Strand-specific miR-28-5p and miR-28-3p have distinct effects in colorectal cancer cells

The authors thank Sue Moreau from the Department of Scientific Publications at The University of Texas MD Anderson Cancer Center for English language editing of the manuscript. Author contributions: Study concept and design: M.I.A., P.A.Z, G.A.C. Acquisition of data: M.I.A., L.Z., X.Z. Drafting of the manuscript: M.I.A., M.N., R.S., M.F., R.M.R., P.A.Z, G.A.C. Analysis and interpretation of data: M.I.A., M.N., R.S., R.M., P.A.Z, G.A.C. Critical revision of the manuscript for important intellectual content: M.I.A., M.N., R.S., M.F., R.M.R., P.A.Z, G.A.C. Statistical analysis: M.I.A., C.I., L.X. Obtained funding: G.A.C. Administrative, technical, or material support: R.G., I.V., F.F., M.F., G.L. Study supervision: G.A.C. Drs Nicoloso and Spizzo are currently at the Division of Experimental Oncology, CRO, National Cancer Institute, Aviano, ItalyBackground & Aims MicroRNAs (miRNAs) can promote or inhibit tumor growth and are therefore being developed as targets for cancer therapies. They are diverse not only in the messenger RNAs (mRNA) they target, but in their production; the same hairpin RNA structure can generate mature products from each strand, termed 5p and 3p, that can bind different mRNAs. We analyzed the expression, functions, and mechanisms of miR-28-5p and miR-28-3p in colorectal cancer (CRC) cells. Methods We measured levels of miR-28-5p and miR-28-3p expression in 108 CRC and 49 normal colorectal samples (47 paired) by reverse transcription, quantitative real-time polymerase chain reaction. The roles of miR-28 in CRC development were studied using cultured HCT116, RKO, and SW480 cells and tumor xenograft analyses in immunodeficient mice; their mRNA targets were also investigated. Results miR-28-5p and miR-28-3p were down-regulated in CRC samples compared with normal colon samples. Overexpression of miRNAs in CRC cells had different effects and the miRNAs interacted with different mRNAs: miR-28-5p altered expression of CCND1 and HOXB3, whereas miR-28-3p bound NM23-H1. Overexpression of miR-28-5p reduced CRC cell proliferation, migration, and invasion in vitro, whereas miR-28-3p increased CRC cell migration and invasion in vitro. CRC cells overexpressing miR-28 developed tumors more slowly in mice compared with control cells, but miR-28 promoted tumor metastasis in mice. Conclusion miR-28-5p and miR-28-3p are transcribed from the same RNA hairpin and are down-regulated in CRC cells. Overexpression of each has different effects on CRC cell proliferation and migration. Such information has a direct application for the design of miRNA gene therapy trials.M.I.A. is supported by a PhD fellowship (SFRH/BD/47031/2008) from Fundação para a Ciência e Tecnologia, Portugal. G.A.C. is supported as a fellow by The University of Texas MD Anderson Cancer Center Research Trust, The University of Texas System Regents Research Scholar, and the Chronic Lymphocytic Leukemia Global Research Foundation. Work in Dr Calin’s laboratory is supported in part by grants from the National Institutes of Health (CA135444), the US Department of Defense, the Pancreatic Cancer Action Network (2009 Seena Magowitz AACR Pilot Grant), and the US-European Alliance for the Therapy of Chronic Lymphoid Leukemia. STR DNA fingerprinting was done by the Cancer Center Support grant funded Characterized Cell Line core, NCI # CA16672

Clinical Outcomes of Primary Posterior Continuous Curvilinear Capsulorhexis in Postvitrectomy Cataract Eyes

Purpose. To evaluate the safety and outcomes of primary posterior continuous curvilinear capsulorhexis (PPCCC) combined with phacoemulsification in postvitrectomy eyes. Design. Retrospective case series. Methods. Twenty-one postvitrectomy eyes of 21 patients with cataract between April 2017 and December 2019 were enrolled. PPCCC through the cornea incision was performed before in-the-bag intraocular lens implantation. All patients were followed up for at least 3 months postoperatively. The outcome measures were corrected distance visual acuity (CDVA), intraocular pressure (IOP), corneal endothelium cell counts (CECC), central macular thickness (CMT), the occurrence of intraoperative or postoperative complications, and the incidence of posterior capsular opacification (PCO). Results. The mean age was 56.14 ± 9.76 years (ranging from 31 to 68). The mean Snellen CDVA was 20/400 preoperatively and improved to 20/67 postoperatively P<0.001. No significant differences were found between IOP P=0.96 and CMT P=0.42 preoperatively and postoperatively. The mean CECC was 2571.8 ± 319.3 cells/mm2 preoperatively and 2498.2 ± 346.3 cells/mm2 postoperatively P<0.05. Lens epithelium cells proliferation was confined to the peripheral capsular bag during a mean follow-up of 12.9 ± 10.5 months (ranging from 3 to 28 months). Intraoperative posterior capsule extension occurred in 1 eye (4%), although it did not affect the patient’s vision. No serious complications, including retinal detachment or endophthalmitis, were detected in any of the 21 cases. Conclusion. PPCCC through cornea incision combined with phacoemulsification may be a safe and practical alternative to prevent PCO in postvitrectomy eyes with cataract

DNA damage response(DDR): a link between cellular senescence and human cytomegalovirus

Abstract The DNA damage response (DDR) is a signaling cascade that is triggered by DNA damage, involving the halting of cell cycle progression and repair. It is a key event leading to senescence, which is characterized by irreversible cell cycle arrest and the senescence-associated secretory phenotype (SASP) that includes the expression of inflammatory cytokines. Human cytomegalovirus (HCMV) is a ubiquitous pathogen that plays an important role in the senescence process. It has been established that DDR is necessary for HCMV to replicate effectively. This paper reviews the relationship between DDR, cellular senescence, and HCMV, providing new sights for virus-induced senescence (VIS)

Optimizing the localization of astaxanthin enzymes for improved productivity

Abstract Background One important metabolic engineering strategy is to localize the enzymes close to their substrates for improved catalytic efficiency. However, localization configurations become more complex the greater the number of enzymes and substrates is involved. Indeed, optimizing synthetic pathways by localizing multiple enzymes remains a challenge. Terpenes are one of the most valuable and abundant natural product groups. Phytoene, lycopene and β-carotene serve as common intermediates for the synthesis of many carotenoids and derivative compounds, which are hydrophobic long-chain terpenoids, insoluble in water and usually accumulate in membrane compartments. Results While β-ionone synthesis by β-carotene cleavage dioxygenase PhCCD1 and astaxanthin synthesis by β-carotene ketolase (CrtW) and β-carotene hydroxylase (CrtZ) differ in complexity (single and multiple step pathways), the productivity of both pathways benefited from controlling enzyme localization to the E. coli cell membrane via a GlpF protein fusion. Especially, the astaxanthin synthesis pathway comprises both CrtW and CrtZ, which perform four interchangeable reactions initiated from β-carotene. Up to four localization strategies of CrtW and CrtZ were exhaustively discussed in this work, and the optimal positioning strategy was achieved. CrtW and CrtZ were linked using a flexible linker and localized to the membrane via a GlpF protein fusion. Enzymes in the optimal localization configuration allowed a 215.4% astaxanthin production increase. Conclusions This work exploits a localization situation involving membrane-bound substrates, intermediates and multiple enzymes for the first time, and provides a workable positioning strategy to solve problems in similar circumstances

Development and validation of a nomogram to better predict hypertension based on a 10-year retrospective cohort study in China

Background: Hypertension is a highly prevalent disorder. A nomogram to estimate the risk of hypertension in Chinese individuals is not available. Methods: 6201 subjects were enrolled in the study and randomly divided into training set and validation set at a ratio of 2:1. The LASSO regression technique was used to select the optimal predictive features, and multivariate logistic regression to construct the nomograms. The performance of the nomograms was assessed and validated by AUC, C-index, calibration curves, DCA, clinical impact curves, NRI, and IDI. Results: The nomogram140/90 was developed with the parameters of family history of hypertension, age, SBP, DBP, BMI, MCHC, MPV, TBIL, and TG. AUCs of nomogram140/90 were 0.750 in the training set and 0.772 in the validation set. C-index of nomogram140/90 were 0.750 in the training set and 0.772 in the validation set. The nomogram130/80 was developed with the parameters of family history of hypertension, age, SBP, DBP, RDWSD, and TBIL. AUCs of nomogram130/80 were 0.705 in the training set and 0.697 in the validation set. C-index of nomogram130/80 were 0.705 in the training set and 0.697 in the validation set. Both nomograms demonstrated favorable clinical consistency. NRI and IDI showed that the nomogram140/90 exhibited superior performance than the nomogram130/80. Therefore, the web-based calculator of nomogram140/90 was built online. Conclusions: We have constructed a nomogram that can be effectively used in the preliminary and in-depth risk prediction of hypertension in a Chinese population based on a 10-year retrospective cohort study. Funding: This study was supported by the Hebei Science and Technology Department Program (no. H2018206110)

DataSheet1_Effects of biochar and vermicompost on microorganisms and enzymatic activities in greenhouse soil.xlsx

The effects of different contents of biochar and vermicompost on the microbial and enzymatic activities of greenhouse soil were determined to provide a theoretical basis for improving the quality of greenhouse soil. The experiment was conducted in a greenhouse using potted tomatoes. Five treatments consisted of different amount ratios of organic amendments: 1% biochar (BC1), 3% biochar (BC3), 5% biochar (BC5), 3% vermicompost (VC3), and 5% vermicompost (VC5), with no addition of organic amendments as the control (CK). Compared with CK, the pH, organic matter content, and DOC concentration increased in treatment groups. The organic matter content of BC3 and BC5 significantly increased by 54.6% and 72.8%, respectively, and DOC concentration of BC3 significantly increased by 43.9%. Biochar and vermicompost significantly increased the diversity of bacterial and fungal communities in soil, as well as the abundance of Actinomycetes, Acidobacteria, Ascomycetes, and Aspergillus, and reduced the abundance of Aspergillus. The activities of urease and alkaline phosphatase were significantly increased, and the activity of nitrate reductase was inhibited in all treatment groups compared with CK. In addition, a highly significant positive correlation was observed among pH, Acidobacteria phylum abundance, and alkaline phosphatase activity in all treatments. DOC concentration was positively correlated with pH, organic matter content, Acidobacteria phylum abundance and alkaline phosphatase activity. Biochar and vermicompost were effective in improving the physicochemical properties of greenhouse soil, enhancing microbial diversity, and affecting enzymatic activities. Therefore, BC3 (3% biochar) had the most significant effect on community diversity and alkaline phosphatase and nitrate reductase activities. VC5 (5% vermicompost) had the best promotion effect on urease activity. This study highlights that biochar and vermicompost as organic amendments are recommended to improve the quality of greenhouse soils.</p

A Novel Aptamer-Imprinted Polymer-Based Electrochemical Biosensor for the Detection of Lead in Aquatic Products

Lead contamination in aquatic products is one of the main hazard factors. The aptasensor is a promising detection method for lead ion (Pb(II)) because of its selectivity, but it is easily affected by pH. The combination of ion-imprinted polymers(IIP) with aptamers may improve their stability in different pH conditions. This paper developed a novel electrochemical biosensor for Pb(II) detection by using aptamer-imprinted polymer as a recognition element. The glassy carbon electrode was modified with gold nanoparticles and aptamers. After the aptamer was induced by Pb(II) to form a G-quadruplex conformation, a chitosan-graphene oxide was electrodeposited and cross-linked with glutaraldehyde to form an imprint layer, improving the stability of the biosensor. Under the optimal experimental conditions, the current signal change (&#8710;I) showed a linear correlation of the content of Pb(II) in the range of 0.1&ndash;2.0 &mu;g/mL with a detection limit of 0.0796 &mu;g/mL (S/N = 3). The biosensor also exhibited high selectivity for the determination of Pb(II) in the presence of other interfering metal ion. At the same time, the stability of the imprinted layer made the sensor applicable to the detection environment with a pH of 6.4&ndash;8.0. Moreover, the sensor was successfully applied to the detection of Pb(II) in mantis shrimp