Hepatocellular carcinoma(HCC)remains the third leading cause of cancer-related deaths worldwide;however,its therapeutic options are limited.Understanding the molecular mechanisms of HCC could provide insight into new ...Hepatocellular carcinoma(HCC)remains the third leading cause of cancer-related deaths worldwide;however,its therapeutic options are limited.Understanding the molecular mechanisms of HCC could provide insight into new therapies.Emerging studies indicate the important role of long-noncoding RNAs(lncRNAs)in the pathogenesis of HCC.The expression of the well-studied lncRNA taurine upregulated gene 1(TUG1)is upregulated in HCC tissues,but its transcriptomic effects in HCC cells remain unexplored.We established TUG1-knockdown and control HCC cells for RNA-seq experiments.KEGG analysis revealed glycolysis as the top enriched pathway upon TUG1 silencing.Accordingly,TUG1-depleted HCC cells showed impairments in glucose uptake,ATP synthesis,and lactate production.Clinical HCC tissue data revealed positive gene expression correlations between TUG1 and several glycolysis-related genes.To identify a molecular function of TUG1 in glycolysis,we explored the competing endogenous model and used bioinformatic tools to find the five microRNAs(miRNAs)that had the most binding sites for TUG1.Among these miRNAs,miR-122-5p exhibited an inverse correlation in gene expression with most TUG1-regulated glycolysis genes,including PKM,ALDOA,ENO2,and PFKM.Dual-luciferase assays demonstrated the direct interaction between TUG1 and miR-122-5p and between miR-122-5p and the 3ʹuntranslated regions of both PKM and ALDOA.We further showed that inhibition of miR-122-5p alleviated the suppression of glycolysis induced by TUG1 depletion.Together,our RNA-seq analysis of TUG1-depleted HCC cells,combined with clinical data,reveals a critical role of TUG1 in regulating glycolysis and provides new insight into its oncogenic function in HCC.展开更多
Hepatocellular carcinoma (HCC) is the leading cause of cancer death worldwide;nevertheless, currenttherapeutic options are limited or ineffective for many patients. Therefore, elucidation of molecular mechanisms inHCC...Hepatocellular carcinoma (HCC) is the leading cause of cancer death worldwide;nevertheless, currenttherapeutic options are limited or ineffective for many patients. Therefore, elucidation of molecular mechanisms inHCC biology could yield important insights for the intervention of novel therapies. Recently, various studies havereported dysregulation of long non-coding RNAs (lncRNAs) in the initiation and progression of HCC, including H19;however, the biological function of H19 in HCC remains unclear. Here, we show that knockdown of H19 disruptedHCC cell growth, impaired the G1-to-S phase transition, and promoted apoptosis, while overexpression of H19yielded the opposite results. Screening for expression of cell cycle-related genes revealed a significant downregulationof CDK6 at both RNA and protein levels upon H19 suppression. Bioinformatic analysis of the H19 sequence and the3′ untranslated region (3′ UTR) of CDK6 transcripts showed several binding sites for microRNA-107 (miR-107), andthe dual luciferase reporter assay confirmed their direct interaction with miR-107. Consistently, blockage of miR-107activity alleviated the growth suppression phenotypes induced by H19 downregulation, suggesting that H19 serves asa molecular sponge for miR-107 to promote CDK6 expression and cell cycle progression. Together, this studydemonstrates a mechanistic function of H19 in driving the proliferation of HCC cells and suggests H19 suppressionas a novel approach for HCC treatment.展开更多
Objective:To systematically map the stepwise events leading to deoxyelephantopin-induced cell death of HCT116 human colorectal cancer cells and evaluate the effectiveness of deoxyelephantopin in vivo.Methods:HCT116 ce...Objective:To systematically map the stepwise events leading to deoxyelephantopin-induced cell death of HCT116 human colorectal cancer cells and evaluate the effectiveness of deoxyelephantopin in vivo.Methods:HCT116 cells were treated with deoxyelephantopin at various concentrations and time points.Autophagy was confirmed by the detection of autophagosomes and autophagosomal proteins by electron microscopy and Western blotting assays,respectively,and then validated by siRNA knockdown.In addition,apoptosis was confirmed by the detection of apoptosis-related proteins.The intracellular reactive oxygen species(ROS)level was measured using flow cytometry.The growth inhibitory effect of deoxyelephantopin was further evaluated in vivo using a mouse xenograft model.Results:Deoxyelephantopin firstly elevated ROS production,which then triggered autophagic flux with the accumulation of autophagosomal proteins including LC3 A/B,ATG5,and ATG7,followed by the induction of apoptosis via the intrinsic and extrinsic pathways.Pre-treatment with N-acetyl-L-cysteine,a ROS inhibitor,reversed both apoptosis and autophagy.The knockdown of LC3 prevented apoptosis induction which confirmed that deoxyelephantopin induced autophagy-dependent apoptosis in HCT116 cells.Accumulation of ROS also activated apoptosis via the mitogen-activated protein kinases signaling pathway.Furthermore,deoxyelephantopin also inhibited the PI3 K/AKT/mTOR pathway,which then released the inhibition of autophagy.In vivo study further showed that deoxyelephantopin significantly suppressed the growth of HCT116 subcutaneous xenograft in nude mice.Conclusions:Our findings revealed that deoxyelephantopin elevates oxidative stress and induces ROS-dependent autophagy followed by apoptosis in HCT116 cells via the concerted modulation of multiple signaling pathways.These findings further support the development of deoxyelephantopin as a therapeutic agent for colorectal cancer.展开更多
Background: Continuous exposure to millimolar (mM) Vitamin C (AA) in vitro kills cancer cells. For superficial bladder carcinoma (SBC), standard chemotherapy is instillation of Bacillus Calmette-Guerin. The recurrence...Background: Continuous exposure to millimolar (mM) Vitamin C (AA) in vitro kills cancer cells. For superficial bladder carcinoma (SBC), standard chemotherapy is instillation of Bacillus Calmette-Guerin. The recurrence rate with this therapy is 91%. But high dosage vitamins including AA reduced the recurrence to 41%. Aim: To determine the oral dosage of AA that causes the highest concentration of AA [AA] in the bladder. Method: We conducted a clinical trial of 14 people who took various dosages of AA, and analyzed the [AA] in their urine. Results: AA above 2 g twice a day was not absorbed. But that intake produced a bladder [AA] above 1 mM in all participants. Conclusion: Taking 2 g of AA BID will increase [AA] in the bladder to a level likely to kill cancer cells that cause SBC. Taking that dosage 2 consecutive days a week is likely to reduce the recurrence rate of SBC substantially.展开更多
Breast cancer is the second leading cause of cancerrelated deaths in women worldwide;a prime cancer biomarker to aid in the diagnosis, directed treatment, clinical management, and reoccurrence of this cancer is a MUC1...Breast cancer is the second leading cause of cancerrelated deaths in women worldwide;a prime cancer biomarker to aid in the diagnosis, directed treatment, clinical management, and reoccurrence of this cancer is a MUC1 peptide fragment: cancer antigen 15-3 (CA 15-3). Herein, an immuno-fluorescence assay for CA 15-3 was developed;this ALYGNSA system consists of a protein biolinker (Protein G’) adsorbed onto Poly (methyl methacrylate) (PMMA). The unique interaction of Protein G’ with PMMA, a thermo-plastic polymer has been demonstrated to improve human IgG capture antibody alignment/ orientation and result in greater assay sensitivity. Indeed a previous report (HEALTH 1 325 - 329, 2009) on the shed extracellular domain of HER-2/neu revealed a 10-fold increase in sensitivity of the ALYGSNA assay over a control ELISA assay. Results from this ALYGNSA assay study revealed that a 16-fold increase in detection (≤0.94 U/mL) of CA 15-3 was found in comparison to a commercial control ELISA kit (≤15 U/mL). In conclusion, this enhanced sensitivity of the ALYGNSA assay for CA 15-3, may provide insights into the role/function of this biomarker in normal, as well as, breast cancer and other epithelial cancers.展开更多
Internal solvation of protein was studied by site-directed mutagenesis, with which an intrinsically fluorescent probe,tryptophan, is inserted into the desired position inside a protein molecule for ultrafast spectrosc...Internal solvation of protein was studied by site-directed mutagenesis, with which an intrinsically fluorescent probe,tryptophan, is inserted into the desired position inside a protein molecule for ultrafast spectroscopic study. Here we review this unique method for protein dynamics research. We first introduce the frontiers of protein solvation, site-directed mutagenesis, protein stability and characteristics, and the spectroscopic methods. Then we present time-resolved spectroscopic dynamics of solvation dynamics inside cavities of active sites. The studies are carried out on a globular protein, staphylococcal nuclease. The solvation at sites inside the protein molecule＇s cavities clearly reveals characteristics of the local environment. These solvation behaviors are directly correlated to enzyme activity.展开更多
Noxious mechanical information is transmitted through molecularly distinct nociceptors,with pinprickevoked sharp sensitivity via A-fiber nociceptors marked by developmental expression of the neuropeptide Y receptor 2(...Noxious mechanical information is transmitted through molecularly distinct nociceptors,with pinprickevoked sharp sensitivity via A-fiber nociceptors marked by developmental expression of the neuropeptide Y receptor 2(Npy2 r)and von Frey filament-evoked punctate pressure information via unmyelinated C fiber nociceptors marked by MrgprD.However,the molecular programs controlling their development are only beginning to be understood.Here we demonstrate that Npy2 r-expressing sensory neurons are in fact divided into two groups,based on transient or persistent Npy2 r expression.Npy2 r-transient neurons are myelinated,likely including A-fiber nociceptors,whereas Npy2 r-persistent ones belong to unmyelinated pruriceptors that co-express Nppb.We then showed that the transcription factors NFIA and Runx1 are necessary for the development of Npy2 r-transient A-fiber nociceptors and MrgprD^+C-fiber nociceptors,respectively.Behaviorally,mice with conditional knockout of Nfia,but not Runx1 showed a marked attenuation of pinprick-evoked nocifensive responses.Our studies therefore identify a transcription factor controlling the development of myelinated nociceptors.展开更多
C omprehensive identification of driver mutations in prostate cancer can serve to enhance our understanding of the disease and expand the use of available treatment options. Two recent and comple- mentary studies from...C omprehensive identification of driver mutations in prostate cancer can serve to enhance our understanding of the disease and expand the use of available treatment options. Two recent and comple- mentary studies from Barbieri et al. and Grasso et al.展开更多
As part of the determination of the possible impact of human activities on surface waters, case of Lake Sonfonia, six sampling sites were selected according to their solicitation by the population and their exposure t...As part of the determination of the possible impact of human activities on surface waters, case of Lake Sonfonia, six sampling sites were selected according to their solicitation by the population and their exposure to probable sources of pollution. The objective of this work is to monitor the physico-chemical quality of the waters of Lake Sonfonia during the low water level (March) and during the flood period (August) of the year 2021. Two sampling campaigns of water were carried out in dry weather and two others in cold weather. Parameters such as temperature, pH, EC (Electrical Conductivity), dissolved oxygen, TDS (Total Dissolved Solids) were measured in situ. Suspended matter, phosphate, nitrate, nitrite, sulphates, total iron, COD (Chemical Oxygen Demand) were measured in the laboratory by the colorimetric method. Stata 15 software was applied for the statistical analysis of the data and the correlation test between the parameters gave highly significant correlations. It has been noted that the situation is not very good and that this pollution comes mainly from human activities.展开更多
Huntington’s disease(HD)is a fatal neurodegenerative disorder characterized by a triad of behavioral symptoms:involuntary movement,emotional change,and cognitive dysfunction.Although alterations in WNT signaling have...Huntington’s disease(HD)is a fatal neurodegenerative disorder characterized by a triad of behavioral symptoms:involuntary movement,emotional change,and cognitive dysfunction.Although alterations in WNT signaling have been reported in HD,its precise role in pathogenesis remains unclear.In this study,we found that astrocytic WNT5B mRNA and protein levels are elevated in the striatum of both HD patients and HD model mice.The noncanonical WNT5B signaling pathway induced sustained expression of matrix metallopeptidase 14(MMP14),an extracellular matrix(ECM)-degrading enzyme,via activation of the NFATc2 transcription factor in both human and primary mouse astrocytes.Robust upregulation of MMP14 led to ECM degradation,medium spiny neuron(MSN)damage,and increased mutant huntingtin aggregation in N171-82Q HD transgenic mice.Furthermore,WNT5B gain-offunction exacerbated neuropathology,impaired motor coordination,and shortened the lifespan of N171-82Q mice.We further demonstrated that the overexpression of the estrogen receptorα(ERα)suppresses NFATc2 transcriptional activity in vitro.A targeted therapy for the WNT5B-NFATc2-MMP14 signaling pathway by genistein,a phytoestrogen,reduced MMP14 transcription by antagonizing NFATc2 activity and preventing ECM degradation in N171-82Q mice.Genistein treatment also ameliorated neuropathology and motor deficits and prolonged the lifespan of HD mice.Together,these findings define a molecular pathological mechanism in which astrocytic MMP14 transcription,driven by the noncanonical WNT5B signaling pathway,promotes ECM degradation and MSN damage and accelerates neurodegeneration in HD.Modulation of the noncell-autonomous WNT5BNFATc2-MMP14 signaling pathway by genistein may serve as a potential therapeutic strategy for mitigating HD pathogenesis.展开更多
Cytotoxic T cells targeting cancer neoantigens harboring driver mutations can lead to durable tumor regression in an HLAIdependent manner.However,it is difficult to extend the population of patients who are eligible f...Cytotoxic T cells targeting cancer neoantigens harboring driver mutations can lead to durable tumor regression in an HLAIdependent manner.However,it is difficult to extend the population of patients who are eligible for neoantigen-based immunotherapy,as immunogenic neoantigen-HLA pairs are rarely shared across different patients.Thus,a way to find other human leukocyte antigen(HLA)alleles that can also present a clinically effective neoantigen is needed.Recently,neoantigen-based immunotherapy targeting the KRAS G12D mutation in patients with HLA-C*08:02 has shown effectiveness.In a proof-ofconcept study,we proposed a combinatorial strategy(the combination of phylogenetic and structural analyses)to find potential HLA alleles that could also present KRAS G12D neoantigen.Compared to in silico binding prediction,this strategy avoids the uneven accuracy across different HLA alleles.Our findings extend the population of patients who are potentially eligible for immunotherapy targeting the KRAS G12D mutation.Additionally,we provide an alternative way to predict neoantigen-HLA pairs,which maximizes the clinical usage of shared neoantigens.展开更多
Ovarian cancer(OC)is associated with poor outcomes and challenges scientists and clinicians.It is usually diagnosed in advanced stages when it is frequently aggressive,chemoresistant,and metastatic.The most prevalent ...Ovarian cancer(OC)is associated with poor outcomes and challenges scientists and clinicians.It is usually diagnosed in advanced stages when it is frequently aggressive,chemoresistant,and metastatic.The most prevalent form of OC is epithelial ovarian cancer(EOC),which displays significant heterogeneity,enhancing the difficulty in managing the disease.Several factors have been associated with the disease’s development and progression,especially those related to the tumor microenvironment(TME).Here,we highlight components of the ovarian TME in the disease development process,including pro-inflammatory pathways activated by interleukins,cytokines and chemokines,cancer-associated fibroblasts,tumor-associated macrophages,and epithelial-mesenchymal transition.We compiled evidence identifying TME factors promoting the development,chemoresistance,and metastasis,including cytokines,chemokines,growth factors,and tumor-associated cells.We identify potential targets for treatment and improving outcomes.These targets block or alter pathways associated with OC(especially EOC)progression.展开更多
ABC transporters are molecular machines which power the solute transport using ATP hydrolysis.The structural biology of ABC transporters has been exploding for the last few years,and this study explores timelines and ...ABC transporters are molecular machines which power the solute transport using ATP hydrolysis.The structural biology of ABC transporters has been exploding for the last few years,and this study explores timelines and trends for various attributes such as structural tools,resolution,fold,sources,and group leaders.This study also evidences the significance of mammalian expression systems,advancements in structural biology tools,and the developing interest of group leaders across the world in the remarkably progressing field.The field started in 2002 and bloomed in 2016,and COVID years were really productive to the field.Specifically,the study explores 337 structures of 58 unique ABC transporters deposited in the PDB database from which P-glycoprotein has the largest number of structures.Approximately,62%of total structures are determined at the resolution of 3-4Åand 53%of structures belong to fold IV type.With progressive advancements in the field,the field is shifting from prokaryotic to eukaryotic sources and X-ray crystallography to cryoelectron microscopy.In the nutshell,this study uniquely provides the detailed snapshot of the field of structural biology of ABC transporters with real-time data.展开更多
The pathogenesis of non-communicable diseases(NCDs)worldwide is closely linked to the global nutrition transition.Functional foods play a crucial role in the prevention and control of NCDs,making them an active area o...The pathogenesis of non-communicable diseases(NCDs)worldwide is closely linked to the global nutrition transition.Functional foods play a crucial role in the prevention and control of NCDs,making them an active area of research.Fermentation,which involves the biotransformation of food,enhances its digestibility and nutritional properties by releasing bioactive molecules.The increased bioactivity during fermentation can be attributed to the liberation of compounds trapped in the food matrix,the generation of metabolites,or the metabolic products of the microorganisms involved.Additionally,fermented foods can serve as a vehicle to deliver live beneficial microbes to the gastrointestinal tract,promoting gut homeostasis.While most studies demonstrate an increase in bioactivity during fermentation,some investigations yield contradictory results,likely due to the complexity of the food matrix,microbial strains utilized,and environmental conditions during the fermentation process.Further research is needed to address conflicting findings,and epidemiological studies are recommended to examine the impact of fermented vegetables on human health.This review discusses changes in antioxidant,antidiabetic,antihyperlipidemic,anticancer,and antihypertensive activities of fermented vegetables,both in vitro and in vivo using animal models.Moreover,the drawbacks associated with vegetable fermentation,their management,and the future prospects of vegetable fermentation are also discussed.展开更多
基金supported by the Thailand Science Research and Innovation Fund Chulalongkorn University(Grant No.HEAF67300078)the 90th Anniversary Scholarship,Chulalongkorn University Ratchadapisek Sompoch Fund(Grant No.Batch#55,T.Boonto)the Center of Excellence in Hepatitis and Liver Cancer,Faculty of Medicine,Chulalongkorn University.T.Boonto was supported by the scholarship from the Graduate School,Chulalongkorn University,to commemorate the 72^(nd) anniversary of His Majesty King Bhumibol Adulyadej(Grant No.Batch#22).
文摘Hepatocellular carcinoma(HCC)remains the third leading cause of cancer-related deaths worldwide;however,its therapeutic options are limited.Understanding the molecular mechanisms of HCC could provide insight into new therapies.Emerging studies indicate the important role of long-noncoding RNAs(lncRNAs)in the pathogenesis of HCC.The expression of the well-studied lncRNA taurine upregulated gene 1(TUG1)is upregulated in HCC tissues,but its transcriptomic effects in HCC cells remain unexplored.We established TUG1-knockdown and control HCC cells for RNA-seq experiments.KEGG analysis revealed glycolysis as the top enriched pathway upon TUG1 silencing.Accordingly,TUG1-depleted HCC cells showed impairments in glucose uptake,ATP synthesis,and lactate production.Clinical HCC tissue data revealed positive gene expression correlations between TUG1 and several glycolysis-related genes.To identify a molecular function of TUG1 in glycolysis,we explored the competing endogenous model and used bioinformatic tools to find the five microRNAs(miRNAs)that had the most binding sites for TUG1.Among these miRNAs,miR-122-5p exhibited an inverse correlation in gene expression with most TUG1-regulated glycolysis genes,including PKM,ALDOA,ENO2,and PFKM.Dual-luciferase assays demonstrated the direct interaction between TUG1 and miR-122-5p and between miR-122-5p and the 3ʹuntranslated regions of both PKM and ALDOA.We further showed that inhibition of miR-122-5p alleviated the suppression of glycolysis induced by TUG1 depletion.Together,our RNA-seq analysis of TUG1-depleted HCC cells,combined with clinical data,reveals a critical role of TUG1 in regulating glycolysis and provides new insight into its oncogenic function in HCC.
基金financially supported by Thailand Science Research and Innovation Fund Chulalongkorn University(CU_FRB65_hea(46)_053_30_34)Ratchadapiseksompotch Fund,Faculty of Medicine,Chulalongkorn University(Grant No.RA 66/017)+1 种基金Thailand Research Fund(TRF)Senior Research Scholar(Grant No.RTA6280004)the Center of Excellence in Hepatitis and Liver Cancer,Faculty of Medicine,Chulalongkorn University.
文摘Hepatocellular carcinoma (HCC) is the leading cause of cancer death worldwide;nevertheless, currenttherapeutic options are limited or ineffective for many patients. Therefore, elucidation of molecular mechanisms inHCC biology could yield important insights for the intervention of novel therapies. Recently, various studies havereported dysregulation of long non-coding RNAs (lncRNAs) in the initiation and progression of HCC, including H19;however, the biological function of H19 in HCC remains unclear. Here, we show that knockdown of H19 disruptedHCC cell growth, impaired the G1-to-S phase transition, and promoted apoptosis, while overexpression of H19yielded the opposite results. Screening for expression of cell cycle-related genes revealed a significant downregulationof CDK6 at both RNA and protein levels upon H19 suppression. Bioinformatic analysis of the H19 sequence and the3′ untranslated region (3′ UTR) of CDK6 transcripts showed several binding sites for microRNA-107 (miR-107), andthe dual luciferase reporter assay confirmed their direct interaction with miR-107. Consistently, blockage of miR-107activity alleviated the growth suppression phenotypes induced by H19 downregulation, suggesting that H19 serves asa molecular sponge for miR-107 to promote CDK6 expression and cell cycle progression. Together, this studydemonstrates a mechanistic function of H19 in driving the proliferation of HCC cells and suggests H19 suppressionas a novel approach for HCC treatment.
文摘Objective:To systematically map the stepwise events leading to deoxyelephantopin-induced cell death of HCT116 human colorectal cancer cells and evaluate the effectiveness of deoxyelephantopin in vivo.Methods:HCT116 cells were treated with deoxyelephantopin at various concentrations and time points.Autophagy was confirmed by the detection of autophagosomes and autophagosomal proteins by electron microscopy and Western blotting assays,respectively,and then validated by siRNA knockdown.In addition,apoptosis was confirmed by the detection of apoptosis-related proteins.The intracellular reactive oxygen species(ROS)level was measured using flow cytometry.The growth inhibitory effect of deoxyelephantopin was further evaluated in vivo using a mouse xenograft model.Results:Deoxyelephantopin firstly elevated ROS production,which then triggered autophagic flux with the accumulation of autophagosomal proteins including LC3 A/B,ATG5,and ATG7,followed by the induction of apoptosis via the intrinsic and extrinsic pathways.Pre-treatment with N-acetyl-L-cysteine,a ROS inhibitor,reversed both apoptosis and autophagy.The knockdown of LC3 prevented apoptosis induction which confirmed that deoxyelephantopin induced autophagy-dependent apoptosis in HCT116 cells.Accumulation of ROS also activated apoptosis via the mitogen-activated protein kinases signaling pathway.Furthermore,deoxyelephantopin also inhibited the PI3 K/AKT/mTOR pathway,which then released the inhibition of autophagy.In vivo study further showed that deoxyelephantopin significantly suppressed the growth of HCT116 subcutaneous xenograft in nude mice.Conclusions:Our findings revealed that deoxyelephantopin elevates oxidative stress and induces ROS-dependent autophagy followed by apoptosis in HCT116 cells via the concerted modulation of multiple signaling pathways.These findings further support the development of deoxyelephantopin as a therapeutic agent for colorectal cancer.
文摘Background: Continuous exposure to millimolar (mM) Vitamin C (AA) in vitro kills cancer cells. For superficial bladder carcinoma (SBC), standard chemotherapy is instillation of Bacillus Calmette-Guerin. The recurrence rate with this therapy is 91%. But high dosage vitamins including AA reduced the recurrence to 41%. Aim: To determine the oral dosage of AA that causes the highest concentration of AA [AA] in the bladder. Method: We conducted a clinical trial of 14 people who took various dosages of AA, and analyzed the [AA] in their urine. Results: AA above 2 g twice a day was not absorbed. But that intake produced a bladder [AA] above 1 mM in all participants. Conclusion: Taking 2 g of AA BID will increase [AA] in the bladder to a level likely to kill cancer cells that cause SBC. Taking that dosage 2 consecutive days a week is likely to reduce the recurrence rate of SBC substantially.
文摘Breast cancer is the second leading cause of cancerrelated deaths in women worldwide;a prime cancer biomarker to aid in the diagnosis, directed treatment, clinical management, and reoccurrence of this cancer is a MUC1 peptide fragment: cancer antigen 15-3 (CA 15-3). Herein, an immuno-fluorescence assay for CA 15-3 was developed;this ALYGNSA system consists of a protein biolinker (Protein G’) adsorbed onto Poly (methyl methacrylate) (PMMA). The unique interaction of Protein G’ with PMMA, a thermo-plastic polymer has been demonstrated to improve human IgG capture antibody alignment/ orientation and result in greater assay sensitivity. Indeed a previous report (HEALTH 1 325 - 329, 2009) on the shed extracellular domain of HER-2/neu revealed a 10-fold increase in sensitivity of the ALYGSNA assay over a control ELISA assay. Results from this ALYGNSA assay study revealed that a 16-fold increase in detection (≤0.94 U/mL) of CA 15-3 was found in comparison to a commercial control ELISA kit (≤15 U/mL). In conclusion, this enhanced sensitivity of the ALYGNSA assay for CA 15-3, may provide insights into the role/function of this biomarker in normal, as well as, breast cancer and other epithelial cancers.
基金Project supported by the National Basic Research Program of China(Grant Nos.2013CB921904,2009CB930504,and 2013CB328700)the National Natural Science Foundation of China(Grant Nos.11074016,11121091,10934001,61177020,11134001,and 10828407)
文摘Internal solvation of protein was studied by site-directed mutagenesis, with which an intrinsically fluorescent probe,tryptophan, is inserted into the desired position inside a protein molecule for ultrafast spectroscopic study. Here we review this unique method for protein dynamics research. We first introduce the frontiers of protein solvation, site-directed mutagenesis, protein stability and characteristics, and the spectroscopic methods. Then we present time-resolved spectroscopic dynamics of solvation dynamics inside cavities of active sites. The studies are carried out on a globular protein, staphylococcal nuclease. The solvation at sites inside the protein molecule＇s cavities clearly reveals characteristics of the local environment. These solvation behaviors are directly correlated to enzyme activity.
基金supported by the National Natural Science Foundation of China (31771621,31171071 and 31671093)the Research Foundation for Advanced Talents from Hangzhou Normal University and the New York State Stem Cell Science contracts C026429 and C030133。
文摘Noxious mechanical information is transmitted through molecularly distinct nociceptors,with pinprickevoked sharp sensitivity via A-fiber nociceptors marked by developmental expression of the neuropeptide Y receptor 2(Npy2 r)and von Frey filament-evoked punctate pressure information via unmyelinated C fiber nociceptors marked by MrgprD.However,the molecular programs controlling their development are only beginning to be understood.Here we demonstrate that Npy2 r-expressing sensory neurons are in fact divided into two groups,based on transient or persistent Npy2 r expression.Npy2 r-transient neurons are myelinated,likely including A-fiber nociceptors,whereas Npy2 r-persistent ones belong to unmyelinated pruriceptors that co-express Nppb.We then showed that the transcription factors NFIA and Runx1 are necessary for the development of Npy2 r-transient A-fiber nociceptors and MrgprD^+C-fiber nociceptors,respectively.Behaviorally,mice with conditional knockout of Nfia,but not Runx1 showed a marked attenuation of pinprick-evoked nocifensive responses.Our studies therefore identify a transcription factor controlling the development of myelinated nociceptors.
文摘C omprehensive identification of driver mutations in prostate cancer can serve to enhance our understanding of the disease and expand the use of available treatment options. Two recent and comple- mentary studies from Barbieri et al. and Grasso et al.
文摘As part of the determination of the possible impact of human activities on surface waters, case of Lake Sonfonia, six sampling sites were selected according to their solicitation by the population and their exposure to probable sources of pollution. The objective of this work is to monitor the physico-chemical quality of the waters of Lake Sonfonia during the low water level (March) and during the flood period (August) of the year 2021. Two sampling campaigns of water were carried out in dry weather and two others in cold weather. Parameters such as temperature, pH, EC (Electrical Conductivity), dissolved oxygen, TDS (Total Dissolved Solids) were measured in situ. Suspended matter, phosphate, nitrate, nitrite, sulphates, total iron, COD (Chemical Oxygen Demand) were measured in the laboratory by the colorimetric method. Stata 15 software was applied for the statistical analysis of the data and the correlation test between the parameters gave highly significant correlations. It has been noted that the situation is not very good and that this pollution comes mainly from human activities.
基金funded by multiple grants from the National Research Foundation of Korea(NRF),supported by the Ministry of Education,Science and Technology,including the Sejong Science Fellowship(2021R1C1C2095827,S.J.H.)grants 2020M3E5D9079742,2021M3A9G1015618,and RS-2022-NR070632 awarded to H.R+3 种基金Additional support was provided through the Korea Dementia Research Project(RS2023-KH137130 to H.R.)under the Korea Dementia Research Center(KDRC),funded by the Ministry of Health&Welfare and the Ministry of Science and ICTThis work was also partially supported by the Korea Institute of Science and Technology(KIST)through grants 2E33701,2E33722,2Z07276,and 2Z07368(H.R.)We further acknowledge support from the U.S.National Institutes of Health(NIH R01NS109537,J.L.)English language and stylistic editing were assisted by AI-based proofreading tools(ChatGPT,OpenAI).
文摘Huntington’s disease(HD)is a fatal neurodegenerative disorder characterized by a triad of behavioral symptoms:involuntary movement,emotional change,and cognitive dysfunction.Although alterations in WNT signaling have been reported in HD,its precise role in pathogenesis remains unclear.In this study,we found that astrocytic WNT5B mRNA and protein levels are elevated in the striatum of both HD patients and HD model mice.The noncanonical WNT5B signaling pathway induced sustained expression of matrix metallopeptidase 14(MMP14),an extracellular matrix(ECM)-degrading enzyme,via activation of the NFATc2 transcription factor in both human and primary mouse astrocytes.Robust upregulation of MMP14 led to ECM degradation,medium spiny neuron(MSN)damage,and increased mutant huntingtin aggregation in N171-82Q HD transgenic mice.Furthermore,WNT5B gain-offunction exacerbated neuropathology,impaired motor coordination,and shortened the lifespan of N171-82Q mice.We further demonstrated that the overexpression of the estrogen receptorα(ERα)suppresses NFATc2 transcriptional activity in vitro.A targeted therapy for the WNT5B-NFATc2-MMP14 signaling pathway by genistein,a phytoestrogen,reduced MMP14 transcription by antagonizing NFATc2 activity and preventing ECM degradation in N171-82Q mice.Genistein treatment also ameliorated neuropathology and motor deficits and prolonged the lifespan of HD mice.Together,these findings define a molecular pathological mechanism in which astrocytic MMP14 transcription,driven by the noncanonical WNT5B signaling pathway,promotes ECM degradation and MSN damage and accelerates neurodegeneration in HD.Modulation of the noncell-autonomous WNT5BNFATc2-MMP14 signaling pathway by genistein may serve as a potential therapeutic strategy for mitigating HD pathogenesis.
基金supported by the National Natural Science Foundation of China(31870728,31470738,and 32000611)the National Basic Research Program of China(2014CB910103)+1 种基金the Fundamental Research Funds for the Central Universities(2042020kfxg02)the China Postdoctoral Science Foundation(2018M642918)。
文摘Cytotoxic T cells targeting cancer neoantigens harboring driver mutations can lead to durable tumor regression in an HLAIdependent manner.However,it is difficult to extend the population of patients who are eligible for neoantigen-based immunotherapy,as immunogenic neoantigen-HLA pairs are rarely shared across different patients.Thus,a way to find other human leukocyte antigen(HLA)alleles that can also present a clinically effective neoantigen is needed.Recently,neoantigen-based immunotherapy targeting the KRAS G12D mutation in patients with HLA-C*08:02 has shown effectiveness.In a proof-ofconcept study,we proposed a combinatorial strategy(the combination of phylogenetic and structural analyses)to find potential HLA alleles that could also present KRAS G12D neoantigen.Compared to in silico binding prediction,this strategy avoids the uneven accuracy across different HLA alleles.Our findings extend the population of patients who are potentially eligible for immunotherapy targeting the KRAS G12D mutation.Additionally,we provide an alternative way to predict neoantigen-HLA pairs,which maximizes the clinical usage of shared neoantigens.
文摘Ovarian cancer(OC)is associated with poor outcomes and challenges scientists and clinicians.It is usually diagnosed in advanced stages when it is frequently aggressive,chemoresistant,and metastatic.The most prevalent form of OC is epithelial ovarian cancer(EOC),which displays significant heterogeneity,enhancing the difficulty in managing the disease.Several factors have been associated with the disease’s development and progression,especially those related to the tumor microenvironment(TME).Here,we highlight components of the ovarian TME in the disease development process,including pro-inflammatory pathways activated by interleukins,cytokines and chemokines,cancer-associated fibroblasts,tumor-associated macrophages,and epithelial-mesenchymal transition.We compiled evidence identifying TME factors promoting the development,chemoresistance,and metastasis,including cytokines,chemokines,growth factors,and tumor-associated cells.We identify potential targets for treatment and improving outcomes.These targets block or alter pathways associated with OC(especially EOC)progression.
基金the American Heart Association postdoctoral fellowship Award no.19POST34370101.
文摘ABC transporters are molecular machines which power the solute transport using ATP hydrolysis.The structural biology of ABC transporters has been exploding for the last few years,and this study explores timelines and trends for various attributes such as structural tools,resolution,fold,sources,and group leaders.This study also evidences the significance of mammalian expression systems,advancements in structural biology tools,and the developing interest of group leaders across the world in the remarkably progressing field.The field started in 2002 and bloomed in 2016,and COVID years were really productive to the field.Specifically,the study explores 337 structures of 58 unique ABC transporters deposited in the PDB database from which P-glycoprotein has the largest number of structures.Approximately,62%of total structures are determined at the resolution of 3-4Åand 53%of structures belong to fold IV type.With progressive advancements in the field,the field is shifting from prokaryotic to eukaryotic sources and X-ray crystallography to cryoelectron microscopy.In the nutshell,this study uniquely provides the detailed snapshot of the field of structural biology of ABC transporters with real-time data.
文摘The pathogenesis of non-communicable diseases(NCDs)worldwide is closely linked to the global nutrition transition.Functional foods play a crucial role in the prevention and control of NCDs,making them an active area of research.Fermentation,which involves the biotransformation of food,enhances its digestibility and nutritional properties by releasing bioactive molecules.The increased bioactivity during fermentation can be attributed to the liberation of compounds trapped in the food matrix,the generation of metabolites,or the metabolic products of the microorganisms involved.Additionally,fermented foods can serve as a vehicle to deliver live beneficial microbes to the gastrointestinal tract,promoting gut homeostasis.While most studies demonstrate an increase in bioactivity during fermentation,some investigations yield contradictory results,likely due to the complexity of the food matrix,microbial strains utilized,and environmental conditions during the fermentation process.Further research is needed to address conflicting findings,and epidemiological studies are recommended to examine the impact of fermented vegetables on human health.This review discusses changes in antioxidant,antidiabetic,antihyperlipidemic,anticancer,and antihypertensive activities of fermented vegetables,both in vitro and in vivo using animal models.Moreover,the drawbacks associated with vegetable fermentation,their management,and the future prospects of vegetable fermentation are also discussed.
