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We aimed to identify serum exosomal microRNAs (miRNAs) associated with the transition from atrial fibrillation (AF) to sinus rhythm (SR) and investigate their potential as biomarkers for the early recurrence of AF within three months post-treatment. We collected blood samples from eight AF patients at Chang Gung Memorial Hospital in Taiwan both immediately before and within 14 days following rhythm control treatment. Exosomes were isolated from these samples, and small RNA sequencing was performed. Using DESeq2 analysis, we identified nine miRNAs (16-2-3p, 22-3p, 23a-3p, 23b-3p, 125a-5p, 328-3p, 423-5p, 504-5p, and 582-3p) associated with restoration to SR. Further analysis using the DIABLO model revealed a correlation between the decreased expression of miR-125a-5p and miR-328-3p and the early recurrence of AF. Furthermore, early recurrence is associated with a longer duration of AF, presumably indicating a more extensive state of underlying cardiac remodeling. In addition, the reads were mapped to mRNA sequences, leading to the identification of 14 mRNAs (AC005041.1, ARHGEF12, AMT, ANO8, BCL11A, DIO3OS, EIF4ENIF1, G2E3-AS1, HERC3, LARS, NT5E, PITX1, SLC16A12, and ZBTB21) associated with restoration to SR. Monitoring these serum exosomal miRNA and mRNA expression patterns may be beneficial for optimizing treatment outcomes in AF patients. [ABSTRACT FROM AUTHOR]

Background: Patients with type 2 diabetes are at increased risk for cardiovascular diseases. Sodium-glucose transport 2 inhibitors (SGLT2i) have been shown to enhance cardiovascular health since their debut as a second-line therapy for diabetes. Acute coronary syndrome (ACS), peripheral arterial occlusive disease (PAOD), and ischemic stroke (IS) are types of atherosclerotic cardiovascular disease (ASCVD), although the benefits of treating these disorders have not been shown consistently. Methods: We searched four databases (PubMed, Embase, the Cochrane library, and clinicaltrial.gov) for randomized clinical trials (RCTs) until November of 2022. Comparisons were made between SGLT2i-treated and control individuals with type 2 diabetes. Primary outcomes were ACS, PAOD, and IS; secondary outcomes included cardiovascular mortality and all-cause mortality. Risk ratio (RR) and 95% confidence intervals (CI) were determined using a fixed effects model. Cochrane's risk-of-bias (RoB2) instrument was used to assess the validity of each study that met the inclusion criteria. Results: We enrolled 79,504 patients with type 2 diabetes from 43 RCTs. There was no difference in the risk of ACS (RR = 0.97, 95% CI 0.89–1.05), PAOD (RR = 0.98, 95% CI 0.78–1.24), or IS (RR = 0.95, 95% CI 0.79–1.14) among patients who took an SGLT2i compared to those who took a placebo or oral hypoglycemic drugs. Subgroup analysis revealed that none of the SGLT2i treatments (canagliflozin, dapagliflozin, empagliflozin, and ertugliflozin) significantly altered outcomes when analyzed separately. Consistent with prior findings, SGLT2i reduced the risk of cardiovascular mortality (RR = 0.85, 95% CI 0.77–0.93) and all-cause mortality (RR = 0.88, 95% CI 0.82–0.94). Conclusion: Our results appear to contradict the mainstream concepts regarding the cardiovascular effects of SGLT2i since we found no significant therapeutic benefits in SGLT2i to reduce the incidence of ACS, PAOD, or IS when compared to placebo or oral hypoglycemic drugs. [ABSTRACT FROM AUTHOR]

Diabetes mellitus (DM) is known to increase the risk of hepatocellular carcinoma (HCC) among individuals with chronic hepatitis C (CHC). We aimed to evaluate whether metformin reduces HCC risk among individuals with DM and CHC after successful antiviral therapy. Individuals with CHC who achieved a sustained virological response (SVR) after interferon-based therapy were enrolled in a large-scale, multicenter cohort in Taiwan (T-COACH). Cases of HCC at least 1 year after SVR were identified through linkage to the catastrophic illness and cancer registry databases. Of 7,249 individuals with CHC enrolled in the study, 781 (10.8%) had diabetes and 647 (82.8%) were metformin users. During a median follow-up of 4.4 years, 227 patients developed new-onset HCC. The 5-year cumulative HCC incidence was 10.9% in non-metformin users and 2.6% in metformin users, compared to 3.0% in individuals without DM (adjusted hazard ratio [aHR] 2.83; 95% CI 1.57-5.08 and aHR 1.46; 95% CI 0.98-2.19, respectively). Cirrhosis was the most important factor significantly associated with higher HCC risk in Cox regression analysis, followed by DM non-metformin use, older age, male sex, and obesity; whereas hyperlipidemia with statin use was associated with a lower HCC risk. Using the two most crucial risk factors, cirrhosis and DM non-metformin use, we constructed a simple risk model that could predict HCC risk among individuals with CHC after SVR. Metformin use was shown to reduce the risk of all liver-related complications. Metformin use greatly reduced HCC risk after successful antiviral therapy in individuals with diabetes and CHC. A simple risk stratification model comprising cirrhosis and DM non-metformin use could predict long-term outcomes in individuals with CHC after SVR. The current study provides evidence that metformin could reduce hepatocellular carcinoma (HCC) incidence after successful antiviral therapy among those with diabetes and chronic hepatitis C in a large-scale nationwide cohort study. Although successful antiviral therapy greatly reduces HCC risk in individuals with chronic hepatitis C, those with cirrhosis, diabetes, obesity, and the elderly remain at high risk of HCC development. We demonstrated that a simple risk model composed of two crucial unfavorable factors, cirrhosis and diabetes without metformin use, predicts the risk of HCC and major liver-related complications after successful antiviral therapy in individuals with chronic hepatitis C. Metformin use is highly recommended for individuals with diabetes and chronic hepatitis C after viral eradication to reduce the risk of HCC. [Display omitted] • Diabetes mellitus is associated with an increased risk of HCC in individuals with CHC. • Diabetes mellitus still increases the risk of HCC after successful antiviral therapy. • Use of metformin for DM greatly reduces HCC risk after successful antiviral therapy. • A simple risk stratification model could predict the risk of HCC and other major liver-related complications. [ABSTRACT FROM AUTHOR]

Objective: Mutations in the colony‐stimulating factor 1 receptor gene (CSF1R) were identified as a cause of adult‐onset inherited leukoencephalopathy. The present study aims at investigating the frequency, clinical characteristics, and functional effects of CSF1R mutations in Taiwanese patients with adult‐onset leukoencephalopathy. Methods: Mutational analysis of CSF1R was performed in 149 unrelated individuals with leukoencephalopathy by a targeted resequencing panel covering the entire coding regions of CSF1R. In vitro analysis of the CSF1‐induced autophosphorylation activities of mutant CSF1R proteins was conducted to assess the pathogenicity of the CSF1R mutations. Results: Among the eight CSF1R variants identified in this study, five mutations led to a loss of CSF1‐induced autophosphorylation of CSF1R proteins. Four mutations (p.K586*, p.G589R, p.R777Q, and p.R782C) located within the tyrosine kinase domain of CSF1R, whereas the p.T79M mutation resided in the immunoglobulin‐like domain. The five patients carrying the CSF1R mutations developed cognitive decline at age 41, 43, 50, 79, and 86 years, respectively. Psychiatric symptoms and behavior changes were observed in four of the five patients. The executive function and processing speed were severely impaired at an early stage, and their cognitive function deteriorated rapidly within 3–4 years. Diffusion‐restricted lesions at the subcortical regions and bilateral corticospinal tracts were found in three patients. Interpretation: CSF1R mutations account for 3.5% (5/149) of the adult‐onset leukoencephalopathy in Taiwan. CSF1R mutations outside the tyrosine kinase domain may also disturb the CSF1R function and lead to the clinical phenotype. Molecular functional validation is important to determine the pathogenicity of novel CSF1R variants. [ABSTRACT FROM AUTHOR]

• Developed a sensitive and fast analytical methodology for monitoring Remdesivir. • An improved vortex-assisted salt-induced liquid-liquid microextraction technique. • Short total run time in both UHPLC-PDA and UHPLC-MS/MS methods. • This method is simple, efficient, low-cost and eco-friendly sample preparation process. • This method can be used for accurate monitoring of Remdesivir in human plasma. • As a standard analytical tool for PK/TDM during clinical trials/COVID-19 treatment. To tackle the harmful consequences of the widespread COVID-19 pandemic, a broad-spectrum anti-viral drug remdesivir (RDV) has gained the utmost attention recently due to its promising application in treating COVID-19 patients. However, a fast and sensitive analytical methodology is important to monitor RDV drug profile in human plasma for pharmacokinetics (PK) and therapeutic drug monitoring (TDM). In this study, we demonstrate an improved vortex-assisted salt-induced liquid-liquid microextraction (VA-SI-LLME) technique coupled with UHPLC-PDA and UHPLC-MS/MS for rapid determination of RDV in human plasma. This technique involves simple one-step protein precipitation with hydrochloric acid and subsequent extraction with acetonitrile for analysis. Under the optimal VA-SI-LLME conditions (500 μL of acetonitrile with 2.5 g ammonium sulfate under 2 min vortex extraction), method validation results indicated an excellent correlation coefficient of 0.9969 for UHPLC-PDA (monitored at 254 nm) and 0.9990 for UHPLC-MS/MS (monitored at electrospray ionization with + ion mode transitions of m/z 603.1→ m/z 402.20 and m/z 603.1→ m/z 199.90). The detection and quantification limits were 1.5 and 5 ng/mL for UHPLC/PDA, and 0.3 and 1 ng/mL for UHPLC-MS/MS, respectively. The developed method showed excellent extraction recoveries between 90.79–116.74 % and 85.68–101.34 % with intraday and interday precision ≤ 9.59 for both methods. These results proved that the developed method is a simple, fast, and sensitive analytical method that can be applied as a standard analytical tool for PK and TDM studies of RDV in clinical trials during the current worldwide outbreak. [ABSTRACT FROM AUTHOR]

Rationale: Organophosphate pesticides (OPPs) are the most commonly used insecticides around the world in various agricultural and domestic practices, and humans are frequently exposed to these hazardous insecticides that can lead to several chronic health effects. Therefore, a fast and sensitive analytical method is required for biomonitoring the markers of OPPs in humans for exposure estimation. In this study, a fast and sensitive analytical procedure was developed for the determination of the metabolites of OPPs in human urine samples. Methods: Metabolites of OPPs were extracted from 2 mL of urine sample using a novel vortex‐assisted salt‐induced liquid–liquid microextraction (VA‐SI‐LLME) technique, and the preconcentrated metabolites were analyzed by ultra‐high‐performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS). Various factors affecting the efficiency of VA‐SI‐LLME were thoroughly investigated. Results: The metabolites of OPPs exhibited very good linearity over the concentration range between 0.05 and 50 ng mL−1 with coefficient (r2) values ranging between 0.9986 and 0.9999. The method showed excellent sensitivity with detection limits ranging from 0.01 to 0.03 ng mL−1 and quantification limits from 0.03 to 0.05 ng mL−1. The developed method was applied to the analysis of real samples and the recoveries ranged between 85.0 and 114.1% with related standard deviations <5%. Conclusions: The results showed the VA‐SI‐LLME/UHPLC/MS/MS method to be a simple, rapid, sensitive, and selective analytical procedure for the biomonitoring of the metabolites of OPPs in humans. This efficient and cost‐effective analytical method could be a potential alternative method for the biomonitoring of the metabolites of pesticides in humans. [ABSTRACT FROM AUTHOR]

Summary: Background: Racial and ethnic disparities exist for hepatitis C virus (HCV) treatment and hepatocellular carcinoma (HCC) survival. Aim: To evaluate the impact of HCV treatment on such disparities. Methods: In a retrospective cohort study, we analysed 6069 patients with HCV‐related HCC (54.2% Asian, 30.1% White, 8.5% Black, and 7.3% Hispanic) from centres in the United States and Asia. Results: The mean age was 61, 60, 59 and 68, respectively, for White, Black, Hispanic and Asian patients. Black patients were most likely to have Barcelona Clinic Liver Cancer stage D, vascular invasion and distant metastasis (23% vs. 5%–15%, 20% vs. 10%–17% and 10% vs. 5%–7%, respectively; all p < 0.0001). Treatment rate with direct‐acting antiviral agents (DAA) was 35.9% for Asian, 34.9% for White, 30.3% for Hispanic (30.3%), and 18.7% for Black patients (p < 0.0001). Among those untreated or without sustained virologic response (SVR), 10‐year survival rates were 35.4, 27.5, 19.3 and 14.0, respectively, for Asian, Hispanic, White and Black patients (p < 0.0001). There were no statistically significant differences among those with SVR (p = 0.44). On multivariable analysis adjusted for relevant confounders, there was no statistically significant association between survival and being Hispanic (aHR: 0.68, p = 0.26) or Black (aHR: 1.18, p = 0.60) versus White. There was a significant association between being Asian American and survival (aHR: 0.24, p = 0.001; non‐U.S. Asian: aHR: 0.66, p = 0.05), and for SVR (aHR: 0.30, p < 0.0001). Conclusion: DAA treatment rates were suboptimal. Racial and ethnic disparities resolved with HCV cure. Early diagnosis and improved access to HCV treatment is needed for all patients with HCV infection. [ABSTRACT FROM AUTHOR]

In this paper, we report a simple, fast, and sensitive analytical procedure for monitoring of synthetic pyrethroid pesticides' (SPPs') metabolites in environmental water samples using a novel vortex-assisted salt-induced liquid-liquid microextraction (VA-SI-LLME) method coupled with liquid chromatography/tandem mass spectrometric (LC-MS/MS) analysis. VA-SI-LLME was carried out by taking 4 mL of sample solution followed by the addition of isopropanol and ethyl acetate (0.4 mL, 1:2, v/v) as a binary extraction solvent, and 4 g of ammonium sulfate for phase separation. Then, the mixture solution was vortex for 2 min and centrifuged for 3 min to collect the extractant and inject into LC-MS/MS for analysis. Experimental factors affecting the VA-SI-LLME including extraction solvent, extraction solvent volume, salt type, amount of salt, extraction time, and sample pH were completely optimized. The developed analytical method was validated, and the results exhibited good linearity in the concentration ranges between 0.01–100 ng mL−1 with the correlation coefficient (r2) >0.992 for the target SPPs' metabolites. Excellent limit of quantification and precision were achieved in the range between 0.01–0.1 ng mL−1, and 1.55–5.06% (in terms relative standard deviation), respectively. The developed method was employed to determine the target SPPs' metabolites in tap, lake, river water, and influent wastewater samples collected in Kaohsiung city area, Taiwan. The extraction recoveries of spiked samples were satisfactory. Therefore, the presented method can be used as a potential alternative analytical procedure for monitoring of pesticides' metabolites in aqueous samples. • Rapid analytical procedure for monitoring and analysis of pyrethroid pesticides' metabolites • A new vortex-assisted salt-induced liquid-liquid microextraction technique • Liquid chromatography/tandem mass spectrometric analysis of SPPs' metabolites • The method was employed to analysis the target SPPs' metabolites in environmental water samples. • Method is simple, fast, sensitive, eco-friendly and efficient method. [ABSTRACT FROM AUTHOR]

Background and Aim: The prevalence of metabolic dysfunction‐associated fatty liver disease (MAFLD) and its interplay with hepatitis B virus (HBV) and hepatitis C virus (HCV) in terms of liver disease severity is elusive. Methods: A mass surveillance program was conducted in a viral hepatitis endemic area. The objective was to identify MAFLD/non‐MAFLD subjects with advanced liver disease. Results: Two thousand two hundred and forty‐two (41.7%) of the 5378 subjects were identified as having MAFLD, and 375 (7.0%) had advanced liver disease. The proportions of anti‐HCV and HBsAg seropositivity were 19.3% and 9.7%, respectively. The proportions of advanced fibrosis in subjects with non‐viral hepatitis (NBNC), HBV and HCV infection were 2.8%, 5.7% and 23.4%, respectively. Subjects with MAFLD had a significantly higher proportion of advanced fibrosis (8.7% vs 5.7%, P < 0.001). Factors associated with advanced fibrosis included age (odds ratio [OR]/95% confidence interval [CI]: 4.8/3.7–6.0, P < 0.001), male sex (OR/CI: 1.3/1.0–1.7, P = 0.019), anti‐HCV seropositivity (OR/CI: 5.9/4.6–7.5, P = 0.019), MAFLD‐lean metabolic dysregulation (MS) (OR/CI: 2.6/1.3–5.2, P = 0.005; compared with the non‐MAFLD group) and MAFLD‐diabetes (OR/CI: 1.5/1.1–2.1, P = 0.008; compared with the non‐MAFLD group). MAFLD did not aggravate liver disease severity in patients with viral hepatitis. However, among NBNC subjects, factors associated with advanced liver disease included MAFLD‐lean MS group (OR/CI: 9.1/2.4–34.6, P = 0.001; compared with non‐MAFLD group) and MAFLD‐DM group (OR/CI: 2.0/1.2–3.2, P = 0.004; compared with non‐MAFLD group). Conclusions: MAFLD patients with diabetes and metabolic dysregulation had a higher risk of advanced liver disease. The effect was more significant in non‐viral hepatitis subjects in a community level. [ABSTRACT FROM AUTHOR]

Objective Charcot-Marie-Tooth disease type X1 ( CMTX1), which is caused by mutations in the gap junction ( GJ) protein beta-1 gene ( GJB1), is the second most common form of Charcot-Marie-Tooth disease ( CMT). GJB1 encodes the GJ beta-1 protein ( GJB1), which forms GJs within the myelin sheaths of peripheral nerves. The process by which GJB1 mutants cause neuropathy has not been fully elucidated. This study evaluated the biophysical characteristics of GJB1 mutants and their correlations with the clinical features of CMTX1 patients. Methods All patients with a validated GJB1 mutation were assessed using the Charcot-Marie-Tooth disease neuropathy score version 2 ( CMTNS). The impacts of the mutations on the biophysical functions of GJB1 were characterized by assessing intracellular localization, expression patterns, and GJ Ca2+ permeability. Result Nineteen GJB1 mutations were identified in 24 patients with a clinical diagnosis of CMT. Six are novel mutations: p.L6S, p.I20F, p.I101Rfs*8, p.F153L, p.R215P, and p.D278V. Diverse pathological effects of the mutations were demonstrated, including reduced GJB1 expression, intracellular mislocalization, and altered GJ functions. GJB1 mutations that caused a complete loss of GJ Ca2+ permeability appeared to be associated with an earlier disease onset, whereas those resulting in preservation of GJ permeability and with predominant cell membrane expression tended to have a later onset and a milder phenotype. Interpretation This study demonstrated that the degree of loss of GJ function caused by the GJB1 mutations might contribute to the onset and severity of neuropathic symptoms in CMTX1. [ABSTRACT FROM AUTHOR]

The epithelial-to-mesenchymal transition is the first step required for breast cancer to initiate metastasis. The aberrant activation of proto-oncogene tyrosine-protein kinase Src regulates multiple functions during tumor progression. Cardiotoxin III, a basic polypeptide isolated fromNaja naja atravenom, has been shown to exhibit anticancer activity; however, the effect of cardiotoxin III on the epithelial-to-mesenchymal transition of cancer cells remains elusive. Exposure of MDA-MB-231 cells to cardiotoxin III resulted in morphological changes and upregulation of E-cadherin with a concomitant decrease in N-cadherin and vimentin protein levels, resulting in the inhibition of cell migration and invasion. Cardiotoxin III induced downregulation of snail and slug expression. Simultaneously, cardiotoxin III suppressed Src phosphorylation and downstream activation of focal adhesion kinase, of the docking protein p130cas, and of paxillin. In addition, cardiotoxin III inhibited the phosphorylation of extracellular signal-regulated kinase and phosphatidylinositol 3-kinase. The Src specific inhibitor 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine decreased in the phosphorylation and the expression changes of epithelial-to-mesenchymal transition markers in a similar way. Thus, cardiotoxin III has a novel anticancer effect by suppressing proto-oncogene tyrosine-protein kinase activity, reversing epithelial-to-mesenchymal transition. [ABSTRACT FROM AUTHOR]

Breast cancer is a highly malignant carcinoma and most deaths of breast cancer are caused by metastasis. The epithelial-to-mesenchymal transition (EMT) has emerged as a pivotal event in the development of the invasive and metastatic potentials of cancer progression. Epidermal growth factor (EGF) and its receptor, EGFR, play roles in cancer metastasis. CTX III, a basic polypeptide isolated from Naja naja atra venom, has been shown to exhibit anticancer activity; however, the effect of CTX III on the EMT of cancer cells remains elusive. CTX III treatment resulted in morphological changes from elongated and spindle shape to rounded and epithelial-like shape, induced upregulation of E-cadherin and concurrent downregulation of N-cadherin and Vimentin protein levels, corresponding to observed decreases in cell migration and invasion. CTX III treatment also decreased the expression of Snail and Twist in EGF-induced MDA-MB-231 cells. Concurrently, CTX III efficiently inhibited the EGFR phosphorylation and downstream activation of phosphatidylinositol 3-kinase (PI3K)/Akt and ERK1/2. The EGFR specific inhibitor AG1478 significantly suppressed ERK1/2 and Akt phosphorylation, cell migration and invasion, as well as the expressional changes associated with EMT markers in EGF-induced MDA-MB-231 cells. CTX III inhibitory effect on EGF-evoked invasion of MDA-MB-231 cells is mediated through suppressing EGF/EGFR activation and EMT process. [ABSTRACT FROM AUTHOR]

Background: A small group of patients with inherited neuropathy that has been shown to be caused by mutations in the BSCL2 gene. However, little information is available about the role of BSCL2 mutations in inherited neuropathies in Taiwan. Methodology and Principal Findings: Utilizing targeted sequencing, 76 patients with molecularly unassigned Charcot-Marie-Tooth disease type 2 (CMT2) and 8 with distal hereditary motor neuropathy (dHMN), who were selected from 348 unrelated patients with inherited neuropathies, were screened for mutations in the coding regions of BSCL2. Two heterozygous BSCL2 mutations, p.S90L and p.R96H, were identified, of which the p.R96H mutation is novel. The p.S90L was identified in a pedigree with CMT2 while the p.R96H was identified in a patient with apparently sporadic dHMN. In vitro studies demonstrated that the p.R96H mutation results in a remarkably low seipin expression and reduced cell viability. Conclusion: BSCL2 mutations account for a small number of patients with inherited neuropathies in Taiwan. The p.R96H mutation is associated with dHMN. This study expands the molecular spectrum of BSCL2 mutations and also emphasizes the pathogenic role of BSCL2 mutations in molecularly unassigned hereditary neuropathies. [ABSTRACT FROM AUTHOR]

Low birth weight (LBW) can have an impact on health outcomes in later life, especially in relation to pre-disposition to metabolic disease. Several studies suggest that LBW resulting from restricted intrauterine growth leaves a footprint on DNA methylation in utero, and this influence likely persists into adulthood. To investigate this further, we performed epigenome-wide association analyses of blood DNA methylation using Infinium HumanMethylation450 BeadChip profiles in 71 adult monozygotic (MZ) twin pairs who were extremely discordant for birth weight. A signal mapping to the IGF1R gene (cg12562232, p = 2.62 × 10−8), was significantly associated with birth weight discordance at a genome-wide false-discovery rate (FDR) of 0.05. We pursued replication in three additional independent datasets of birth weight discordant MZ pairs and observed the same direction of association, but the results were not significant. However, a meta-analysis across the four independent samples, in total 216 birth-weight discordant MZ twin pairs, showed a significant positive association between birth weight and DNA methylation differences at IGF1R (random-effects meta-analysis p = .04), and the effect was particularly pronounced in older twins (random-effects meta-analysis p = .008, 98 older birth-weight discordant MZ twin pairs). The results suggest that severe intra-uterine growth differences (birth weight discordance >20%) are associated with methylation changes in the IGF1R gene in adulthood, independent of genetic effects. [ABSTRACT FROM PUBLISHER]

Naphtho[1,2- b]furan-4,5-dione ( NFD), a bioactive component of Avicennia marina, has been shown to exhibit anticancer activity. The aim of the present study was to explore the effect of NFD on hepatocyte growth factor ( HGF)-induced cell migration and invasion of MDA- MB-231 human breast cancer cells, as well as the underlying mechanism of action. Cell viability was determined using the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay, western blot analysis was used to measure protein expression and cell migration and invasion were evaluated by the cell wound healing assay, Boyden chamber assay and gelatin zymography. When cells were treated with non-toxic concentrations of NFD (1-3 μmol/L, 24 h), NFD concentration-dependently inhibited HGF-promoted cell migration and invasion. Simultaneously, NFD efficiently suppressed c-Met phosphorylation and downstream activation of phosphatidylinositol 3-kinase ( PI3K) and Akt. In addition, NFD inhibited the phosphorylation of I κB kinases and I κB α and nuclear translocation of nuclear factor ( NF)- κB, as well as matrix metalloproteinase ( MMP)-9 activity. Furthermore, the c-Met inhibitor PHA665752 (10 μmol/L) inhibited HGF-induced MMP-9 expression, cell migration and invasion, as well as the activation of PI3K/Akt, suggesting that PI3K/Akt activation occur downstream of c-Met activation. In conclusion, the results of the present study suggest that NFD inhibits HGF-induced invasion and migration of MDA- MB-231 cells via HGF- and/or c-Met-mediated PI3K/Akt and NF- κB signalling pathways, leading to downregulation of MMP-9 expression and cell migration. [ABSTRACT FROM AUTHOR]

Background: Gastric cancer is common cancer. Discovering novel genetic biomarkers might help to identify high-risk individuals. Copy number variation (CNV) has recently been shown to influence risk for several cancers. The aim of the present study was sought to test the association between copy number at a variant region and GC. Methods: A total of 110 gastric cancer patients and 325 healthy volunteers were enrolled in this study. We searched for a CNV and found a CNV (Variation 7468) containing part of the APC gene, the SRP19 gene and the REEP5 gene. We chose four probes targeting at APC-intron8, APC-exon9, SRP19 and REEP5 to interrogate this CNV. Specific Taqman probes labeled by different reporter fluorophores were used in a real-time PCR platform to obtain copy number. Both the original non-integer data and transformed integer data on copy number were used for analyses. Results: Gastric caner patients had a lower non-integer copy number than controls for the APC-exon9 probe (Adjusted p = 0.026) and SRP19 probe (Adjusted p = 0.002). The analysis of integer copy number yielded a similar pattern although less significant (Adjusted p = 0.07 for APC-exon9 probe and Adjusted p = 0.02 for SRP19 probe). Conclusions: Losses of a CNV at 5q22, especially in the DNA region surrounding APC-exon 9, may be associated with a higher risk of gastric cancer. [ABSTRACT FROM AUTHOR]

The hepatocyte growth factor (HGF)/c-Met signalling pathway is deregulated in most cancers and associated with a poor prognosis in breast cancer. Cardiotoxin III (CTX III), a basic polypeptide isolated from Naja naja atra venom, has been shown to exhibit anticancer activity. In this study, we use HGF as an invasive inducer to investigate the effect of CTX III on MDA-MB-231 cells. When cells were treated with non-toxic doses of CTX III, CTX III inhibited the HGF-promoted cell migration and invasion. CTX III significantly suppressed the HGF-induced c-Met phosphorylation and downstream activation of phosphatidylinositol 3-kinase (PI3k)/Akt and extracellular signal-regulated kinase (ERK) 1/2. Additionally, CTX III similar to wortmannin (a PI3K inhibitor) and U0126 (an upstream kinase regulating ERK1/2 inhibitor) attenuated cell migration and invasion induced by HGF. This effect was paralleled by a significant reduction in phosphorylation of I κB α kinase and I κB α and nuclear translocation of nuclear factor κB (NF- κB) as well as a reduction of matrix metalloproteinase-9 (MMP-9) activity. Furthermore, the c-Met inhibitor PHA665752 inhibited HGF-induced MMP-9 expression, cell migration and invasion, as well as the activation of ERK1/2 and PI3K/Akt, suggesting that ERK1/2 and PI3K/Akt activation occurs downstream of c-Met activation. Taken together, these findings suggest that CTX III inhibits the HGF-induced invasion and migration of MDA-MB-231 cells via HGF/c-Met-dependent PI3K/Akt, ERK1/2 and NF- κB signalling pathways, leading to the downregulation of MMP-9 expression. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Polyunsaturated fatty acids (PUFAs) consumption indicates beneficial effects on cardiovascular disease (CVD) and physiological processes in humans. However, the inappropriate ratio of omega-(ω)-PUFA levels in human blood is considered as raising the risk of CVD. Therefore, monitoring dietary ω-FAs in human serum is vital for early diagnosis for individuals to predict CVD risk. This work reports a fast green sample pre-treatment protocol for sensitive and simultaneous monitoring of ω-3-FAs and ω-6-FAs in serum by novel in-syringe-based ultrasonication-assisted alkaline hydrolysis coupled with vortex-induced liquid-liquid microextraction (IS-USAH-VI-LLME) technique connected with UHPLC–MS/MS analysis. Factors affecting extraction recoveries of ten ω-PUFAs by the presented method were well-studied. ω-3 and ω-6 PUFAs demonstrated excellent linearities between the concentrations between 0.1–10,000 ng mL−1 with good regression coefficients between 0.9910–0.9997. The detection and quantification limits were between 0.05–0.35 and 0.16–1.07 ng mL−1, demonstrating that the presented method is highly sensitive and versatile. The precision of the technique was <8.2% that deemed acceptable in clinical analysis. Further, the proposed method was applied for ω-PUFAs analysis in human blood samples, and spiked recoveries showed between 80.32–119.34% with <9.82% precision. Results proved that the developed method is green, sensitive, and reliable to simultaneously determine ten ω-PUFAs in human blood samples for clinical diagnosis applications for predicting health hazards. [Display omitted] • Trace level anaysis of environmentally healthy poly-unsaturated fatty acids in serum. • Sensitive and simultaneous biomonitoring of ω-PUFAs in human blood by LC-MS/MS. • In-syringe ultrasonication-assisted vortex-induced liquid-liquid microextraction. • This technique was employed for the real-time biomonitoring of human serum samples. • This method is green, and efficient for real-time analysis for health hazard assessment. [ABSTRACT FROM AUTHOR]

Highlights: [•] Development of a class II PHA synthase protein model for structure predition. [•] Identification of beneficial sites Gln481 and Ala547 for enhancing PHA production. [•] The residue Leu484 changes the substrate specificity of PhaC1 Pp from 8C shift to 4C. [ABSTRACT FROM AUTHOR]

Abstract: Cardiotoxin III (CTX III), a basic polypeptide isolated from Naja naja atra venom, has been demonstrated to display anticancer activity. Breast cancer is a highly malignant carcinoma and most deaths of breast cancer are caused by metastasis. In this study, we show that CTX III blocks migration and invasion of MDA-MB-231 breast cancer cells without affecting apoptosis or cell cycle arrest. CTX III caused significant block of Src kinase activity in MDA-MB-231 cells. Moreover, CTX III treatment was correlated with reduced phosphorylation of FAK at Tyr576, 861 and 925 sites, p130Cas at Tyr410, and paxillin at Tyr118. CTX III also suppressed the activation of extracellular signal-regulated kinase1/2 and phosphatidylinositol 3-kinase/Akt. Consistent with inhibition of these signaling pathways and invasion, CTX III inhibited the expression of matrix metalloproteinase-9. In addition, Src specific inhibitor PP2 caused a significant decrease in the phosphorylation of FAK, p130Cas, paxillin, PI3K/Akt, and ERK1/2. Taken together, CTX III significantly inhibited phosphorylation of Src and downstream molecules as well as cell migration and invasion. Our findings provide evidences that CTX III inhibits Src-mediated signaling pathways involved in controlling MDA-MB-231 cell migration and invasion, suggesting that it has therapeutic potential in breast cancer treatment. [Copyright &y& Elsevier]

Abstract: Mutations in the 5′ non-coding region of GJB1 are rarely reported in patients with Charcot–Marie–Tooth disease (CMT). We therefore aimed to assess the frequency and identities of the GJB1 5′ non-coding region mutations in a cohort of CMT. We analyzed the 5′ non-coding region of GJB1 (including the promoter P2 and exon 1b) in 91 unrelated CMT patients without an identified genetic cause. Two mutations, c.-529T>C, and c.-459C>T, were identified in one patient each. One polymorphism, c.-713G>A, was also identified in 53 patients and 73 of the 100 control subjects. The luciferase reporter assays showed that c.-459C>T significantly reduced the luciferase expression with or without SOX10 activation, whereas c.-529T>C impaired the expression only with SOX10 co-expression. c.-713G>A had no apparent functional effect. Mutations in the 5′ non-coding region of GJB1 account for 0.8% (2 of 251) of CMT and 2.2% (2 of 91) of genetically unassigned CMT in a Taiwanese cohort. As previously demonstrated, c.-459C>T and c.-529T>C may cause CMT through compromising GJB1 expression whereas c.-713G>A is a benign variant. This study highlights the pathogenic role of the GJB1 5′ non-coding region mutations in CMT, and suggests that their identification should be considered for CMT patients without commonly observed mutations. [Copyright &y& Elsevier]

Background/Aims: MicroRNA miR-21, miR-221 and miR-145 have been implicated in the cardiovascular system. We aimed to compare the serum levels of the three microRNAs (miRNAs) in different severities of cerebrovascular diseases and evaluate the feasibility of using these miRNAs as biomarkers for stroke. Methods: We enrolled 167 subjects with ischemic stroke, 66 atherosclerosis subjects with any carotid plaque score and 157 healthy controls. These three types of subjects represent three levels of severity in cerebrovascular diseases. Analysis of covariance was used to evaluate the relationship between miRNAs and disease severity with adjustment for conventional risk factors. To test the prediction for stroke, we built regression models containing the serum miRNA levels and risk factors. Prediction capabilities were compared by the receiver operating characteristic curves. Results: Stroke patients and atherosclerosis subjects had significantly higher miR-21 and lower miR-221 serum levels than healthy controls, while the miR-145 expression was too low to provide useful information in this regard. The best model showed that miR-21 and miR-221 were independent predictors. There was a 6.2-fold increase for stroke risk when miR-21 levels increase by log102-ΔCt = 1, while a 10.4-fold increase was observed as miR-221 decreases by log102-ΔCt = 1. Conclusions: Serum miR-145 was not detected in over 50% of the patients and it may not be an ideal marker to predict stroke. MiR-21 and miR-221 are novel biomarkers for atherosclerosis and stroke. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Highlights: [•] We first noted that brazilein inhibited MDA-MB-231 cell migration and invasion. [•] Brazilein suppressed the expression, activation, and mRNA of MMP-2. [•] Brazilein decreased the nuclear protein level of NF-κB. [•] Brazilein suppressed the phosphorylation of p38 MAPK, PI3K and Akt. [Copyright &y& Elsevier]

Abstract: Naphtho[1,2-b]furan-4,5-dione (NFD), a bioactive component of Avicennia marina, has been demonstrated to display anti-cancer activity. Activation of epidermal growth factor receptor (EGFR)-induced signaling pathway has been correlated with cancer metastasis in various tumors, including breast carcinoma. We use EGF as a metastatic inducer of MDA-MB-231 cells to investigate the effect of NFD on cell migration and invasion. NFD suppressed EGF-mediated protein levels of c-Jun and c-Fos, and reduced MMP-9 expression and activity, concomitantly with a marked inhibition on cell migration and invasion without obvious cellular cytotoxicity. NFD abrogated EGF-induced phosphorylation of EGF receptor (EGFR) and phosphatidylinositol 3-kinase (PI3K)/Akt. The specific PI3K inhibitor, wortmannin, blocked significantly EGF-induced cell migration and invasion. Furthermore, the EGFR inhibitor AG1478 inhibited EGF-induced MMP-9 expression, cell migration and invasion, as well as the activation of PI3K/Akt, suggesting that PI3K/Akt activation occur downstream of EGFR activation. These findings suggest that NFD inhibited the EGF-induced invasion and migration of MDA-MB-231 cells via EGFR-dependent PI3K/Akt signaling, leading to the down-regulation of MMP-9 expression. These results provide a novel mechanism to explain the role of NFD as a potent anti-metastatic agent in MDA-MB-231 cells. [Copyright &y& Elsevier]

Abstract: Background: In 1979, approximately 2,000 people in central Taiwan were accidentally exposed to polychlorinated biphenyls and dibenzofurans due to ingestion of contaminated cooking oil. This event was called Yucheng, “oil-syndrome” in Chinese. We followed the exposed persons and compared their cause-specific mortality with that of neighborhood referents 30 years after the accident. Methods: We obtained age- and gender-matched referents from the 1979 neighborhoods of the exposed people. Cause-specific mortality was compared between exposed subjects (N=1803) and their neighborhood referents (N=5170) using standardized mortality ratios (SMR). Total person-years for the Yucheng subjects and neighborhood referents were 48,751 and 141,774, respectively. Results: The SMR for all causes (SMR=1.2, 95% CI: 1.1–1.3), diseases of the circulatory system (SMR=1.3, 95% CI: 1.0–1.6), and diseases of the musculoskeletal system and connective tissue (SMR=6.4, 95% CI: 2.8–12.7) were elevated in Yucheng subjects. Among Yucheng males, the SMRs for diseases of the digestive system (SMR=1.9, 95% CI: 1.2–2.8), malignant neoplasm of stomach (SMR=3.5, 95% CI: 1.5–7.0), and malignant neoplasm of lymphatic and hematopoietic tissue (SMR=3.0, 95% CI: 1.1–6.6) were increased. The SMR for total neoplasms was increased (SMR=1.3, 95% CI: 0.9–1.7). Conclusion: We conclude that exposure to PCBs/PCDFs at levels that produced symptoms in many affects mortality patterns 3 decades after exposure. [Copyright &y& Elsevier]

Abstract: Cardiotoxin III (CTX III), a basic polypeptide isolated from Naja naja atra venom, has been shown to exhibit anticancer activity. Epidermal growth factor (EGF) and its receptor, EGFR, play roles in cancer metastasis in various tumors. We use EGF as a metastatic inducer of MDA-MB-231 cells to investigate the effect of CTX III on cell migration. CTX III inhibited the EGF-induced activation of matrix metalloproteinase-9 (MMP-9), and further suppressed cell invasion and migration without obvious cellular cytotoxicity. CTX III suppressed EGF-induced nuclear factor-kappaB (NF-κB) nuclear translocation and also abrogated the EGF-induced phosphorylation of EGFR, phosphatidylinositol 3-kinase (PI3K)/Akt, and extracellular regulated kinase (ERK)1/2. In addition, CTX III similar to wortmannin (a PI3K inhibitor) and U0126 (an up-stream kinase regulating ERK1/2 inhibitor) attenuated cell migration and invasion induced by EGF. Furthermore, the EGFR inhibitor AG1478 inhibited EGF-induced MMP-9 expression, cell migration and invasion, as well as the activation of ERK1/2 and PI3K/Akt, suggesting that ERK1/2 and PI3K/Akt activation occur downstream of EGFR activation. These findings suggest that CTX III inhibited the EGF-induced invasion and migration of MDA-MB-231 cells via EGFR-dependent PI3K/Akt, ERK1/2, and NF-κB signaling, leading to the down-regulation of MMP-9 expression. These results provide a novel mechanism to explain the role of CTX III as a potent anti-metastatic agent in MDA-MB-231 cells. [Copyright &y& Elsevier]

Abstract: Cancer metastasis is a primary cause of cancer death. Ovatodiolide, a bioactive cembrane-type diterpenoid isolated from Anisomeles indica (L.) Kuntze (Labiatae), has been shown to inhibit the growth and proliferation of cancer cells. However, the anti-metastatic effects of ovatodiolide on highly metastatic human breast cancer MDA-MB-231 cells remain unclear. In this study, we first noted that ovatodiolide inhibited MDA-MB-231 cell migration and invasion by wound-healing assay and Boyden chamber assay. Western blot, gelatin zymography and reversed transcription-PCR analysis showed that ovatodiolide significantly and selectively suppressed the expression, activation, and mRNA of matrix metalloproteinase-9 (MMP-9) in a concentration-dependent manner. Ovatodiolide significantly decreased the nuclear level of nuclear factor kappaB (NF-κB), increased inhibitor of kappaBα (IκBα) through preventing phosphorylation of upstream signal IκB kinase (IKK). Pretreatment with a specific NF-κB inhibitor (PDTC) and an IκB protease inhibitor (TPCK) also reduced MMP-9 activity, cell migration and cell invasion. Moreover, ovatodiolide can suppress activation of c-Jun N-terminal kinase, p38 mitogen-activated protein kinase, phosphatidylinositol 3-kinase and Akt, while it did not affect phosphorylation of extracellular signal regulating kinases (ERK)1/2. Additionally, the treatment of inhibitors specific for PI3K (wortmannin), JNK (SP600125) or p38 MAPK (SB203580) to MDA-MB-231 cells could result in a reduced activation of MMP-9, concomitantly with a marked inhibition on cell migration and invasion. Taken together, these results demonstrate that ovatodiolide inhibits the metastatic ability of MDA-MB-231 cells by reducing MMP-9 activity through suppressing JNK, p38 MAPK and PI3K/Akt signaling pathways and inhibiting NF-κB activity. These results are the first to reveal the function of ovatodiolide in tumor metastasis and its underlying molecular mechanism, thus suggesting ovatodiolide to be a promising antimetastatic agent. [Copyright &y& Elsevier]

Abstract: Polychlorinated biphenyls (PCBs) and polychlorinated dibenzofurans (PCDFs) are ubiquitous pollutants found in the environment and human tissues. A cohort in Taiwan has undergone follow-up for 24 years after exposure to high levels of PCBs and PCDFs. The incidence of chloracne, hyperkeratosis, and abnormal nail was increased among exposed people. We conducted a study to identify the genes whose expressions were affected by such exposure. A cDNA microarray system consisting of 908 genes was used for pooled serum samples from non-smoking men exposed to PCBs and PCDFs (n=15) and their matched referents (n=15) in triplicate. After adjusting for background and housekeeping genes, genes with different expressions between the exposure and reference groups were determined by both regression and cluster analysis, and further confirmed by real-time RT-PCR. The tumor suppressor gene von Hippel–Lindau (VHL) was found to be down-regulated in the microarray analysis. VHL gene expression levels were also found to be positively associated with age, shown by real-time RT-PCR. Upon age adjustment, VHL gene expression was reduced in Yucheng (“oil disease”) subjects as compared to referents. Among Yucheng people, those with abnormal nails had lower VHL expressions than those without abnormal nails. These findings provide new insights into the potential role of VHL in health conditions associated with PCB and PCDF exposures. [Copyright &y& Elsevier]

Heavy metal ions (HMIs) pollution is always a serious issue worldwide. Therefore, monitoring HMIs in environmental water is an important and challenging step to ensure environmental health and human safety. In this study, we spotlight an effortless, single-step in-situ electrochemical polymerization deposition technique to fabricate a novel, low-cost, efficient, nano-engineered poly(melamine)/graphitic-carbon nitride nanonetwork (PM/g-C 3 N 4) modified screen-printed carbon electrode (SPE) for sensitive, selective, and simultaneous electrochemical monitoring of toxic HMIs in environmental waters. g-C 3 N 4 nanomaterial was prepared using melamine as a precursor via pyrolysis technique. As-prepared g-C 3 N 4 and melamine monomer were electrochemically in-situ polymerized/deposited over pre-anodized SPE (ASPE) using cyclic voltammetry technique. XRD, XPS, and SEM were engaged to characterize the developed electrode. The fabricated PM/g-C 3 N 4 /ASPE was applied as an environmental sensor to selective and simultaneous electrochemical detection of Pb2+ and Cd2+ ions using differential pulse voltammetry technique. The developed sensor displayed excellent selectivity and sensitivity towards Pb2+ and Cd2+ with limit of detections of 0.008 µM and 0.02 µM, respectively. The fabricated PM/g-C 3 N 4 /ASPE sensor exhibits superior stability, repeatability, good anti-interference, and applicability for recognition of Pb2+ and Cd2+ ions in real water samples. These results proved that developed environmental sensor is low-cost, efficient, practical platform for rapid, selective, simultaneous monitoring of HMIs in the environment. [Display omitted] • New nano-engineered poly(melamine)/graphitic-carbon nitride based nanohybrid electrode. • Applied as environmental sensor for sensitive and simultaneous monitoring of heavy metals. • The developed sensor is low-cost and efficient for real-time analysis and monitoring. [ABSTRACT FROM AUTHOR]

In this work, a new semi-automated syringe infusion-pump assisted graphene nanosheets (GNSs) based pipette-tip micro-solid phase extraction (PT-μSPE) as a green sample preparation technique was demonstrated for the sensitive analysis of emerging environmental pollutant in environmental waters using HPLC-UV. Microwave-assisted synthesized GNSs powder was packed into a 100 μL pipette-tip (as PT-μSPE cartridge) connected with a commercial plastic syringe (contains water sample). This setup was attached to a programmable auto-syringe infusion pump for the GNSs-PT-μSPE process. Triclosan (TCS) is an emerging environmental pollutant chosen as a target analyte to examine the extraction capacity and feasibility of GNSs as a sorbent material for PT-μSPE. Parameters affecting the extraction capability were systematically evaluated and thoroughly optimized. At optimized experimental parameters, excellent linearity (r2 = 0.9979) was achieved over the concentration range of 2–250 ng mL−1 for TCS, with a detection limit of 0.5 ng mL−1. Applicability of the presented method was examined with real water samples, and extraction recoveries obtained were ranged between 94.6–102.4% with RSD less than 7.8%. The presented protocol is a simple, semi-automated, eco-friendly, low-cost, and efficient sample pretreatment technique for quick analysis of TCS in environmental wastewaters. ✓ New semi-automated graphene based pipette-tip assisted micro-solid phase extraction. ✓ This eco-friendly technique was applied for rapid analysis of triclosan by HPLC-UV. ✓ This method is easy, low-cost, efficient, and sensitive technique for water analysis. ✓ This method can be applied to real-time analysis of emerging pollutants in waters. [ABSTRACT FROM AUTHOR]

OBJECTIVE -- Polychlorinated biphenyls (PCBs) and polychlorinated dibenzofurans (PCDFs) are important and persistent organic pollutants (POPs) in humans. Recent cross-sectional studies have detected increased concentrations of serum POPs in diabetic patients. We aimed to examine the association between previous high exposures to PCBs and PCDFs and the cumulative incidence of type 2 diabetes and hypertension. RESEARCH DESIGN AND METHODS -- During the late 1970s, the consumption of rice-bran oil laced with PCBs poisoned thousands of Taiwanese. Between 1993 and 2003, we examined 1,054 Yucheng ("oil disease") victims against neighborhood reference subjects using a protocol blinded for POP exposure. Here, we report the results derived from 378 Yucheng subjects and 370 matched references. RESULTS -- The diabetes risk to members of the Yucheng cohort relative to their reference subjects was significantly increased for women (odds ratio [OR] 2.1 [95% CI 1.1-4.5]) but not for men after considering age, BMI, cigarette smoking, and alcohol intake. Yucheng women diagnosed with chloracne had adjusted ORs of 5.5 (95% CI 2.3-13.4) for diabetes and 3.5 (1.7-7.2) for hypertension compared with those who were chloracne free. CONCLUSIONS -- Yucheng women, who had endured previous exposure to PCBs and PCDFs, suffered from increased incidences of diabetes, particularly those who had retained significant levels of pollutant as evident from chloracne. When planning treatments against diabetes, the body burden of PCBs and dioxins should be carefully considered, especially for women. [ABSTRACT FROM AUTHOR]

A follow-up study was designed to compare the 24-year overall and disease-specific mortality in Yucheng people who were highly exposed to polychlorinated biphenyls/dibenzofurans (PCBs/PCDFs) to that in the background population in Taiwan. In 1979, the Yucheng (oil-disease in Chinese) incident occurred in central Taiwan involving approximately 2000 victims due to ingestion of rice oil contaminated with PCBs/PCDFs. Long-term follow-up of these people has been continued for 24 years. Methods: Standardized mortality ratios were calculated using the Taiwan population as comparison group. Overall and disease-specific mortality was compared between Yucheng and background populations. Results: Mortality from chronic liver disease and cirrhoses was increased in the Yucheng men, but not in women, in the early period after exposure. Cancer mortality was not increased in the Yucheng population up to 24 years after exposure. SLE in females was highly increased in the later period after PCB/PCDF exposure. Mortality from disease in any other organ system was not significantly different between Yucheng and background populations. Conclusions: The study provided a long-term mortality picture after the Taiwanese PCB/PCDF exposure incident. In addition to re-confirming the increase in liver mortality, we found high mortality of SLE among exposed population. This finding highlights the importance of further investigating the immunological effects associated with PCB/PCDF exposure. [Copyright &y& Elsevier]

Abstract: Introduction: Polychlorinated biphenyls (PCBs) and dibenzofurans (PCDFs) are ubiquitous persistent pollutants in humans. Whether people with different genotypes are with different susceptibility to these chemicals are unknown. In a group of people highly exposed to PCBs/PCDFs, we tested the hypothesis that genotypic polymorphisms affected susceptibility for development of skin manifestations. Methods: In 1979, approximately 2000 people in central Taiwan ingested cooking oil contaminated with PCBs/PCDFs. Skin disorder such as chloracne, abnormal nail, hyperkeratosis and skin allergy were found in PCBs/PCDFs exposed group. We recruited exposed and community background exposure subjects for blood testing and telephone-interview. Single nucleotide polymorphisms, AhR Arg554Lys, CYP1A1 Ile462Val, CYP1A1 T6235C, and GSTM1/T1 deletion, were determined. Occurrence of skin manifestations was compared among people with different genotypes while stratified by PCB exposure levels by logistic regression. Results: Data on exposure, medical history, and genotypes were obtained from 393 exposed and 181 background exposure groups. Skin manifestations including chloracne, allergy, abnormal nail, and hyperkeratosis were more prevalent in exposed people in a dose-related manner. Among highly exposed individuals, combined CYP1A1-MspI mutant genotype and GSTM1-null genotype were associated with increased risk of chloracne (odds ratio 2.8, 95% confidence interval 1.1–7.6). Among intermediately exposed individuals, GSTM1 null genotype was associated with skin allergy. AhR Arg554Lys genotype and GSTT1 null genotype were not related to susceptibility to skin manifestations in PCB/PCDF-exposed population. Conclusion: CYP1A1 and GSTM1 genotypic polymorphisms might be related to the susceptibility to PCB/PCDF-induced skin manifestations. [Copyright &y& Elsevier]

The GGGGCC hexanucleotide expansion in the C9ORF72 gene is the most common cause of familial amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) in Caucasian populations. The phenotypic spectrum of C9ORF72 hexanucleotide repeat expansion mutation has been reported to include parkinsonian syndrome, Huntington's disease-like syndrome and dementia syndrome. Although few individuals with cerebellar ataxia have also anecdotally been found to harbor the mutation, the relationship between the mutation and cerebellar ataxia awaits further clarification. We hereby screened for the presence of the C9ORF72 hexanucleotide repeat expansion in 331 patients with multiple system atrophy-cerebellar variant and 98 unrelated patients with molecularly un-assigned spinocerebellar ataxia in Taiwan utilizing a repeat-primed polymerase chain reaction assay. We found that none of the 429 patients had the C9ORF72 hexanucleotide repeat expansion mutation. Therefore, our study does not support that the mutation plays a significant role in cerebellar ataxia. [ABSTRACT FROM AUTHOR]

Objective Methods Results Interpretation TFG mutations have previously been implicated in autosomal recessive hereditary spastic paraplegia (HSP), also known as SPG57. This study aimed to investigate the clinical and molecular features of TFG mutations in a Taiwanese HSP cohort.Genetic analysis of TFG was conducted in 242 unrelated Taiwanese HSP patients using a targeted resequencing panel covering the entire coding regions of TFG. Functional assays were performed using an in vitro cell model to assess the impact of TFG variants on protein function. Additionally, other representative TFG mutant proteins were examined to understand the broader implications of TFG mutations in HSP.The study identified a novel homozygous TFG c.177A>C (p.(Lys59Asn)) variant in a family with adolescent‐onset, pure form HSP. Functional analysis revealed that the Lys59Asn TFG variant, similar to other HSP‐associated TFG mutants, exhibited a low affinity between TFG monomers and abnormal assembly of TFG homo‐oligomers. These structural alterations led to aberrant intracellular distribution, compromising TFG's protein secretion function and resulting in decreased cellular viability.These findings confirm that the homozygous TFG c.177A>C (p.(Lys59Asn)) variant is a novel cause of SPG57. The study expands our understanding of the clinical and mutational spectrum of TFG‐associated diseases, highlighting the functional defects associated with this specific TFG variant. Overall, this research contributes to the broader comprehension of the genetic and molecular mechanisms underlying HSP. [ABSTRACT FROM AUTHOR]

Parahydroxybenzoates (parabens) are considered as emerging environmental contaminants because of their extensive usage in our daily life products, causing parabens contamination into environmental water systems and lead to toxic effects on environmental health. This study describes a greener extraction method using a new cationic polymer poly (ethyleneimine) functionalized acid-treated carbon nanofibers (PEI-CNFs) coated cellulose paper (CP) as solid-phase extraction (SPE) sorbent material for the extraction of parabens from environmental water samples. The fabrication of PEI-CNFs modified CP was confirmed using field-emission scanning electron microscope, transmission electron microscopy, and fourier-transformer infrared spectroscopy techniques. Various factors affecting the adsorption and desorption of parabens on PEI-CNFs@CP and its extraction efficiencies were studied using HPLC-UV analysis. Under the optimal experimental conditions, maximum extraction efficiencies were achieved for four target parabens, and PEI-CNFs@CP/HPLC-UV method exhibited excellent linearities ranged from 0.5–50 ng mL−1 with regression coefficient values were between 0.9952–0.9970. The presented method showed good sensitivity with quantification limits between 0.5–0.75 ng mL−1 and detection limits between 0.1–0.25 ng mL−1. The developed technique was applied for the real sample analysis (river, lake, domestic sewage water, and drinking tap water). The spiked recovery revealed good recoveries between 86.8–116.0% with RSD less than 8.8% for all the water samples. These results proved that it a simple, fast, efficient, low-cost, and eco-friendly method for the extraction and determination of parabens in environmental water samples and can be applied as a routine analytical tool in environmental monitoring and quality control laboratories. Image 1 • Poly (ethyleneimine)-Functionalized Carbon Nanofibers Coated Cellulose Paper was developed. • Applied as Low-Cost Disposable Efficient Solid Phase Extraction Sorbent Material. • Maximum Extraction Recovery of Parahydroxybenzoates were achieved from Environmental Waters. • Eco-friendly method can be employed as a routine method in wastewater treatment. [ABSTRACT FROM AUTHOR]

Fabrication of g-C 3 N 4 /Fe 2 O 3 /PdNPs/SPE nanocomposite modified electrode by electrochemical deposition method for the ethylene glycol oxidation. • Simple one-step fabrication of novel nanocomposite by electrodeposition technique. • Graphitic carbon nitride/iron oxide/palladium nanocomposite modified electrode. • Nanocomposite exhibited a three-dimensional nanoflower like structure. • Excellent catalytic activity over ethylene glycol oxidation compared with Pd/C catalyst. • Developed electrocatalyst is highly suitable for direct alcohol fuel cell applications. The design and development of highly efficient and environmental-friendly methods for the synthesis of electro-catalysts are highly significant for fuel cell applications. Herein, we demonstrate a facile technique for the synthesis of graphitic carbon nitride/iron oxide/palladium nanoparticles (g-C 3 N 4 /Fe 2 O 3 /PdNPs) composite towards efficient electro-catalytic oxidation of ethylene glycol. g-C 3 N 4 /Fe 2 O 3 /PdNPs nanocomposite was prepared using a one-step simultaneous electrochemical co-deposition process. The electro-deposition was performed on a single-disk carbon screen-printed electrode (SPE) under acidic electrolyte solution containing g-C 3 N 4 nanosheets, Fe 2 O 3 nanoparticles, and palladium chloride as precursors. The structural and elemental composition of the electrochemically fabricated g-C 3 N 4 /Fe 2 O 3 /PdNPs nanocomposites modified SPE was investigated using fourier-transform infrared spectroscopy, field-emission scanning electron microscopy, and X-ray photoelectron spectroscopy techniques and the results indicated that three-dimensional nanoflower-like Fe 2 O 3 and spherical shaped PdNPs were electro-deposited on the g-C 3 N 4 nanosheets to form the nanocomposite. The voltammetric behavior of the developed g-C 3 N 4 /Fe 2 O 3 /PdNPs/SPE showed excellent electro-catalytic response towards ethylene glycol and superior electro-oxidation with high durability when compared to commercial Pd/C and other recently reported electro-catalyst materials. Therefore, the proposed g-C 3 N 4 /Fe 2 O 3 /PdNPs nanocomposite is proved to be an efficient electro-catalyst and can be highly suitable for direct alcohol fuel cell applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Cost-effective, Hepatocellular carcinoma (HCC) risk-based surveillance strategies should be established after achieving sustained virologic response (SVR). Circulating microRNAs are considered stable serum markers for early cancer diagnosis and prognosis and treatment response prediction. The aim of our longitudinal follow-up study was to assess whether Let-7 family members can predict HCC risk in CHC patients. We assessed the sera of 54 patients with CHC who developed HCC and 173 patients with CHC who did not develop HCC after antiviral therapy. Cox's regression model revealed the independent role of let-7i as an effective surrogate biomarker for predicting HCC development in patients with CHC. HCC, a leading cause of cancer-related mortality, is diagnosed at advanced stages. Although antiviral therapy has reduced the risk of HCC among chronic hepatitis C (CHC) patients, the risk of HCC remains, thus, highlighting the unmet need for continuous surveillance. Therefore, stable and cost-effective biomarkers, such as circulating microRNAs, must be identified. We aimed to clarify whether serum levels of the Let-7 family can predict HCC risk in patients with CHC using univariate and multivariate Cox's proportional hazards model. We analyzed the sera of 54 patients with CHC who developed HCC after antiviral therapy and compared the data with those of 173 patients without HCC development. The Let-7 family (except for let-7c) exhibited significant negative correlations with the fibrosis score (r = −0.2736 to −0.34, p = 0.0002 to <0.0001). After Cox's regression model was used to adjust for age, sex, HCV genotype, and FIB-4 ≥ 3.25, patients with CHC with let-7i median ≥ −1.696 (adjusted hazard ratio [aHR] = 0.31, 95% CI: 0.08–0.94, p = 0.0372) in the sustained virologic response (SVR) groups and ≥−1.696 (aHR = 0.09, 95% CI: 0.08–0.94, p = 0.0022) in the non-SVR group were less likely to develop HCC. Thus, circulating let-7i can be used for early CHC surveillance in patients with HCC risk after antiviral treatment. [ABSTRACT FROM AUTHOR]

The facile fabrication is reported of highly electrochemically active Ti3C2Tx MXene/MWCNT (3D/1D)-modified screen-printed carbon electrode (SPE) for the efficient simultaneous electrochemical detection of paracetamol, theophylline, and caffeine in human blood samples. 3D/1D Ti3C2Tx MXene/MWCNT nanocomposite was synthesized using microwave irradiation and ultrasonication processes. Then, the Ti3C2Tx/MWCNT-modified SPE electrode was fabricated and thoroughly characterized towards its physicochemical and electrochemical properties using XPS, TEM, FESEM, XRD, electrochemical impedance spectroscopy, cyclic voltammetry, and differential pulse voltammetry techniques. As-constructed Ti3C2Tx-MWCNT/SPE offers excellent electrochemical sensing performance with good detection limits (0.23, 0.57, and 0.43 µM) and wide linear ranges (1.0 ~ 90.1, 2.0 ~ 62.0, and 2.0–90.9 µM) for paracetamol, caffeine, and theophylline, respectively, in the human samples. Notably, the non-enzymatic electroactive nanocomposite-modified electrode has depicted a semicircle Nyquist plot with low charge transfer resistance (Rct∼95 Ω), leading to high ionic diffusion and facilitating an excellent electron transfer path. All the above results in efficient stability, reproducibility, repeatability, and sensitivity compared with other reported works, and thus, it claims its practical utilization in realistic clinical applications. [ABSTRACT FROM AUTHOR]

Background and Aim: Chronic hepatitis C virus (HCV) infection is associated with impaired renal function. The aim of this study is to explore the risk of and factors associated with end‐stage renal diseases (ESRD) under maintenance dialysis among HCV patients after anti‐HCV therapy. Methods: A total of 12 696 HCV‐infected patients with interferon‐based therapy, including 9679 (76.2%) achieving sustained virological response (SVR), were enrolled from 23 hospitals in Taiwan. Results: During a mean follow‐up period of 5.3 years (67 554 person‐years), the annual incidence of 4.1/10 000 person‐years, 4.0/10 000 and 4.7/10 000 person‐years among SVR patients and non‐SVR patients, respectively. History of diabetes and baseline estimated glomerular filtration rate < 60 mL/min/m2, instead of SVR, were the significant risk factors for developing ESRD with maintenance dialysis after anti‐HCV therapy (adjusted hazard ratio 7.75 and 9.78). Conclusion: Diabetes and baseline impaired renal function were strongly associated with progression to ESRD with maintenance dialysis among chronic HCV‐infected patients after antiviral therapy. [ABSTRACT FROM AUTHOR]

Intermediate-length CAG repeats in ATXN2 have been well recognized as a genetic risk factor for amyotrophic lateral sclerosis (ALS). However, the role of similar trinucleotide repeat expansions in the TATA-box binding protein gene (TBP), another disease-associated gene for inherited ataxia, in ALS remains elusive. To assess the association between TBP trinucleotide repeat expansions and ALS, we investigated the repeat lengths in 325 unrelated ALS patients and 1500 controls in the Taiwanese population. The most common size of repeats in the patients and controls were both 36. The repeat lengths ranged from 29 to 46 repeats in the ALS patients and 27 to 43 repeats in the controls. Two ALS patients carried a TBP allele with a repeat number equal or greater than 44 (44 and 46). The patient with the 46 trinucleotide repeats also had a C9ORF72 GGGGCC hexanucleotide repeat expansion. The odds ratio of an individual carrying the CAG/CAA repeats ≥ 44 to have ALS is 23.2 (95% confidence interval: 1.11–484.24; p = 0.04). Our findings suggest that intermediate-length CAG/CAA repeat expansions in TBP may associate with ALS risk. [ABSTRACT FROM AUTHOR]

Background: Genetic and environmental risk factors contribute to periodontal disease, but the underlying susceptibility pathways are not fully understood. Epigenetic mechanisms are malleable regulators of gene function that can change in response to genetic and environmental stimuli, thereby providing a potential mechanism for mediating risk effects in periodontitis. The aim of this study is to identify epigenetic changes across tissues that are associated with periodontal disease. Methods: Self-reported gingival bleeding and history of gum disease, or tooth mobility, were used as indicators of periodontal disease. DNA methylation profiles were generated using the Infinium HumanMethylation450 BeadChip in whole blood, buccal, and adipose tissue samples from predominantly older female twins (mean age 58) from the TwinsUK cohort. Epigenome-wide association scans (EWAS) of gingival bleeding and tooth mobility were conducted in whole blood in 528 and 492 twins, respectively. Subsequently, targeted candidate gene analysis at 28 genomic regions was carried out testing for phenotype-methylation associations in 41 (tooth mobility) and 43 (gingival bleeding) buccal, and 501 (tooth mobility) and 556 (gingival bleeding) adipose DNA samples. Results: Epigenome-wide analyses in blood identified one CpG-site (cg21245277 in ZNF804A) associated with gingival bleeding (FDR = 0.03, nominal p value = 7.17e−8) and 58 sites associated with tooth mobility (FDR < 0.05) with the top signals in IQCE and XKR6. Epigenetic variation at 28 candidate regions (247 CpG-sites) for chronic periodontitis showed an enrichment for association with periodontal traits, and signals in eight genes (VDR, IL6ST, TMCO6, IL1RN, CD44, IL1B, WHAMM, and CXCL1) were significant in both traits. The methylation-phenotype association signals validated in buccal samples, and a subset (25%) also validated in adipose tissue. Conclusions: Epigenome-wide analyses in adult female twins identified specific DNA methylation changes linked to self-reported periodontal disease. Future work will explore the environmental basis and functional impact of these results to infer potential for strategic personalized treatments and prevention of chronic periodontitis. [ABSTRACT FROM AUTHOR]

Abstract Mutations in the annexin A11 gene (ANXA11) have been recently identified in British patients and Italian patients with amyotrophic lateral sclerosis (ALS), and their role in other ALS populations remains unclear. The aim of this study was to investigate the ANXA11 mutations in a Taiwanese ALS cohort. Mutational analysis of ANXA11 was performed in 286 unrelated Taiwanese patients with ALS by Sanger sequencing. Eight ANXA11 missense variants were identified initially, and only one of them was absent from population databases. This missense variant, p.Q362L, was identified in 1 single patient with apparently sporadic ALS, and no further strong evidence was available to support its pathogenicity. Therefore, it is classified as a variant of uncertain significance. Our data indicate that pathogenic ANXA11 mutations are absent or rare in ALS patients in Taiwan. [ABSTRACT FROM AUTHOR]

Background: Tobacco smoking is a risk factor for multiple diseases, including cardiovascular disease and diabetes. Many smoking-associated signals have been detected in the blood methylome, but the extent to which these changes are widespread to metabolically relevant tissues, and impact gene expression or metabolic health, remains unclear. Methods: We investigated smoking-associated DNA methylation and gene expression variation in adipose tissue biopsies from 542 healthy female twins. Replication, tissue specificity, and longitudinal stability of the smoking-associated effects were explored in additional adipose, blood, skin, and lung samples. We characterized the impact of adipose tissue smoking methylation and expression signals on metabolic disease risk phenotypes, including visceral fat. Results: We identified 42 smoking-methylation and 42 smoking-expression signals, where five genes (AHRR, CYP1A1, CYP1B1, CYTL1, F2RL3) were both hypo-methylated and upregulated in current smokers. CYP1A1 gene expression achieved 95% prediction performance of current smoking status. We validated and replicated a proportion of the signals in additional primary tissue samples, identifying tissue-shared effects. Smoking leaves systemic imprints on DNA methylation after smoking cessation, with stronger but shorter-lived effects on gene expression. Metabolic disease risk traits such as visceral fat and android-to-gynoid ratio showed association with methylation at smoking markers with functional impacts on expression, such as CYP1A1, and at tissue-shared smoking signals, such as NOTCH1. At smoking-signals, BHLHE40 and AHRR DNA methylation and gene expression levels in current smokers were predictive of future gain in visceral fat upon smoking cessation. Conclusions: Our results provide the first comprehensive characterization of coordinated DNA methylation and gene expression markers of smoking in adipose tissue. The findings relate to human metabolic health and give insights into understanding the widespread health consequence of smoking outside of the lung. [ABSTRACT FROM AUTHOR]

Background The World Health Organization has established interim guidance for hepatitis C virus (HCV) elimination. We aimed to prove the concept of "treatment as prevention" by conducting a prospective HCV elimination program for hemodialysis (HD) patients. Methods A universal HCV screen was launched in 22 HD centers in 2019. HCV-viremic patients were linked to care with direct-acting antivirals (DAAs). The second screen was performed in 2021 to evaluate the effect of link-to-care in lowering the prevalence of HCV viremia and the incidence of HCV new/re-infections. Results Of 2336 patients enrolled in the first screening in 2019, 320 (13.7%) were seropositive for anti-HCV and 181 (7.7%) were HCV-viremic. Of 152 patients successfully linked to treat with DAA, 140 (92.1%) patients achieved a sustained virological response. Of them, 1733 patients participated in the second surveillance. Five anti-HCV-negative patients experienced anti-HCV seroconversion. Of 119 DAA-cured patients and 102 spontaneous HCV clearance patients, none had HCV reinfection. The annual incidence of HCV new infection was 0.1%. Sixty-one of the 620 (9.8%) newly enrolled patients were anti-HCV-seropositive in the second survey. The overall HCV-viremic rate decreased from 7.7% in 2019 to 0.6% (15/2353) in 2021. At the institutional level, 45.5% (10/22) eradicated HCV and 82% (18/22) of HD units had no HCV new infections or reinfections. Conclusions The link-to-care project proved the concept of "treatment as prevention" by which HCV microelimination helps to prevent reinfection and new infections in the HD population. Trial registration: ClinicalTrials.gov identifier: NCT03803410 and NCT03891550. [ABSTRACT FROM AUTHOR]

Mutations in the cyclin F gene ( CCNF ) have been recently identified in a small number of patients with amyotrophic lateral sclerosis (ALS) and/or frontotemporal dementia, and their role in patients with ALS in Taiwan remains elusive. The aim of this study was to elucidate the frequency and spectrum of CCNF mutations in a Taiwanese ALS cohort of Han Chinese origin. Mutational analyses of the CCNF gene were performed using Sanger sequencing in a cohort of 255 unrelated patients with ALS. Among these patients, the genetic diagnoses of 204 patients remained unclear after mutations in SOD1 , C9ORF72 , TARDBP , FUS , ATXN2 , OPTN , VCP , UBQLN2 , SQSTM1 , PFN1 , HNRNPA1 , HNRNPA2B1 , MATR3 , CHCHD10 , TUBA4A , and TKB1 had been investigated. Two novel heterozygous missense mutations in CCNF , p.S222P (c.664T>C) and p.S532R (c.1596C>T), were identified; 1 in each patient with apparently sporadic ALS. In vitro functional study demonstrated that both mutations result in a general and cyclin F-mediated ubiquitin-proteasome pathway dysfunction. The frequency of CCNF mutations in ALS patients in Taiwan is, therefore, approximately 0.8% (2/255). These findings expand the mutational spectrum of CCNF and also emphasize the pathogenic role of CCNF mutations in ALS. [ABSTRACT FROM AUTHOR]

The GGC repeat expansion in the 5′-untranslated region of NOTCH2NLC was recently identified as the cause of neuronal intranuclear inclusion disease (NIID). To determine if the NIID repeat expansion contributes to amyotrophic lateral sclerosis (ALS), we screened 304 unrelated ALS patients and 637 healthy controls for the GGC repeat expansion in NOTCH2NLC using repeat-primed PCR and fragment analysis. None of the ALS patients carried the GGC repeat expansion. The sizes of the GGC repeats ranged from 7 to 36 in the ALS patients and 4 to 46 in the controls. The distribution of the GGC repeat sizes did not differ between the two groups. Our findings indicate that the NOTCH2NLC GGC repeat expansion is absent or extremely rare in Taiwanese patients with ALS. [ABSTRACT FROM AUTHOR]

• Review reports on physiological and biochemical reaction to N-stress in microalgae. • Lipid and carbohydrate increased and protein decreased in N-stressed cells. • During nitrogen deprivation, radicals are overproduced creating oxidative stress. • The increase in cellular oxidative stress causes the accumulation of lipids. Microalgae have higher productivity of biomass than the conventional crops of fuel and are therefore, considered a potential biofuel source. Lipid, an important precursor of biodiesel, can be overproduced in microalgae by nitrogen deprivation. During nitrogen deficiency, radicals are overproduced, and the antioxidant levels are insufficient to counteract the radicals. Thus, the increase in cellular oxidative stress level, consequently acts as a stimulus for lipid accumulation. Lipid accumulation requires an excess of acetyl CoA and NADPH that is made possible by the following mechanism. Glycolysis upregulation overproduces pyruvate, which could be further transformed into acetyl CoA by the pyruvate dehydrogenase complex; while the upregulation of the oxidative pentose phosphate cycle generates a high amount of NADPH. In addition to lipid overproduction, the lack of nitrogen often causes the accumulation of carbohydrates in selected species of microalgae, which could be used to generate biogas and bioethanol from the defatted biomass. By providing details on the differential regulation of the biochemical pathways leading to lipid and carbohydrate accumulation in nitrogen starved microalgae, the review opens up new possibilities in the microalgal biofuel production. [ABSTRACT FROM AUTHOR]

Microalgae are a group of microorganisms considered to be a potential source of next-generation biofuel owing to high photosynthetic rate. However, biofuel compounds including lipid, carbohydrate, and hydrogen are not sufficiently produced by microalgae under normal conditions; therefore economical viability of microalgal biofuel production has not reached the ultimate stage of commercial viability. In order to overcome the above limitation, various stress conditions have been applied on microalgae to boost lipid, carbohydrate and hydrogen production. Even though stress conditions increases the production of biofuel compounds, microalgae growth is severely affected. In this regard, a two-stage cultivation mode wherein first stage involving high biomass production under media optimized heterotrophic condition and second stage involving lipid, carbohydrate and hydrogen accumulation under stress integrated phototrophic mode is considered as a potential route for biofuel production in microalgae. The present review critically evaluates various two-stage strategies adopted in microalgal biofuel production by comparing lipid, carbohydrate and hydrogen productivities with that of single stage cultivation. In addition, advantages of two-stage cultivation including co-product generation, less bacterial contamination, improved biodiesel quality, auto-flocculation, and bioremediation potential are also discussed. Finally, the paper summarizes the two-stage cultivation modes by identifying the bottlenecks and suggesting the future potentialities of the process. [ABSTRACT FROM AUTHOR]

• Passive lipid extraction techniques that skip conventional cell lysis step for microalgal biofuel production. • Energy consumption by passive methods are compared with physical methods. • In-situ transesterification, saponification-acidification, and switchable solvents have low energy consumption. • Passive methods suitable for lipid-based biorefinery is presented. The conventional method of lipid extraction mandatorily includes cell lysis which is considered an essential process for the enhancement of extraction efficiency. However, cell lysis method especially those incorporating mechanical and physical methods such as homogenization, sonication, microwave techniques, etc., are energy intensive and drastically escalates the biofuel production cost. Therefore, the development of an alternate route of extraction skipping the conventional mechanical cell lysis step remains a challenge to be accomplished. In this context, we discuss the reports involving passive techniques that are able to extract lipid from microalgae without significant cell wall shearing. This includes methods such as in-situ transesterification, direct saponification, supercritical fluid extraction, organic solvent extraction, etc. Moreover, the review discusses the employment of passive lipid extraction methods in a biorefinery set-up, outlining the various bioproducts that could be generated along with lipid. The review concludes with the analysis of the economics of microalgal lipid extraction using passive disruption methods and compares with the processes incorporating cell lysis steps. [ABSTRACT FROM AUTHOR]