Antisense inhibition of ATM gene enhances the radiosensitivity of head and neck squamous cell carcinoma in mice

<p>Abstract</p> <p>Background</p> <p>Treatment failure after radiotherapy of head and neck squamous cell carcinoma (HNSCC) could be a significant problem. Our objective is to sensitize SCCVII cells to ionizing radiation <it>in vitro </it>and <it>in vivo </it>through inhibiting ATM expression using antisense oligodeoxynucleotides (AS-ODNs), and investigate the potential mechanism of radiosensitization.</p> <p>Methods</p> <p>We designed and synthesized AS-ODNs that target ATM mRNA to reduce the ATM expression. The influence on the expression of ATM mRNA and protein in SCCVII cells were analysed by real-time quantitative PCR and western blotting respectively. Clonogenic survival assay was performed to detect the survival ability of SCCVII cells after irradiation, while flow cytometry used to analyse the cell cycle and apoptosis. The volume of solid tumors generated with SCCVII cells was measured, and cell apoptosis was analysed by TUNEL assay after irradiation.</p> <p>Results</p> <p>The relative ATM mRNA and protein expression in SCCVII cells treated with ATM AS-ODNs were decreased to 25.7 ± 3.1% and 24.1 ± 2.8% of that in untreated cells respectively (<it>P </it>< 0.05). After irradiation, the survival fraction (SF) of cells treated with ATM AS-ODNs was lower than that of other groups at the same dose of radiation (<it>P </it>< 0.05), while the percentage of cells in G2/M phase decreased and apoptotic rate of cells increased(<it>P </it>< 0.05). The inhibition rate in SCCVII cells solid tumor exposed to X-ray alone was 23.2 ± 2.7%, while it was 56.1 ± 3.8% in the group which irradiated in combination with the treatment of ATM AS-ODNs (<it>P </it>< 0.05). The apoptotic index for the group irradiated in combination with ATM AS-ODNs injection was 19.6 ± 3.2, which was significantly higher than that of others (<it>P </it>< 0.05)</p> <p>Conclusion</p> <p>Inhibition of ATM expression sensitized SCCVII cells to ionizing radiation <it>in vitro </it>and <it>in vivo</it>. The potential mechanism should be the defective G2/M cell cycle checkpoint control and enhanced radiation-induced apoptosis.</p

ESG performance and corporate value: Analysis from the stakeholders’ perspective

Based on the panel data of China’s A-share non-financial listed enterprises from 2011 to 2020, we empirically explore whether EGS performance can significantly promote corporate value and how to promote it, from the stakeholders’ perspective. We find that: 1) ESG performance significantly improves corporate value. 2) Both media attention and analyst coverage play an intermediary role in the impact of ESG performance on corporate value. 3) Further analysis of the single dimension of ESG illustrates that Environmental (E) and Social (S) have a positive impact on corporate value, but the effect size of Social (S) is smaller, and there is no evidence for a significant relationship between Governance (G) and corporate value. 4) The heterogeneity analysis shows that ESG performance of non-heavily polluting enterprises has a significant positive effect on corporate value, but not on heavily polluting enterprises. Meanwhile, ESG performance of enterprises with a low percentage of institutional investor ownership has a significant positive effect on corporate value, but not with a high percentage. Overall, our study shows that high-quality ESG performance triggers the attention of media and analysts, which in turn promotes corporate value by raising stakeholder pressure. We also analyze the possible causes of heterogeneous results from the perspective of stakeholders, and put forward reasonable suggestions to promote ESG performance and corporate value, as well as protect the interests of stakeholders

Effect of single-patient room design on the incidence of nosocomial infection in the intensive care unit: a systematic review and meta-analysis

BackgroundPrevious studies have yielded varying conclusions regarding the impact of single-patient room design on nosocomial infection in the intensive care unit (ICU). We aimed to examine the impact of ICU single-patient room design on infection control.MethodsWe conducted a comprehensive search of PubMed, Embase, the Cochrane Library, Web of Science, CNKI, WanFang Data, and CBM databases from inception to October 2023, without language restrictions. We included observational cohort and quasi-experimental studies assessing the effect of single- versus multi-patient rooms on infection control in the ICU. Outcomes measured included the nosocomial infection rate, incidence density of nosocomial infection, nosocomial colonization and infection rate, acquisition rate of multidrug-resistant organisms (MDROs), and nosocomial bacteremia rate. The choice of effect model was determined by heterogeneity.ResultsOur final analysis incorporated 12 studies involving 12,719 patients. Compared with multi-patient rooms in the ICU, single-patient rooms demonstrated a significant benefit in reducing the nosocomial infection rate (odds ratio [OR]: 0.68; 95% confidence interval [CI]: 0.59, 0.79; p &lt; 0.00001). Analysis based on nosocomial infection incidence density revealed a statistically significant reduction in single-patient rooms (OR: 0.64; 95% CI: 0.44, 0.92; p = 0.02). Single-patient rooms were associated with a marked decrease in nosocomial colonization and infection rate (OR: 0.44; 95% CI: 0.32, 0.62; p &lt; 0.00001). Furthermore, patients in single-patient rooms experienced lower nosocomial bacteremia rate (OR: 0.73; 95% CI: 0.59, 0.89; p = 0.002) and lower acquisition rate of MDROs (OR: 0.41; 95% CI: 0.23, 0.73; p = 0.002) than those in multi-patient rooms.ConclusionImplementation of single-patient rooms represents an effective strategy for reducing nosocomial infections in the ICU.Systematic review registrationhttps://www.crd.york.ac.uk/PROSPERO/)

Effect of Exogenous Nitric Oxide on Postharvest Storage Quality of Hyacinth Bean

In order to study the effect of nitric oxide (NO) on the storage quality of hyacinth bean after harvest, sodium nitroprusside (SNP) was used as an exogenous NO donor in this study. Hyacinth bean was soaked in 0.2 mmol/L SNP solution or distilled water as control for 10 min and then stored at (20 ± 1) ℃ and 80%–90% relative humidity. Decay incidence, rust incidence, hardness, the contents of total soluble solids (TSS), malondialdehyde (MDA), flavonoids, total phenols and chlorophyll, and the activities of antioxidant enzymes (peroxidase (POD), polyphenol oxidase (PPO), phenylalanine ammoniase (PAL), catalase (CAT) and ascorbate peroxidase (APX)) were observed during the storage period. The results showed that exogenous NO treatment could inhibit the rot and rust, keep the color and hardness, and inhibit the degradation of TSS and chlorophyll in hyacinth bean, so that hyacinth bean could maintain good sensory quality. Exogenous NO treatment could also prevent the accumulation of MDA and increase the contents of total phenols and flavonoids. In addition, exogenous NO treatment maintained the activities of PAL, CAT and APX during storage, and inhibited the increase in the activities of POD and PPO, thereby enhancing the antioxidant capacity and delaying the maturation and senescence of hyacinth bean. In conclusion, exogenous NO treatment can delay the postharvest maturation and senescence, maintain the physiological quality during storage, and effectively prolong the shelf life of hyacinth bean

Anopheles sinensis mosquito insecticide resistance: comparison of three mosquito sample collection and preparation methods and mosquito age in resistance measurements

BACKGROUND: Insecticide resistance monitoring in malaria mosquitoes is essential for guiding the rational use of insecticides in vector control programs. Resistance bioassay is the first step for insecticide monitoring and it lays an important foundation for molecular examination of resistance mechanisms. In the literature, various mosquito sample collection and preparation methods have been used, but how mosquito sample collection and preparation methods affect insecticide susceptibility bioassay results is largely unknown. The objectives of this study were to determine whether mosquito sample collection and preparation methods affected bioassay results, which may cause incorrect classification of mosquito resistance status. METHODS: The study was conducted in Anopheles sinensis mosquitoes in two study sites in central China. Three mosquito sample collection and preparation methods were compared for insecticide susceptibility, kdr frequencies and metabolic enzyme activities: 1) adult mosquitoes collected from the field; 2) F1 adults from field collected, blood-fed mosquitoes; and 3) adult mosquitoes reared from field collected larvae. RESULTS: Mosquito sample collection and preparation methods significantly affected mortality rates in the standard WHO tube resistance bioassay. Mortality rate of field-collected female adults was 10-15% higher than in mosquitoes reared from field-collected larvae and F1 adults from field collected blood-fed females. This pattern was consistent in mosquitoes from the two study sites. High kdr mutation frequency (85-95%) with L1014F allele as the predominant mutation was found in our study populations. Field-collected female adults consistently exhibited the highest monooxygenase and GST activities. The higher mortality rate observed in the field-collected female mosquitoes may have been caused by a mixture of mosquitoes of different ages, as older mosquitoes were more susceptible to deltamethrin than younger mosquitoes. CONCLUSIONS: Female adults reared from field-collected larvae in resistance bioassays are recommended to minimize the effect of confounding factors such as mosquito age and blood feeding status so that more reliable and reproducible mortality may be obtained

High Human Bocavirus Viral Load Is Associated with Disease Severity in Children under Five Years of Age

Human bocavirus (HBoV) is a parvovirus and detected worldwide in lower respiratory tract infections (LRTIs), but its pathogenic role in respiratory illness is still debatable due to high incidence of co-infection with other respiratory viruses. To determine the prevalence of HBoV infection in patients with LRTI in Shanghai and its correlation with disease severity, we performed a 3-year prospective study of HBoV in healthy controls, outpatients and inpatients under five years of age with X-ray diagnosed LRTIs. Nasopharyngeal aspirates were tested by PCR for common respiratory viruses and by real time PCR for HBoV subtypes 1–4. Nasopharyngeal swabs from healthy controls and serum samples and stools from inpatients were also tested for HBoV1-4 by real time PCR. Viral loads were determined by quantitative real time PCR in all HBoV positive samples. HBoV1 was detected in 7.0% of inpatients, with annual rates of 5.1%, 8.0% and 4.8% in 2010, 2011 and 2012, respectively. Respiratory syncytial virus (RSV) subtype A was the most frequent co-infection detected; HBoV1 and RSVA appeared to co-circulate with similar seasonal variations. High HBoV viral loads (>10(6) copies/ml) were significantly more frequent in inpatients and outpatients than in healthy controls. There was a direct correlation of high viral load with increasing disease severity in patients co-infected with HBoV1 and at least one other respiratory virus. In summary, our data suggest that HBoV1 can cause LRTIs, but symptomatic HBoV infection is only observed in the context of high viral load

Optimizing Proof of Aliveness in cyber-physical systems

At ACSAC 2019, we introduced a new cryptographic primitive called proof of aliveness (PoA), allowing us to remotely and automatically track the running status (aliveness) of devices in the fields in cyber-physical systems. We proposed to use a one-way function (OWF) chain structure to build an efficient proof of aliveness, such that the prover sends every node on the OWF chain in a reverse order periodically, and it can be verified by a remote verifier with the possession of the tail node (last node) of the OWF chain. However, the practicality of this initial construction is limited by the finite number of nodes on an OWF chain. We enhance our first PoA construction by linking multiple OWF chains together using a pseudo-random generator chain in our second PoA scheme. This enhancement allows us to integrate one-time signature (OTS) schemes into the structure of the second construction to realize the auto-replenishment of the aliveness proofs. This implies that securely an initialized PoA instance can be used forever without interruption for reinitialization. In this work, our primary motivation is to further improve our secondary PoA and auto-replenishment schemes. Instead of storing the tail nodes of multiple OWF chains on the verifier side, we use a Bloom Filter to compress them. This saves 4.7 times the storage cost compared to our previous version at ACSAC 2019. Moreover, the OTS-based auto-replenishment solution cannot be applied to our first scheme solely based on OWFs, and it is not so efficient despite its standard model security. To overcome these limitations, we design a new auto-replenishment scheme from a hash-based commitment under the random oracle model in this work, which is much faster and can be used by both PoA schemes. Additionally, we implement and evaluate our PoA constructions on Raspberry Pis to demonstrate their performance. Considering the implementation on a storage/memory-constrained device, we particularly study the strategies for efficiently generating proofs

A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation

The relative ease, speed and biological scope of CRISPR/Cas9-based reagents for genomic manipulations are revolutionizing virtually all areas of molecular biosciences, including functional genomics, genetics, applied biomedical research and agricultural biotechnology. In plant systems, however, a number of hurdles currently exist that limit this technology from reaching its full potential. For example, significant plant molecular biology expertise and effort is still required to generate functional expression constructs that allow simultaneous editing, and especially transcriptional regulation, of multiple different genomic loci or "multiplexing", which is a significant advantage of CRISPR/Cas9 versus other genome editing systems. In order to streamline and facilitate rapid and wide-scale use of CRISPR/Cas9-based technologies for plant research, we developed and implemented a comprehensive molecular toolbox for multifaceted CRISPR/Cas9 applications in plants. This toolbox provides researchers with a protocol and reagents to quickly and efficiently assemble functional CRISPR/Cas9 T-DNA constructs for monocots and dicots using Golden Gate and Gateway cloning methods. It comes with a full suite of capabilities, including multiplexed gene editing and transcriptional activation or repression of plant endogenous genes. We report the functionality and effectiveness of this toolbox in model plants such as tobacco, Arabidopsis and rice, demonstrating its utility for basic and applied plant research.ECU Open Access Publishing Support Fun

Mechanistic Insight Into the Interaction Between Helicobacter pylori Urease Subunit α and Its Molecular Chaperone Hsp60

Helicobacter pylori is the etiologic agent in a variety of gastroduodenal diseases. As its key pathogenic factors, both urease and Hsp60 play important roles in the pathogenesis of H. pylori. Previous studies have suggested that there is close relationship between urease and Hsp60, which implied that Hsp60 may act as a chaperone in urease stabilization and assembly. However, how these two proteins interact remains unclear. In this study, the impact of Hsp60 on urease activity of H. pylori lysate was first detected to confirm the interaction between urease and Hsp60. Pull-down assays further indicated that Hsp60 could bind to UreA subunit but not UreB. Then, the 3D structure of Hsp60 was modeled using I-TASSER to simulate the binding complex with UreA by molecular docking. The results showed that UreA is a perfect fit for the cavity of Hsp60. Analysis of the resulting model demonstrated that at least seven residues of UreA, located on two interfaces, participate in the interaction. Site-directed mutagenesis of these potential residues showed reduced affinity with Hsp60 than the wild type UreA through surface plasmon resonance (SPR) experiments, and D68 appears to have an important role in the affinity. Further analysis also showed that mutation of E25 and K26 caused a more rapid association and dissociation than with wild UreA, implying that they have roles in stabilizing the interaction complex. These affinity comparisons suggested that the interfaces predicted by molecular docking are credible. Our study indicated a direct interaction between Hsp60 and urease and revealed the binding interfaces and key residues involved in the interaction. These results provide further evidence for the chaperone activity of Hsp60 toward urease and lay a foundation to better understand the maturation mechanism of urease in H. pylori

Autopalmitoylation of TEAD Proteins Regulates Transcriptional Output of Hippo Pathway

TEA domain (TEAD) transcription factors bind to the co-activator YAP/TAZ, and regulate the transcriptional output of Hippo pathway, playing critical roles in organ size control and tumorigenesis. Protein S-palmitoylation attaches fatty acid (palmitate) to cysteine residues, and regulates protein trafficking, membrane localization and signaling activities. Using activity-based chemical probes, we discovered that human TEADs possess intrinsic palmitoylating enzyme-like activities, and undergo autopalmitoylation at evolutionarily conserved cysteine residues under physiological conditions. We determined the crystal structures of lipid-bound TEADs, and found that the lipid chain of palmitate inserts into a conserved deep hydrophobic pocket. Strikingly, palmitoylation is required for TEAD’s binding to YAP/TAZ, but dispensable for the binding to Vgll4 tumor suppressor. In addition, palmitoylation does not alter TEAD’s localization. Moreover, TEAD palmitoylation-deficient mutants impaired TAZ-mediated muscle differentiation in vitro, and Yorkie-mediated tissue overgrowth in Drosophila in vivo. Our study directly linked autopalmitoylation to the transcriptional regulation of Hippo pathway