and their evaluation as irreversible inhibitors of S-adenosylmethionine decarboxylase from Escherichia coli

The conformationally restricted S-adenosylmethionine analogue AdoMac (S-(5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2-cyclopentene has been shown to act as an enzyme activated, irreversible inhibitor of theEscherichia coli form of the enzyme S-adenosylmethionine decarboxylase. Inactivation of the enzyme is presumably initiated by formation of an imine linkage between the inhibitor and the terminal pyruvate of the enzyme, followed by base-catalyzed elimination of methylthioadenosine and generation of a latent electrophile. Removal of the driving force for the elimination of methylthioadenosine resulted in a reversibly binding inhibitor. Thus, the thioether analogue corresponding to AdoMac, and the corresponding dihydro derivative (H₂-AdoMac), reversibly inhibit the enzyme. AdoMac was resolved into its four pure diastereomeric forms, and each diastereomer was evaluated as an irreversible inhibitor of the enzyme. The K_I values for the individual diastereomers range between 3.83 and 39.6 μM, with the cis-1S,4R diastereomer being the most potent inhibitor. However, the k_inact values for the four diastereomers are not significantly different, suggesting that the binding of each diastereomer to the enzyme is configuration-dependent, while the subsequent inactivation likely proceeds through a single intermediate which is formed from each of the four diastereomers. Since each pure diastereomer represents a distinct conformational mimic exhibiting restricted sidechain rotation, the data suggests that these and related analogues may be useful as conformational probes for the catalytic site of AdoMet-DC.     

Reduced Application of Nitrogen Fertilizer Affects the Carbon Metabolism of Leaves and Maintains the Number of Flowers in Coreopsis tinctoria

This study examined the effects of nitrogen (N) fertilizer reduction on the carbon (C) metabolism and yield of Coreopsis tinctoria. A two-year (2020–2021) hydroponic experiment was conducted in accordance with a randomized complete group design with five N levels [0.875 mM Ca(NO₃)₂ (N1), 1.750 mM Ca(NO₃)₂ (N2), 3.500 mM Ca(NO₃)₂ (N3), 7.000 mM Ca(NO₃)₂ (N4), and 14.000 mM Ca(NO₃)₂ (N5)] and three replications. The results showed that low N significantly affected the functional leaf weight, C metabolism, and flower bud (or flower) numbers of C. tinctoria at harvest. Lower-N levels, especially those of the N2 treatment, significantly increased Rubisco, sucrose synthase (SS), sucrose phosphate synthase (SPS), soluble acid invertase (SAI), glucose 6-phosphate dehydrogenase (G6PDH), and 6-phosphogluconate dehydrogenase (6PGDH) activity and maintained the flower number of C. tinctoria. In addition, the balance of carbohydrates (sucrose, starch, glucose, and fructose) and ATP contents was more efficiently maintained under relatively low-N levels. These findings might suggest that reduced application of N fertilizer affects the C metabolism of leaves and maintains the number of flowers in Coreopsis tinctoria. Applying relatively low-N fertilizer levels is also a promising cultivation strategy for C. tinctoria.

     

The Effect of High-Temperature Stress on the Physiological Indexes,Chloroplast Ultrastructure,and Photosystems of two Herbaceous Peony Cultivars

In this study, two herbaceous peony cultivars with different heat tolerances (‘Fenyunu’ FYN low sensitivity and ‘Qiaoling’ QL high sensitivity) were used as research materials. An integrated view of the factors underlying the decrease in photosynthetic rate under high-temperature (HT) stress was provided by analyzing the biochemical parameters, chloroplast ultrastructure, gas-exchange parameters, chlorophyll fluorescence, and modulated 820 nm reflection of herbaceous peony leaves. The results showed that hydrogen peroxide, superoxide anion, malondialdehyde, and electrical conductivity increased significantly, while the photosynthetic pigments content and photosynthetic capacity decreased significantly in QL than in FYN under HT. The contents of soluble sugars and proline increased greatly in FYN than in QL, while the activity of SOD decreased markedly in QL than in FYN after HT. Compared with FYN, the ultrastructure of QL was more seriously disrupted under HT. Chlorophyll fluorescence analysis showed that HT changed the shapes of OJIP curve, resulting in the increase of K phase and J phase. The PSII acceptor side was more damaged than the donor side, and the electron transfer was seriously blocked. The energy flow in the process of light energy absorption, capture, and electron transfer were significantly changed after HT stress. Meanwhile, PSI was also significantly inhibited, and the coordination of both photosystems decreased. The variation of these parameters in FYN was less than that in QL. These results suggested that FYN featured a more heat-tolerance ability as evidenced by the good performances on the antioxidant system, osmoregulatory capacity, and the thermostability of membranes and photosystems.

     

声学指数在城市森林鸟类多样性评估中的应用

鸣声是鸟类之间进行沟通和传递信息的重要方式,这为通过声学监测评估鸟类多样性提供了独特的机会。利用声学指数快速评估生物多样性是一种新兴的调查方法,但城市森林中的复杂声环境可能会导致声学指数的指示结果出现偏差。为了解声学指数在城市森林中应用的可行性,本研究在北京市东郊森林公园设置了50个矩阵式调查样点,于2021年4–6月每月进行1次鸟类传统观测和同步鸣声采集,通过比较两种方法的结果来探究声学监测的有效性。采用Spearman相关分析和广义线性混合模型评估6个常用声学指数与鸟类丰富度和多度的关系,并衡量了每个指数的性能。结果表明:(1)本研究共记录到鸟类10目23科35种,通过声学监听识别的总物种数与传统鸟类观测相等,但具体鸟种存在差异;(2)不同月份间声学指数与鸟类丰富度和多度的相关性有明显差别,声学复杂度指数(ACI)和标准化声景差异指数(NDSI)优于其他指数,是评估鸟类多样性的关键变量;(3)声学指数对鸟类多度的预测能力(R²m=0.32,R2c=0.80)要高于丰富度(R²m=0.12,R2c=0.18)。声学指数为快速评估生物多样性提...     

Insulin-like growth factors: Putative muscle-derived trophic agents that promote motoneuron survival

Treatment of chick embryos in ovo with IGF-I during the period of normal, developmentally regulated neuronal death (embryonic days 5–10) resulted in a dose-dependent rescue of a significant number of lumbar motoneurons from degeneration and death. IGF-II and two variants of IGF-I with reduced affinity for IGF binding proteins, des(1-3) IGF-I and long R³ IGF-I, also elicited enhanced survival of motoneurons equal to that seen in IGF-I-treated embryos. IGF-I did not enhance mitogenic activity in motoneuronal populations when applied to embryos during the period of normal neuronal proliferation (E2-5). Treatment of embryos with IGF-I also reduced two types of injury-induced neuronal death. Following either deafferentation or axotomy, treatment of embryos with IGF-I rescued approximately 75% and 50%, respectively, of the motoneurons that die in control embryos as a result of these procedures. Consistent with the survival-promoting activity on motoneurons in ovo, IGF-I, -II, and des(1-3) IGF-I elevated choline acetyltransferase activity in embryonic rat spinal cord cultures, with des (1-3) IGF-I demonstrating 2.5 times greater potency than did IGF-I. A single addition of IGF-I at culture initiation resulted in the maintenance of 80% of the initial ChAT activity for up to 5 days, during which time ChAT activity in untreated control cultures fell to 9%. In summary, these results demonstrate clear motoneuronal trophic activity for the IGFs. These findings, together with previous reports that IGFs are synthesized in muscle and may participate in motoneuron axonal regeneration and sprouting, indicate that these growth factors may have an important role in motoneuron development, maintenance, and recovery from injury. © 1993 John Wiley & Sons, Inc.