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Microbial genetic circuits are vital for regulating gene expression and synthesizing bioactive compounds. However, assessing their strength and timing, especially in multicellular fungi, remains challenging. Here, an advanced microfluidic platform is combined with a mathematical model enabling precise characterization of fungal gene regulatory circuits (GRCs) at the single‐cell level. Utilizing this platform, the expression intensity and timing of 30 transcription factor‐promoter combinations derived from two representative fungal GRCs, using the model fungus Aspergillus nidulans are determined. As a proof of concept, the selected GRC combination is utilized to successfully refactor the biosynthetic pathways of bioactive molecules, precisely control their production, and activate the expression of the silenced biosynthetic gene clusters (BGCs). This study provides insights into microbial gene regulation and highlights the potential of platform in fungal synthetic biology applications and the discovery of novel natural products. [ABSTRACT FROM AUTHOR]

Objective: To assess the effects of amoxicillin and metronidazole with scaling and root planing (SRP) on periodontal parameters and glycemic control in patients with severe periodontitis and diabetes mellitus. Background: Adjunctive antibiotics use is advantageous for treating periodontitis in patients with severe periodontitis and diabetes. However, the effects of adjunctive antibiotic use on hemoglobin A1c (HbA1c) levels remain unclear. Methods: This short‐term, randomized controlled trial enrolled patients with severe periodontitis and type 2 diabetes. The patients were randomly allocated to SPR only (i.e., control) or SPR + antibiotics (500 mg of amoxicillin and 200 mg of metronidazole, three times daily for 7 days) groups. Periodontal and hematological parameters were assessed at baseline and 3 months after treatment. Inter‐ and intra‐group analyses were performed using Student's t‐tests, Mann–Whitney U tests, and the binary logistic regression models. p‐values of <.05 were considered statistically significant. Results: This study enrolled 49 patients, with 23 and 26 patients in the SRP‐only and SRP + antibiotics groups, respectively. The periodontal parameters improved significantly and similarly in both groups after treatment (p <.05). The SRP + antibiotics group had more sites of improvement than the SRP‐only group when the initial probing depth was >6 mm. (698 [78.96%] vs. 545 [73.35%], p =.008). The HbA1c levels decreased in the SRP‐only and SRP + antibiotics groups after treatment (0.39% and 0.53%, respectively). The multivariable binary logistic regression model demonstrated that antibiotics administration and a high baseline HbA1c level were associated with a greater reduction in the HbA1c level (odds ratio = 4.551, 95% confidence interval: 1.012–20.463; odds ratio = 7.162, 95% confidence interval: 1.359–37.753, respectively). Conclusions: SRP and SRP plus systemic antibiotics were beneficial for glycemic control. Adjunctive antibiotic use slightly improved the outcome for patients with severe periodontitis and poorly controlled diabetes. [ABSTRACT FROM AUTHOR]

A previous genome‐wide association study (GWAS) revealed an association of the noncoding SNP rs1663689 with susceptibility to lung cancer in the Chinese population. However, the underlying mechanism is unknown. In this study, using allele‐specific 4C‐seq in heterozygous lung cancer cells combined with epigenetic information from CRISPR/Cas9‐edited cell lines, we show that the rs1663689 C/C variant represses the expression of ADGRG6, a gene located on a separate chromosome, through an interchromosomal interaction of the rs1663689 bearing region with the ADGRG6 promoter. This reduces downstream cAMP‐PKA signaling and subsequently tumor growth both in vitro and in xenograft models. Using patient‐derived organoids, we show that rs1663689 T/T—but not C/C—bearing lung tumors are sensitive to the PKA inhibitor H89, potentially informing therapeutic strategies. Our study identifies a genetic variant‐mediated interchromosomal interaction underlying ADGRG6 regulation and suggests that targeting the cAMP‐PKA signaling pathway may be beneficial in lung cancer patients bearing the homozygous risk genotype at rs1663689. Synopsis: This study shows a trans‐effect of a noncoding genetic variant and suggests a personalized treatment strategy for lung cancer patients harboring the T/T genotype at rs1663689. rs1663689 regulates lung cancer progression.rs1663689 regulates ADGRG6 expression through interchromosomal interaction.Organoids derived from lung cancer patients carrying the risk allele at rs1663689 respond to PKA inhibitor. [ABSTRACT FROM AUTHOR]

Background: To investigate the relation of established glucose and lipid metabolism indexes and blood inflammatory biomarkers with severe periodontitis in systemically healthy patients.Methods: Systemically healthy stage III/IV periodontitis patients (case group) (n = 397), stage II periodontitis patients (n = 36), and periodontally healthy subjects (control group) (n = 285) were recruited. A periodontal examination, complete blood cell examination, and blood biochemical examination were conducted for all participants. Full-mouth apical films were taken for the case group. Both the case and control groups were divided by age into younger (≤35 years) and elder subjects. Multiple logistic regression analysis and Pearson correlation analysis were conducted. A logistic least absolute shrinkage and selection operator (LASSO) model was constructed for the younger subgroups.Results: Various glucose and lipid metabolism indexes and blood inflammatory biomarkers significantly differed between severe periodontitis patients and healthy controls, and the younger subgroups presented a greater degree of statistical differences than the elder ones. More pairs of periodontal parameters and blood indexes with significantly fair linear correlations were found in the younger patient subgroup. A logistic LASSO regression model containing eight blood indexes to assess a severe periodontitis outcome in younger subgroups showed satisfactory predictive ability.Conclusion: The present study revealed various glucose and lipid metabolism indexes and blood inflammatory biomarkers significantly differ between severe periodontitis patients and healthy controls, especially in the younger subgroups. A LASSO regression model could be a viable option to assess severe periodontitis risk for younger patients.Summary: The present study revealed significant differences in glucose and lipid metabolism indexes and blood inflammatory biomarkers between patients with severe periodontitis and periodontally healthy individuals, especially in younger subjects. This article is protected by copyright. All rights reserved. [ABSTRACT FROM AUTHOR]

The regulation of the catalytic activities of DNAzymes is of great significance for their biosensing and biomedical applications. Herein, thermoregulation of the catalytic activity of DNAzyme is achieved through the immobilization of DNAzymes into the crosslinked polymeric matrix of poly‐N‐isopropylacrylamide (pNIPAM) microgels prepared with a microfluidic device. At a temperature lower than the lower critical solution temperature (LCST) of pNIPAM, the immobilized DNAzymes exhibit lower catalytic performance due to the restricted entry of DNA substrates into the polymeric networks; while at a temperature higher than the LCST, the hydrophilic DNAzyme structures are exposed to exhibit high catalytic activity due to the shrinking of the pNIPAM networks. Results show a more than 60% of increase in catalytic activity by raising the temperature from 29 to 34 °C and reversible regulation of the DNAzyme catalytic activity is achieved by cyclic raising and lowering of the solution temperature. Through a simple heating‐centrifugation process, the DNAzyme/pNIPAM microgels can be recycled for further applications. Moreover, by introducing photothermal MXene nanosheets, the remote control of the catalytic activity of the DNAzyme by near‐infrared light irradiation is achieved. These DNAzyme/pNIPAM microgels as low‐cost, stable, reversible, and recyclable biocatalyst may have potential in biosensing and biomedical applications. [ABSTRACT FROM AUTHOR]

Aim: Recently, microRNA‐149 (miR‐149) has been indicated to act as an oncogene or a tumor suppressor in various malignant tumors, while its inner mechanisms in recurrent miscarriage (RM) are still in infancy. Therein, this study intends to decode the mechanism of miR‐149 in RM. Methods: miR‐149 and RUNX2 expression in the chorionic tissues of normal pregnant women and RM patients were first examined, and the correlation between miR‐149 and RUNX2 was analyzed. Subsequently, miR‐149 was upregulated in HTR‐8 cells or downregulated in BEWO cells, and then the changes in biological functions of trophoblasts in RM were detected. Furthermore, the expression of PTEN/Akt signaling pathway‐related factors in trophoblasts was detected by western blot analysis. Results: miR‐149 expression was increased while RUNX2 expression was suppressed in RM patients, and miR‐149 was negatively correlated with RUNX2. Overexpressed miR‐149 induced cell apoptosis and inhibited cell activity, while reduced miR‐149 in trophoblasts contributed to opposite experimental results. Moreover, miR‐149 promoted the expression of PTEN and inhibited Akt phosphorylation by targeting RUNX2, thereby inhibiting trophoblast activity and promoting their apoptosis. Conclusion: Our study demonstrates that miR‐149 knockdown halted the RM development through upregulating RUNX2 and activation of the PTEN/Akt signaling pathway. [ABSTRACT FROM AUTHOR]

Background: The authors hypothesized that epigenetic changes may help to clarify the underlying biologic mechanism linking aspirin use to breast cancer prognosis. To the authors' knowledge, this is the first epidemiologic study to examine whether global methylation and/or tumor promoter methylation of breast cancer‐related genes interact with aspirin use to impact mortality after breast cancer. Methods: Prediagnosis aspirin use was assessed through in‐person interviews within a population‐based cohort of 1508 women diagnosed with a first primary breast cancer in 1996 and 1997. Global methylation in peripheral blood was assessed by long interspersed elements‐1 (LINE‐1) and the luminometric methylation assay. Promoter methylation of 13 breast cancer‐related genes was measured in tumor by methylation‐specific polymerase chain reaction and the MethyLight assay. Vital status was determined by the National Death Index through December 31, 2014 (N = 202/476 breast cancer‐specific/all‐cause deaths identified among 1266 women with any methylation assessment and complete aspirin data). Cox proportional hazards regression was used to estimate hazard ratios (HRs) and 95% CIs, and the likelihood ratio test was used to evaluate multiplicative interactions. Results: All‐cause mortality was elevated among aspirin users who had methylated promotor of BRCA1 (HR, 1.67; 95% CI, 1.26‐2.22), but not among those with unmethylated promoter of BRCA1 (HR, 0.99; 95% CI, 0.67‐1.45; P for interaction ≤.05). Decreased breast cancer‐specific mortality was observed among aspirin users who had unmethylated promotor of BRCA1 and PR and global hypermethylation of LINE‐1 (HR, 0.60, 0.78, and 0.63, respectively; P for interaction ≤.05), although the 95% CIs included the null. Conclusions: The current study suggests that the LINE‐1 global methylation and promoter methylation of BRCA1 and PR in tumor may interact with aspirin use to influence mortality after breast cancer. Findings from this study suggest that the association between aspirin use and mortality after breast cancer may depend upon patients' DNA methylation profiles. The study may help to identify certain women who may benefit from aspirin use to improve survival after a breast cancer diagnosis. [ABSTRACT FROM AUTHOR]

Blue light plays an important role during the growth of Ganoderma lucidum, one of the best-known medicinal macrofungi in China. In the present study, we cloned Glwc-1 and Glwc-2, the homologue of the blue light photoreceptors Ncwc-1 and Ncwc-2 of Neurospora crassa, from G. lucidum. The deduced amino acid sequence of Glwc-1 contained the similar function domains as NcWC-1 including LOV, PAS B, PAS C, and PAC domains. The deduced amino acid sequence of Glwc-2 contained PAS domain and GATA-type zinc finger (Znf) domain as well as NcWC-2. Phylogenetic analysis based on fungal WC-1 and WC-2 supported GlWC-1 and GlWC-2 were blue light receptors. The expression of Glwc-1 and Glwc-2 indicated that they might play an important role during the primordium differentiation process of G. lucidum, and the external blue light stimulation increased the expression of Glwc-1 and Glwc-2. [ABSTRACT FROM AUTHOR]

Pseudocercospora is an important plant pathogenic genus causing leaf spots on grapevine leaves and other hosts worldwide. A survey of vineyards in four provinces of China was carried out to identify the causal agent of Pseudocercospora leaf spot. A phylogenetic analysis based on a combination of LSU, ITS and ACT loci, coupled with morphology, showed that Pseudocercospora vitis is the sole causal agent of this leaf spot disease. This study represents the first attempt to identify and characterize the cercosporoid species causing Pseudocercospora leaf spot in China using morphological and molecular approaches. [ABSTRACT FROM AUTHOR]

Tetralogy of Fallot ( TOF) is a complex congenital heart defect and the micro RNAs regulation in TOF development is largely unknown. Herein, we explored the role of mi RNAs in TOF. Among 75 dysregulated mi RNAs identified from human heart tissues, mi RNA-940 was the most down-regulated one. Interestingly, mi RNA-940 was most highly expressed in normal human right ventricular out-flow tract comparing to other heart chambers. As TOF is caused by altered proliferation, migration and/or differentiation of the progenitor cells of the secondary heart field, we isolated Sca-1+ human cardiomyocyte progenitor cells (h CMPC) for mi RNA-940 function analysis. mi RNA-940 reduction significantly promoted h CMPCs proliferation and inhibited h CMPCs migration. We found that JARID2 is an endogenous target regulated by mi RNA-940. Functional analyses showed that JARID2 also affected h CMPCs proliferation and migration. Thus, decreased mi RNA-940 affects the proliferation and migration of the progenitor cells of the secondary heart field by targeting JARID2 and potentially leads to TOF development. [ABSTRACT FROM AUTHOR]

Sarcolemmal Na+/ H+ exchanger 1 ( NHE1) activity is essential for the intracellular pH (pHi) homeostasis in cardiac myocytes. Emerging evidence indicates that sarcolemmal NHE1 dysfunction was closely related to cardiomyocyte death, but it remains unclear whether defective trafficking of NHE1 plays a role in the vital cellular signalling processes. Dynamin ( DNM), a large guanosine triphosphatase ( GTPase), is best known for its roles in membrane trafficking events. Herein, using co-immunoprecipitation, cell surface biotinylation and confocal microscopy techniques, we investigated the potential regulation on cardiac NHE1 activity by DNM. We identified that DNM2, a cardiac isoform of DNM, directly binds to NHE1. Overexpression of a wild-type DNM2 or a dominant-negative DNM2 mutant with defective GTPase activity in adult rat ventricular myocytes ( ARVMs) facilitated or retarded the internalization of sarcolemmal NHE1, whereby reducing or increasing its activity respectively. Importantly, the increased NHE1 activity associated with DNM2 deficiency led to ARVMs apoptosis, as demonstrated by cell viability, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling assay, Bcl-1/Bax expression and caspase-3 activity, which were effectively rescued by pharmacological inhibition of NHE1 with zoniporide. Thus, our results demonstrate that disruption of the DNM2-dependent retrograde trafficking of NHE1 contributes to cardiomyocyte apoptosis. [ABSTRACT FROM AUTHOR]

Tetralogy of Fallot (TOF) is a complex congenital heart defect and the microRNAs regulation in TOF development is largely unknown. Herein, we explored the role of miRNAs in TOF. Among 75 dysregulated miRNAs identified from human heart tissues, miRNA-940 was the most down-regulated one. Interestingly, miRNA-940 was most highly expressed in normal human right ventricular out-flow tract comparing to other heart chambers. As TOF is caused by altered proliferation, migration and/or differentiation of the progenitor cells of the secondary heart field, we isolated Sca-1(+) human cardiomyocyte progenitor cells (hCMPC) for miRNA-940 function analysis. miRNA-940 reduction significantly promoted hCMPCs proliferation and inhibited hCMPCs migration. We found that JARID2 is an endogenous target regulated by miRNA-940. Functional analyses showed that JARID2 also affected hCMPCs proliferation and migration. Thus, decreased miRNA-940 affects the proliferation and migration of the progenitor cells of the secondary heart field by targeting JARID2 and potentially leads to TOF development., (© 2014 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

Sarcolemmal Na(+) /H(+) exchanger 1 (NHE1) activity is essential for the intracellular pH (pHi ) homeostasis in cardiac myocytes. Emerging evidence indicates that sarcolemmal NHE1 dysfunction was closely related to cardiomyocyte death, but it remains unclear whether defective trafficking of NHE1 plays a role in the vital cellular signalling processes. Dynamin (DNM), a large guanosine triphosphatase (GTPase), is best known for its roles in membrane trafficking events. Herein, using co-immunoprecipitation, cell surface biotinylation and confocal microscopy techniques, we investigated the potential regulation on cardiac NHE1 activity by DNM. We identified that DNM2, a cardiac isoform of DNM, directly binds to NHE1. Overexpression of a wild-type DNM2 or a dominant-negative DNM2 mutant with defective GTPase activity in adult rat ventricular myocytes (ARVMs) facilitated or retarded the internalization of sarcolemmal NHE1, whereby reducing or increasing its activity respectively. Importantly, the increased NHE1 activity associated with DNM2 deficiency led to ARVMs apoptosis, as demonstrated by cell viability, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling assay, Bcl-1/Bax expression and caspase-3 activity, which were effectively rescued by pharmacological inhibition of NHE1 with zoniporide. Thus, our results demonstrate that disruption of the DNM2-dependent retrograde trafficking of NHE1 contributes to cardiomyocyte apoptosis., (© 2013 The Authors. Journal of Cellular and Molecular Medicine Published by Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.)