Specific identification and molecular typing analysis of Lactobacillus johnsonii by using PCR-based methods and pulsed-field gel electrophoresis

A fast and reliable Multiplex-PCR assay was established to identify the species Lactobacillus johnsonii. Two opposing rRNA gene-targeted primers have been designed for this specific PCR detection. Specificity was verified with DNA samples isolated from different lactic acid bacteria. Out of 47 Lactobacillus strains isolated from different environments, 16 were identified as L. johnsonii by PCR. The same set of strains was investigated with five alternative molecular typing methods: enterobacterial repetitive intergenic consensus PCR (ERIC-PCR), repetitive extragenic palindromic PCR (REP-PCR), amplified fragment length polymorphism, single triplicate arbitrarily primed PCR, and pulsed-field gel electrophoresis in order to compare the discriminatory power of these methods. The reported data strongly support the highly significant heterogeneity among all L. johnsonii isolates, potentially linked to their origin of isolation. The use of species-specific primers as well as rapid and highly powerful PCR-based molecular typing tools (namely ERIC- and REP-PCR techniques) should be respectively envisaged for identifying, differentiating and monitoring L. johnsonii strains from various environmental samples, for product monitoring, for species tracing in clinical studies as well as bacterial profiling of various microecological or gastrointestinal environments.     

Inheritance of Fusarium wilt resistance introgressed from Solanum aethiopicum Gilo and Aculeatum groups into cultivated eggplant (S. melongena) and development of associated PCR-based markers

The two eggplant relatives Solanum aethiopicum gr. Gilo and Solanum aethiopicum gr. Aculeatum (=Solanum integrifolium) carry resistance to the fungal wilt disease caused by Fusarium oxysporum f. sp. melongenae, a worldwide soil-borne disease of eggplant. To introgress the resistance trait into cultivated eggplant, the tetraploid somatic hybrids S. melongena + S. aethiopicum and S. melongena + S. integrifolium were used. An inheritance study of the resistance was performed on advanced anther culture-derived androgenetic backcross progenies from the two somatic hybrids. The segregation fitted a 3 resistant (R): 1 susceptible (S) ratio in the selfed populations and a 1R:1S ratio in the backcross progenies for the trait derived from S. aethiopicum and S. integrifolium. These ratios are consistent with a single gene, which we designated as Rfo-sa1, controlling the resistance to Fusarium oxysporum f. sp. melongenae. The allelic relationship between the resistance genes from S. aethiopicum and S. integrifolium indicate that these two genes are alleles of the same locus. Bulked Segregant Analysis (BSA) was performed with RAPD markers on the BC₃/BC₅ resistant advanced backcross progenies, and three RAPD markers associated with the resistance trait were identified. Cleaved Amplified Polymorphic Sequences (CAPSs) were subsequently obtained on the basis of the amplicon sequences. The evaluation of the efficiency of these markers in predicting the resistant phenotype in segregating progenies revealed that they represent useful tools for indirect selection of Fusarium resistance in eggplant.     

Variations in Chemical Composition,Vasorelaxant and Angiotensin I‐Converting Enzyme Inhibitory Activities of Essential Oil from Aerial Parts of Seseli pallasii Besser (Apiaceae)

The present paper describes environmental and seasonal‐related chemical composition variations, vasorelaxant and angiotensin I‐converting enzyme (ACE) activities of essential oil from aerial parts of Seseli pallasii Besser . The composition was analyzed by GC and GC/MS. Monoterpenes were found to be the most abundant chemical class with α ‐pinene (42.7 – 48.2%) as the most prevalent component. Seseli pallasi essential oil relaxed isolated endothelium‐intact mesenteric arteries rings precontracted with phenylephrine with IC ₅₀ = 3.10 nl/ml (IC ₅₀ = 2.70 μg/ml). Also, S. pallasii essential oil was found to exhibit a dose‐dependent ACE inhibitory activity with an IC ₅₀ value of 0.33 mg/ml. In silico evaluation of ACE inhibitory activity of the individual components showed that spathulenol exhibited the best binding affinity with ACE, and the lowest binding energy of ?7.5 kcal/mol. The results suggested that combination of vasorelaxing and ACE inhibitory effects of the analyzed S. pallasii essential oil might have the potential therapeutic significance in hypertension.     

Do large‐seeded herbs have a small range size? The seed mass–distribution range trade‐off hypothesis

We aimed to introduce and test the “seed mass–distribution range trade‐off” hypothesis, that is, that range size is negatively related to seed mass due to the generally better dispersal ability of smaller seeds. Studying the effects of environmental factors on the seed mass and range size of species, we also aimed to identify habitats where species may be at risk and need extra conservation effort to avoid local extinctions. We collected data for seed mass, global range size, and indicators for environmental factors of the habitat for 1,600 species of the Pannonian Ecoregion (Central Europe) from the literature. We tested the relationship between species’ seed mass, range size, and indicator values for soil moisture, light intensity, and nutrient supply. We found that seed mass is negatively correlated with range size; thus, a seed mass–distribution range trade‐off was validated based on the studied large species pool. We found increasing seed mass with decreasing light intensity and increasing nutrient availability, but decreasing seed mass with increasing soil moisture. Range size increased with increasing soil moisture and nutrient supply, but decreased with increasing light intensity. Our results supported the hypothesis that there is a trade‐off between seed mass and distribution range. We found that species of habitats characterized by low soil moisture and nutrient values but high light intensity values have small range size. This emphasizes that species of dry, infertile habitats, such as dry grasslands, could be more vulnerable to habitat fragmentation or degradation than species of wet and fertile habitats. The remarkably high number of species and the use of global distribution range in our study support our understanding of global biogeographic processes and patterns that are essential in defining conservation priorities.     

Phytochemicals with NO inhibitory effects and interactions with iNOS protein from Trigonostemon howii

A phytochemical investigation to obtain new NO inhibitors led to the isolation of nine compounds including one new guaiane-type sesquiterpenoid (1) and two new cleistanthane diterpenoids (2 and 3) from the stems of Trigonostemon howii. Their structures were elucidated on the basis of extensive 1D and 2D NMR spectroscopic data analysis, and the absolute configurations of new compounds 1–3 were established via comparison of experimental and calculated electronic circular dichroism (ECD) spectra. Compounds 2 and 3 possess a rare 3,4-seco-cleistanthane diterpenoid skeleton. All of the compounds showed inhibitory effects on lipopolysaccharide-induced NO production in murine microglial BV-2 cells. The further molecular docking studies indicated the strong interactions between some bioactive compounds with the iNOS protein, which revealed the possible and potential mechanism of NO inhibition of bioactive compounds.     

鸡腿蘑胞外多糖的形貌结构及分子量动态变化与抗氧化的相关性研究

对鸡腿蘑多糖的结构进行检测,并在此基础上探讨结构与活性的关系,对深度发掘鸡腿蘑多糖的功效具有重要意义.制备发酵时间为72 h、96 h和120 h的鸡腿蘑胞外粗多糖,采用PMP柱前衍生化-HPLC法分析其单糖组成,结果表明发酵72 h、96 h和120 h胞外多糖均由D-甘露糖、L-鼠李糖、D-葡萄糖、D-半乳糖、D-...     

The second phantom aquatic leaf beetle in Japan: Macroplea mutica rediscovery in the wetlands (Coleoptera: Chrysomelidae)

Wetland biodiversity is currently declining on a global scale. Wetland biodiversity understanding is critical for determining the wetlands' conservation value. In this study, Macroplea Samouelle, 1819 (Coleoptera: Chrysomelidae) was discovered in Aomori Prefecture, Honshu Island, Japan. Only two Macroplea species have been recorded in Japan, M. japana (Jacoby, 1885) and M. mutica (Fabricius, 1792). Macroplea japana had been unrecorded for 60 years before being rediscovered in Honshu Island in 2022, and a single adult M. mutica female was discovered in Hokkaido Prefecture in 2003. The discovered individuals were concluded to be M. mutica based on morphological and molecular analyses. Although morphological differences were observed with the Eurasian M. mutica individuals, the male genitalia was nearly identical to M. mutica. For the molecular phylogenetic analysis based on COI and 28S sequences, Macroplea individuals in Japan were clustered with M. mutica on the Eurasian Continent. This is the first record of this species on Honshu Island (and the second in Japan), as well as the first record of adult males. This species would require conservation policies and additional distributional surveys.     

Integrating experimental data with molecular simulations to investigate RNA structural dynamics

Conformational dynamics is crucial for ribonucleic acid (RNA) function. Techniques such as nuclear magnetic resonance, cryo-electron microscopy, small- and wide-angle X-ray scattering, chemical probing, single-molecule Förster resonance energy transfer, or even thermal or mechanical denaturation experiments probe RNA dynamics at different time and space resolutions. Their combination with accurate atomistic molecular dynamics (MD) simulations paves the way for quantitative and detailed studies of RNA dynamics. First, experiments provide a quantitative validation tool for MD simulations. Second, available data can be used to refine simulated structural ensembles to match experiments. Finally, comparison with experiments allows for improving MD force fields that are transferable to new systems for which data is not available. Here we review the recent literature and provide our perspective on this field.     

Morphophylogenetic study revealed that Erysiphe gracilis (powdery mildew of evergreen oaks,Erysiphales) is a species complex consisting of six different species

Erysiphe gracilis is a powdery mildew species that occurs on evergreen oak species belonging to Quercus subgen. Cyclobalanopsis in East Asia (China and Japan). In a previous report, we found that E. gracilis var. gracilis is divided into four genotypes each of them forming a separate clade with strong bootstrap support. In this study, we further investigated genotype speciation in E. gracilis var. longissima occurring on Q. acuta and Q. sessilifolia, and found that this variety is also divided into two distinct genotypes. These results suggested that E. gracilis represents a species complex consisting of six different species. Based on detailed morphological examinations correlating with results of molecular sequence analyses, we propose to divide E. gracilis into six species, encompassing three new species (E. uncinuloides, E. pseudogracilis, and E. longiappendiculata), one new name (E. longifilamentosa), and two known species (E. gracilis s. str. and E. hiratae). A key to the species concerned is provided.     

Quantitative Biomarkers for Prediction of Epidermal Growth Factor Receptor Mutation in Non-Small Cell Lung Cancer

OBJECTIVES: To predict epidermal growth factor receptor (EGFR) mutation status using quantitative radiomic biomarkers and representative clinical variables. METHODS: The study included 180 patients diagnosed as of non-small cell lung cancer (NSCLC) with their pre-therapy computed tomography (CT) scans. Using a radiomic method, 485 features that reflect the heterogeneity and phenotype of tumors were extracted. Afterwards, these radiomic features were used for predicting epidermal growth factor receptor (EGFR) mutation status by a least absolute shrinkage and selection operator (LASSO) based on multivariable logistic regression. As a result, we found that radiomic features have prognostic ability in EGFR mutation status prediction. In addition, we used radiomic nomogram and calibration curve to test the performance of the model. RESULTS: Multivariate analysis revealed that the radiomic features had the potential to build a prediction model for EGFR mutation. The area under the receiver operating characteristic curve (AUC) for the training cohort was 0.8618, and the AUC for the validation cohort was 0.8725, which were superior to prediction model that used clinical variables alone. CONCLUSION: Radiomic features are better predictors of EGFR mutation status than conventional semantic CT image features or clinical variables to help doctors to decide who need EGFR tyrosine kinase inhibitor (TKI) treatment.