Arabidopsis root responses to salinity depend on pectin modification and cell wall sensing.

Owing to its detrimental effect on plant growth, salinity is an increasing worldwide problem for agriculture. To understand the molecular mechanisms activated in response to salt in Arabidopsis thaliana, we investigated the Catharanthus roseus receptor-like kinase 1-like family, which contains sensors that were previously shown to be involved in sensing the structural integrity of the cell walls. We found that herk1 the1-4 double mutants, lacking the function of HERKULES1 (HERK1) and combined with a gain-of-function allele of THESEUS1 (THE1), strongly respond to salt application, resulting in an intense activation of stress responses, similarly to plants lacking FERONIA (FER) function. We report that salt triggers pectin methyl esterase (PME) activation and show its requirement for the activation of several salt-dependent responses. Because chemical inhibition of PMEs alleviates these salt-induced responses, we hypothesize a model in which salt directly leads to cell wall modifications through the activation of PMEs. Responses to salt partly require the functionality of FER alone or HERK1/THE1 to attenuate salt effects, highlighting the complexity of the salt-sensing mechanisms that rely on cell wall integrity.

KLF4 expression enhances the efficacy of chemotherapy drugs in ovarian cancer cells.

KLF4 is a transcriptional factor that can function either as a tumor suppressor or oncogene in cancer based on its cellular context. We recently demonstrated that KLF4 was a tumor suppressor in ovarian cancer cells by inhibiting the epithelial to mesenchymal transition. Here we report that KLF4 expression was downregulated in ovarian cancer tissue compared to normal ovarian tissue, and low KLF4 expression correlated with high risk ovarian carcinoma and poor patient survival. Enforced KLF4 expression by lentiviral transduction sensitized ovarian cancer cells to the effects of the chemotherapy drugs, paclitaxel and cisplatin. Treatment of ovarian cancer cells with APTO-253, a small molecule inducer of KLF4, enhanced the efficacy of both chemotherapy drugs. KLF4 expression mediated by lentiviral vector or induced by APTO-253 resulted in G1 phase arrest in ovarian cancer cells. Our results demonstrate that for the first time that inducing KLF4 expression with APTO-253 is a novel therapeutic strategy for treating ovarian cancer.

Using multi-focus group method as an effective tool for eliciting business system requirements: Verified by a case study.

This research aims to explore the multi-focus group method as an effective tool for systematically eliciting business requirements for business information system (BIS) projects. During the COVID-19 crisis, many businesses plan to transform their businesses into digital businesses. Business managers face a critical challenge: they do not know much about detailed system requirements and what they want for digital transformation requirements. Among many approaches used for understanding business requirements, the focus group method has been used to help elicit BIS needs over the past 30 years. However, most focus group studies about research practices mainly focus on a particular disciplinary field, such as social, biomedical, and health research. Limited research reported using the multi-focus group method to elicit business system requirements. There is a need to fill this research gap. A case study is conducted to verify that the multi-focus group method might effectively explore detailed system requirements to cover the Case Study business's needs from transforming the existing systems into a visual warning system. The research outcomes verify that the multi-focus group method might effectively explore the detailed system requirements to cover the business's needs. This research identifies that the multi-focus group method is especially suitable for investigating less well-studied, no previous evidence, or unstudied research topics. As a result, an innovative visual warning system was successfully deployed based on the multi-focus studies for user acceptance testing in the Case Study mine in Feb 2022. The main contribution is that this research verifies the multi-focus group method might be an effective tool for systematically eliciting business requirements. Another contribution is to develop a flowchart for adding to Systems Analysis & Design course in information system education, which may guide BIS students step by step on using the multi-focus group method to explore business system requirements in practice.

On growth performance, nutrient digestibility, blood T lymphocyte subsets, and cardiac antioxidant status of broilers.

Different lipid sources differ in the fatty acid profiles and differently affect growth performance as well as immune function of broilers. The influences of different dietary lipid sources on growth performance, nutrient digestibility, blood T lymphocyte population, and cardiac antioxidant status were investigated of broilers. A total of 360 one-day-old male broilers (BW = 44 ± 3 g) were randomized into 3 treatment groups, consisting of 6 replicates with 20 birds in each group. Broilers received standard diets supplemented with 5% (wt/wt) of lard (LD, as a control diet), sesame oil (SO), or flaxseed oil (FO). Broilers in both SO and FO treatment groups had lower (P < 0.05) feed conversion ratios from 22 to 42 d and during the overall phase compared to those in LD treatment group. Meanwhile, the apparent total tract nutrient digestibility of crude fat in SO and FO treatment groups was higher than that in LD treatment group. Both FO and SO treatments decreased (P < 0.05) abdominal fat percentage compared to LD treatment. Total triglycerides and total cholesterol in chicken blood were decreased (P < 0.05) by SO and FO treatments compared to LD treatment. Feeding broilers with FO and SO led to a decrease (P < 0.05) in blood CD4+ T lymphocyte count and in CD4+:CD8+ ratio compared to LD treatment. Sesame oil and FO treatments increased cardiac glutathione peroxidase (P < 0.05) compared to LD treatment. It is concluded that addition of 5% SO and FO to the standard corn-soybean meal diet improved feed efficiency, increased the activities of cardiac glutathione peroxidase, and affected the T lymphocytes ratio of fast growing broilers.

Knockout of MTF1 Inhibits the Epithelial to Mesenchymal Transition in Ovarian Cancer Cells.

Due to peritoneal metastasis and frequent recurrence, ovarian cancer has the highest mortality among gynecological cancers. Epithelial to mesenchymal transition (EMT) contributes to ovarian tumor metastasis. In this study, we report for the first time that metal regulatory transcription factor 1 (MTF1) was upregulated in ovarian cancer, and its high expression was associated with poor patient survival and disease relapse. Knockout of MTF1 using lentiviral CRISPR/Cas9 nickase vector-mediated gene editing inhibited EMT by upregulating epithelial cell markers E-cadherin and cytokeratin 7, and downregulating mesenchymal markers Snai2 and ß-catenin in ovarian cancer SKOV3 and OVCAR3 cells. Loss of MTF1 reduced cell proliferation, migration, and invasion in both SKOV3 and OVCAR3 cells. Knockout of MTF1 upregulated the expression of the KLF4 transcription factor, and attenuated two cellular survival pathways, ERK1/2 and AKT. Our studies demonstrated that MTF1 plays an oncogenic role and contributes to ovarian tumor metastasis by promoting EMT. MTF1 may be a novel biomarker for early diagnosis as well as a drug target for clinical therapy.

Lentiviral CRISPR/Cas9 vector mediated miR-21 gene editing inhibits the epithelial to mesenchymal transition in ovarian cancer cells.

CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats) mediated genome editing is a powerful approach for loss of function studies. Here we report that lentiviral CRISPR/Cas9 vectors are highly efficient in introducing mutations in the precursor miRNA sequence, thus leading to the loss of miRNA expression and function. We constructed four different lentiviral CRISPR/Cas9 vectors that target different regions of the precursor miR-21 sequence and found that these lentiviral CRISPR/Cas9 miR-21 gRNA vectors induced mutations in the precursor sequences as shown by DNA surveyor mutation assay and Sanger sequencing. Two miR-21 lentiviral CRISPR/Cas9 gRNA vectors were selected to probe miR-21 function in ovarian cancer SKOV3 and OVCAR3 cell lines. Our data demonstrate that disruption of pre-miR-21 sequences leads to reduced cell proliferation, migration and invasion. Moreover, CRISPR/Cas9-mediated miR-21 gene editing sensitizes both SKOV3 and OVCAR3 cells to chemotherapeutic drug treatment. Disruption of miR-21 leads to the inhibition of epithelial to mesenchymal transition (EMT) in both SKOV3 and OVCAR3 cells as evidenced by the upregulation of epithelial cell marker E-cadherin and downregulation of mesenchymal marker genes, vimentin and Snai2. The miR-21 target genes PDCD4 and SPRY2 were upregulated in cells transduced with miR-21gRNAs compared to controls. Our study indicates that lentiviral CRISPR/Cas9-mediated miRNA gene editing is an effective approach to address miRNA function, and disruption of miR-21 inhibits EMT in ovarian cancer cells.