Transforming Gene Research Articles

The FveFT2 florigen/FveTFL1 antiflorigen balance is critical for the control of seasonal flowering in strawberry while FveFT3 modulates axillary meristem fate and yield.

Summary Plant architecture is central in determining crop yield. In the short‐day species strawberry, a crop vegetatively propagated by daughter‐plants produced by stolons, fruit yield is further dependent on the trade‐off between sexual reproduction (fruits) and asexual reproduction (daughter‐plants). Both are largely dependent on meristem identity, which establishes the development of branches, stolons and inflorescences.Floral initiation and plant architecture are modulated by the balance between two related proteins, FLOWERING LOCUS T (FT) and TERMINAL FLOWER 1 (TFL1). We explored in woodland strawberry the role of the uncharacterised FveFT2 and FveFT3 genes and of the floral repressor FveTFL1 through gene expression analyses, grafting and genetic transformation (overexpression and gene editing).We demonstrate the unusual properties of these genes. FveFT2 is a nonphotoperiodic florigen permitting short‐day (SD) flowering and FveTFL1 is the long‐hypothesised long‐day systemic antiflorigen that contributes, together with FveFT2, to the photoperiodic regulation of flowering. We additionally show that FveFT3 is not a florigen but promotes plant branching when overexpressed, that is likely to be through changing axillary meristem fate, therefore resulting in a 3.5‐fold increase in fruit yield at the expense of stolons.We show that our findings can be translated into improvement of cultivated strawberry in which FveFT2 overexpression significantly accelerates flowering.

Leukemogenic Pathway of MLL-AF4 Fusion-Positive Acute Lymphoblastic Leukemia

Background Many hematologic malignancies, including leukemia, are caused by genetic abnormalities such as chromosomal translocation. These gene defects usually perturb differentiation and proliferation of hematopoietic stem/progenitor cells. As observed in most cancers, multiple gene defects are required to develop overt leukemia, and multi-step tumorigenesis is also the rule in leukemogenesis. Leukemia-specific fusion genes are usually generated early in leukemogenesis as essential genetic defects (1st hit) producing pre-leukemic stem cells. Additional genetic defect(s) occur in pre-leukemic stem cells, finally generating leukemic stem cells that expand to present overt leukemia. Infant acute lymphoblastic leukemia (ALL) are frequently characterized by chromosome translocations involving 11q23, resulting in the rearrangement of the mixed-lineage leukemia (MLL) gene and subsequent generation of MLL fusion genes. Amongst the > 50 genes that have been identified as fusion partners of MLL, fusion with AF4 is characteristically and commonly observed in infant ALL, representing a hallmark of poor prognosis. In most cases of ALL with MLL-AF4, the fusion gene originates in utero as a primary event. Although mutations in RAS and FLT3 are frequently observed in MLL-AF4 -positive ALL, recent genetic data showed significantly lower number of accompanying mutations, which suggested that 2nd hit mutations were not necessary for developing MLL fusion-positive leukemia. The necessity of 2nd hit mutations in leukemogenesis with MLL-AF4 is still under debate. Materials and Methods Several previous studies have modeled leukemic transformation by introducing fusion genes into hematopoietic stem/progenitor cells; however, reproduction of certain forms of ALL, such as MLL-AF4 -positive infant ALL, has been challenging. We used the immature hematopoietic cells derived from mouse embryonic stem (ES) cells as targets for MLL-AF4 -mediated transformation. The MLL-AF4 fusion gene was introduced into mouse ES cells to obtain ES cell clones with stable expression of MLL-AF4 . These ES cells were conditioned to generate embryoid bodies (EBs) and subsequently differentiated into hematopoietic lineages. Tie2-positive cells, which contain immature hematopoietic stem cells, were separated and transplanted into immunodeficient non-obese diabetic/Shi-scid, IL-2Rγ null (NOG) mice to investigate leukemia development. Retroviral insertional mutagenesis was used to introduce additional genetic defect(s) as candidates for tumor-promoting 2nd hit mutations. Purified Tie2-positive cells were transduced using the murine stem cell virus (MSCV) and transplanted into NOG mice. Results Mouse ES cells with stable expression of the MLL-AF4 fusion were viable and could proliferate and differentiate to produce EBs. However, EBs derived from MLL-AF4 -positive ES cells differentiated via the mesenchymal pathway rather than into the Tie2-positive hematopoietic lineage. Mice transplanted with Tie2-positive cells with MLL-AF4 expression did not develop leukemia within the observed period. However, addition of an additional genetic change by retroviral insertional mutagenesis accelerated tumorigenesis in the recipient mice. The MLL-AF4 -expressing Tie2-positive cells with insertional mutagenesis showed clonal expansion and formed mostly extra-medullary tumors with B220 expression, but did not fully develop into tumors of hematopoietic organs. Viral integration sites of these tumors were analyzed to determine additional genetic events inducing tumor formation. In the MLL-AF4 model, viral integration often caused overexpression of gene(s) located near the integration site. Conclusion Considering the in utero formation of MLL-AF4 harboring pre-leukemic stem cells in ALL, hematopoietic progenitor cells or immature fetal cells may be potential targets of leukemic transformation. Immature hematopoietic cells such as the Tie2-positive cells derived from ES cells could be suitable candidates for transformation with MLL-AF4 . Clonal expansion of MLL-AF4 -positive cells after introduction of retroviral insertional mutagenesis suggests the necessity of additional genetic change(s) that activate certain target gene(s) in leukemogenesis with MLL-AF4 . We are currently analyzing the roles of these genes in the leukemic transformation of MLL-AF4 -positive hematopoietic cells. Disclosures No relevant conflicts of interest to declare.

A transgenic female killing system for the genetic control of Drosophila suzukii

The spotted wing Drosophila (Drosophila suzukii) is an invasive pest of soft-skinned fruit crops. It is rapidly transmitted in Europe and North America, causing widespread agricultural losses. Genetic control strategies such as the sterile insect technique (SIT) have been proposed as environment-friendly and species-restricted approaches for this pest. However, females are inefficient agents in SIT programs. Here we report a conditional female-killing (FK) strategy based on the tetracycline-off system. We assembled sixteen genetic constructs for testing in vitro and in vivo. Twenty-four independent transgenic strains of D. suzukii were generated and tested for female-specific lethality. The strongest FK effect in the absence of tetracycline was achieved by the construct containing D. suzukii nullo promoter for early gene expression, D. suzukii pro-apoptotic gene hidAla4 for lethality, and the transformer gene intron from the Mediterranean fruit fly Ceratitis capitata for female-specific splicing. One strain carrying this construct eliminated 100% of the female offspring during embryogenesis and produced only males. However, homozygous females from these FK strains were not viable on a tetracycline-supplemented diet, possibly due to the basal expression of hidAla4. Potential improvements to the gene constructs and the use of such FK strains in an SIT program are discussed.

What It Takes to Transform a T Cell.

The role of fusion genes and cancer driver genes in malignant transformation has traditionally been explored using transgenic or chimeric mouse models. It has been challenging to develop models that fully resemble the characteristics and morphology of human cancers. This applies to anaplastic large-cell lymphoma (ALCL), a malignancy classified as a peripheral T-cell lymphoma. It is still unclear at which stage of T-cell development ALCL can occur, as well as the early molecular events required for malignant transformation. In this issue of Cancer Research, Pawlicki and colleagues introduced the NPM-ALK fusion gene and mutant variants into primary T cells from healthy donors. By monitoring transduced T-cell clones over time, they demonstrated that transformed T cells undergo a progressive loss of T-cell identity accompanied with upregulation of epithelial-to-mesenchymal transition program and reemergence of an immature, thymic profile. Introduction of NPM-ALK was, however, not sufficient to convert healthy T cells to malignant clones, as this process required activation of T-cell receptor signaling. The study sets the stage for modeling early genetic changes in human tumors.See related article by Pawlicki et al., p. 3241.

Dissolved oxygen disturbs nitrate transformation by modifying microbial community, co-occurrence networks, and functional genes during aerobic-anoxic transition

No consensus has been achieved among researchers on the effect of dissolved oxygen (DO) on nitrate (NO3−-N) transformation and the microbial community, especially during aerobic-anoxic transition. To supplement this knowledge, NO3−-N transformation, microbial communities, co-occurrence networks, and functional genes were investigated during aerobic-anoxic transition via microcosm simulation. NO3−-N transformation rate in the early stage (DO ≥2 mg/L) was always significantly higher than that in the later stage (DO <2 mg/L) during aerobic-anoxic transition, and NO2−-N accumulation was more significant during the anoxic stage, consistent with the result obtained under constant DO conditions. These NO3−-N transformation characteristics were not affected by other environmental factors, indicating the important role of DO in NO3−-N transformation during aerobic-anoxic transition. Changes in DO provoked significant alterations in microbial diversity and abundance of functional bacteria dominated by Massilia, Bacillus, and Pseudomonas, leading to the variation in NO3−-N transformation. Co-occurrence network analysis revealed that NO3−-N transformation was performed by the interactions between functional bacteria including symbiotic and competitive relationship. In the presence of oxygen, these interactions accelerated the NO3−-N transformation rate, and bacterial metabolization proceeded via increasingly varied pathways including aerobic and anoxic respiration, which was demonstrated through predicted genes. The higher relative abundance of genes narG, narH, and napA suggested the occurrence of coupled aerobic-anoxic denitrification in the early stage. NO3−-N transformation rate decreased accompanied by a significant NO2−-N accumulation with the weakening of coupled aerobic-anoxic denitrification during aerobic-anoxic transition. Structural equation modeling further demonstrated the relationship between DO and NO3−-N transformation. DO affects NO3−-N transformation by modifying microbial community, bacterial co-occurrence, and functional genes during aerobic-anoxic transition.

Enhanced polyethylene glycol (PEG)-mediated protoplast transformation system for the phytopathogenic fungus, Ganoderma boninense.

The basidiomycete fungus, Ganoderma boninense, has been identified as the main causal agent of oil palm basal stem rot (BSR) disease which has caused significant economic losses to the industry especially in Malaysia and Indonesia. Various efforts have been initiated to understand the disease and this plant pathogen especially at the molecular level. This is the first study of its kind on the development of a polyethylene glycol (PEG)-mediated protoplast transformation system for G. boninense. Based on the minimal inhibitory concentration study, 60µg/mL and above of hygromycin were effective to completely inhibit G. boninense growth. Approximately 5.145×107cells/mL of protoplasts with the viability of 97.24% was successfully obtained from G. boninense mycelium tissue. The PEG-mediated G. boninense protoplast transformation using 1µg of transformation vector, 25% of PEG solution, 10min of pre-transformation incubation, and 30min of post-transformation incubation has improved the transformation rate as compared with the previous reported protocols for other basidiomycete fungi. Optimization of four transformation parameters has improved the transformation efficiency of G. boninense from an average of 2 to 67 putative transformants. The presence of hygromycin phosphotransferase (hpt) and enhanced green fluorescent protein (eGFP) genes in the putative transformants was detected by PCR and verified by gene sequence analysis. Southern hybridization result further confirmed the integration of hpt gene in G. boninense transformants, and the green fluorescent signal was detected in the G. boninense transformants under the microscopic analysis. The establishment of this transformation system will accelerate the gene function studies of G. boninense especially those genes that may contribute to the pathogenesis of this fungus in oil palm.

Effects of hydrostatic pressure on phosphorus transformation at the water–sediment interface of a deep reservoir: novel insights into bacterial community and functional genes

The release of endogenous phosphorus from sediments leads to the eutrophication of water bodies. Bacteria in sediments play a vital role in phosphorus cycling. A high hydrostatic pressure can alter the microecology and affect bacterial functions at the water–sediment interface. The aim of this study was to analyze the expression patterns of functional genes in bacteria associated with phosphorus transformation at the water–sediment interface of a deep reservoir under different hydrostatic pressures. Furthermore, the effects of hydrostatic pressure on phosphorus cycling pathways mediated by bacteria were explored. A device was designed to simulate the microecological environment in sediments from a reservoir in Xi’an, China, at atmospheric pressure, 0.2 MPa, 0.5 MPa, and 0.8 MPa. Phosphorus transformation genes, phoD, ppk, and pqqC, were quantified using real-time PCR. The bacterial community composition and diversity were analyzed using high-throughput sequencing of the ppk gene. The total phosphorus contents in the sediments were 724.72, 646.97, 630.92, and 682.88 mg kg–1 at the four respective hydrostatic pressures, respectively. Of this, inorganic phosphorus accounted for 68.8–79.0%, and Fe/Al-P accounted for 47.4–57.2%. The expression of phoD facilitated the transformation of organic to inorganic phosphorus and of Ca-P to Fe/Al-P, whereas the expression of ppk and pqqC regulated organic phosphorus hydrolysis by affecting phosphatase activity. The abundance and diversity of ppk-harboring bacteria were highest at 0.2 MPa, while the dominant genera did not change at the four pressures. The five most abundant genera were Pseudomonas, Bordetella, Cupriavidus, Achromobacter, and Rhizobium. High hydrostatic pressure facilitates the transformation of Fe/Al-P to Ca-P in, and phosphorus release from, reservoir sediments by regulating phoD, ppk, and pqqC expression in bacteria. Changing the hydrostatic pressure can alter the bacterial community structure at the genus level, but not the dominant populations harboring the ppk gene.

Downregulation of lncRNA XR_429159.1 Linked to Brain Metastasis in Patients With Limited-Stage Small-Cell Lung Cancer.

Purpose: The purpose of this study was to identify aberrant long non-coding RNAs (lncRNAs) and explore the predictive value of lncRNA expression patterns on the risk of brain metastases (BMs) in patients with limited-stage small-cell lung cancer (SCLC).Patients and Methods: We executed an array of lncRNA and mRNA chip assays to examine the expression in peripheral blood mononuclear cells obtained from SCLC patients with BMs and compared the expression patterns against those from patients without BMs to identify lncRNAs associated with BMs. Validation was performed against clinical data to further confirm the relationship between lncRNAs and BM. Kaplan–Meier analysis was applied to estimate the cumulative incidence of BMs, and differences between the groups were analyzed using the log-rank test.Results: The expression of 67 lncRNAs (27 upregulated and 40 downregulated) and 47 mRNAs (20 upregulated and 27 downregulated) was significantly different between the BM and non-BM groups (fold change ≥ 2.0, p-value ≤ 0.05), based on the lncRNA and mRNA chip assays. Four lncRNAs were verified by quantitative real-time polymerase chain reaction (qRT-PCR) to confirm the accuracy of the microarray data, and the results of 11 patient pairs (11 patients with BM and 11 patients without BM) revealed that low LncRNA XR_429159.1 expression was a high-risk factor for BM. Further clinical data showed that the incidence of BM among 25 patients with low-level LncRNA XR_429159.1 expression was 31% at 1 year, compared with 14.3% among the 18 patients with high-level LncRNA XR_429159.1 expression (p = 0.035).Conclusion: Our present study identified the low-level expression of lncRNA XR_429159.1 as a high-risk factor among BM in patients with limited-stage SCLC. LncRNA XR_429159.1 is a critical molecule that regulates SCLC metastasis, involved in the neuroepithelial transforming gene 1 (NET1) pathway, and serum levels of this lncRNA are significantly associated with the risk of BM.

Tissue Culture- and Selection-Independent Agrobacterium tumefaciens-Mediated Transformation of a Recalcitrant Grain Legume, Cowpea (Vigna unguiculata L. Walp).

A simple and generally fast Agrobacterium-mediated transformation system with no tissue culture and selection steps has been developed for the first time in a recalcitrant food legume, cowpea. The approach involves wounding of 1-day-old germinated seeds with a needle or sonication either alone or in combination of vacuum infiltration with A. tumefaciens EH105 (pCAMBIA2301) carrying a β-glucuronidase (GUS) gene (uidA) and a neomycin phosphotransferase (nptII) gene for stable transformation. Sonicated and vacuum infiltrated seedlings showed the highest transient GUS activity in 90% of the explants. The sprouted co-cultured seeds directly established in soil and without selection were allowed to develop into plants which on maturity produced T0 seeds. The presence of the alien genes, nptII and uidA in T0 plants and their integration into the genome of T1 plants were confirmed by polymerase chain reaction (PCR) and Southern blot analyses, respectively. The transgenes were inherited in the subsequent T2 generation in a Mendelian fashion and their expression was confirmed by semi-quantitative PCR. The transformation frequency of 1.90% was obtained with sonication followed by vacuum infiltration with Agrobacterium. This approach provides favorable circumstances for the rapid meristem transformation and likely makes translational research ease in an important recalcitrant food legume, cowpea.

Pilot-scale two-stage constructed wetlands based on novel solid carbon for rural wastewater treatment in southern China: Enhanced nitrogen removal and mechanism

Constructed wetlands (CWs) have been proved to be an alternative to the treatment of various wastewater. However, there are few studies focused on the removal performance and mechanisms of pollutants in pilot-scale CWs packed with novel solid carbon. In this study, we investigated the effect of poly-3-hydroxybutyrate-co-3-hydroxyvalerate/polyacetic acid (PHBV/PLA) blends as carbon source on pollutant's transformation, microbial communities and functional genes in pilot-scale aeration-anoxic two-stage CWs for polishing rural runoff in southern China. Results showed a striking improvement of TN removal in CWs with PHBV/PLA blends (64.5%) compared to that in CWs with ceramsite (52.9%). NH4+-N (61.3–64.6%), COD (40.4–53.8%) and TP (43.6–47.1%) were also removed effectively in both two CWs. In addition, the strains of Rhodocyclaceae and Bacteroidetes were the primary denitrifiers on the surface of PHBV/PLA blends. Further, the aerobic stage induced gathering of 16 S and amoA genes and the anoxic zone with PHBV/PLA blends increased the nirS genes, which fundamentally explained the better denitrification performance in CW based on PHBV/PLA blends. Consequently, this study will provide straightforward guidance for the operation of engineering CWs packed with polymers to govern the low-C/N rural wastewater.

Falcarindiol Enhances Cisplatin Chemosensitivity of Hepatocellular Carcinoma via Down-Regulating the STAT3-Modulated PTTG1 Pathway.

Hepatocellular carcinoma (HCC) is the most frequent primary liver malignancy globally and the third leading cause of cancer-related death. Chemotherapy is one of the main methods in treating HCC, while recent studies have found that the resistance of HCC to chemotherapeutic drugs reduces the efficacy of the chemotherapy. Falcarindiol (FAD) is a cytotoxic and anti-inflammatory polyacetylenic oxylipin found in food plants of the carrot family (Apiaceae), while its role in HCC remains to be explored. Here, HCC cells (Huh7 and LM3) were treated with FAD at different doses. Cell proliferation was tested by the cell counting kit-8 (CCK-8) method and colony formation assay, while the apoptosis was monitored by flow cytometry. The profiles of apoptosis-related proteins (Bax, bcl2, and Caspase-3), DNA repair proteins (Rad51, BRCA1, and MDC1), and the signal transducer and activator of transcription 3 (STAT3)/Pituitary Tumor Transforming Gene 1 (PTTG1) were verified by western blot (WB) or quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The interaction between STAT3 and PTTG1 was verified by immunoprecipitation (IP). In addition, a xenograft tumor model was constructed in mice to explore the anti-tumor effects of FAD in vivo, and immunohistochemistry (IHC) was performed to count the number of Ki67-stained cells. As a result, FAD inhibited HCC cell proliferation and DNA repair, facilitated their apoptosis, and also enhanced cisplatin (DDP) chemosensitivity. The Combination Index (CI) evaluation showed that FAD and DDP had synergistic effects in repressing HCC cell proliferation. Besides, FAD dampened the STAT3/PTTG1 pathway expression. Further studies revealed that inhibiting STAT3 enhanced the inhibitive effect of FAD on HCC cells, whereas overexpressing PTTG1 attenuated the anti-tumor effect of FAD. Overall, our study illustrated that FAD is a potential anticancer drug and strengthens the chemosensitivity of HCC cells to DDP by inhibiting the STAT3/PTTG1 pathway.

NET1 promotes HCC growth and metastasis in vitro and in vivo via activating the Akt signaling pathway.

Neuroepithelial cell transforming gene 1 (NET1), a member of the guanine nucleotide exchange factor family, is involved in various cancers, including gastric cancer, breast cancer and glioma. However, the role of NET1 in hepatocellular carcinoma (HCC) remains largely uncovered. In this study, we found that NET1 expression was upregulated in HCC, and that upregulated NET1 expression was closely associated with poor prognosis and some clinical characteristics in HCC patients. Whilst forced expression of NET1 in HCC cells was observed to significantly promote cell growth and metastasis in vitro and in vivo; downregulation of NET1 was shown to exhibit an opposite inhibitory effect. RNA-seq analysis and gene set enrichment analysis demonstrated that knockdown of NET1 significantly suppressed the level of Akt phosphorylation level and the expression of Akt downstream genes in HCC cells. Moreover, MK2206, a potent Akt inhibitor was shown to block the NET1-induced effects in HCC. Taken together, this study demonstrated that, through the Akt signaling pathway, NET1 plays an oncogenic role in HCC progression and metastasis. Hence, NET1 may potentially be used as a potential therapeutic target and prognostic marker of HCC.

Nitrogen transformation and dynamic changes in related functional genes during functional-membrane covered aerobic composting

The effects of functional membrane covering (FMC) on nitrogen transformation and related functional genes during aerobic composting were investigated by performing a comparable experiment. The FMC increased the pile temperature, promoted compost maturity, and decreased nitrogen loss. The FMC reduced NH3 and N2O emissions by 7.34% and 26.27%, respectively. The water film and the micro-positive pressure environment under the membrane effectively prevented NH3 escaping. The FMC up-regulated the amoA gene copy number (promoting NH3/NH4+ oxidation). The reduction of N2O emission by the FMC was mainly related to denitrifying genes (nirK, nirS, and nosZ). The FMC down-regulated the nirK and nirS gene copy numbers, but up-regulated the nosZ gene copy number. Pearson correlation analysis indicated that the functional membrane characteristics and differences between the composting pile environments caused by the FMC significantly affected the nitrogen forms and the related functional genes. The FMC strongly decreased nitrogen emissions and therefore conserved nitrogen.

Methylation Status of Arabidopsis DNA Repair Gene Promoters During Agrobacterium Infection Reveals Epigenetic Changes in Three Generations

Agrobacterium tumefaciens is a unique pathogen with the ability to transfer a portion of its DNA, the T-DNA, to other organisms. The role of DNA repair genes in Agrobacterium transformation remains controversial. In order to understand if the host DNA repair response and dynamics was specific to bacterial factors such as Vir proteins, T-DNA, and oncogenes, we profiled the expression and promoter methylation of various DNA repair genes. These genes belonged to nucleotide excision repair (NER), base excision repair (BER), mismatch repair (MMR), homologous recombination (HR), and non-homologous end joining (NHEJ) pathways. We infected Arabidopsis plants with different Agrobacterium strains that lacked one or more of the above components so that the influence of the respective factors could be analysed. Our results revealed that the expression and promoter methylation of most DNA repair genes was affected by Agrobacterium, and it was specific to Vir proteins, T-DNA, oncogenes, or the mere presence of bacteria. In order to determine if Agrobacterium induced any transgenerational epigenetic effect on the DNA repair gene promoters, we studied the promoter methylation in two subsequent generations of the infected plants. Promoters of at least three genes, CEN2, RAD51, and LIG4 exhibited transgenerational memory in response to different bacterial factors. We believe that this is the first report of Agrobacterium-induced transgenerational epigenetic memory of DNA repair genes in plants. In addition, we show that Agrobacterium induces short-lived DNA strand breaks in Arabidopsis cells, irrespective of the presence or absence of virulence genes and T-DNA.

GPx1-rs1050450 Gene Polymorphism Association with End Stage Renal Disease in Type2 Diabetic Patients of Babylon Province/Iraq

Background: Glutathione Peroxidase-1 is an antioxidant enzyme, play an important role in defense againstoxidative stress. In Pro198Leu (C/T) polymorphism of Glutathione gene variation which result in loweringenzyme activity increasing microvascular complications of diabetic patients. Aim: This study was aimed toestimate the relationship between GPX1 gene variation and the risk of develop and progression of end stagerenal disease in type I diabetic nephropathy patients. In this study, it has been examined 140 type I diabetesmellitus patients with nephropathy and 40 healthy control. Results: All samples have been genotyped byusing polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques.In diabetic nephropathy patients, there were a significant elevation in in the variation (TT) genotype and (T)allele frequencies compared with control group.Conclusions: Using major Iraqi diabetic patients of Babylon province we studied the transforming effect ofGPX1 gene polymorphism on the risk of development diabetic nephropathy, this study was found that the(TT) genotype of GPX1 SNPs increases the risk of development diabetic nephropathy, while (CC) genotypedecreases the chance for developing disease. These results will gave the way for better perception of thegenetic influencing on the development and progression of diabetic nephropathy.

Expression of PTTG1 in gastric adenocarcinoma tissues and its relationship with prognosis

Objective: To identify new prognostic markers and therapeutic targets for gastric adenocarcinoma. Methods: The public datasets of gastric adenocarcinoma collected from GEO database (GSE33335 and GSE63089) were downloaded for analysis. There were 70 GC tissues and paired normal tissues in the two profile datasets. Differentially expressed genes (DEGs) between GC tissues and normal stomach tissues were selected by the R software. Protein-protein interaction (PPI) of these DEGs were visualized by the Search Tool for the Retrieval of Interacting Genes (STRING). The key gene sets were analyzed by Cytoscape and Molecular Complex Detection (MCODE). The mRNA and protein expression levels of prognosis related genes identified by public database were confirmed by using GC tissues and paired normal tissues collected from July 2019 to September 2019 in Affiliated Hospital of Nantong University. Results: DEGs were identified in the two datasets by using R software. A total of 128 DEGs were detected, including 85 up-regulated genes (log(2)FC>1.2 and FDR<0.01) and 43 down-regulated genes (log(2)FC<-1.2 and FDR<0.01) in the GC tissues. PPI network model and MCODE model were established by using the Online String tool and Cytoscape software, and 27 key genes were obtained, including 25 genes related with prognosis of GC patients (P<0.05). We identified 14 significant DEGs in GC tissues, including cyclin B1 (CCNB1), polo-like kinase 1 (PLK1) and pituitary-tumor transforming gene (PTTG1), which were significantly enriched in the cell cycle pathway through KEGG pathway enrichment analysis. The positive expression rate of PTTG1 in GC tissues was 68.8% (22/32), significantly higher than 18.8% (6/32) in normal gastric tissues (P<0.05). Conclusions: The expression of PTTG1 is different in GC and gastric tissues, implicates it is the key gene in gastric carcinogenesis. The prognoses of GC patients with higher PTTG1 expression are worse. PTTG1 might participate in the development of gastric adenocarcinoma by regulating cell cycle.

Role of Small GTPase RhoA in DNA Damage Response.

Accumulating evidence has suggested a role of the small GTPase Ras homolog gene family member A (RhoA) in DNA damage response (DDR) in addition to its traditional function of regulating cell morphology. In DDR, 2 key components of DNA repair, ataxia telangiectasia-mutated (ATM) and flap structure-specific endonuclease 1 (FEN1), along with intracellular reactive oxygen species (ROS) have been shown to regulate RhoA activation. In addition, Rho-specific guanine exchange factors (GEFs), neuroepithelial transforming gene 1 (Net1) and epithelial cell transforming sequence 2 (Ect2), have specific functions in DDR, and they also participate in Ras-related C3 botulinum toxin substrate 1 (Rac1)/RhoA interaction, a process which is largely unappreciated yet possibly of significance in DDR. Downstream of RhoA, current evidence has highlighted its role in mediating cell cycle arrest, which is an important step in DNA repair. Unraveling the mechanism by which RhoA modulates DDR may provide more insight into DDR itself and may aid in the future development of cancer therapies.

Enhanced Lutein Production in Chlamydomonas reinhardtii by Overexpression of the Lycopene Epsilon Cyclase Gene.

Chlamydomonas reinhardtii is a well-established microalgal model species with a shorter doubling time, which is a promising natural source for the efficient production of high-value carotenoids. In the microalgal carotenoid biosynthetic pathway, lycopene is converted either into β-carotene by lycopene β-cyclase or into α-carotene by lycopene ε-cyclase (LCYE) and lycopene β-cyclase. In this study, we overexpressed the LCYE gene in C. reinhardtii to estimate its effect on lycopene metabolism and lutein production. Chlamydomonas transformants (CrLCYE#L1, #L5, and #L6) produced significantly increased amounts of lutein per culture (up to 2.6-fold) without a decrease in cell yields. Likewise, the expression levels of LCYE gene in transformants showed a significant increase compared with that of the wild-type strain. These results suggest that LCYE overexpression enhances the conversion of lycopene to α-carotene, which in turn improves lutein productivity. Interestingly, their β-carotene productivity appeared to increase slightly rather than decrease. Considering that the inhibition of the lycopene cyclization steps often induces higher expression in genes upstream of metabolic branches, this result implies that the redirection from β-carotene to α-carotene by LCYE overexpression might also enhance upstream gene expression, thereby leading to auxiliary β-carotene production.

High concentration and high dose of disinfectants and antibiotics used during the COVID-19 pandemic threaten human health

The issue of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has created enormous threat to global health. In an effort to contain the spread of COVID-19, a huge amount of disinfectants and antibiotics have been utilized on public health. Accordingly, the concentration of disinfectants and antibiotics is increasing rapidly in various environments, including wastewater, surface waters, soils and sediments. The aims of this study were to analyze the potential ecological environment impacts of disinfectants and antibiotics by summarizing their utilization, environmental occurrence, distribution and toxicity. The paper highlights the promoting effects of disinfectants and antibiotics on antibiotic resistance genes (ARGs) and even antibiotic resistant bacteria (ARB). The scientific evidences indicate that the high concentration and high dose of disinfectants and antibiotics promote the evolution toward antimicrobial resistance through horizontal gene transformation and vertical gene transformation, which threaten human health. Further concerns should be focused more on the enrichment, bioaccumulation and biomagnification of disinfectants, antibiotics, antibiotic resistance genes (ARGs) and even antibiotic resistant bacteria (ARB) in human bodies.

Alteration of Pituitary Tumor Transforming Gene 1 by MicroRNA-186 and 655 Regulates Invasion Ability of Human Oral Squamous Cell Carcinoma.

Background: Pituitary tumor-transforming gene 1 (PTTG1) was recently shown to be involved in the progression as well as the metastasis of cancers. However, their expression and function in the invasion of oral squamous cell carcinoma (SCC) remain unclear. Methods: The expressions of PTTG1 and PTTG1-targeted miRNA in oral SCC cell lines and their invasion capability depended on PTTG1 expression were analyzed by quantitative RT-PCR, Western blots, the transwell insert system and Zymography. Results: Invasion abilities were decreased in oral SCC cells treated with siRNA-PTTG1. When PTTG1 were downregulated in oral SCC cells treated with microRNA-186 and -655 inhibited their invasion abilities via MMP-9 activity. Conclusions: These results indicate that alteration of expression of PTTG1 in oral SCC cells by newly identified microRNA-186 and -655 can regulate invasion activity. Therefore, these data offer new insights into further understanding PTTG1 function in oral SCC and should provide new strategies for diagnostic markers for oral SCC.