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In this work, we demonstrate the bioavailability of dietary sialyllactose by concentration–time profiles in an osteoarthritis mini-pig model and suggest a potential therapeutic effect of sialyllactose on osteoarthritis.

Extracellular signal-regulated kinase 3 (ERK3) is an atypical member of the mitogen-activated protein kinase (MAPK) family, members of which play essential roles in diverse cellular processes during carcinogenesis, including cell proliferation, differentiation, migration, and invasion. Unlike other MAPKs, ERK3 is an unstable protein with a short half-life. Although deubiquitination of ERK3 has been suggested to regulate the activity, its ubiquitination has not been described in the literature. Here, we report that FBXW7 (F-box and WD repeat domain-containing 7) acts as a ubiquitination E3 ligase for ERK3. Mammalian two-hybrid assay and immunoprecipitation results demonstrated that ERK3 is a novel binding partner of FBXW7. Furthermore, complex formation between ERK3 and the S-phase kinase-associated protein 1 (SKP1)-cullin 1-F-box protein (SCF) E3 ligase resulted in the destabilization of ERK3 via a ubiquitination-mediated proteasomal degradation pathway, and FBXW7 depletion restored ERK3 protein levels by inhibiting this ubiquitination. The interaction between ERK3 and FBXW7 was driven by binding between the C34D of ERK3, especially at Thr417 and Thr421, and the WD40 domain of FBXW7. A double mutant of ERK3 (Thr417 and Thr421 to alanine) abrogated FBXW7-mediated ubiquitination. Importantly, ERK3 knockdown inhibited the proliferation of lung cancer cells by regulating the G1/S-phase transition of the cell cycle. These results show that FBXW7-mediated ERK3 destabilization suppresses lung cancer cell proliferation in vitro.

Resolution to chemoresistance is a major challenge in patients with advanced-stage malignancies. Thus, identification of action points and elucidation of molecular mechanisms for chemoresist human cancer are necessary to overcome this challenge. In this study, we provide important evidence that kaempferol targeting RSKs might be a strategy to reduce the oxaliplatin-resistant colon cancer cells. We found that MAPK and PI3K-AKT signaling were increased in oxaliplatin (Ox)-resistant HCT116 (HCT116-OxR) cells compared to Ox-sensitive HCT116 (HCT116-OxS) cells. Comparison of cell sensitivities using SP600125 (JNK inhibitor), SB206580 (p38 kinase inhibitor), or MK-2206 (AKT inhibitor) revealed that cell proliferation inhibition was strongly observed in HT29 cells compared to that in HCT116 cells in both OxS and OxR cells. Interestingly, SP600125, SB206580, and MK-2206 treatment showed higher cell proliferation inhibition in OxS cells than that in OxR cells in both HCT116 and HT29 cells, except following treatments with 10 µM of SP600125, and 30 µM of SB206580. In comparison to magnolin and aschantin, kaempferol showed the strongest inhibitory effect on cell proliferation in both HCT116 and HT29 cells. Importantly, HCT116- and HT29-OxR cells showed higher sensitivities to cell proliferation inhibition than those of HCT116- and HT29-OxS cells, resulting in the accumulation of cells at the G2/M-phases of the cell cycle. Finally, we showed that AP-1 transactivation activity was markedly decreased by kaempferol in HCT116- and HT29-OxR cells compared to the activity levels in HCT116- and HT29-OxS cells. Taken together, the results demonstrate that kaempferol-mediated AP-1 inhibition might be an important signaling mechanism to resolve the chemoresistance of Ox-resistant colon cancer cells.

Dasatinib is an inhibitor of Src that has anti‐tumour effects on many haematological and solid cancers. However, the anti‐tumour effects of dasatinib on human oral cancers remain unclear. In this study, we investigated the effects of dasatinib on different types of human oral cancer cells: the non‐tumorigenic YD‐8 and YD‐38 and the tumorigenic YD‐10B and HSC‐3 cells. Strikingly, dasatinib at 10 µM strongly suppressed the growth and induced apoptosis of YD‐38 cells and inhibited the phosphorylation of Src, EGFR, STAT‐3, STAT‐5, PKB and ERK‐1/2. In contrast, knockdown of Src blocked the phosphorylation of EGFR, STAT‐5, PKB and ERK‐1/2, but not STAT‐3, in YD‐38 cells. Dasatinib induced activation of the intrinsic caspase pathway, which was inhibited by z‐VAD‐fmk, a pan‐caspase inhibitor. Dasatinib also decreased Mcl‐1 expression and S6 phosphorylation while increased GRP78 expression and eIF‐2α phosphorylation in YD‐38 cells. In addition, to its direct effects on YD‐38 cells, dasatinib also exhibited anti‐angiogenic properties. Dasatinib‐treated YD‐38 or HUVEC showed reduced HIF‐1α expression and stability. Dasatinib alone or conditioned media from dasatinib‐treated YD‐38 cells inhibited HUVEC tube formation on Matrigel without affecting HUVEC viability. Importantly, dasatinib's anti‐growth, anti‐angiogenic and pro‐apoptotic effects were additionally seen in tumorigenic HSC‐3 cells. Together, these results demonstrate that dasatinib has strong anti‐growth, anti‐angiogenic and pro‐apoptotic effects on human oral cancer cells, which are mediated through the regulation of multiple targets, including Src, EGFR, STAT‐3, STAT‐5, PKB, ERK‐1/2, S6, eIF‐2α, GRP78, caspase‐9/3, Mcl‐1 and HIF‐1α.

Extracellular matrix (ECM) proteins play a pivotal role in cell growth and differentiation. To characterize aged ECM proteins, we compared the proteomes by shotgun method of young (passage #15) and late senescent (passage #40) human dermal fibroblasts (HDFs) using SDS-PAGE coupled with LC–MS/MS. The relative abundance of identified proteins was determined using mol% of individual proteins as a semi-quantitative index. Fifteen ECM proteins including apolipoprotein B (APOB) and high-temperature requirement factor 1 (HTRA1) were up-regulated, whereas 50 proteins including fibronectin 1 (FN1) and vitronectin (VTN) were down-regulated in late senescent HDFs. The identified ECM proteins combined with plasma membrane were queried to construct the protein–protein interaction network using Ingenuity Pathways Analysis, resulting in a distinct FN1-centered network. Of differentially abundant ECM proteins in shotgun proteomics, the protein levels of FN1, VTN, APOB, and HTRA1 were verified by immunoblot analysis. The results suggest that the aging process in HDFs might be finally involved in the impaired FN1 regulatory ECM network combined with altered interaction of neighboring proteins. Shotgun proteomics of highly aged HDFs provides insight for further studies of late senescence-related alterations in ECM proteins.

A novel and facile post-mortem interval (PMI) biosensor was fabricated using a double-label strategy to detect the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) biomarker. A monoclonal anti-GAPDH antibody was immobilized on a surface label containing cadmium selenide quantum dots (CdSe QDs) on a cysteamine graphene oxide (Cys-GO) self-assembled monolayer. Glucose oxidase (GOx) was used as a signal label to conjugate with GAPDH. GAPDH recognition was achieved through the dissolution of the surface-attached CdSe QDs by hydrogen peroxide generated through GAPDH-conjugated GOx-catalyzed β-glucose oxidation. To enhance sensitivity, a competitive interaction was introduced between free and conjugated GAPDH to the active site of the anti-GAPDH antibody. The electrochemical response due to CdSe dissolution decreased proportionally with the concentration of free GAPDH. Differential pulsed voltammetry was conducted to determine the analytical characteristics of the immunosensor, including the limit of detection, linear dynamic range, target selectivity, system stability, and applicability toward the analysis of real samples.

The present study aimed to analyze the metaproteome of the microbial community comprising harmful algal bloom (HAB) in the Daechung reservoir, Korea. HAB samples located at GPS coordinates of 36°29’N latitude and 127°28’E longitude were harvested in October 2013. Microscopic observation of the HAB samples revealed red signals that were presumably caused by the autofluorescence of chlorophyll and phycocyanin in viable cyanobacteria. Metaproteomic analysis was performed by a gelbased shotgun proteomic method. Protein identification was conducted through a two-step analysis including a forward search strategy (FSS) (random search with the National Center for Biotechnology Information (NCBI), Cyanobase, and Phytozome), and a subsequent reverse search strategy (RSS) (additional Cyanobase search with a decoy database). The total number of proteins identified by the two-step analysis (FSS and RSS) was 1.8-fold higher than that by one-step analysis (FSS only). A total of 194 proteins were assigned to 12 cyanobacterial species (99 mol%) and one green algae species (1 mol%). Among the species identified, the toxic microcystin-producing Microcystis aeruginosa NIES-843 (62.3%) species was the most dominant. The largest functional category was proteins belonging to the energy category (39%), followed by metabolism (15%), and translation (12%). This study will be a good reference for monitoring ecological variations at the meta-protein level of aquatic microalgae for understanding HAB.

Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)+/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD+/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD+/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria. Keywords: Akt-mTOR-sirtuin signaling, Ginsenoside Rg3(S), Human dermal fibroblast, Reversal, senescence

The antigen-based rapid diagnostic test (Ag-RDT) using saliva specimens is fast, noninvasive, and suitable for SARS-CoV-2 self-testing, unlike nasopharyngeal swab (NPS) testing. We evaluated a novel Beanguard gargle (BG)-based virus collection method that can be applied to Ag-RDT as an alternative to the current RT-PCR with an NPS for early diagnosis of COVID-19. This clinical trial comprised 102 COVID-19-positive patients hospitalized after a governmental screening process and 100 healthy individuals. Paired NPS and BG-based saliva specimens from COVID-19 patients and healthy individuals were analyzed using NPS-RT-PCR, BG-RT-PCR, and BG-Ag-RDTs, whose diagnostic performance for detecting SARS-CoV-2 was compared. BG-Ag-RDTs showed high sensitivity (97.8%) and specificity (100%) in 45 patients within 6 days of illness and detected all cases of SARS-CoV-2 Alpha and Delta variants. In 11 asymptomatic active COVID-19 cases, both BG-Ag-RDTs and BG-RT-PCR showed sensitivities and specificities of 100%. Sensitivities of BG-Ag-RDT and BG-RT-PCR toward salivary viral detection were highly concordant, with no discrimination between symptomatic (97.0%), asymptomatic (100%), or SARS-CoV-2 variant (100%) cases. The intermolecular interactions between SARS-CoV-2 spike proteins and truncated canavalin, an active ingredient from the bean extract (BE), were observed in terms of physicochemical properties. The detachment of the SARS-CoV-2 receptor-binding domain from hACE2 increased as the BE concentration increased, allowing the release of the virus from hACE2 for early diagnosis. Using BG-based saliva specimens remarkably enhances the Ag-RDT diagnostic performance as an alternative to NPS and enables noninvasive, rapid, and accurate COVID-19 self-testing and mass screening, supporting efficient COVID-19 management.

ImportanceThe antigen-based rapid diagnostic test (Ag-RDT), using saliva specimens, is fast, non-invasive and suitable for SARS-CoV-2 self-testing, unlike nasopharyngeal swab (NPS) testing.ObjectiveTo assess the diagnostic sensitivity of a novel Beanguard gargle™ (BG)-based virus detection method for early diagnosis of COVID-19.DesignThis clinical trial was conducted at Gunsan Medical Center, Namwon Medical Center, and Jeonbuk National University Hospital, between May 7 and July 7, 2021.SettingPaired NPS and BG-based saliva specimens collected from COVID-19 patients and healthy individuals were analyzed using NPS-RT-PCR, BG-RT-PCR, and BG-Ag-RDTs.ParticipantsThe study comprised 102 COVID-19-positive patients hospitalized after governmental screening process and 100 healthy individuals. Forty-five COVID-19 patients were sampled within 6 days of illness and 57 within 7–15 days; 27 were categorized as asymptomatic and 75, as symptomatic. Eight and 2 patients carried the SARS-CoV-2 Alpha and Delta variants, respectively.InterventionThe diagnostic performances of BG-Ag-RDT, BG-RT-PCR, and NPS-RT-PCR for detecting SARS-CoV-2 were compared.Main outcomesThe sensitivities of BG-Ag-RDT and BG-RT-PCR towards salivary viral detection were highly concordant, with no discrimination between symptomatic, asymptomatic, or SARS-CoV-2 variant cases.ResultsAmong total participants (mean age, 43.7 years), 51% were women. BG-Ag-RDTs showed high sensitivity (97.8%, [95% CI, 88.4% to 99.6%]) and specificity (100%, [95% CI, 96.3% to 100%) in 45 patients within 6 days of illness and could detect all cases of SARS-CoV-2 Alpha and Delta variants. In 11 asymptomatic early-stage cases, both BG-Ag-RDTs and BG-RT-PCR showed excellent sensitivity and specificity of 100% (95% CI, 74.1% to 100% and 95% CI, 20.7% to 100%, respectively). The interaction between SARS-CoV-2 spike proteins and truncated canavalin, an active ingredient from bean extract (BE) and the ultrastructural features of SARS-CoV-2 particles coated with BE were observed. The detachment of the SARS-CoV-2 receptor-binding domain from hACE2 increased as the BE concentration increased, allowing the release of the virus from hACE2 for early diagnosis.Conclusions and RelevanceUsing BG-based saliva remarkably enhances the Ag-RDT diagnostic performance as an alternative to NPS and enables rapid and accurate COVID-19 self-testing and mass screening, supporting efficient COVID-19 management.Trial RegistrationKCT0006438Key PointsQuestionHow can we collect SARS-CoV-2 from oral cavity to improve the sensitivity of antigen-based rapid diagnostic test (Ag-RDT)?FindingsIn this clinical study involving 102 hospitalized COVID-19 patients, the Ag-RDT test using Beanguard gargle™-based saliva specimens showed significantly enhanced sensitivity and specificity towards detection of SARS-CoV-2 along with Alpha and Delta variants in all patients tested within 6 days of illness.MeaningOur self-testing method represents an attractive alternative to nasopharyngeal swab RT-PCR for the early diagnosis of symptomatic and asymptomatic COVID-19 cases.

Nectandrin B (NecB) is a bioactive lignan compound isolated from Myristica fragrans (nutmeg), which functions as an activator of AMP-activated protein kinase (AMPK). Because we recently found that treatment with NecB increased the cell viability of old human diploid fibroblasts (HDFs), the underlying molecular mechanism was investigated. NecB treatment in old HDFs reduced the activity staining of senescence-associated β-galactosidase and the levels of senescence markers, such as the Ser15 phosphorylated p53, caveolin-1, p21waf1, p16ink4a, p27kip1, and cyclin D1. NecB treatment increased that in S phase, indicating a enhancement of cell cycle entry. Interestingly, NecB treatment ameliorated age-dependent activation of AMPK in old HDFs. Moreover, NecB reversed the age-dependent expression and/or activity changes of certain sirtuins (SIRT1-5), and cell survival/death-related proteins. The transcriptional activity of Yin-Yang 1 and the expression of downstream proteins were elevated in NecB-treated old HDFs. In addition, NecB treatment exerted a radical scavenging effect in vitro, reduced cellular ROS levels, and increased antioxidant enzymes in old HDFs. Moreover, NecB-mediated activation of the AMPK pathway reduced intracellular ROS levels. These results suggest that NecB-induced protection against cellular senescence is mediated by ROS-scavenging through activation of AMPK. NecB might be useful in ameliorating age-related diseases and extending human lifespan.

Chelidonium majus has been used as a traditional medicine in China and western countries for various diseases, including inflammation and cancer. However, the anti-cancer effect of chelidonine, a major compound of C. majus extracts, on pancreatic cancer remains poorly understood. In this study, we found that treatment with chelidonine inhibited proliferation of BxPC-3 and MIA PaCa-2 human pancreatic cancer cells. Annexin-V/propidium iodide staining assay showed that this growth inhibitory effect of chelidonine was induced through apoptosis. We found that chelidonine treatment upregulated mRNA levels and transcription factor activity in both cell lines. Increases in protein expression levels of p53, GADD45A, p21 and cleaved caspase-3 were also observed, with more distinct changes in MIA PaCa-2 cells compared to the BxPC-3 cells. These results suggest that chelidonine induces pancreatic cancer apoptosis through the p53 and GADD45A pathways. Our findings provide new insights into the use of chelidonine for the treatment of pancreatic cancer.

Background: Antigen based rapid diagnostic test (Ag-RDT) is faster and applicable for self-testing of SARS-CoV-2. Nasopharyngeal swab (NPS) is not technically feasible for self-testing due to requirement of heath professional and risk of contaminated aerosol during sample collection. Thus, saliva specimen can be appropriate for self-sampling technique required for Ag-RDT. However, the detection of virus by Ag-RDT in saliva could be less sensitive. Therefore, we aimed to evaluate novel Beanguard gargleTM (BG)-based virus collection method that can be applied to antigen-rapid detection test (Ag-RDT) as an alternative to the current RT-PCR with a nasopharyngeal swab (NPS) for early diagnosis of COVID-19. Methods: Paired BG-based saliva and NPS specimens were collected from 102 patients with COVID-19 and 100 healthy subjects. The diagnostic performance of Ag-RDT (BG-Ag-RDT) and RT-PCR (BG-RT-PCR) of BG-based saliva specimens for SARS-CoV-2 detection was compared to NPS-RT-PCR. Physical interactions between the bean extract (BE) in BG and SARS-CoV-2 were evaluated using cryo-electron microscopy (cryo-EM), isothermal titration calorimetry, and ELISA. Findings: BG-Ag-RDTs showed high sensitivity (97·8%) and specificity (100%) in all patients within 6 days of illness associated with the infection of SARS-CoV-2 along with Alpha and Delta variants. Notably, in 11 asymptomatic early-stage cases, both BG-Ag-RDTs and BG-RT-PCR showed excellent performance with sensitivity and specificity of 100%. The interaction between SARS-CoV-2 spike proteins and truncated canavalin, an active ingredient from BE, was shown by calorimetry. The ultrastructural features of SARS-CoV-2 particles coated by BE were well observed in cryo-EM, and the detachment of the receptor binding domain of SARS-CoV-2 from hACE2 was increased with increasing concentration of BE in ELISA, allowing us to apply for releasing the virus from hACE2 in the oral cavity to early diagnosis. Interpretation: BG-based saliva specimens applied to Ag-RDT can be an alternative to NPS-RT-PCR for early diagnosis of COVID-19. The method can facilitate self-testing without trained healthcare workers for mass screening, surveillance, and efficient quarantine. Funding: BIO3S, Inc., Korea Basic Science Institute, and Jeonbuk National University Hospital. Declaration of Interest: DSK received grant support for clinical study from BIO3S, Inc. DK, JK and BV were involved in developing Beanguard gargleTM. All disclosures are unrelated to present clinical study. All other authors declare no competing interests. Ethical Approval: Ethical approval for the study was granted by the Institutional Review Board of Jeonbuk National University Hospital (CUH 2021-04-036-002) and written informed consent statements were obtained from all participants.

We identified optimal conditions for the disposal of high concentration of organic contaminants within a short time using a hybrid advanced oxidation process (AOP) combining various oxidizing agents. Plasma-treated water (PTW) containing many active species, that play dominant roles in the degradation of organic substances like hydroxyl radicals, atomic oxygen, ozone, and hydrogen peroxide, was used in this study as a strategy to improve degradation performance without the use of expensive chemical reagents like hydrogen peroxide. In particular, the optimal decomposition conditions using PTW, which were combined with 10 mg/h ozone, 2 g/L iron oxide, and 4 W UV light, demonstrated excellent removal abilities of a high concentration of reactive black 5 (RB5; 100 mg/L, >99%, [k] = 4.15 h−1) and tetracycline (TC; 10 mg/L, >96.5%, [k] = 3.35 h−1) for 25 min, approximately 1.5 times higher than that without PTW (RB5; 100 mg/L, 94%, [k] = 2.80 h−1). These results confirmed that the production of strong reactive hydroxyl radicals from the decomposition process, as well as various reactive species included in PTW efficiently attacked pollutant substances, resulting in a higher removal rate. This suggests that a water treatment system with this optimal condition based on complex AOP systems using PTW could be useful in critical environmental and biomedical applications.

Meridianin C is a marine natural product known for its anti‐cancer activity. At present, the anti‐tumour effects of meridianin C on oral squamous cell carcinoma are unknown. Here, we investigated the effect of meridianin C on the proliferation of four different human tongue cancer cells, YD‐8, YD‐10B, YD‐38 and HSC‐3. Among the cells tested, meridianin C most strongly reduced the growth of YD‐10B cells; the most aggressive and tumorigenic of the cell lines tested. Strikingly, meridianin C induced a significant accumulation of macropinosomes in the YD‐10B cells; confirmed by the microscopic and TEM analysis as well as the entry of FITC‐dextran, which was sensitive to the macropinocytosis inhibitor amiloride. SEM data also revealed abundant long and thin membrane extensions that resemble lamellipodia on the surface of YD‐10B cells treated with meridianin C, pointing out that meridianin C‐induced macropinosomes was the result of macropinocytosis. In addition, meridianin C reduced cellular levels of Dickkopf‐related protein‐3 (DKK‐3), a known negative regulator of macropinocytosis. A role for DKK‐3 in regulating macropinocytosis in the YD‐10B cells was confirmed by siRNA knockdown of endogenous DKK‐3, which led to a partial accumulation of vacuoles and a reduction in cell proliferation, and by exogenous DKK‐3 overexpression, which resulted in a considerable inhibition of the meridianin C‐induced vacuole formation and decrease in cell survival. In summary, this is the first study reporting meridianin C has novel anti‐proliferative effects via macropinocytosis in the highly tumorigenic YD‐10B cell line and the effects are mediated in part through down‐regulation of DKK‐3.

Background: The cellular senescence of primary cultured cells is an irreversible process characterized by growth arrest. Restoration of senescence by ginsenosides has not been explored so far. Rg3(S) treatment markedly decreased senescence-associated β-galactosidase activity and intracellular reactive oxygen species levels in senescent human dermal fibroblasts (HDFs). However, the underlying mechanism of this effect of Rg3(S) on the senescent HDFs remains unknown. Methods: We performed a label-free quantitative proteomics to identify the altered proteins in Rg3(S)-treated senescent HDFs. Upregulated proteins induced by Rg3(S) were validated by real-time polymerase chain reaction and immunoblot analyses. Results: Finally, 157 human proteins were identified, and variable peroxiredoxin (PRDX) isotypes were highly implicated by network analyses. Among them, the mitochondrial PRDX3 was transcriptionally and translationally increased in response to Rg3(S) treatment in senescent HDFs in a time-dependent manner. Conclusion: Our proteomic approach provides insights into the partial reversing effect of Rg3 on senescent HDFs through induction of antioxidant enzymes, particularly PRDX3. Keywords: Ginsenoside Rg3(S), Human dermal fibroblast, Label-free quantitative proteomics, Restoration, Senescence

In recent times, the treatment of harmful algal blooms (HABs) became an important environmental issue to preserve and remediate water resources globally. In the present study, the adsorptive removal of harmful algal species Microcystis aeruginosa directly from an aqueous medium was attempted. Waste biomass (Escherichia coli) was immobilized using polysulfone and coated using the cationic polymer polyethylenimine (PEI) to generate PEI-coated polysulfone-biomass composite fiber (PEI-PSBF). The density of M. aeruginosa in an aqueous medium (BG11) was significantly decreased by treatment with PEI-PSBF. additionally, analysis using FE-SEM, confirmed that the removal of M. aeruginosa algal cells by PEI-PSBF was caused by the adsorption mechanism. According to the profiles of phosphorus for the algal cell growth in M. aeruginosa cultivating samples, we found that the adsorbed M. aeruginosa onto the PEI-PSBF lost their biological activity compared to the non-treated M. aeruginosa cells.

The proviral integration moloney murine leukaemia virus (Pim) kinases, consisting of Pim‐1, Pim‐2 and Pim‐3, are involved in the control of cell growth, metabolism and differentiation. Pim kinases are emerging as important mediators of adipocyte differentiation. AZD1208 is a pan‐Pim kinase inhibitor and is known for its anti‐cancer activity. In this study, we investigated the effect of AZD1208 on adipogenesis and lipolysis in 3T3‐L1 cells, a murine preadipocyte cell line. AZD1208 markedly suppressed lipid accumulation and reduced triglyceride contents in differentiating 3T3‐L1 cells, suggesting the drug's anti‐adipogenic effect. On mechanistic levels, AZD1208 reduced not only the expressions of CCAAT/enhancer‐binding protein‐α (C/EBP‐α), peroxisome proliferator‐activated receptor‐γ (PPAR‐γ), fatty acid synthase (FAS), acetyl‐CoA carboxylase (ACC) and perilipin A but also the phosphorylation of signal transducer and activator of transcription‐3 (STAT‐3) in differentiating 3T3‐L1 cells. Remarkably, AZD1208 increased cAMP‐activated protein kinase (AMPK) and LKB‐1 phosphorylation while decreased intracellular ATP contents in differentiating 3T3‐L1 cells. Furthermore, in differentiated 3T3‐L1 adipocytes, AZD1208 also partially promoted lipolysis and enhanced the phosphorylation of hormone‐sensitive lipase (HSL), a key lipolytic enzyme, indicating the drug's HSL‐dependent lipolysis. In summary, the findings show that AZD1208 has anti‐adipogenic and lipolytic effects on 3T3‐L1 adipocytes. These effects are mediated by the expression and/or phosphorylation levels of C/EBP‐α, PPAR‐γ, FAS, ACC, perilipin A, STAT‐3, AMPK and HSL.

Unwanted, rapid increases in the algal populations of water systems cause harmful algal blooms, which have recently become a major environmental problem. The cyanobacterium Microcystis aeruginosa is the most prevalent bloom species and is responsible for the majority of blooms in freshwater environments. In this study, we attempted to develop an eco-friendly method to suppress M. aeruginosa bloom based on a biological control using bacteria newly isolated from the soil. In a screen for bacteria with strong lethal activity toward Microcystis, we isolated Bacillus sp. T4 and characterised its algicidal activity. Microcystis aeruginosa cells were killed via indirect attack by compound(s) secreted by T4 bacteria. ELISA revealed a dramatic increase in extracellular microcystins in M. aeruginosa cultures upon treatment with T4. Therefore, we screened for bacteria that could degrade these toxins, and three new isolates (R12, S42 and S65) were identified. Simultaneous application of both T4 as a lytic agent and R12 or S42 as toxin-degrading bacteria could eliminate both Microcystis cells and its problematic toxin. Our eco-friendly approach, based on the application of newly isolated bacteria, provides a novel method to control harmful algal blooms.

Rapid methods for the detection and clinical treatment of human norovirus (HuNoV) are needed to control foodborne disease outbreaks, but reliable techniques that are fast and sensitive enough to detect small amounts of HuNoV in food and aquatic environments are not yet available. We explore the interactions between HuNoV and concanavalin A (Con A), which could facilitate the development of a sensitive detection tool for HuNoV. Biophysical studies including hydrogen/deuterium exchange (HDX) mass spectrometry and surface plasmon resonance (SPR) revealed that when the metal coordinated region of Con A, which spans Asp16 to His24, is converted to nine alanine residues (mCon AMCR), the affinity for HuNoV (GII.4) diminishes, demonstrating that this Ca2+ and Mn2+ coordinated region is responsible for the observed virus-protein interaction. The mutated carbohydrate binding region of Con A (mCon ACBR) does not affect binding affinity significantly, indicating that MCR of Con A is a major region of interaction to HuNoV (GII.4). The results further contribute to the development of a HuNoV concentration tool, Con A-immobilized polyacrylate beads (Con A-PAB), for rapid detection of genotypes from genogroups I and II (GI and GII). This method offers many advantages over currently available methods, including a short concentration time. HuNov (GI and GII) can be detected in just 15 min with 90% recovery through Con A-PAB application. In addition, this method can be used over a wide range of pH values (pH 3.0 – 10.0). Overall, this rapid and sensitive detection of HuNoV (GI and GII) will aid in the prevention of virus transmission pathways, and the method developed here may have applicability for other foodborne viral infections.

Estimating the age of a bloodstain is important for tracking down the true perpetrator of a crime from among the suspects. The current methods available for this estimation suffer from several shortcomings, including the time required and low accuracy. A mobile detection system, named Smart Forensic Phone, developed in this study enabled us to rapidly and precisely estimate the age of a bloodstain. We placed blood on five different materials (wallpaper, fabric, glass, wood, and A4 paper), monitored the RGB (Red, Green, and Blue) values per pixel of the bloodstain picture at 6-h intervals, and estimated the approximate age of the bloodstain using a smartphone application. The RGB values were converted into the V value of HSV (hue, saturation, and brightness). A rapid decline in RGB and V values was observed over the first 42 h and thereafter remained relatively unchanged. The age of the bloodstain could be calculated from the plot of V vs. time. This method enables the estimation of the age of the bloodstain if it is less than 42 h old. We will be continuing to work toward optimizing the method, so that the age of the bloodstain older than 42 h can be determined using the Smart Forensic Phone . Our detection system offers a novel method to both criminal investigators and researchers for predicting the time elapsed after the victim’s death, which is also called postmortem interval (PMI) or the approximate time when the crime was committed.

A novel highly sensitive dendrimer coupled impedimetric immunosensor was developed for the label-free and reagent-free detection of cardiac troponin I (TnI) in serum samples. The immunosensor probe was fabricated by covalently attaching carboxylic acid-functionalized third generation (G3) poly (amidoamine) (PAMAM) dendrimer (Den) on the 3, 3', 5, 5'-tetramethyl benzidine (TMB) modified 6-mercaptohexanoic acid (MHA) self-assembled monolayer (SAM) on a gold (Au) electrode. Monoclonal anti-TnI antibody was then covalently immobilized on the Den and TMB attached MHA SAM modified surface. TMB was used as an internal surface redox couple for generating signal which also allowed to avoid the use of an external one (i.e. ferricyanide couple) in solution during the impedance measurement for monitoring the antibody-antigen binding. On the other hand, Den was used as a signal enhancer by immobilizing more anti-body on the immunosensor probe. The immunosensor probe was characterized using X-ray photoelectron spectroscopy (XPS), quartz crystal microbalance (QCM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) techniques. The TnI detection in diluted serum was based on the measurement of charge transfer resistance (Rct) of the electron transfer process of the surface-attached TMB before and after immunobinding. Under the optimized condition, the proposed immunosensor could detect human TnI in diluted serum samples as low as 11.7 fM with a wide linear dynamic range, good stability, and excellent specificity. The validity of the proposed method was tested in various TnI spiked human undiluted serum samples and was compared with the enzyme-linked immunosorbent assay (ELISA). The results suggested that the proposed immunosensor could be a useful tool for practical applications in clinical diagnosis.

We report a new paper-based multiplex analytical device (mPAD) for simultaneous screening of three analytes (glutamate dehydrogenase, toxin A, and toxin B) known as biomarkers for Clostridioides difficile infection (CDI). To overcome the limitation of common rapid assays (e.g. lateral flow immunochromatographic and enzyme immunoassays) in terms of multiplexing, sensitivity, simplicity, and ease-of-use, the mPAD is constructed with a three dimensional (3D) configuration of paper components with a multi-channel design. Multiple fluidic paths developed with wax-patterned paper allow the simultaneous detection of glutamate dehydrogenase, toxin A, and toxin B without any cross-reactivity. The 3D fluidic network on the mPAD facilitates a self-operating test procedure for the mixing and addition of amplification reagents with a one-step sliding operation. The results of the multiplex CDI assay can be easily interpreted by the naked eye within 10 min, and are visually intensified over time resulting in up to 3-fold signal amplification. Our device exhibited remarkable analytical performances for the simultaneous detection of three CDI biomarkers, providing a sensitivity of 97%, specificity of 88%, accuracy of 95%, and limits of detection for glutamate dehydrogenase, toxin A, and toxin B of 0.16 ng mL−1, 0.09 ng mL−1, and 0.03 ng mL−1, respectively. These results indicate the high applicability and feasibility of mPAD for multiplex testing for CDI with the advantages of being simple, sensitive, inexpensive, user-friendly, and equipment-free.

While the importance of α-Ga2O3 crystals is increasingly gaining much interest, the structural behavior of halide vapor phase epitaxy (HVPE) α-Ga2O3 crystals in wet chemical etching has not been explored yet. In this study, we investigate the structural characterization of HVPE α-Ga2O3 materials via alkali KOH solution etching. Further, the activation energy of the etch rate for the materials, etch pit, and surface roughness after wet chemical treatment have been examined. The cross-sectional TEM analysis demonstrates that the triangular-shaped etch pits with (11 2 ‾ 6) plane are caused by the propagation of threading dislocations in the HVPE α-Ga2O3 crystals.

Background: Fatigue is a common problem in breast cancer patients and the use of health-functional foods has been one of the popular choices for the management of fatigue and the recovery of health status. The purpose of this study was to investigate the prevalence of health-functional food consumption and its relationship with fatigue in breast cancer patients. Methods: In total, 270 women with breast cancer and without distant metastases (M=0 in tumor node metastasis staging) who underwent postoperative radiation therapy at a Kosin University Gospel Hospital in Busan between January and December 2011 were enrolled. The patients completed a questionnaire for the assessment of the social factors, lifestyle, health-functional food consumption status, and disease status. The subjects were stratified into groups according to the severity of their fatigue. Results: Among the 270 subjects, 218 (80.7%) consumed health-functional foods daily. Most of them used health-functional foods to recover from fatigue and improve health through recommendations from other individuals or following the mass media. There were significant differences between the fatigue and the non-fatigue group according to occupation, chronic illness status, quality of sleep, and activity level. The prevalence of fatigue was 58% among the study subjects. The health-functional food consumption significantly increased with increasing severity of fatigue (P=0.013). Conclusion: The consumption of health-functional food is high in breast cancer patients with fatigue compared to the general population. Therefore, we suggest that these patients should consult with their doctor concerning the consumption of health-functional food as a prescription.

This study analyzed eleven genomic segments of three Korean porcine G8P[7] group A rotavirus (RVA) strains. Phylogenetically, these strains contained two bovine-like and nine porcine-like genomic segments. Eight genes (VP1, VP2, VP6 and NSP1-NSP5) of strains 156-1 and 42-1 and seven genes (VP1, VP2, VP6 and NSP2-NSP5) of strain C-1 clustered closely with porcine and porcine-like animal strains and distantly from typical human Wa-like strains. The VP3-M2 genotype of these strains clustered closely with bovine-like strains, but distantly with typical human DS-1-like strains. These data indicate that multiple reassortments involving porcine and bovine RVA strains in Korea must have occurred. ispartof: Archives of Virology vol:161 issue:10 pages:2835-2841 ispartof: location:Austria status: published

An ultrasensitive electrochemical nanostructured immunosensor for a breast cancer biomarker carbohydrate antigen 15-3 (CA 15-3) was fabricated using non-covalent functionalized graphene oxides (GO/Py-COOH) as sensor probe and multiwalled carbon nanotube (MWCNTs)-supported numerous ferritin as labels. The immunosensor was constructed by immobilizing a monoclonal anti-CA 15-3 antibody on the GO modified cysteamine (Cys) self-assembled monolayer (SAM) on an Au electrode (Au/Cys) through the amide bond formation between the carboxylic acid groups of GO/Py-COOH and amine groups of anti-CA 15-3. Secondary antibody conjugated MWCNT-supported ferritin labels (Ab2-MWCNT-Ferritin) were prepared through the amide bond formation between amine groups of Ab2 and ferritin and carboxylic acid groups of MWCNTs. The detection of CA 15-3 was based on the enhanced bioelectrocatalytic reduction of hydrogen peroxide mediated by hydroquinone (HQ) at the GO/Py-COOH-based sensor probe. The GO/Py-COOH-based sensor probe and Ab2-MWCNT-Ferritin labels were characterized using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scanning electron microscope (SEM), transmission electron microscope (TEM), and x-ray photoelectron spectroscopy (XPS) techniques. Using differential pulse voltammetry (DPV) technique, CA 15-3 can be selectively detected as low as 0.01 ± 0.07 U/mL in human serum samples. Additionally, the proposed CA 15-3 immunosensor showed excellent selectivity and better stability in human serum samples, which demonstrated that the proposed immunosensor has potentials in proteomic researches and diagnostics.

Diabetes mellitus is a complex and heterogeneous disease, which has β-cell dysfunction at its core. Glucotoxicity affects pancreatic islets, causing β-cell apoptosis. However, the role of JNK/β-catenin signaling in glucotoxic β-cell apoptosis is not well understood. Recently, we identified tetraspanin-2 (TSPAN2) protein as a proapoptotic β-cell factor induced by glucose, suggesting that TSPAN2 might contribute to pancreatic β-cell glucotoxicity. To investigate the effects of glucose concentration on TSPAN2 expression and apoptosis, we used reverted immortalized RNAKT-15 human pancreatic β cells. High TSPAN2 levels up-regulated phosphorylated (p) JNK and induced apoptosis. p-JNK enhanced the phosphorylation of β-catenin and Dickkopf-1 (Dkk1). Dkk1 knockdown by small interfering (si)RNA up-regulated nuclear β-catenin, suggesting that it is a JNK/β-catenin-dependent pathway. siRNA-mediated TSPAN2 depletion in RNAKT-15 cells increased nuclear β-catenin. This decreased BCL2-associated X protein (Bax) activation, leading to marked protection against high glucose-induced apoptosis. Bax subfamily proteins induced apoptosis through caspase-3. Thus, TSPAN2 might have induced Bax translocation and caspase-3 activation in pancreatic β cells, thereby promoting the apoptosis of RNAKT-15 cells by regulating the JNK/β-catenin pathway in response to high glucose concentrations. Targeting TSPAN2 could be a potential therapeutic strategy to treat glucose toxicity-induced β-cell failure.-Hwang, I.-H., Park, J., Kim, J. M., Kim, S. I., Choi, J.-S., Lee, K.-B., Yun, S. H., Lee, M.-G., Park, S. J., Jang, I.-S. Tetraspanin-2 promotes glucotoxic apoptosis by regulating the JNK/β-catenin signaling pathway in human pancreatic β cells.

Haematococcus pluvialis is a green microalga of particular interest, since it is considered the best potential natural source of astaxanthin, which is widely used as an additive for natural pigmentation. In addition, astaxanthin has recently garnered commercial interest as a nutraceutical, cosmetic, and pharmaceutical. However, producing astaxanthin from H. pluvialis necessitates separation with distinctive culture conditions, dividing between the microalgae growth and the astaxanthin production stages. Light-emitting diodes (LEDs) have emerged as a replacement for traditional light sources, and LED applications are now rapidly expanding to multiple areas in fields such as biotechnology. However, further detail application into microalgae biotechnology remains limited. In this study, we have attempted to establish new protocols based on the specific wavelength of LEDs for the cultivation and production of astaxanthin using H. pluvialis. Specifically, we applied red LEDs for microalgae cell growth and then switched to blue LEDs to induce astaxanthin biosynthesis. The result showed that astaxanthin productions based on a wavelength shift from red to blue were significantly increased, compared to those with continuous illumination using red LEDs. Furthermore, additional increase of astaxanthin production was achieved with simultaneous application of exogenous carbon with blue LED illumination. Our approach based on the proper manipulation of LED wavelengths upon H. pluvialis cell stages will enable the improvement of biomass and enhance astaxanthin production using H. pluvialis.

Herein, we present a simple readout of the binding between a chemical drug and its target proteins in the cytoplasm by using a two-step bioorthogonal labeling method combined with spatially-localized expression of proteins. Dasatinib was modified with trans-cyclooctene (TCO), and its cytoplasmic target kinases were expressed in intracellular compartments, such as endosomes and F-actins. After bioorthogonal labeling, the colocalization between Dasatinib and its target proteins was observed in intracellular compartments.

Postpartum depression is a severe emotional and mental disorder that involves maternal care defects and psychiatric illness. Postpartum depression is closely associated with a combination of physical changes and physiological stress during pregnancy or after parturition in stress-sensitive women. Although postpartum depression is relatively well known to have deleterious effects on the developing fetus, the influence of genetic risk factors on the development of postpartum depression remains unclear. In this study, we discovered a novel function of T cell death-associated gene 51 (TDAG51/PHLDA1) in the regulation of maternal and depressive-like behavior. After parturition, TDAG51-deficient dams showed impaired maternal behavior in pup retrieving, nursing and nest building tests. In contrast to the normal dams, the TDAG51-deficient dams also exhibited more sensitive depressive-like behaviors after parturition. Furthermore, changes in the expression levels of various maternal and depressive-like behavior-associated genes regulating neuroendocrine factor and monoamine neurotransmitter levels were observed in TDAG51-deficient postpartum brain tissues. These findings indicate that TDAG51 plays a protective role against maternal care defects and depressive-like behavior after parturition. Thus, TDAG51 is a maternal care-associated gene that functions as a crucial regulator of maternal and depressive-like behavior after parturition., Author summary Postpartum depression is a severe emotional and mental disease that can affect women typically after parturition. However, the genetic risk factors associated with the development of postpartum depression are still largely unknown. We discovered a novel function of T cell death-associated gene 51 (TDAG51) in the regulation of maternal behavior and postpartum depression. We report that TDAG51 deficiency induces depressive-like and abnormal maternal behavior after parturition. The loss of TDAG51 in postpartum brain tissues induces changes in the expression levels of various maternal and depressive-like behavior-associated genes that regulate the levels of neuroendocrine factors and monoamine neurotransmitters. TDAG51 is a maternal care-associated gene that functions as a crucial regulator of maternal and depressive-like behavior after parturition.

The oxidative damage initiated by reactive oxygen species (ROS) is a major contributor to the functional decline and disability that characterizes aging. The anti-oxidant flavonoid, quercetin, is a plant polyphenol that may be beneficial for retarding the aging process. We examined the restoring properties of quercetin on human dermal fibroblasts (HDFs). Quercetin directly reduced either intracellular or extracellular ROS levels in aged HDFs. To find the aging-related target genes by quercetin, microarray analysis was performed and two up-regulated genes LPL and KCNE2 were identified. Silencing LPL increased the expression levels of senescence proteins such as p16INK4A and p53 and silencing KCNE2 reversed gene expressions of EGR1 and p-ERK in quercetin-treated aged HDFs. Silencing of LPL and KCNE2 decreased the expression levels of anti-oxidant enzymes such as superoxide dismutase and catalase. Also, the mitochondrial dysfunction in aged HDFs was ameliorated by quercetin treatment. Taken together, these results suggest that quercetin has restoring effect on the cellular senescence by down-regulation of senescence activities and up-regulation of the gene expressions of anti-oxidant enzymes in aged HDFs.

The amino acids found to be abundant in common buckwheat (CB) sprouts was Val (40%), followed by Tyr (28%), whereas Val accounted for 62% in Tartary buckwheat (TB). The buckwheat stem and root commonly contained Gln (40%–42% in stem; 30%–37% in root). Thus, soluble amino nitrogen source is used for Gln in buckwheat. The main difference of amino acid distribution in 3 tissues between CB and TB was Tyr in sprouts. A low level of Tyrin TB presumably resulted from the conversion to other phenolic metabolites. The content of essential free amino acids in TB sprout was 53% higher than that in CB. Thus, the TB sprouts are beneficial to human nutrition.