Genomic, Phylogenetic and Physiological Characterization of the PAH-Degrading Strain Gordonia polyisoprenivorans 135.

The strain Gordonia polyisoprenivorans 135 is able to utilize a wide range of aromatic compounds. The aim of this work was to study the features of genetic organization and biotechnological potential of the strain G. polyisoprenivorans 135 as a degrader of aromatic compounds. The study of the genome of the strain 135 and the pangenome of the G. polyisoprenivorans species revealed that some genes, presumably involved in PAH catabolism, are atypical for Gordonia and belong to the pangenome of Actinobacteria. Analyzing the intergenic regions of strain 135 alongside the "panIGRome" of G. polyisoprenivorans showed that some intergenic regions in strain 135 also differ from those located between the same pairs of genes in related strains. The strain G. polyisoprenivorans 135 in our work utilized naphthalene (degradation degree 39.43%) and grew actively on salicylate. At present, this is the only known strain of G. polyisoprenivorans with experimentally confirmed ability to utilize these compounds.

A Novel Intergenic Region (chr2: 30,316,870)-ALK Fusion in a Patient with Lung Adenocarcinoma Responding to Crizotinib Combined with Pemetrexed Treatment: A Case Report.

Background: Anaplastic lymphoma kinase (ALK) rearrangements have been reported as an important oncogenic driver in 5-7% non-small cell lung cancer (NSCLC) patients. Reports about the intergenic region (IGR) as an ALK fusion partner are rare. In this study, we report a novel IGR (chr2: 30,316,870)-ALK fusion in an advanced lung adenocarcinoma patient that responded effectively to crizotinib combined with pemetrexed. Case Presentation: A 68-year-old Chinese female was diagnosed with stage IV right lung adenocarcinoma (cT3N3M1c). The targeted next-generation sequencing (NGS) of 14 cancer-related genes identified an IGR (chr2: 30,316,870)-ALK fusion. Her lung lesions have been successfully converted from a partial response to a complete response after administrating crizotinib for 1 year combined with 6 cycles of chemotherapy with pemetrexed. So far, her progression-free-survival has reached 21 months. Conclusion: In this case, we firstly report a novel IGR (chr2: 30,316,870)-ALK fusion by using targeted NGS, and highlight the efficacy of crizotinib combined with pemetrexed to reduce unbearable gastrointestinal adverse reactions. It provides valuable clinical guidance for the treatment of similar cases in the future.

Predicting infarction growth rate II using ANFIS-based binary particle swarm optimization technique in ischemic stroke.

Ischemic stroke, a severe medical condition triggered by a blockage of blood flow to the brain, leads to cell death and serious health complications. One key challenge in this field is accurately predicting infarction growth - the progressive expansion of damaged brain tissue post-stroke. Recent advancements in artificial intelligence (AI) have improved this prediction, offering crucial insights into the progression dynamics of ischemic stroke. One such promising technique, the Adaptive Neuro-Fuzzy Inference System (ANFIS), has shown potential, but it faces the 'curse of dimensionality' and long training times as the number of features increased. This paper introduces an innovative, automatic method that combines Binary Particle Swarm Optimization (BPSO) with ANFIS architecture, achieves reduction in dimensionality by reducing the number of rules and training time. By analyzing the Pearson correlation coefficients and P-values, we selected clinically relevant features strongly correlated with the Infarction Growth Rate (IGR II), extracted after one CT scan. We compared our model's performance with conventional ANFIS and other machine learning techniques, including Support Vector Regressor (SVR), shallow Neural Networks, and Linear Regression. •Inputs: Real data about ischemic stroke represented by clinically relevant features.•Output: An innovative model for more accurate and efficient prediction of the second infarction growth after the first CT scan.•Results: The model achieved commendable statistical metrics, which include a Root Mean Square Error of 0.091, a Mean Squared Error of 0.0086, a Mean Absolute Error of 0.064, and a Cosine distance of 0.074.

Debris flow susceptibility assessment based on information value and machine learning coupling method: from the perspective of sustainable development.

Accurately assessing the susceptibility of debris flow disasters is of great significance for reducing the cost of disaster prevention and mitigation, as well as disaster losses. Machine learning (ML) models have been widely used in the susceptibility assessment of debris flow disasters. However, these models often have randomness in the selection of non-disaster data, which can lead to redundant information and poor applicability and accuracy of susceptibility evaluation results. To address this issue, this paper focuses on debris flow disasters in Yongji County, Jilin Province, China; optimizes the sampling method of non-disaster datasets in machine learning susceptibility assessment; and proposes a susceptibility prediction model that couples information value (IV) with artificial neural network (ANN) and logistic regression (LR) models. A debris flow disaster susceptibility distribution map with higher accuracy was drawn based on this model. The model's performance is evaluated using the area under the receiver operating characteristic curve (AUC), information gain ratio (IGR), and typical disaster point verification methods. The results show that the rainfall and topography were found to be decisive factors in the occurrence of debris flow disasters, and the IV-ANN model established in this study had the highest accuracy (AUC = 0.968). Compared to traditional machine learning models, the coupling model produced an increase in economic benefit of about 25% while reducing the average disaster prevention and control investment cost by about 8%. Based on model's susceptibility map, this paper proposes practical disaster prevention and control suggestions that promote sustainable development in the region, such as establishing monitoring systems and information platforms to aid disaster management.

Active or passive maternal smoking increases the risk of low birth weight or preterm delivery: Benefits of cessation and tobacco control policies.

In France, maternal smoking, active or passive, remains one of the highest in Europe. At the same time, there is an increase in the number of low birth weight (<2500 g) and premature (<37 weeks of amenorrhea) newborns. The objective of this narrative review is to examine the impact of active or passive maternal smoking on birth weight or prematurity rates, and to consider the benefits of policies to stop or control smoking. This is a narrative review that analyzes and discusses the major articles published over the past 20 years regarding the role of active or passive maternal smoking on the risk of low birth weight or preterm delivery. Articles were selected using the following keywords: maternal smoking, low birth weight, preterm birth, smoking cessation, passive smoking, exhaled carbon monoxide, tobacco control policies. Active smoking is associated, in a dose-response relationship, with increased risks of low birth weight and preterm delivery. Passive smoking, mainly related to the presence of a smoking spouse, increases the risk of low birth weight and preterm birth. Our review confirmed also the benefits of smoking cessation, even in the third trimester, in reducing the risk of small for gestation age or fetal growth restriction and preterm birth. Several studies of tobacco control policies have been shown to be effective in significantly reducing maternal smoking. There is sufficient evidence to infer a causal link between active or passive maternal smoking and low birth weight or preterm delivery. This causal link is compelling and sufficient to justify intensifying efforts to promote rapid progress in tobacco control policies, with the vision of a tobacco-free generation, and smoking cessation with best practices during preconception or pregnancy.

Effects of Diabetes and Voluntary Exercise on IgA Concentration and Polymeric Immunoglobulin Receptor Expression in the Submandibular Gland of Rats.

Background and Objectives: Patients with diabetes are more susceptible to upper respiratory tract infections (URTIs) because they are easily infected. Salivary IgA (sali-IgA) levels play a major role in transmitting URTIs. Sali-IgA levels are determined by salivary gland IgA production and polymeric immunoglobulin receptor (poly-IgR) expression. However, it is unknown whether salivary gland IgA production and poly-IgR expression are decreased in patients with diabetes. While exercise is reported to increase or decrease the sali-IgA levels, it is unclear how exercise affects the salivary glands of patients with diabetes. This study aimed to determine the effects of diabetes and voluntary exercise on IgA production and poly-IgR expression in the salivary glands of diabetic rats. Materials and Methods: Ten spontaneously diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats (eight-week-old) were divided into two groups of five rats each: a non-exercise group (OLETF-C) and a voluntary wheel-running group (OLETF-E). Five Long-Evans Tokushima Otsuka (LETO) rats without diabetes were bred under the same conditions as the OLETF-C. Sixteen weeks after the study began, the submandibular glands (SGs) were collected and analyzed for IgA and poly-IgR expression levels. Results: IgA concentrations and poly-IgR expression levels in SGs were lower in OLETF-C and OLETF-E than in LETO (p < 0.05). These values did not differ between the OLETF-C and OLETF-E. Conclusions: Diabetes decreases IgA production and poly-IgR expression in the salivary glands of rats. Moreover, voluntary exercise increases sali-IgA levels but does not increase IgA production and poly-IgR expression in the salivary glands of diabetic rats. Increasing IgA production and poly-IgR expression in the salivary glands, which is reduced in diabetes, might require slightly higher-intensity exercise than voluntary exercise under the supervision of a doctor.

Metagenomic 16S rDNA amplicon data of microbial diversity of Cimex hemipterus (F.) (Hemiptera: Cimicidae) treated with insect growth regulators (IGR).

The metagenomics dataset presented here is based on bacterial 16S rDNA gene amplicons of DNA extracted from tropical bed bugs (Cimex hemipterus). Amplicon-based sequencing was performed using the Illumina MiSeq platform, and the raw sequence data were analyzed using QIIME (version 2022.8.3). The metagenome sequence comprised ten samples that include C1 (133 511bps), C2 (108 920bps), CH1 (106 562bps), CH2 (101 778bps), P1 (103 618bps), P2 (133 258bps), T1 (113 558bps), T2 (133 952bps), TM1 (125 335bps), and TM2 (118 345bps). The sequence data is readily accessible at the NCBI SRA under bio project PRJNA918835. The most abundant microbial community present in the C. hemipterus is the Proteobacteria, with more than 99% of the abundance.

Compounds Inhibiting Noppera-bo, a Glutathione S-transferase Involved in Insect Ecdysteroid Biosynthesis: Novel Insect Growth Regulators.

Glutathione S-transferases (GSTs) are conserved in a wide range of organisms, including insects. In 2014, an epsilon GST, known as Noppera-bo (Nobo), was shown to regulate the biosynthesis of ecdysteroid, the principal steroid hormone in insects. Studies on fruit flies, Drosophila melanogaster, and silkworms, Bombyx mori, demonstrated that loss-of-function mutants of nobo fail to synthesize ecdysteroid and die during development, consistent with the essential function of ecdysteroids in insect molting and metamorphosis. This genetic evidence suggests that chemical compounds that inhibit activity of Nobo could be insect growth regulators (IGRs) that kill insects by disrupting their molting and metamorphosis. In addition, because nobo is conserved only in Diptera and Lepidoptera, a Nobo inhibitor could be used to target IGRs in a narrow spectrum of insect taxa. Dipterans include mosquitoes, some of which are vectors of diseases such as malaria and dengue fever. Given that mosquito control is essential to reduce mosquito-borne diseases, new IGRs that specifically kill mosquito vectors are always in demand. We have addressed this issue by identifying and characterizing several chemical compounds that inhibit Nobo protein in both D. melanogaster and the yellow fever mosquito, Aedes aegypti. In this review, we summarize our findings from the search for Nobo inhibitors.

Effect of resistance vs. aerobic exercise in pre-diabetes: an RCT.

BACKGROUND: This study aimed to assess the different impacts of aerobic and resistance exercise intervention on pre-diabetes and its possible influencing factor (obesity) to identify which exercise intervention mode was better for pre-diabetes to control their blood glucose levels. METHODS: Single-blind randomized controlled trial. Participants were recruited from Southwest Hospital between February 2016 and May 2017 and randomly divided into three groups using stratified randomization: aerobic exercise (A), resistance exercise (R), and control (C). The effects of each group were analyzed, and the relationship with obesity was investigated following a 12-week intervention. RESULTS: Eighty participants were enrolled (9 were lost, and 1 was excluded). Finally, 26 participants were included in group A, 23 in group R, and 21 in group C. In groups A and R, FPG, OGTT 2-h PG, and HOMA2-IR decreased by 6.17% (P = 0.001) and 4.81% (P = 0.019), 20.39% (P < 0.001) and 16.50% (P < 0.001), and 8.34% (P = 0.026) and 18.31% (P = 0.001, superior to that in group A), respectively (all P < 0.001 compared with group C, with no significant differences between groups A and R). The ratio of reversal to euglycemia was 69.2% (P = 0.003 compared with group C) in group A and 43.5% (P = 0.213 compared with group C) in group R. The decreased ratio of GSP in group R was greater (65.2%, P = 0.008 compared with group C) compared with group A (38.5%, P = 0.355 compared with group C). Decreases in the parameters BMI (3.1 ± 3.2% P < 0.001, moderately positive correlation with the decreased FPG level, r = 0.498, P = 0.010, two-tailed) and waist circumference (3.1 ± 2.7% P < 0.001) were noted in group A, but no significant correlations were noted between other indicators in group R. CONCLUSIONS: Both resistance and aerobic exercise can control and reverse IGR. Compared with aerobic exercise, resistance exercise may be superior in terms of GSP and IR improvement. Aerobic exercise decreases blood glucose levels through weight loss. However, the effect of resistance exercise might not be mediated via weight loss and obesity control. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2000038304. Registered on September 17, 2020.

Emergence of a novel intergenic region (IGR) IV variant of african swine fever virus genotype II in domestic pigs in Vietnam.

African swine fever virus (ASFV) causes African swine fever (ASF), a deadly disease affecting both domestic pigs and wild boars. ASF has become endemic in Vietnam since its first appearance in early 2019. Our previous molecular surveillance studies revealed that all the ASFV strains circulating in Vietnam belong to p72 genotype II, p54 genotype II, CD2v serogroup 8, and CVR of B602L gene variant type I. However, the genetic analysis based on the tandem repeat sequences located between I73R and I329L genes revealed three different intergenic region (IGR) variants; I, II, and III. In this study, using ASFV field isolates collected from September 24th to December 27th, 2021, we report, for the first time, novel IGR IV variants circulating in the Vietnamese pig population.

Circulating spexin levels are influenced by the glycemic status and correlated with pancreatic ß-cell function in Chinese subjects.

AIMS: Spexin plays a role in regulating glucose metabolism. This study investigated the spexin levels in different glycemic status and its association with insulin secretion in humans. METHODS: A total of 462 subjects were recruited in this study, including 52 healthy subjects, 106 first-degree relatives (FDRs) of type 2 diabetes mellitus (T2DM), 115 impaired glucose regulation (IGR), 80 newly diagnosed T2DM, and 106 established T2DM. Serum spexin was measured using ELISA. The homeostasis model assessment of insulin resistance (HOMA2-IR) and ß-cell function (HOMA2-ß), and Stumvoll index estimating first- and second-phase insulin secretion were calculated. RESULTS: Spexin levels were higher in FDRs [235.53 pg/ml (185.28, 293.95)] and IGR [239.79 pg/ml (191.52, 301.69)], comparable in newly diagnosed T2DM [224.68 pg/ml (187.37, 279.74)], and lower in established T2DM [100.11 pg/ml (78.50, 137.34)], compared with healthy subjects [200.23 pg/ml (160.32, 275.65)]. Spexin levels were negatively correlated with fasting plasma glucose (FPG) (r = - 0.355, P < 0.001), hemoglobin A1C (HbA1c) (r = - 0.379, P < 0.001), and HOMA2-IR (r = - 0.225, P < 0.001), and positively correlated with HOMA2-ß (r = 0.245, P < 0.001) after adjusting for age, sex, and BMI. Multivariate linear regression analysis showed that established T2DM and HOMA2-ß were independently associated with serum spexin levels. CONCLUSIONS: Serum spexin levels represented as a bell-shaped curve along the glycemic continuum and is closely related with insulin secretion in humans.

Irisin as a predictor of bone metabolism in Han Chinese Young Men with pre-diabetic individuals.

BACKGROUND: Irisin is a novel myokine both in mice and humans, and it can also be secreted by adipose tissue and the liver in a small amounts. There are few studies on irisin and bone metabolism. The aim of this study was to assess the relationship between serum irisin levels and bone metabolism and analyze its related factors in Han young male with pre-diabetic individuals. METHODS: This cross-sectional study included 41 pre-diabetes and 45 normal glucose tolerance (NGT). Anthropometric measurements, including height, weight, waist circumference (WC), and bone mineral content (BMC), were performed. All patients underwent an oral glucose tolerance test (OGTT) after 8 h of fasting, and the levels of glucose, insulin, lipids, serum irisin and bone turnover markers were measured. RESULTS: The levels of serum irisin (4.4 ± 1.4 vs. 6.3 ± 1.5 µg/mL), P1NP and OC were significantly lower and CTX was significantly higher in the pre-diabetes group (P < 0.05). BMC did not differ in the two groups (P > 0.05). Serum irisin levels negatively correlated with BMI (r =-0.325), FPG (r =-0.329), TG (r =-0.339) (P < 0.05) in NGT individuals. Serum irisin levels positively correlated with P1NP (r = 0.398), OC (r = 0.351), HDL-C (r = 0.432) and negatively correlated with FPG (r = -0.725), 2 h-PG (r = -0.360) (P < 0.05) in pre-diabetic individuals. Multiple regression analysis revealed that Serum irisin (ß = 9.768, P = 0.025) and WC (ß = -2.355, P = 0.002) were significant independent predictors for P1NP. CONCLUSION: Bone turnover markers were changed rather than bone mineral content in young men with pre-diabetes. In pre-diabetes individuals, serum irisin levels were reduced and close relationship with P1NP. Falling irisin levels may be a predictor of decreased bone formation in Han young men with pre-diabetes individuals.

Alterations in Microbiota and Metabolites Related to Spontaneous Diabetes and Pre-Diabetes in Rhesus Macaques.

Spontaneous type 2 diabetes mellitus (T2DM) macaques are valuable resources for our understanding the pathological mechanism of T2DM. Based on one month's fasting blood glucose survey, we identified seven spontaneous T2DM macaques and five impaired glucose regulation (IGR) macaques from 1408 captive individuals. FPG, HbA1c, FPI and IR values were significant higher in T2DM and IGR than in controls. 16S rRNA sequencing of fecal microbes showed the significantly greater abundance of Oribacterium, bacteria inhibiting the production of secondary bile acids, and Phascolarctobacterium, bacteria producing short-chain fatty acids was significantly lower in T2DM macaques. In addition, several opportunistic pathogens, such as Mogibacterium and Kocuria were significantly more abundant in both T2DM and IGR macaques. Fecal metabolites analysis based on UHPLC-MS identified 50 differential metabolites (DMs) between T2DM and controls, and 26 DMs between IGR and controls. The DMs were significantly enriched in the bile acids metabolism, fatty acids metabolism and amino acids metabolism pathways. Combining results from physiochemical parameters, microbiota and metabolomics, we demonstrate that the imbalance of gut microbial community leading to the dysfunction of glucose, bile acids, fatty acids and amino acids metabolism may contribute to the hyperglycaemia in macaques, and suggest several microbes and metabolites are potential biomarkers for T2DM and IGR macaques.

Rational Design and Identification of Novel Piperine Derivatives as Multichitinase Inhibitors.

Asian corn borer (Ostrinia furnacalis) is one of the most destructive pests in agriculture. Three chitinases OfChtI, OfChtII, and OfChi-h are regarded as potential targets for discovering novel agrochemicals to control O. furnacalis. In this study, piperine (Ki = 43.78∼83.03 µM) was first shown to exhibit inhibitory activities against all three chitinases. Subsequently, 19 novel piperine derivatives were rationally designed based on the conserved aromatic residues of three chitinases and then synthesized. Among them, Compound 5k (Ki = 11.78∼22.82 µM) was identified as the most effective multichitinase inhibitor and indeed displayed higher insecticidal activity against O. furnacalis than dual- or single-chitinase inhibitors. Molecular mechanism studies clarified that Compound 5k interacted with two conserved TRP and TYR of three chitinases in identical modes through hydrogen bonds, hydrophobic, and π-π interactions. Moreover, the microinjection experiment indicated that Compound 5k exhibited substantial sublethal effects against O. furnacalis by regulating its growth and development. This study provides evidence of multichitinase inhibitors to be applied in the control of O. furnacalis.

Novel HIVEP1-ALK fusion in a patient with lung adenocarcinoma demonstrating sensitivity to alectinib: a case report.

Background: Anaplastic lymphoma kinase (ALK) fusion is an important oncogenic driver in non-small cell lung cancer (NSCLC). Reports on the intergenic region (IGR) as an ALK fusion partner are rare. Here, we report the case of a patient with advanced NSCLC harboring a human immunodeficiency virus type I enhancer binding protein 1 (HIVEP1)-ALK fusion that responded effectively to alectinib. Case Description: A 60-year-old non-smoking male was referred with a 3-month history of productive cough secondary to lung adenocarcinoma metastatic to mediastinal lymph nodes, brain, liver, and bone (T2N3M1c, stage IVB). Next-generation sequencing identified an IGR (upstream HIVEP1-) ALK fusion, and immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) results were consistent with an ALK-positive tumor. The patient was subsequently started on alectinib, with no obvious adverse reaction. After 1 month of therapy, the patient achieved significantly remission of the clinical symptoms and had led to an ongoing partial response (PR) lasting >33 months. Conclusions: Our experience highlights the efficacy of alectinib in a patient with HIVEP1-ALK fusion positive NSCLC with multiple metastases including brain disease, and the need for multiple genetic testing methods to verify the oncogenicity of ALK fusions prior to treatment. It could provide useful guidance for the treatment of similar cases in the future.

A new species of anthothelid octocoral (Cnidaria, Alcyonacea) discovered on an algal reef of Taiwan.

A molecular phylogenetic analysis of 132 octocoral species reveals a close relationship between specimens collected from the intertidal pools of the Datan Algal Reef, Taoyuan, Taiwan, and Erythropodiumcaribaeorum (Duchassaing & Michelotti, 1860), but the two species have distinct morphological features. On the basis of morphological differences in polyps and sclerites, we identify and describe a new Erythropodium species: E.taoyuanensis sp. nov. The distinct identifying features of E.taoyuanensis sp. nov. include the upright contractile polyps from thin encrusting membranes and abundant 6-radiate sclerites. Using an integrative approach, we present the findings of morphological comparisons and molecular phylogenetic analyses to demonstrate that E.taoyuanensis sp. nov. is distinct from other Erythropodium species. Our study contributes to the knowledge of octocoral biodiversity in marginal habitats.

Intragenic recombination within the pun allele of the pink-eyed dilution locus in pre-melanocytes and primordial germ cells of embryonic mice treated with N-ethyl-N-nitrosourea.

The forward or reverse processes of intragenic recombination (IGR), which occur through the addition or deletion of duplicated homologous exons of the pun allele in Pun mice, was observed in vivo, after introducing an homozygous pun allele in a C57BL/6 background. We assessed the frequency of IGR upon N-ethyl-N-nitrosourea (ENU) treatment of pre-melanocytes (PMCs: somatic cells) and primordial germ cells (PGCs: germ cells) of embryonic mice at 10.5 days of development (E10.5). We simultaneously examined IGR and other mutations at the p locus of PMCs responsible for coat color in the offspring obtained by crossing pun/pun with pun/P mice. The frequencies of both spontaneous and ENU-induced IGR were markedly higher than that of the recessive mutation (RM) in PMCs obtained from crossing C57BL/6 and PW strains (Shibuya et al., 1982). ENU also induces IGR at a higher frequency in PGCs at E10.5, which was observed in the next generation. These results indicate that ENU, which preferentially induces gene mutations through base substitution, also induces IGR at a high frequency in the pun allele in both somatic and germ cells of embryonic mice at the E10.5 developmental stage.

NS-11021 Modulates Cancer-Associated Processes Independently of BK Channels in Melanoma and Pancreatic Duct Adenocarcinoma Cell Lines.

Potassium channels have emerged as regulators of carcinogenesis, thus introducing possible new therapeutic strategies in the fight against cancer. In particular, the large-conductance Ca2+-activated K+ channel, often referred to as BK channel, is involved in several cancer-associated processes. Here, we investigated the effects of different BK activators, NS-11021, NS-19504, and BMS-191011, in IGR39 (primary melanoma cell line) and Panc-1 (primary pancreatic duct carcinoma cell line), highly expressing the channel, and in IGR37 (metastatic melanoma cell line) that barely express BK. Our data showed that NS-11021 and NS-19504 potently activated BK channels in IGR39 and Panc-1 cells, while no effect on channel activation was detected in IGR37 cells. On the contrary, BK channel activator BMS-191011 was less effective. However, only NS-11021 showed significant effects in cancer-associated processes, such as cell survival, migration, and proliferation in these cancer cell lines. Moreover, NS-11021 led to an increase of intracellular Ca2+ concentration, independent of BK channel activation, thus complicating any interpretation of its role in the regulation of cancer-associated mechanisms. Overall, we conclude that the activation of the BK channel by itself is not sufficient to produce beneficial anti-cancer effects in the melanoma and PDAC cell lines examined. Importantly, our results raise an alarm flag regarding the use of presumably specific BK channel openers as anti-cancer agents.

The molecular feature of macrophages in tumor immune microenvironment of glioma patients.

BACKGROUND: Gliomas are one of the most common types of primary tumors in central nervous system. Previous studies have found that macrophages actively participate in tumor growth. METHODS: Weighted gene co-expression network analysis was used to identify meaningful macrophage-related gene genes for clustering. Pamr, SVM, and neural network were applied for validating clustering results. Somatic mutation and methylation were used for defining the features of identified clusters. Differentially expressed genes (DEGs) between the stratified groups after performing elastic regression and principal component analyses were used for the construction of MScores. The expression of macrophage-specific genes were evaluated in tumor microenvironment based on single cell sequencing analysis. A total of 2365 samples from 15 glioma datasets and 5842 pan-cancer samples were used for external validation of MScore. RESULTS: Macrophages were identified to be negatively associated with the survival of glioma patients. Twenty-six macrophage-specific DEGs obtained by elastic regression and PCA were highly expressed in macrophages at single-cell level. The prognostic value of MScores in glioma was validated by the active proinflammatory and metabolic profile of infiltrating microenvironment and response to immunotherapies of samples with this signature. MScores managed to stratify patient survival probabilities in 15 external glioma datasets and pan-cancer datasets, which predicted worse survival outcome. Sequencing data and immunohistochemistry of Xiangya glioma cohort confirmed the prognostic value of MScores. A prognostic model based on MScores demonstrated high accuracy rate. CONCLUSION: Our findings strongly support a modulatory role of macrophages, especially M2 macrophages in glioma progression and warrants further experimental studies.

In Vitro Reconstitution of Yeast Translation System Capable of Synthesizing Long Polypeptide and Recapitulating Programmed Ribosome Stalling.

The rates of translation elongation or termination in eukaryotes are modulated through cooperative molecular interactions involving mRNA, the ribosome, aminoacyl- and nascent polypeptidyl-tRNAs, and translation factors. To investigate the molecular mechanisms underlying these processes, we developed an in vitro translation system from yeast, reconstituted with purified translation elongation and termination factors, utilizing CrPV IGR IRES-containing mRNA, which functions in the absence of initiation factors. The system is capable of synthesizing not only short oligopeptides but also long reporter proteins such as nanoluciferase. By setting appropriate translation reaction conditions, such as the Mg2+/polyamine concentration, the arrest of translation elongation by known ribosome-stalling sequences (e.g., polyproline and CGA codon repeats) is properly recapitulated in this system. We describe protocols for the preparation of the system components, manipulation of the system, and detection of the translation products. We also mention critical parameters for setting up the translation reaction conditions. This reconstituted translation system not only facilitates biochemical analyses of translation but is also useful for various applications, such as structural and functional studies with the aim of designing drugs that act on eukaryotic ribosomes, and the development of systems for producing novel functional proteins by incorporating unnatural amino acids by eukaryotic ribosomes.