Pseudothrombosis due to blood-contrast level in the infrarenal vena cava.

Filling defects within the inferior vena cava (IVC) are common findings on computed tomography (CT); nevertheless, a majority of these defects are attributed to artifacts. The documentation pertaining to pseudothrombosis specifically affecting the infrarenal vena cava is notably insufficient in current literature. The aim of this study is to present a case demonstrating a blood-contrast level in the infrarenal vena cava, resembling an intraluminal filling defect.

Acute acalculous cholecystitis with portal cavernoma: A case report with literature review.

Portal cavernoma cholangiopathy (PCC) refers to morphological changes in the intrahepatic, extrahepatic biliary system, along with the gallbladder (GB), induced by portal cavernoma (PC). Acute acalculous cholecystitis (AAC) represents an infrequent clinical manifestation of PCC. Given the inadequacy of documentation within medical literature, AAC may go undiagnosed among patients with PC presenting symptoms of right upper quadrant pain. The current study aims to report a case of acute acalculous cholecystitis secondary to portal cavernoma, focusing on radiological findings, with a brief review of literature.

Prevalence and factors associated with malnutrition among hemodialysis patients in a single hemodialysis center in Vietnam: A cross-sectional study.

Malnutrition is a common problem among hemodialysis patients that increases morbidity and mortality and decreases the quality of life. This study aimed to assess the prevalence of malnutrition and associated factors and survey the consumption of energy and several nutrients among hemodialysis patients. A prospective observational study with a cross-sectional design was conducted on 76 patients on hemodialysis therapy at Nguyen Tri Phuong Hospital, Ho Chi Minh City, Vietnam, for 2 months (from May to July 2022). Dialysis malnutrition score was used to determine patients' nutritional status. Data about their biochemical parameters were retrieved from records with the newest results. Among the 76 patients, 38 (50.0%) were female. The mean age of the patients was 55.0 ±â€…13.5 years. Based on the dialysis malnutrition score, 56 (73.7%) patients had mild to moderate malnutrition, while 2 (2.6%) had severe malnutrition. The average energy intake was 21.5 kcal/kg/day, with only 3.9% meeting the recommended intake. The average protein intake was 1.0 g/kg/day, and about 10.5% of participants complied with the recommended protein level. In addition, the majority of patients did not reach the recommendations for sodium (56.6%), potassium (88.2%), phosphate (75.0%), and calcium (82.9%). We found a significant association between patients' occupation (P < .05), dialysis vintage (P < .001), and malnutrition status. Malnutrition is widespread among Vietnamese hemodialysis patients, which necessitates regular assessment and monitoring. We recommend paying more attention to the nutritional status of patients who are unemployed, retired, or stopped working and those with ≥ 5 years of hemodialysis.

Automated Machine Learning for Predicting Diabetic Retinopathy Progression From Ultra-Widefield Retinal Images.

Importance: Machine learning (ML) algorithms have the potential to identify eyes with early diabetic retinopathy (DR) at increased risk for disease progression. Objective: To create and validate automated ML models (autoML) for DR progression from ultra-widefield (UWF) retinal images. Design, Setting and Participants: Deidentified UWF images with mild or moderate nonproliferative DR (NPDR) with 3 years of longitudinal follow-up retinal imaging or evidence of progression within 3 years were used to develop automated ML models for predicting DR progression in UWF images. All images were collected from a tertiary diabetes-specific medical center retinal image dataset. Data were collected from July to September 2022. Exposure: Automated ML models were generated from baseline on-axis 200° UWF retinal images. Baseline retinal images were labeled for progression based on centralized reading center evaluation of baseline and follow-up images according to the clinical Early Treatment Diabetic Retinopathy Study severity scale. Images for model development were split 8-1-1 for training, optimization, and testing to detect 1 or more steps of DR progression. Validation was performed using a 328-image set from the same patient population not used in model development. Main Outcomes and Measures: Area under the precision-recall curve (AUPRC), sensitivity, specificity, and accuracy. Results: A total of 1179 deidentified UWF images with mild (380 [32.2%]) or moderate (799 [67.8%]) NPDR were included. DR progression was present in half of the training set (590 of 1179 [50.0%]). The model's AUPRC was 0.717 for baseline mild NPDR and 0.863 for moderate NPDR. On the validation set for eyes with mild NPDR, sensitivity was 0.72 (95% CI, 0.57-0.83), specificity was 0.63 (95% CI, 0.57-0.69), prevalence was 0.15 (95% CI, 0.12-0.20), and accuracy was 64.3%; for eyes with moderate NPDR, sensitivity was 0.80 (95% CI, 0.70-0.87), specificity was 0.72 (95% CI, 0.66-0.76), prevalence was 0.22 (95% CI, 0.19-0.27), and accuracy was 73.8%. In the validation set, 6 of 9 eyes (75%) with mild NPDR and 35 of 41 eyes (85%) with moderate NPDR progressed 2 steps or more were identified. All 4 eyes with mild NPDR that progressed within 6 months and 1 year were identified, and 8 of 9 (89%) and 17 of 20 (85%) with moderate NPDR that progressed within 6 months and 1 year, respectively, were identified. Conclusions and Relevance: This study demonstrates the accuracy and feasibility of automated ML models for identifying DR progression developed using UWF images, especially for prediction of 2-step or greater DR progression within 1 year. Potentially, the use of ML algorithms may refine the risk of disease progression and identify those at highest short-term risk, thus reducing costs and improving vision-related outcomes.

Maturity of Food Safety Management Systems in the Vietnamese Seafood Processing Industry.

Fifty-nine percent (59%) of the reported food safety issues in Vietnam are related to seafood products, mainly fish and fish products. The international export of seafood products continues to grow due to intensification of the production in the Vietnamese seafood processing industry. To ensure the production of safe food, a company-specific, effective food safety management system is essential. This research explores the maturity of food safety management systems in a convenience sample of the Vietnamese seafood processing industry to identify potential gaps and interventions for improvement. The food safety management system diagnostic instrument was used to assess the context riskiness, maturity of control and assurance activities and food safety performance of 11 companies. Maturity of their food safety management systems was further explored through hierarchical cluster analysis, and the differences in maturity between clusters were statistically tested through Mann-Whitney U tests (nonparametric). The influence of companies' organizational characteristics on the maturity of control and assurance activities was assessed through nonparametric K independent tests. A variability in the maturity of food safety management systems between the eleven Vietnamese companies was measured. Cluster analysis revealed two clusters, Cluster I (six companies) and Cluster II (five companies). The companies in both these clusters operate under a moderate level context riskiness and average to advanced level of food safety performance. However, control and assurance activities are at a lower maturity in Cluster I compared to Cluster II. None of the companies' organizational characteristics (i.e. certification level) have a statistically significant influence on the maturity of control and assurance activities. However, compliance with multiple food safety standards and the presence of physical intervention system(s) have a positive influence on food safety performance.

Mapping alternative polyadenylation in human cells using direct RNA sequencing technology.

Alternative cleavage and polyadenylation (APA) is a widespread mechanism to generate mRNA isoforms with alternative 3' untranslated regions. Here, we detail a protocol for detecting APA genome wide using direct RNA sequencing technology including computational analysis. We describe steps for RNA sample and library preparation, nanopore sequencing, and data analysis. Experiments and data analysis can be performed over a period of 6-8 days and require molecular biology and bioinformatics skills. For complete details on the use and execution of this protocol, please refer to Polenkowski et al.1.

Identifying temporal sensory drivers of liking of biscuit supplemented with brewer's spent grain for young consumers.

Brewer's spent grain (BSG), a by-product of the brewing industry, has great potential as food additive. BSG is particularly rich in protein and fibre content which makes it an ideal nutritional fortifier for biscuits. However, adding BSG to biscuits can lead to changes in sensory perception and consumer acceptance. This study explored the temporal sensory profiles and drivers/inhibitors of liking in BSG-fortified biscuits. Six biscuit formulations were obtained from a design with factors oat flake particle size (three levels: 0.5 mm, small commercial flakes, large commercial flakes) and baking powder (two levels: with, without). Consumers (n = 104) tasted the samples, described their dynamic sensory perception using the Temporal Check-All-That-Apply (TCATA) method, and rated their liking on a 7-point categorical scale. The Clustering around Latent Variables (CLV) approach was used to group consumers into two clusters based on their preferences. The temporal sensory profiles and drivers/inhibitors of liking were investigated within each cluster. Foamy and Easy-to-swallow were sensory drivers of liking for both groups of consumers. However, inhibitors of liking were different in the two clusters: Dense and Hard-to-swallow for one cluster and Chewy, Hard-to-swallow and Hard for the other cluster. These findings give evidence that manipulating oat particle size and presence/absence of baking powder changes BSG-fortified biscuits' sensory profiles and consumer preferences. A complementary analysis of the area-under-curve of the TCATA data and inspection of individual temporal curves showed the dynamics of perception and showed how oat particle size and presence/absence of baking powder affected consumer perception and acceptance of BSG-fortified biscuits. The methods proposed in this paper can be further applied to understand how enriching products with ingredients that would otherwise go to waste affects acceptance in different consumer segments.

TSSUNet-MB - ab initio identification of σ70 promoter transcription start sites in Escherichia coli using deep multitask learning.

MOTIVATION: Computational promoter prediction (CPP) tools designed to classify prokaryotic promoter regions usually assume that a transcription start site (TSS) is located at a predefined position within each promoter region. Such CPP tools are sensitive to any positional shifting of the TSS in a windowed region, and they are unsuitable for determining the boundaries of prokaryotic promoters. RESULTS: TSSUNet-MB is a deep learning model developed to identify the TSSs of σ70 promoters. Mononucleotide and bendability were used to encode input sequences. TSSUNet-MB outperforms other CPP tools when assessed using the sequences obtained from the neighborhood of real promoters. TSSUNet-MB achieved a sensitivity of 0.839 and specificity of 0.768 on sliding sequences, while other CPP tool cannot maintain both sensitivities and specificities in a compatible range. Furthermore, TSSUNet-MB can precisely predict the TSS position of σ70 promoter-containing regions with a 10-base accuracy of 77.6%. By leveraging the sliding window scanning approach, we further computed the confidence score of each predicted TSS, which allows for more accurately identifying TSS locations. Our results suggest that TSSUNet-MB is a robust tool for finding σ70 promoters and identifying TSSs.

Performance of Automated Machine Learning for Diabetic Retinopathy Image Classification from Multi-field Handheld Retinal Images.

PURPOSE: To create and validate code-free automated deep learning models (AutoML) for diabetic retinopathy (DR) classification from handheld retinal images. DESIGN: Prospective development and validation of AutoML models for DR image classification. PARTICIPANTS: A total of 17 829 deidentified retinal images from 3566 eyes with diabetes, acquired using handheld retinal cameras in a community-based DR screening program. METHODS: AutoML models were generated based on previously acquired 5-field (macula-centered, disc-centered, superior, inferior, and temporal macula) handheld retinal images. Each individual image was labeled using the International DR and diabetic macular edema (DME) Classification Scale by 4 certified graders at a centralized reading center under oversight by a senior retina specialist. Images for model development were split 8-1-1 for training, optimization, and testing to detect referable DR ([refDR], defined as moderate nonproliferative DR or worse or any level of DME). Internal validation was performed using a published image set from the same patient population (N = 450 images from 225 eyes). External validation was performed using a publicly available retinal imaging data set from the Asia Pacific Tele-Ophthalmology Society (N = 3662 images). MAIN OUTCOME MEASURES: Area under the precision-recall curve (AUPRC), sensitivity (SN), specificity (SP), positive predictive value (PPV), negative predictive value (NPV), accuracy, and F1 scores. RESULTS: Referable DR was present in 17.3%, 39.1%, and 48.0% of the training set, internal validation, and external validation sets, respectively. The model's AUPRC was 0.995 with a precision and recall of 97% using a score threshold of 0.5. Internal validation showed that SN, SP, PPV, NPV, accuracy, and F1 scores were 0.96 (95% confidence interval [CI], 0.884-0.99), 0.98 (95% CI, 0.937-0.995), 0.96 (95% CI, 0.884-0.99), 0.98 (95% CI, 0.937-0.995), 0.97, and 0.96, respectively. External validation showed that SN, SP, PPV, NPV, accuracy, and F1 scores were 0.94 (95% CI, 0.929-0.951), 0.97 (95% CI, 0.957-0.974), 0.96 (95% CI, 0.952-0.971), 0.95 (95% CI, 0.935-0.956), 0.97, and 0.96, respectively. CONCLUSIONS: This study demonstrates the accuracy and feasibility of code-free AutoML models for identifying refDR developed using handheld retinal imaging in a community-based screening program. Potentially, the use of AutoML may increase access to machine learning models that may be adapted for specific programs that are guided by the clinical need to rapidly address disparities in health care delivery. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

THOC5 complexes with DDX5, DDX17, and CDK12 to regulate R loop structures and transcription elongation rate.

THOC5, a member of the THO complex, is essential for the 3'processing of some inducible genes, the export of a subset of mRNAs and stem cell survival. Here we show that THOC5 depletion results in altered 3'cleavage of >50% of mRNAs and changes in RNA polymerase II binding across genes. THOC5 is recruited close to high-density polymerase II sites, suggesting that THOC5 is involved in transcriptional elongation. Indeed, measurement of elongation rates in vivo demonstrated decreased rates in THOC5-depleted cells. Furthermore, THOC5 is preferentially recruited to its target genes in slow polymerase II cells compared with fast polymerase II cells. Importantly chromatin-associated THOC5 interacts with CDK12 (a modulator of transcription elongation) and RNA helicases DDX5, DDX17, and THOC6 only in slow polymerase II cells. The CDK12/THOC5 interaction promotes CDK12 recruitment to R-loops in a THOC6-dependent manner. These data demonstrate a novel function of THOC5 in transcription elongation.

Genome-Wide Methylation Mapping Using Nanopore Sequencing Technology Identifies Novel Tumor Suppressor Genes in Hepatocellular Carcinoma.

Downregulation of multiple tumor suppressor genes (TSGs) plays an important role in cancer formation. Recent evidence has accumulated that cancer progression involves genome-wide alteration of epigenetic modifications, which may cause downregulation of the tumor suppressor gene. Using hepatocellular carcinoma (HCC) as a system, we mapped 5-methylcytosine signal at a genome-wide scale using nanopore sequencing technology to identify novel TSGs. Integration of methylation data with gene transcription profile of regenerated liver and primary HCCs allowed us to identify 10 potential tumor suppressor gene candidates. Subsequent validation led us to focus on functionally characterizing one candidate-glucokinase (GCK). We show here that overexpression of GCK inhibits the proliferation of HCC cells via induction of intracellular lactate accumulation and subsequently causes energy crisis due to NAD+ depletion. This suggests GCK functions as a tumor suppressor gene and may be involved in HCC development. In conclusion, these data provide valuable clues for further investigations of the process of tumorigenesis in human cancer.

C20orf204, a hepatocellular carcinoma-specific protein interacts with nucleolin and promotes cell proliferation.

In most human cancers, a large number of proteins with driver mutations are involved in tumor development, implying that multiple fine tuners are involved in cancer formation and/or maintenance. A useful strategy for cancer therapy may therefore be to target multiple cancer type-specific fine tuners. Furthermore, genome-wide association studies of tumor samples have identified a large number of long noncoding (lnc)RNA associated with various types of tumor. In this context we have previously found that C20orf204 (a splice variant of Linc00176) RNA contains a 189 amino acid (AA) long open reading frame (C20orf204-189AA) that is expressed predominantly in hepatocellular carcinoma (HCC). We report here that a protein, C20orf204-189AA, was detected in the nucleus of 14 out of 20 primary HCC, but not in control livers. Strikingly, overexpression of C20orf204-189AA enhanced cell proliferation and ribosomal RNA transcription. C20orf204-189AA is co-localized, and interacted with nucleolin via the C-terminal and with ribosomal RNA via the N-terminal domain. Furthermore, the expression of C20orf204-189AA upregulates the protein level of nucleolin. Nucleolin and C20orf204 mRNA levels in HCC are correlated with tumor differentiation grade and patient survival, suggesting that C20orf204-189AA is a cancer type-specific fine tuner in some HCC that presents itself for potential targeting therapy and cancer biomarker. Thus, cancer cells exhibit remarkable transcriptome alterations partly by adopting cancer-specific splicing isoforms of noncoding RNAs and may participate in tumor development.

Food Fingerprinting: Using a Two-Tiered approach to Monitor and Mitigate Food Fraud in Rice.

BACKGROUND: Rice is an important staple food that is consumed around the world. Like many foods, the price of rice varies considerably, from very inexpensive for a low-quality product to premium pricing for highly prized varieties from specific locations. Therefore, like other foods it is vulnerable to economically motivated adulteration through substitution or misrepresentation of inferior-quality rice for more expensive varieties. OBJECTIVE: In this article we describe results of a research project focused on addressing potential food fraud issues related to rice supplies in China, India, Vietnam, and Ghana. Rice fraud manifests differently in each country; therefore, tailored solutions were required. METHOD: Here we describe a two-tiered testing regime of rapid screening using portable Near Infrared technology supported by second tier testing using mass spectrometry-based analysis of suspicious samples. RESULTS: Portable Near Infrared spectroscopy models and laboratory-based Gas Chromatography-Mass Spectrometry methods were developed to differentiate between: high-value Basmati rice varieties and their potential adulterants; six Geographic Indicated protected rice varieties from specific regions within China; various qualities of rice in Ghana and Vietnam; and locally produced and imported rice in Ghana. Furthermore, an Inductively Coupled Plasma-Mass Spectrometry method was developed to support the Chinese rice varieties methods as well as a Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry method for quality differentiation in Vietnam. CONCLUSIONS/HIGHLIGHTS: This two-tier approach can provide a substantially increased level of testing through rapid screening outside of the laboratory with the reassurance of corroborating mass spectrometry-based laboratory analysis to support decision making.

Identification of Novel Micropeptides Derived from Hepatocellular Carcinoma-Specific Long Noncoding RNA.

Identification of cancer-specific target molecules and biomarkers may be useful in the development of novel treatment and immunotherapeutic strategies. We have recently demonstrated that the expression of long noncoding (lnc) RNAs can be cancer-type specific due to abnormal chromatin remodeling and alternative splicing. Furthermore, we identified and determined that the functional small protein C20orf204-189AA encoded by long intergenic noncoding RNA Linc00176 that is expressed predominantly in hepatocellular carcinoma (HCC), enhances transcription of ribosomal RNAs and supports growth of HCC. In this study we combined RNA-sequencing and polysome profiling to identify novel micropeptides that originate from HCC-specific lncRNAs. We identified nine lncRNAs that are expressed exclusively in HCC cells but not in the liver or other normal tissues. Here, DNase-sequencing data revealed that the altered chromatin structure plays a key role in the HCC-specific expression of lncRNAs. Three out of nine HCC-specific lncRNAs contain at least one open reading frame (ORF) longer than 50 amino acid (aa) and enriched in the polysome fraction, suggesting that they are translated. We generated a peptide specific antibody to characterize one candidate, NONHSAT013026.2/Linc013026. We show that Linc013026 encodes a 68 amino acid micropeptide that is mainly localized at the perinuclear region. Linc013026-68AA is expressed in a subset of HCC cells and plays a role in cell proliferation, suggesting that Linc013026-68AA may be used as a HCC-specific target molecule. Our finding also sheds light on the role of the previously ignored 'dark proteome', that originates from noncoding regions in the maintenance of cancer.

Metabolomic fingerprinting of volatile organic compounds for the geographical discrimination of rice samples from China, Vietnam and India.

Rice is one of the most important cereals for human nutrition and is a basic staple food for half of the global population. The assessment of rice geographical origins in terms of its authenticity is of great interest to protect consumers from misleading information and fraud. In the present study, a head space gas chromatography mass spectrometry (HS-GC-MS) strategy for characterising volatile organic compounds (VOCs) profiles to distinguish rice samples from China, India and Vietnam is described. Partial Least Square Discriminant Analysis (PLS-DA) model exhibited a good discrimination (R2 = 0.98182, Q2 = 0.9722, and Accuracy = 1.0) for rice samples from China, India and Vietnam. Moreover, Data-Driven Soft Independent Modelling of Class Analogy (DD-SIMCA) and K-nearest neighbors shown good specificity 100% and accuracy 100% in identifying the origin of samples. The present study established VOC fingerprinting as a highly efficient approach to identify the geographical origin of rice.

Myc/Max dependent intronic long antisense noncoding RNA, EVA1A-AS, suppresses the expression of Myc/Max dependent anti-proliferating gene EVA1A in a U2 dependent manner.

The Myc gene has been implicated in the pathogenesis of most types of human cancerous tumors. Myc/Max activates large numbers of pro-tumor genes; however it also induces anti-proliferation genes. When anti-proliferation genes are activated by Myc, cancer cells can only survive if they are downregulated. Hepatocellular carcinoma (HCC) specific intronic long noncoding antisense (lnc-AS) RNA, the EVA1A-AS gene, is located within the second intron (I2) of the EVA1A gene (EVA-1 homolog A) that encodes an anti-proliferation factor. Indeed, EVA1A, but not EVA1A-AS, is expressed in normal liver. Depletion of EVA1A-AS suppressed cell proliferation of HepG2 cells by upregulation of EVA1A. Overexpression of EVA1A caused cell death at the G2/M phase via microtubule catastrophe. Furthermore, suppressed EVA1A expression levels are negatively correlated with differentiation grade in 365 primary HCCs, while EVA1A-AS expression levels are positively correlated with patient survival. Notably, both EVA1A and EVA1A-AS were activated by the Myc/Max complex. Eva1A-AS is transcribed in the opposite direction near the 3'splice site of EVA1A I2. The second intron did not splice out in a U2 dependent manner and EVA1A mRNA is not exported. Thus, the Myc/Max dependent anti-proliferating gene, EVA1A, is controlled by Myc/Max dependent anti-sense noncoding RNA for HCC survival.

Isolation of Penicillium citrinum from Roots of Clerodendron cyrtophyllum and Application in Biosynthesis of Aglycone Isoflavones from Soybean Waste Fermentation.

Soybeans offer an abundant source of isoflavones, which confer useful bioactivities when existing in aglycone forms. The conversion of isoflavones into aglycones via fermentation of soybean products is often realized by ß-glucosidase, an enzyme produced by fungi. In this study, a filamentous fungus, Clerodendron cyrtophyllum, was isolated from root of Clerodendron cyrtophyllum Turcz, which was able to produce the highest activity of ß-glucosidase up to 33.72 U/mL at 144 h during fermentation on Potato Dextrose Broth (PDB). The obtained fungus was grown on isoflavones-rich soybean extract to produce genistein and daidzein, achieving the conversion rate of 98.7%. Genistein and daidzein were isolated and purified by column chromatography using hexane/acetone (29:1/1:1), reaching purities of over 90% of total isoflavones, as identified and determined by TLC, LC-MS/MS, and 1H and 13C NMR spectroscopy. These results imply that the isolated P. citrinum is a potential fungal strain for industrial-scale production of genistein and daidzein from isoflavones-containing soybean extracts. These products may serve as potential raw materials for manufacture of functional foods that are based on aglycones.

Design of a lightweight magnetizer to enable a portable circumferential magnetic flux leakage detection system.

We proposed a lightweight magnetizer toward the realization of a portable circumferential magnetic flux leakage (MFL) inspection tool for the detection of longitudinal defects with a linear motion between pipes and the inspection tool. The proposed magnetizer consists of four permanent magnetic poles constructing a configuration in which the like poles are separated by a θ angle ranging from 30° to 60° and the opposite poles are separated by a supplemental angle of θ. The distribution of the magnetic field generated by this magnetizer and caused by defects was studied by finite element methods as well as by experimental measurements. The results reveal that the new magnetizer has advantages of high amplitude and good uniformity of magnetic field over the state-of-the-art ones with configurations of two facing poles or quadrupole geometries. With the new magnetizer, the theoretical and experimental results show that the magnetization in the pipe increases slightly with the θ angle, but the uniformity of magnetization in the pipe and the magnetic field on the surface of pipe decrease dramatically with this angle. So, the optimum angle θ is proposed at around 30°-40°. The finite element calculation and the measurements of the MFL signal caused by artificial defects evidence the capabilities in detecting shallow longitudinal defects of the MFL inspection tool with our novel magnetizer.

Multiresidue Pesticides Analysis of Vegetables in Vietnam by Ultrahigh-Performance Liquid Chromatography in Combination with High-Resolution Mass Spectrometry (UPLC-Orbitrap MS).

An ultrahigh-performance liquid chromatography in combination with high-resolution mass spectrometry Thermo Q-Extractive Focus Orbitrap MS has been introduced for analysis of multiclass pesticides in vegetable samples collected in Hanoi, Vietnam. Multiclass pesticides were separated on the Thermo Hypersil Gold PFP column utilizing a gradient of the mobile phase consisting of 5 mM ammonium formate, 0.1% formic acid in deionized water, and methanol. The target analytes were detected in the full-scan mode on Thermo Scientific Q-Exactive Focus Orbitrap MS for quantitation at the optimum operating conditions. These conditions included, but not limit to, the resolution of 70000 at the full width at half maximum in both positive and negative mode, mass range from 80 to 1000 m/z, and optimized parameters for the heated electrospray ionization source. The identification of the analytes in real samples was based on retention times, mass to charge ratios, mass accuracies, and MS/MS spectra at the confirmation mode with the inclusion list of target analytes. The mass accuracies of target analytes were from -4.14 ppm (dinotefuran) to 1.42 ppm (cinosulfuron) in the neat solvent and from -3.91 ppm (spinosad D) to 1.29 ppm (cinosulfuron) in the matrix-matched solution. Target analytes in the vegetable-based matrix were extracted by the QuEChERS method. Some critical parameters of the analytical method such as linearity, repeatability, limit of detection, and limit of quantitation have been evaluated and implemented. Excellent LOD and LOQ of the developed method were achieved at the range of 0.04-0.85 and 0.13-2.9 µg·kg-1, respectively. Intraday and interday repeatability of the analytical signal (peak area, n=6) of the developed method were below 3% and 10%, correspondingly. The matrix effect, extraction recovery, and overall recovery were fully investigated by spiking experiments. Experimental results demonstrated that the ionization suppression or enhancement was the main contribution on the overall recoveries of target analytes. Finally, the in-house validated method was applied to pesticides screening in vegetables samples in local villages in Hanoi, Vietnam. The concentrations of all target analytes were below limit of quantitation and lower than US-FDA or EU maximum residue levels.

Colony-stimulating factor-1 receptor provides a growth advantage in epithelial cancer cell line A431 in the presence of epidermal growth factor receptor inhibitor gefitinib.

Although epidermal growth factor receptor (EGFR) has been identified as a potent "oncogenic driver" in various tumors of epithelial origin, EGFR-targeted therapies are often of limited success. One of the challenges of improving targeted therapies is to overcome bypassing signaling pathways. Analysis of RNA-seq data of 1006 cell lines from the Cancer Cell Line Encyclopedia (CCLE) revealed that more than 12% of carcinoma cell lines expressed markedly elevated mRNA levels of colony-stimulating factor (CSF)-1 receptor (CSF-1R). Since epithelial cells also express CSF-1, elevated levels of CSF-1R may participate in providing alternative growth and survival signals under targeted therapies. To address this question, we ectopically expressed CSF-1R in A431 cells that express EGFR at high levels, but no biologically relevant level of CSF-1R. In the presence of EGFR inhibitor gefitinib, CSF-1R provided a significant growth advantage in A431 cells. As expected, activation of both receptors, EGFR or CSF-1R, induced phosphorylation of extracellular signal-regulated kinase (Erk)1/2, Akt, protein kinase C (PKC) and signal transducer and activator of transcription (STAT)3. However, EGFR, but not CSF-1R, also induced STAT5 phosphorylation. Inhibitor of phosphatidylinositol 3-kinase (PI3K) (AZD8186), MAPK/ERK kinase (MEK)1/2 (U0126), PKCs (Bisindolylmaleimide I or Gö6976) or STAT3 (Stattic) partially reduced proliferation of CSF-1R expressing A431 cells in the presence of gefitinib. Moreover, multi-kinase inhibitor, cabozantinib, suppressed CSF-1R activation and drastically reduced cell growth when combined with gefitinib. These data suggest that CSF-1R has the potential to reduce sensitivity to gefitinib and may be involved in resistance development.