Next available on Tuesday 9 a.m.–5 p.m.

Additional Options

Search Results

240,710 results sorted by

  • Article

    A metabolically stable PET tracer for imaging synaptic vesicle protein 2A: synthesis and preclinical characterization of [18F]SDM-16

    Authors
    Zheng, Chao; Holden, Daniel; Zheng, Ming-Qiang; Pracitto, Richard; Wilcox, Kyle C.; Lindemann, Marcel; Felchner, Zachary; Zhang, Li; Tong, Jie; Fowles, Krista; Finnema, Sjoerd J.; Nabulsi, Nabeel; Carson, Richard E.; Huang, Yiyun; Cai, Zhengxin
    Part of
    European journal of nuclear medicine and molecular imaging, 2022-04, Vol.49 (5), p.1482-1496
    Abstract
    • Purpose To quantify the synaptic vesicle glycoprotein 2A (SV2A) changes in the whole central nervous system (CNS) under pathophysiological conditions, a high affinity SV2A PET radiotracer with improved in vivo stability is desirable to minimize the potential confounding effect of radiometabolites. The aim of this study was to develop such a PET tracer based on the molecular scaffold of UCB-A, and evaluate its pharmacokinetics, in vivo stability, specific binding, and nonspecific binding signals in nonhuman primate brains, in comparison with [ 11 C]UCB-A, [ 11 C]UCB-J, and [ 18 F]SynVesT-1. Methods The racemic SDM-16 (4-(3,5-difluorophenyl)-1-((2-methyl-1 H -imidazol-1-yl)methyl)pyrrolidin-2-one) and its two enantiomers were synthesized and assayed for in vitro binding affinities to human SV2A. We synthesized the enantiopure [ 18 F]SDM-16 using the corresponding enantiopure arylstannane precursor. Nonhuman primate brain PET scans were performed on FOCUS 220 scanners. Arterial blood was drawn for the measurement of plasma free fraction ( f P ), radiometabolite analysis, and construction of the plasma input function. Regional time-activity curves (TACs) were fitted with the one-tissue compartment (1TC) model to obtain the volume of distribution ( V T ). Nondisplaceable binding potential ( BP ND ) was calculated using either the nondisplaceable volume of distribution ( V ND ) or the centrum semiovale (CS) as the reference region. Results SDM-16 was synthesized in 3 steps with 44% overall yield and has the highest affinity ( K i  = 0.9 nM) to human SV2A among all reported SV2A ligands. [ 18 F]SDM-16 was prepared in about 20% decay-corrected radiochemical yield within 90 min, with greater than 99% radiochemical and enantiomeric purity. This radiotracer displayed high specific binding in monkey brains and was metabolically more stable than the other SV2A PET tracers. The f P of [ 18 F]SDM-16 was 69%, which was higher than those of [ 11 C]UCB-J (46%), [ 18 F]SynVesT-1 (43%), [ 18 F]SynVesT-2 (41%), and [ 18 F]UCB-H (43%). The TACs were well described with the 1TC. The averaged test–retest variability (TRV) was 7 ± 3%, and averaged absolute TRV (aTRV) was 14 ± 7% for the analyzed brain regions. Conclusion We have successfully synthesized a novel SV2A PET tracer [ 18 F]SDM-16, which has the highest SV2A binding affinity and metabolical stability among published SV2A PET tracers. The [ 18 F]SDM-16 brain PET images showed superb contrast between gray matter and white matter. Moreover, [ 18 F]SDM-16 showed high specific and reversible binding in the NHP brains, allowing for the reliable and sensitive quantification of SV2A, and has potential applications in the visualization and quantification of SV2A beyond the brain.
    Subjects
    Affinity; Aminoacridines; Animals; Binding; Brain; Brain - diagnostic imaging; Brain - metabolism; Cardiology; Central nervous system; Enantiomers; Fluorine isotopes; Glycoproteins; Humans; Image contrast; Imaging; In vivo methods and tests; Medicine; Medicine & Public Health; Membrane Glycoproteins - metabolism; Neuroimaging; Neurology; Nuclear Medicine; Oncology; Original; Original Article; Orthopedics; Pharmacokinetics; Positron emission; Positron emission tomography; Positron-Emission Tomography - methods; Primates; Protein biosynthesis; Radioactive tracers; Radiochemical analysis; Radiology; Radiopharmaceuticals - chemistry; Stability analysis; Substantia alba; Substantia grisea; Synaptic Vesicles - metabolism; Synthesis; Tomography; Tracers
  • Article

    Microbiome recovery in adult females with uncomplicated urinary tract infections in a randomised phase 2A trial of the novel antibiotic gepotidacin (GSK140944)

    Authors
    Nuzzo, Andrea; Van Horn, Stephanie; Traini, Christopher; Perry, Caroline R; Dumont, Etienne F; Scangarella-Oman, Nicole E; Gardiner, David F; Brown, James R
    Part of
    BMC microbiology, 2021-06, Vol.21 (1), p.181-11, Article 181
    Abstract
    • With increasing concerns about the impact of frequent antibiotic usage on the human microbiome, it is important to characterize the potential for such effects in early antibiotic drug development clinical trials. In a randomised Phase 2a clinical trial study that evaluated the pharmacokinetics of repeated oral doses of gepotidacin, a first-in-chemical-class triazaacenaphthylene antibiotic with a distinct mechanism of action, in adult females with uncomplicated urinary tract infections for gepotidacin (GSK2140944) we evaluated the potential changes in microbiome composition across multiple time points and body-sites ( ClinicalTrials.gov : NCT03568942). Samples of gastrointestinal tract (GIT), pharyngeal cavity and vaginal microbiota were collected with consent from 22 patients at three time points relative to the gepotidacin dosing regimen; Day 1 (pre-dose), Day 5 (end of dosing) and Follow-up (Day 28 ± 3 days). Microbiota composition was determined by DNA sequencing of 16S rRNA gene variable region 4 amplicons. By Day 5, significant changes were observed in the microbiome diversity relative to pre-dose across the tested body-sites. However, by the Follow-up visit, microbiome diversity changes were reverted to compositions comparable to Day 1. The greatest range of microbiome changes by body-site were GIT followed by the pharyngeal cavity then vagina. In Follow-up visit samples we found no statistically significant occurrences of pathogenic taxa. Our findings suggest that gepotidacin alteration of the human microbiome after 5 days of dosing is temporary and rebound to pre-dosing states is evident within the first month post-treatment. We recommend that future antibiotic drug trials include similar exploratory investigations into the duration and context of microbiome modification and recovery. NCT03568942 . Registered 26 June 2018.
    Subjects
    Acenaphthenes - administration & dosage; Acenaphthenes - pharmacokinetics; Adult; Anti-Bacterial Agents - administration & dosage; Anti-Bacterial Agents - pharmacokinetics; Antibiotic; Antibiotics; Bacteria; Bacteria - classification; Bacteria - drug effects; Bacteria - genetics; Bacteria - isolation & purification; Bacterial infections; Biodiversity; Clinical trial; Clinical trials; Composition; Deoxyribonucleic acid; DNA; DNA sequencing; Dosage; Drug development; Drug therapy; Female; Females; Gastrointestinal system; Gastrointestinal tract; Gastrointestinal Tract - microbiology; Gepotidacin; Health aspects; Heterocyclic Compounds, 3-Ring - administration & dosage; Heterocyclic Compounds, 3-Ring - pharmacokinetics; Humans; Infections; Microbiome; Microbiomes; Microbiota; Microbiota (Symbiotic organisms); Microbiota - drug effects; Middle Aged; Pathogens; Patients; Pharmacokinetics; Pharynx; Pharynx - microbiology; Recovery; rRNA 16S; Statistical analysis; Statistical methods; Testing; Trends; Urinary tract; Urinary tract infection; Urinary tract infections; Urinary Tract Infections - drug therapy; Urinary Tract Infections - microbiology; Urogenital system; Vagina; Vagina - microbiology; Variable region; Women patients
  • Article

    A phase 2a randomised, double-blind, placebo-controlled, parallel-group, add-on clinical trial of ebselen (SPI-1005) as a novel treatment for mania or hypomania

    Authors
    Sharpley, Ann L; Williams, Clare; Holder, Adele A; Godlewska, Beata R; Singh, Nisha; Shanyinde, Milensu; MacDonald, Orla; Cowen, Philip J
    Part of
    PSYCHOPHARMACOLOGY, 2020-12, Vol.237 (12), p.3773-3782
    Abstract
    • Rationale Lithium is an effective prophylactic and anti-manic treatment in bipolar disorder; however, its use is declining through perceived poor tolerance and toxicity. Lithium inhibits inositol monophosphatase (IMPase), a probable key therapeutic mechanism. The anti-inflammatory drug, ebselen, also inhibits IMPase and appears well-tolerated and safe. Objectives To assess the efficacy of adjunctive ebselen in mania using the Young Mania Rating Scale (YMRS) (primary outcome) and the Altman Self-Rating Mania (ASRM) Scale and Clinical Global Impression-Severity Scale (CGI-S) among the secondary outcomes. Methods Randomised, double-blind, placebo-controlled, parallel-group trial conducted between October 2017 and June 2019, at Oxford Health NHS Foundation Trust. Pharmacy-controlled randomisation was computer-generated, with full allocation concealment. In/outpatients ( n  = 68) aged 18–70, experiencing mania or hypomania, were assigned to 3 weeks ebselen (600 mg bd) ( n  = 33) or placebo ( n  = 35). Participants received usual clinical care and psychotropic medication. Results Ebselen was numerically, but not statistically, superior to placebo in lowering scores on the YMRS (adjusted mean difference and 95% confidence interval, − 1.71 (− 5.34 to 1.91), p  = 0.35) and ASRM (− 1.36 (− 3.75 to 1.17), p  = 0.29). However, scores on the CGI-S were significantly lower at week 3 in ebselen-treated participants (adjusted mean difference, − 0.58 (− 1.14 to − 0.03), p  = 0.04). A post hoc analysis excluding patients taking concomitant valproate treatment magnified the difference between ebselen and placebo on the YMRS. Adverse events were comparable between groups, and mild. Conclusions Ebselen merits further investigation where concomitant psychotropic medication is better controlled and participants taking valproate are excluded. If effective, ebselen’s superior tolerance and safety could make it a useful alternative to lithium. Trial registration Trial Registry: www.clinicaltrials.gov , Identifier: NCT03013400.
    Subjects
    Adult; Affective disorders; Aged; Anti-inflammatory agents; Antimanic Agents - administration & dosage; Azoles - administration & dosage; Biomedical and Life Sciences; Biomedicine; Bipolar disorder; Bipolar Disorder - diagnosis; Bipolar Disorder - drug therapy; Bipolar Disorder - psychology; Care and treatment; Clinical trials; Double-Blind Method; Drug Therapy, Combination; Female; Humans; Inflammation; Isoindoles; Life Sciences & Biomedicine; Lithium; Male; Mania; Mania - diagnosis; Mania - drug therapy; Mania - psychology; Middle Aged; myo-Inositol-1 (or 4)-monophosphatase; Neuroprotective Agents - administration & dosage; Neurosciences; Neurosciences & Neurology; Organoselenium Compounds - administration & dosage; Original Investigation; Pharmacology & Pharmacy; Pharmacology/Toxicology; Psychiatry; Psychotropic drugs; Science & Technology; Toxicity; Treatment Outcome; Valproic acid; Valproic Acid - administration & dosage; Young Adult
  • Article

    Synaptic Vesicle Glycoprotein 2A Ligands in the Treatment of Epilepsy and Beyond

    Authors
    Löscher, Wolfgang; Gillard, Michel; Sands, Zara A.; Kaminski, Rafal M.; Klitgaard, Henrik
    Part of
    CNS drugs, 2016-11, Vol.30 (11), p.1055-1077
    Abstract
    • The synaptic vesicle glycoprotein SV2A belongs to the major facilitator superfamily (MFS) of transporters and is an integral constituent of synaptic vesicle membranes. SV2A has been demonstrated to be involved in vesicle trafficking and exocytosis, processes crucial for neurotransmission. The anti-seizure drug levetiracetam was the first ligand to target SV2A and displays a broad spectrum of anti-seizure activity in various preclinical models. Several lines of preclinical and clinical evidence, including genetics and protein expression changes, support an important role of SV2A in epilepsy pathophysiology. While the functional consequences of SV2A ligand binding are not fully elucidated, studies suggest that subsequent SV2A conformational changes may contribute to seizure protection. Conversely, the recently discovered negative SV2A modulators, such as UCB0255, counteract the anti-seizure effect of levetiracetam and display procognitive properties in preclinical models. More broadly, dysfunction of SV2A may also be involved in Alzheimer’s disease and other types of cognitive impairment, suggesting potential novel therapies for levetiracetam and its congeners. Furthermore, emerging data indicate that there may be important roles for two other SV2 isoforms (SV2B and SV2C) in the pathogenesis of epilepsy, as well as other neurodegenerative diseases. Utilization of recently developed SV2A positron emission tomography ligands will strengthen and reinforce the pharmacological evidence that SV2A is a druggable target, and will provide a better understanding of its role in epilepsy and other neurological diseases, aiding in further defining the full therapeutic potential of SV2A modulation.
    Subjects
    Animals; Anticonvulsants - pharmacology; Anticonvulsants - therapeutic use; Brain research; Cognitive enhancement; Conflicts of interest; Disease; Drug dosages; Epilepsy; Epilepsy - drug therapy; Epilepsy - metabolism; Funding; Humans; Ligands; Medicine; Medicine & Public Health; Membrane Glycoproteins - metabolism; Neurology; Neurosciences; Open access; Pharmacotherapy; Psychiatry; Psychopharmacology; Review; Review Article; Rodents; Synaptic Vesicles - metabolism
  • Article

    Safety, tolerability and efficacy of the glutaminyl cyclase inhibitor PQ912 in Alzheimer's disease: results of a randomized, double-blind, placebo-controlled phase 2a study

    Authors
    Scheltens, Philip; Hallikainen, Merja; Grimmer, Timo; Duning, Thomas; Gouw, Alida A; Teunissen, Charlotte E; Wink, Alle Meije; Maruff, Paul; Harrison, John; van Baal, Caroline M; Bruins, Suzanne; Lues, Inge; Prins, Niels D
    Part of
    Alzheimer's research & therapy, 2018-10, Vol.10 (1), p.107-14, Article 107
    Abstract
    • PQ912 is an inhibitor of the glutaminyl cyclase enzyme that plays a central role in the formation of synaptotoxic pyroglutamate-A-beta oligomers. We report on the first clinical study with PQ912 in subjects with biomarker-proven Alzheimer's disease (AD). The aim was to determine the maximal tolerated dose, target occupancy and treatment-related pharmacodynamic effects. The exploratory efficacy readouts selected were tailored to the patient population with early AD. The therapeutic approach focuses on synaptic dysfunction as captured by various measures such as electroencephalography (EEG), synaptic biomarkers and sensitive cognitive tests. This was a randomized, double-blind, placebo-controlled trial evaluating the safety, tolerability and efficacy of PQ912 800 mg twice daily (bid) for 12 weeks in subjects with mild cognitive impairment or mild dementia due to AD. The 120 enrolled subjects were treatment-naïve at the start of the study, had confirmed AD biomarkers in their cerebrospinal fluid at screening and had a Mini Mental State Examination score between 21 and 30. After 1 week of treatment with 400 mg bid, patients were up-titrated to 800 mg bid for 11 weeks. Patients were randomized 1:1 to either PQ912 or placebo. The primary composite endpoints were to assess safety and tolerability based on the number of patients who discontinued due to (serious) adverse events (safety), and based on dose adjustment during the treatment period and/or nonadherence to randomized treatment (tolerability). All randomized subjects who took at least one dose of the study treatment or placebo were used for safety analyses. There was no significant difference between treatments in the number of subjects with (serious) adverse events, although there were slightly more patients with a serious adverse event in the PQ912 group compared to placebo. More subjects treated with PQ912 discontinued treatment due to adverse events, mostly related to gastrointestinal and skin/subcutaneous tissue disorders. PQ912 treatment resulted in a significant reduction in glutaminyl cyclase activity, which resulted in an average target occupancy of > 90%. A significant reduction of theta power in the EEG frequency analysis and a significant improvement in the One Back test of our Neuropsychological Test Battery was observed. The exploratory biomarker readouts, neurogranin for synaptic toxicity and YKL-40 as a marker of inflammation, appear to be sensitive enough to serve as efficacy markers in the next phase 2b study. The maximal tolerated dose of PQ912 has been identified and the results support future studies at still lower doses reaching > 50% target occupancy, a longer up-titration phase to potentially induce adaptation and longer treatment periods to confirm the early signals of efficacy as seen in this study. Clinicaltrials.gov, NCT 02389413 . Registered on 17 March 2015.
    Subjects
    Aging; Alzheimer's disease; Alzheimers disease; Animal cognition; Biomarkers; Brain research; Cognitive ability; Dementia; Diagnosis; Electroencephalography; Enzymes; Glutaminyl cyclase inhibitor; Multivariate analysis; Neurotoxicity; Phase 2a; PQ912; Risk factors; Systematic review
  • Article

    Combined RNA interference and gene replacement therapy targeting MFN2 as proof of principle for the treatment of Charcot–Marie–Tooth type 2A

    Authors
    Rizzo, Federica; Bono, Silvia; Ruepp, Marc David; Salani, Sabrina; Ottoboni, Linda; Abati, Elena; Melzi, Valentina; Cordiglieri, Chiara; Pagliarani, Serena; De Gioia, Roberta; Anastasia, Alessia; Taiana, Michela; Garbellini, Manuela; Lodato, Simona; Kunderfranco, Paolo; Cazzato, Daniele; Cartelli, Daniele; Lonati, Caterina; Bresolin, Nereo; Comi, Giacomo; Nizzardo, Monica; Corti, Stefania
    Part of
    Cellular and molecular life sciences : CMLS, 2023-12, Vol.80 (12), p.373-373, Article 373
    Abstract
    • Mitofusin-2 (MFN2) is an outer mitochondrial membrane protein essential for mitochondrial networking in most cells. Autosomal dominant mutations in the MFN2 gene cause Charcot–Marie–Tooth type 2A disease (CMT2A), a severe and disabling sensory-motor neuropathy that impacts the entire nervous system. Here, we propose a novel therapeutic strategy tailored to correcting the root genetic defect of CMT2A. Though mutant and wild-type MFN2 mRNA are inhibited by RNA interference (RNAi), the wild-type protein is restored by overexpressing cDNA encoding functional MFN2 modified to be resistant to RNAi. We tested this strategy in CMT2A patient-specific human induced pluripotent stem cell (iPSC)-differentiated motor neurons (MNs), demonstrating the correct silencing of endogenous MFN2 and replacement with an exogenous copy of the functional wild-type gene. This approach significantly rescues the CMT2A MN phenotype in vitro, stabilizing the altered axonal mitochondrial distribution and correcting abnormal mitophagic processes. The MFN2 molecular correction was also properly confirmed in vivo in the MitoCharc1 CMT2A transgenic mouse model after cerebrospinal fluid (CSF) delivery of the constructs into newborn mice using adeno-associated virus 9 (AAV9). Altogether, our data support the feasibility of a combined RNAi and gene therapy strategy for treating the broad spectrum of human diseases associated with MFN2 mutations.
    Subjects
    Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; Cell Biology; Cell differentiation; Cerebrospinal fluid; Charcot-Marie-Tooth disease; Charcot-Marie-Tooth Disease - genetics; Charcot-Marie-Tooth Disease - metabolism; Charcot-Marie-Tooth Disease - therapy; Gene expression; Gene therapy; GTP Phosphohydrolases - genetics; GTP Phosphohydrolases - metabolism; Humans; Hydrolases - genetics; Induced Pluripotent Stem Cells - metabolism; Interference; Life Sciences; Membrane proteins; Mice; Mice, Transgenic; Mitochondria; Mitochondrial Proteins - genetics; Mitochondrial Proteins - metabolism; Motor neurons; mRNA; Mutation; Nervous system; Neuropathy; Original Article; Phenotypes; Pluripotency; Proteins; Ribonucleic acid; RNA; RNA Interference; RNA-mediated interference; Stem cells; Transgenic mice; Viruses
  • Article

    Protein phosphatase 2A-B55δ enhances chemotherapy sensitivity of human hepatocellular carcinoma under the regulation of microRNA-133b

    Authors
    Zhuang, Qunying; Zhou, Tengjian; He, Chengyong; Zhang, Shili; Qiu, Yang; Luo, Bing; Zhao, Ran; Liu, Hengchuan; Lin, Yuchun; Lin, Zhongning
    Part of
    Journal of experimental & clinical cancer research, 2016-04, Vol.35 (67), p.67-67, Article 67
    Abstract
    • Hepatocellular carcinoma (HCC) remains a major public health problem worldwide. The identification of effective chemotherapeutic targets for advanced HCC patients is urgently required. In this study, we investigated the role of protein phosphatase 2A-B55δ subunit (PP2A-B55δ, encoded by the PPP2R2D gene) and related mechanisms affecting chemotherapy sensitivity of HCC. Experimental approaches for measuring the levels of PPP2R2D mRNA and B55δ protein in HCC included bioinformatics analyses, quantitative real-time polymerase chain reaction (qRT-PCR), western blotting (WB), immunofluorescence and immunohistochemistry assays. Cell cycle, migration, colony formation, apoptosis, and cell proliferation assays in stable PPP2R2D-knockdown and -overexpression cell lines in vitro, and tumorigenicity assays in vivo, were performed to explore the function of B55δ in cisplatin (cDDP) chemotherapy of HCC. Bioinformatics prediction, luciferase reporter assays, qRT-PCR, WB, and cell cycle analyses were used to reveal the regulatory relationship between microRNA-133b (miR-133b) and PPP2R2D expression. miR-133b mimic and inhibitor were used to elucidate the regulatory mechanism. Our studies showed that PPP2R2D expression was down-regulated in both HCC tumors and HCC cell lines. Treatment with cDDP increased the amount of B55δ protein. Artificially increasing the expression of B55δ counteracted cyclin-dependent kinase 1 activation, modulated transitions of the cell cycle, and increased the suppressive effect of cDDP on cell migration, colony formation, apoptosis, and proliferation in vitro and tumor growth in vivo, thus enhancing therapeutic efficiency. In contrast, knockdown of B55δ partially inhibited the effect of cDDP chemotherapy. miR-133b was shown to regulate PPP2R2D expression by binding to the 3'-untranslated region of PPP2R2D mRNA. The miR-133b/PPP2R2D signaling pathway affects the effectiveness of cDDP chemotherapy. PP2A-B55δ, regulated by miR-133b, enhances the sensitivity of HCC to cDDP chemotherapy. Our data indicate that PP2A-B55δ might be a novel and attractive target for increasing chemotherapy sensitivity of HCC.
    Subjects
    Animals; Antineoplastic Agents - administration & dosage; Antineoplastic Agents - pharmacology; Cancer; Carcinoma, Hepatocellular - drug therapy; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - metabolism; Care and treatment; Cell Cycle - drug effects; Cell Line, Tumor; Cell Movement - drug effects; Chemotherapy; Cisplatin - administration & dosage; Cisplatin - pharmacology; Complications and side effects; Gene Expression Regulation, Neoplastic - drug effects; Health aspects; Hep G2 Cells; Hepatoma; Humans; Influence; Liver Neoplasms - drug therapy; Liver Neoplasms - genetics; Mice; MicroRNA; MicroRNAs - genetics; Protein Phosphatase 2 - genetics; Protein Phosphatase 2 - metabolism; Xenograft Model Antitumor Assays
  • Article

    Boosting expression level of plectasin in recombinant Pichia pastoris via 2A self-processing peptide assembly

    Authors
    Liang, Xingxing; Jiang, Hong; Si, Xiandong; Xin, Qi; Meng, Di; Chen, Peng; Mao, Xiangzhao
    Part of
    Applied microbiology and biotechnology, 2022-05, Vol.106 (9-10), p.3669-3678
    Abstract
    • Plectasin is a promising and potent antimicrobial peptide isolated from the fungus Pseudoplectania nigrella which has been heterologously expressed in various hosts. In this study, a four-copy cassette of plectasin was constructed via 2A peptide assembly to further increase its expression level in recombinant Pichia pastoris . The yeast transformant 4Ple-61 harboring four-copy cassette of plectasin could secrete 183.2 mg/L total protein containing 60.8% of plectasin at the flask level within 120 h, which was 2.3 times higher than that of the yeast transformant Ple-6 carrying one-copy cassette of plectasin. Western blot confirmed the significant peptide expression level in the transformant 4Ple-61. Furthermore, it yielded as high as 426.3 mg/L total protein within 120 h during a 5-L fermentation. The purified plectasin shows superior stability and good antimicrobial activity against conventional Staphylococcus aureus ATCC 26,001 and some food-borne antibiotic-resistant S. aureus strains with the MICs ranging from 8 to 32 μg/mL. Therefore, the strategy based on 2A peptide assembly can enhance the expression of plectasin and further expand its application prospect. Key points
      • A yeast transformant 4Ple-61 with four-copy cassette of plectasin was constructed.
      • The plectasin level yield by the transformant 4Ple-61 was boosted by 2.3 times.
      • The plectasin showed good activity against food-borne antibiotic-resistant S. aureus.
    Subjects
    Antibiotic resistance; Antibiotics; Antiinfectives and antibacterials; Antimicrobial activity; Antimicrobial agents; Antimicrobial peptides; Assembly; Biomedical and Life Sciences; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; Drug resistance; Fermentation; Food; Gene expression; Genetic aspects; Life Sciences; Microbial Genetics and Genomics; Microbiology; Peptides; Physiological aspects; Pichia pastoris; Proteins; Staphylococcus aureus; Yeast; Yeast fungi; Yeasts
  • Article

    Tamoxifen induces apoptosis through cancerous inhibitor of protein phosphatase 2A–dependent phospho-Akt inactivation in estrogen receptor–negative human breast cancer cells

    Authors
    Liu, Chun-Yu; Hung, Man-Hsin; Wang, Duen-Shian; Chu, Pei-Yi; Su, Jung-Chen; Teng, Tsung-Han; Huang, Chun-Teng; Chao, Ting-Ting; Wang, Cheng-Yi; Shiau, Chung-Wai; Tseng, Ling-Ming; Chen, Kuen-Feng
    Part of
    Breast Cancer Research, 2014-09, Vol.16 (5), p.431-431, Article 431
    Abstract
    • Tamoxifen, a selective estrogen receptor (ER) modulator, may affect cancer cell survival through mechanisms other than ER antagonism. In the present study, we tested the efficacy of tamoxifen in a panel of ER-negative breast cancer cell lines and examined the drug mechanism. In total, five ER-negative breast cancer cell lines (HCC-1937, MDA-MB-231, MDA-MB-468, MDA-MB-453 and SK-BR-3) were used for in vitro studies. Cellular apoptosis was examined by flow cytometry and Western blot analysis. Signal transduction pathways in cells were assessed by Western blot analysis. The in vivo efficacy of tamoxifen was tested in xenograft nude mice. Tamoxifen induced significant apoptosis in MDA-MB-231, MDA-MB-468, MDA-MB-453 and SK-BR-3 cells, but not in HCC-1937 cells. Tamoxifen-induced apoptosis was associated with inhibition of cancerous inhibitor of protein phosphatase 2A (CIP2A) and phospho-Akt (p-Akt) in a dose-dependent manner. Ectopic expression of either CIP2A or Akt protected MDA-MB-231 cells from tamoxifen-induced apoptosis. In addition, tamoxifen increased protein phosphatase 2A (PP2A) activity, and tamoxifen-induced apoptosis was attenuated by the PP2A antagonist okadaic acid in the sensitive cell lines, but not in resistant HCC-1937 cells. Moreover, silencing CIP2A by small interfering RNA sensitized HCC-1937 cells to tamoxifen-induced apoptosis. Furthermore, tamoxifen regulated CIP2A protein expression by downregulating CIP2A mRNA. Importantly, tamoxifen inhibited the in vivo growth of MDA-MB-468 xenograft tumors in association with CIP2A downregulation, whereas tamoxifen had no significant effect on CIP2A expression and anti-tumor growth in HCC-1937 tumors. Inhibition of CIP2A determines the effects of tamoxifen-induced apoptosis in ER-negative breast cancer cells. Our data suggest a novel "off-target" mechanism of tamoxifen and suggest that CIP2A/PP2A/p-Akt signaling may be a feasible anti-cancer pathway.
    Subjects
    Aged; Analysis; Animals; Antineoplastic Agents, Hormonal - pharmacology; Apoptosis - drug effects; Autoantigens - genetics; Autoantigens - metabolism; Breast Neoplasms - drug therapy; Breast Neoplasms - metabolism; Breast Neoplasms - mortality; Breast Neoplasms - pathology; Cancer; Cell Line, Tumor; Down-Regulation; Female; Humans; Kaplan-Meier Estimate; Male; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice, Nude; Middle Aged; Oncology, Experimental; Physiological aspects; Proto-Oncogene Proteins c-akt - metabolism; Receptors, Estrogen - metabolism; Tamoxifen - pharmacology; Transcription, Genetic; Tumor Burden - drug effects; Xenograft Model Antitumor Assays
  • Article

    Protein phosphatase 2A to lung cancer

    Authors
    Nader, Cassandra P; Cidem, Aylin; Verrills, Nicole M; Ammit, Alaina J
    Part of
    Respiratory research, 2019-10, Vol.20 (1)
    Abstract
    • Lung cancer (LC) has the highest relative risk of development as a comorbidity of chronic obstructive pulmonary disease (COPD). The molecular mechanisms that mediate chronic inflammation and lung function impairment in COPD have been identified in LC. This suggests the two diseases are more linked than once thought. Emerging data in relation to a key phosphatase, protein phosphatase 2A (PP2A), and its regulatory role in inflammatory and tumour suppression in both disease settings suggests that it may be critical in the progression of COPD to LC. In this review, we uncover the importance of the functional and active PP2A holoenzyme in the context of both diseases. We describe PP2A inactivation via direct and indirect means and explore the actions of two key PP2A endogenous inhibitors, cancerous inhibitor of PP2A (CIP2A) and inhibitor 2 of PP2A (SET), and the role they play in COPD and LC. We explain how dysregulation of PP2A in COPD creates a favourable inflammatory micro-environment and promotes the initiation and progression of tumour pathogenesis. Finally, we highlight PP2A as a druggable target in the treatment of COPD and LC and demonstrate the potential of PP2A re-activation as a strategy to halt COPD disease progression to LC. Although further studies are required to elucidate if PP2A activity in COPD is a causal link for LC progression, studies focused on the potential of PP2A reactivating agents to reduce the risk of LC formation in COPD patients will be pivotal in improving clinical outcomes for both COPD and LC patients in the future. Keywords: CIP2A, COPD, Inflammation, Lung Cancer, PP2A, SET
    Subjects
    Chronic obstructive lung disease; Comorbidity; Development and progression; Enzymes; Inflammation; Lung cancer; Lung diseases; Phosphatases; Respiratory system agents; Risk factors; Tumors
  • Article

    The neuropharmacology of sleep paralysis hallucinations: serotonin 2A activation and a novel therapeutic drug

    Author
    Jalal, Baland
    Part of
    Psychopharmacology, 2018-11, Vol.235 (11), p.3083-3091
    Abstract
    • Sleep paralysis is a state of involuntary immobility occurring at sleep onset or offset, often accompanied by uncanny “ghost-like” hallucinations and extreme fear reactions. I provide here a neuropharmacological account for these hallucinatory experiences by evoking the role of the serotonin 2A receptor (5-HT 2A R). Research has shown that 5-HT 2A R activation can induce visual hallucinations, “mystical” subjective states, and out-of-body experiences (OBEs), and modulate fear circuits. Hallucinatory experiences triggered by serotonin—serotonergic (“pseudo”) hallucinations, induced by hallucinogenic drugs—tend to be “dream-like” with the experiencer having insight (“meta-awareness”) that he is hallucinating, unlike dopaminergic (“psychotic” and “life-like”) hallucinations where such insight is lost. Indeed, hallucinatory experiences during sleep paralysis have the classic features of serotonergic hallucinations, and are strikingly similar to perceptual and subjective states induced by hallucinogenic drugs (e.g., lysergic acid diethylamide [LSD] and psilocybin), i.e., they entail visual hallucinations, mystical experiences, OBEs, and extreme fear reactions. I propose a possible mechanism whereby serotonin could be functionally implicated in generating sleep paralysis hallucinations and fear reactions through 5-HT 2A R activity. Moreover, I speculate on the role of 5-HT 2C receptors vis-à-vis anxiety and panic during sleep paralysis, and the orbitofrontal cortex—rich with 5-HT 2A receptors—in influencing visual pathways during sleep paralysis, and, in effect, hallucinations. Finally, I propose, for the first time, a drug to target sleep paralysis hallucinations and fear reactions, namely the selective 5-HT 2A R inverse agonist, pimavanserin. This account implicates gene HTR2A on chromosome 13q as the underlying cause of sleep paralysis hallucinations and could be explored using positron emission tomography.
    Subjects
    Activation; Animals; Anxiety; Biomedical and Life Sciences; Biomedicine; Chromosome 13; Dopamine; Dopamine - metabolism; Dopamine - pharmacology; Dopamine receptors; Drug therapy; Fear; Hallucinations; Hallucinations - chemically induced; Hallucinations - drug therapy; Hallucinations - metabolism; Hallucinogens - metabolism; Hallucinogens - pharmacology; Health aspects; Humans; LSD; Lysergic acid diethylamide; Lysergic Acid Diethylamide - adverse effects; Lysergic Acid Diethylamide - metabolism; Lysergic Acid Diethylamide - pharmacology; Lysergide; Neuropharmacology; Neurosciences; Paralysis; Pharmacology/Toxicology; Piperidines - metabolism; Piperidines - pharmacology; Piperidines - therapeutic use; Positron emission tomography; Psilocybin; Psilocybin - metabolism; Psilocybin - pharmacology; Psychiatry; Receptor, Serotonin, 5-HT2A - metabolism; Serotonin; Serotonin - metabolism; Serotonin - pharmacology; Serotonin 5-HT2 Receptor Agonists - adverse effects; Serotonin 5-HT2 Receptor Agonists - metabolism; Serotonin 5-HT2 Receptor Agonists - pharmacology; Serotonin 5-HT2 Receptor Antagonists - metabolism; Serotonin 5-HT2 Receptor Antagonists - pharmacology; Serotonin 5-HT2 Receptor Antagonists - therapeutic use; Serotonin receptors; Serotonin S2 receptors; Sleep; Sleep - drug effects; Sleep - physiology; Sleep Paralysis - chemically induced; Sleep Paralysis - drug therapy; Sleep Paralysis - metabolism; Theoretical and Methodological; Theoretical and Methodological Perspective; Urea - analogs & derivatives; Urea - metabolism; Urea - pharmacology; Urea - therapeutic use; Visual cortex; Visual pathways
  • Article

    P53, Somatostatin receptor 2a and Chromogranin A immunostaining as prognostic markers in high grade gastroenteropancreatic neuroendocrine neoplasms

    Authors
    Nielsen, Kirstine; Binderup, Tina; Langer, Seppo W.; Kjaer, Andreas; Knigge, Pauline; Grondahl, Veronica; Melchior, Linea; Federspiel, Birgitte; Knigge, Ulrich
    Part of
    BMC Cancer, 2020-01, Vol.20 (1), p.27--27, Article 27
    Abstract
    • AbstractBackgroundHigh grade gastroenteropancreatic (GEP) neuroendocrine neoplasms (NEN) with a Ki67 proliferation index > 20%, include well-differentiated tumours grade 3 (NET G3) and poorly differentiated (PD) neuroendocrine carcinomas (NEC). Abnormal p53-expression is a feature of PD tumours, while expression of chromogranin A (CgA) and somatostatin-receptor 2a (SSTR-2a) may be a feature of well-differentiated tumours. The aim of this study was to elucidate the expression and prognostic value of these three markers in 163 GEP-NEN patients with a Ki67-index > 20%.MethodClinical data, histopathology and overall survival were analysed according to Kaplan-Meier’s method and Cox regression. The expression of SSTR-2a, CgA and synaptophysin was analysed in tumour specimens by immunohistochemistry, and semi-quantitatively scored as negative (< 5%), heterogeneously positive (5–30%) or strongly positive (> 30%). P53 was defined as normal when scored as heterogeneously positive (1–30%), and abnormal when negative (0%) or strongly positive (> 30%).ResultsIn multivariate analysis, better survival was observed among patients with heterogeneously positive p53 compared to strongly positive (p < 0.001). When dichotomised, tumours with a heterogeneously positive p53 vs. negative and strongly positive p53 also showed a significantly better survival (p = 0.002).Survival was significantly worse for negative CgA compared to heterogeneously positive CgA (p = 0.02). Strongly positive SSTR-2a expression was found in 26% of the 163 included patients. Well-differentiated morphology correlated with strong expression of SSTR-2a and CgA, and heterogeneously positive p53-staining, and was more frequent in pancreatic primaries. In pancreatic primaries, strongly positive SSTR-2a was associated with longer survival (univariate analysis, p = 0.02). A significantly lower Ki67 proliferation index was found in patients with a heterogeneously positive p53, a positive SSTR-2a and CgA expression.ConclusionOur results suggest that abnormal p53-expression is an independent negative prognostic marker in GEP-NEN with a Ki67-index > 20%. Patients with heterogeneously positive p53 had the best prognosis. SSTR-2a was a positive prognostic marker in pancreatic NEN. Negative CgA was associated with a significantly worse OS compared to heterogeneously positive CgA-expression in a multivariate sub-analysis. Lower Ki67 index correlated significantly with heterogeneously positive p53, positive SSTR-2a and CgA expression.
    Subjects
    Age; Analysis; Cancer; Cancer Research; Carcinoma; Chemotherapy; Chromogranin A; Classification; Gastroenteropancreatic neuroendocrine neoplasms; Genetics; Histochemistry; Immunohistochemistry; Life Sciences & Biomedicine; Medical prognosis; Metastasis; Morphology; Movement disorders; Multivariate analysis; Mutation; NEC; Neuroendocrine carcinomas; Neuroendocrine tumors; Oncology; p53; p53 Protein; Pancreas; Pathology; Patients; Prognosis; Science & Technology; Somatostatin; Somatostatin receptor 2a; Surgery; Survival; Synaptophysin; Tumor proteins; Tumors
  • Article

    Phase 1b/2a study of trastuzumab emtansine (T-DM1), paclitaxel, and pertuzumab in HER2-positive metastatic breast cancer

    Authors
    Krop, Ian E; Modi, Shanu; LoRusso, Patricia M; Pegram, Mark; Guardino, Ellie; Althaus, Betsy; Lu, Dan; Strasak, Alexander; Elias, Anthony
    Part of
    Breast Cancer Research, 2016-03, Vol.18 (1), p.34--34, Article 34
    Abstract
    • In pre-clinical studies, the anti-tumor activity of T-DM1 was enhanced when combined with taxanes or pertuzumab. This phase 1b/2a study evaluated the safety/tolerability of T-DM1 + paclitaxel ± pertuzumab in HER2-positive advanced breast cancer. In phase 1b (n = 60), a 3 + 3 dose-escalation approach was used to determine the maximum tolerated dose (MTD) of T-DM1 + paclitaxel ± pertuzumab. The primary objective of phase 2a was feasibility, with 44 patients randomized to T-DM1 + paclitaxel ± pertuzumab at the MTD identified in phase 1b. The MTD was T-DM1 3.6 mg/kg every three weeks (q3w) or 2.4 mg/kg weekly + paclitaxel 80 mg/m(2) weekly ± pertuzumab 840 mg loading dose followed by 420 mg q3w. Phase 2a patients had received a median of 5.0 (range: 0-10) prior therapies for advanced cancer. In phase 2a, 51.2 % received ≥12 paclitaxel doses within 15 weeks, and 14.0 % received 12 paclitaxel doses by week 12. Common all-grade adverse events (AEs) were peripheral neuropathy (90.9 %) and fatigue (79.5 %). A total of 77.3 % experienced grade ≥3 AEs, most commonly neutropenia (25.0 %) and peripheral neuropathy (18.2 %). Among the 42 phase 2a patients with measurable disease, the objective response rate (ORR) was 50.0 % (95 % confidence interval (CI) 34.6-65.4); the clinical benefit rate (CBR) was 56.8 % (95 % CI 41.6-71.0). No pharmacokinetic interactions were observed between T-DM1 and paclitaxel. This regimen showed clinical activity. Although there is potential for paclitaxel to be added to T-DM1 ± pertuzumab, peripheral neuropathy was common in this heavily pretreated population. ClinicalTrials.gov NCT00951665 . Registered August 3, 2009.
    Subjects
    Adult; Aged; Antibodies, Monoclonal, Humanized - administration & dosage; Antibodies, Monoclonal, Humanized - adverse effects; Antineoplastic Combined Chemotherapy Protocols - administration & dosage; Antineoplastic Combined Chemotherapy Protocols - adverse effects; Breast cancer; Breast Neoplasms - drug therapy; Breast Neoplasms - pathology; Care and treatment; Clinical trials; Complications and side effects; Female; Health aspects; Humans; Maximum Tolerated Dose; Maytansine - administration & dosage; Maytansine - adverse effects; Maytansine - analogs & derivatives; Metastasis; Middle Aged; Paclitaxel - administration & dosage; Paclitaxel - adverse effects; Risk factors; Trastuzumab
  • Article

    Metallothionein 2A (MT2A) controls cell proliferation and liver metastasis by controlling the MST1/LATS2/YAP1 signaling pathway in colorectal cancer

    Authors
    Liu, Xi; Quan, Jun; Shen, Zhaolong; Zhang, Zequn; Chen, Zhijian; Li, Liang; Li, Xiaorong; Hu, Gui; Deng, Xiaofeng
    Part of
    Cancer cell international, 2022-05, Vol.22 (1), p.205-205, Article 205
    Abstract
    • Colorectal cancer (CRC) is one of the three major cancers in the world and is the cancer with the most liver metastasis. The present study aimed to investigate the role of metallothionein 2A (MT2A) in the modulation of CRC cell proliferation and liver metastasis, as well as its molecular mechanisms. The expression profile of metallothionein 2A (MT2A) in colorectal cancer retrieved from TCGA, GEO and Oncomine database. The biological effect of MT2A overexpression was investigated mainly involving cell proliferation and migration in CRC cells as well as growth and metastasis in CRC animal models. To explore the specific mechanism of MT2A metastasis in CRC, transcriptome sequencing was used to compare the overall expression difference between the control group and the MT2A overexpression group. Metallothionein 2A (MT2A) was downregulated in the tumor tissues of patients with CRC compared to adjacent normal tissues and was related to the tumor M stage of patients. MT2A overexpression inhibited CRC cell proliferation and migration in cells, as well as growth and metastasis in CRC animal models. While knockdown of MT2A had the opposite effect in cells. Western blotting confirmed that MT2A overexpression promoted the phosphorylation of MST1, LAST2 and YAP1, thereby inhibiting the Hippo signaling pathway. Additionally, specific inhibitors of MST1/2 inhibited MT2A overexpression-mediated phosphorylation and relieved the inhibition of the Hippo signaling pathway, thus promoting cell proliferation. Immunohistochemistry in subcutaneous grafts and liver metastases further confirmed this result. Our results suggested that MT2A is involved in CRC growth and liver metastasis. Therefore, MT2A and MST1 may be potential therapeutic targets for patients with CRC, especially those with liver metastases.
    Subjects
    Allografts; Animal models; Cell growth; Cell migration; Cell proliferation; Colorectal cancer; Colorectal carcinoma; Esophageal cancer; Genes; Hepatocytes; Hippo signaling pathway; Immunohistochemistry; Liver; Liver cancer; Liver metastasis; Liver transplantation; Medical prognosis; Metallothionein; Metastases; Metastasis; Molecular modelling; MT2A; Mutation; Phosphorylation; Proteins; Signal transduction; Stomach cancer; Syngeneic grafts; Therapeutic targets; Transcriptomes; Tumors; Western blotting; Yes-associated protein
  • Article

    Cancerous Inhibitor of Protein Phosphatase 2A (CIP2A): Could It Be a Promising Biomarker and Therapeutic Target in Parkinson’s Disease?

    Authors
    Yin, Sijia; Han, Chao; Xia, Yun; Wan, Fang; Hu, Junjie; Kou, Liang; Sun, Yadi; Wu, Jiawei; Li, Yunna; Zhou, Qiulu; Xiong, Nian; Huang, Jinsha; Wang, Tao
    Part of
    Molecular neurobiology, 2022-02, Vol.59 (2), p.1333-1344
    Abstract
    • Parkinson’s disease (PD) is an incurable neurodegenerative disease characterized by aggregation of pathological alpha-synuclein (α-syn) and loss of dopaminergic neuron in the substantia nigra. Inhibition of phosphorylation of the α-syn has been shown to mediate alleviation of PD-related pathology. Protein phosphatase 2A (PP2A), an important serine/threonine phosphatase, plays an essential role in catalyzing dephosphorylation of the α-syn. Here, we identified and validated cancerous inhibitor of PP2A (CIP2A), as a potential diagnostic biomarker for PD. Our data showed that plasma CIP2A concentrations in PD patients were significantly lower compared to age- and sex-matched controls, 1.721 (1.435–2.428) ng/ml vs 3.051(2.36–5.475) ng/ml, p  < 0.0001. The area under the curve of the plasma CIP2A in distinguishing PD from the age- and sex-matched controls was 0.776. In addition, we evaluated the role of CIP2A in PD-related pathogenesis in PD cellular and MPTP-induced mouse model. The results demonstrated that CIP2A is upregulated in PD cellular and MPTP-induced mouse models. Besides, suppression of the CIP2A expression alleviates rotenone induced aggregation of the α-syn as well as phosphorylation of the α-syn in SH-SY5Y cells, which is associated with increased PP2A activity. Taken together, our data demonstrated that CIP2A plays an essential role in the mechanisms related to PD development and might be a novel PD biomarker.
    Subjects
    alpha-Synuclein - metabolism; Animal models; Animals; Autoantigens; Biomarkers; Biomarkers - metabolism; Biomedical and Life Sciences; Biomedicine; Cell Biology; Dephosphorylation; Dopamine receptors; Humans; Intracellular Signaling Peptides and Proteins; Membrane Proteins; Mice; Movement disorders; MPTP; Neurobiology; Neurodegenerative diseases; Neurodegenerative Diseases - metabolism; Neurology; Neurosciences; Parkinson Disease - diagnosis; Parkinson Disease - drug therapy; Parkinson Disease - metabolism; Parkinson's disease; Parkinsons disease; Phosphatase; Phosphoprotein phosphatase; Phosphorylation; Protein phosphatase; Protein Phosphatase 2 - metabolism; Protein-serine/threonine phosphatase; Rotenone; Substantia nigra; Substantia Nigra - pathology; Synuclein; Therapeutic targets; Threonine; Threonine phosphatase
  • Article

    Betulinic Acid Hydroxamate is Neuroprotective and Induces Protein Phosphatase 2A-Dependent HIF-1α Stabilization and Post-transcriptional Dephosphorylation of Prolyl Hydrolase 2

    Authors
    Prados, María E.; Correa-Sáez, Alejandro; Unciti-Broceta, Juan D.; Garrido-Rodríguez, Martín; Jimenez-Jimenez, Carla; Mazzone, Massimiliano; Minassi, Alberto; Appendino, Giovanni; Calzado, Marco A.; Muñoz, Eduardo
    Part of
    Neurotherapeutics, 2021-07, Vol.18 (3), p.1849-1861
    Abstract
    • Huntington’s disease (HD) is a neurodegenerative disorder characterized by unwanted choreatic movements, behavioral and psychiatric disturbances, and dementia. The activation of the hypoxic response pathway through the pharmacological inhibition of hypoxia-inducing factor (HIF) prolyl-hydroxylases (PHDs) is a promising approach for neurodegenerative diseases, including HD. Herein, we have studied the mechanism of action of the compound Betulinic acid hydroxamate (BAH), a hypoximimetic derivative of betulinic acid, and its efficacy against striatal neurodegeneration using complementary approaches. Firstly, we showed the molecular mechanisms through which BAH modifies the activity of the PHD2 prolyl hydroxylase, thus directly affecting HIF-1α stability. BAH treatment reduces PHD2 phosphorylation on Ser-125 residue, responsible for the control of its hydrolase activity. HIF activation by BAH is inhibited by okadaic acid and LB-100 indicating that a protein phosphatase 2A (PP2A) is implicated in the mechanism of action of BAH. Furthermore, in striatal cells bearing a mutated form of the huntingtin protein, BAH stabilized HIF-1α protein, induced Vegf and Bnip3 gene expression and protected against mitochondrial toxin-induced cytotoxicity. Pharmacokinetic analyses showed that BAH has a good brain penetrability and experiments performed in a mouse model of striatal neurodegeneration induced by 3-nitropropionic acid showed that BAH improved the clinical symptoms. In addition, BAH also prevented neuronal loss, decreased reactive astrogliosis and microglial activation, inhibited the upregulation of proinflammatory markers, and improved antioxidant defenses in the brain. Taken together, our results show BAH’s ability to activate the PP2A/PHD2/HIF pathway, which may have important implications in the treatment of HD and perhaps other neurodegenerative diseases.
    Subjects
    3-Nitropropionic acid; Acids; Animals; Antioxidants; Betulinic Acid; Biomedical and Life Sciences; Biomedicine; BNIP3 gene; BNIP3 protein; Corpus Striatum - drug effects; Corpus Striatum - metabolism; Corpus Striatum - pathology; Cytotoxicity; Dementia disorders; Dephosphorylation; Gene expression; Gliosis; HEK293 Cells; Humans; Huntingtin; Huntington's disease; Hydrolase; Hydroxylase; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Hypoxia-inducible factor 1a; Hypoxia-Inducible Factor-Proline Dioxygenases - metabolism; Inflammation; Mice; Mice, Inbred C57BL; Mitochondria; Molecular modelling; Neostriatum; Neurobiology; Neurodegeneration; Neurodegenerative diseases; Neurology; Neuroprotection; Neuroprotective Agents - pharmacology; Neurosciences; Neurosurgery; NIH 3T3 Cells; Nitro Compounds - toxicity; Original; Original Article; Pentacyclic Triterpenes - pharmacology; Pharmacokinetics; Phosphatase; Phosphorylation - drug effects; Phosphorylation - physiology; Propionates - toxicity; Protein Phosphatase 2 - metabolism; Proteins; Vascular endothelial growth factor
  • Article

    Loci on chromosomes 1A and 2A affect resistance to tan (yellow) spot in wheat populations not segregating for tsn1

    Authors
    Shankar, Manisha; Jorgensen, Dorthe; Taylor, Julian; Chalmers, Ken J.; Fox, Rebecca; Hollaway, Grant J.; Neate, Stephen M.; McLean, Mark S.; Vassos, Elysia; Golzar, Hossein; Loughman, Robert; Mather, Diane E.
    Part of
    Theoretical and applied genetics, 2017-12, Vol.130 (12), p.2637-2654
    Abstract
    • Key message QTL for tan spot resistance were mapped on wheat chromosomes 1A and 2A. Lines were developed with resistance alleles at these loci and at the tsn1 locus on chromosome 5B. These lines expressed significantly higher resistance than the parent with tsn1 only. Tan spot (syn. yellow spot and yellow leaf spot) caused by Pyrenophora tritici - repentis is an important foliar disease of wheat in Australia. Few resistance genes have been mapped in Australian germplasm and only one, known as tsn1 located on chromosome 5B, is known in Australian breeding programs. This gene confers insensitivity to the fungal effector ToxA. The main aim of this study was to map novel resistance loci in two populations: Calingiri/Wyalkatchem, which is fixed for the ToxA-insensitivity allele tsn1 , and IGW2574/Annuello, which is fixed for the ToxA-sensitivity allele Tsn1 . A second aim was to combine new loci with tsn1 to develop lines with improved resistance. Tan spot severity was evaluated at various growth stages and in multiple environments. Symptom severity traits exhibited quantitative variation. The most significant quantitative trait loci (QTL) were detected on chromosomes 2A and 1A. The QTL on 2A explained up to 29.2% of the genotypic variation in the Calingiri/Wyalkatchem population with the resistance allele contributed by Wyalkatchem. The QTL on 1A explained up to 28.1% of the genotypic variation in the IGW2574/Annuello population with the resistance allele contributed by Annuello. The resistance alleles at both QTL were successfully combined with tsn1 to develop lines that express significantly better resistance at both seedling and adult plant stages than Calingiri which has tsn1 only.
    Subjects
    Agriculture; Alleles; Ascomycota; Australia; Biochemistry; Biomedical and Life Sciences; Biotechnology; Breeding; Chromosome Mapping; Chromosomes; Chromosomes, Plant; Disease Resistance - genetics; Genes; Genotype & phenotype; Germplasm; Leafspot; Life Sciences; Linear Models; Models, Genetic; Original; Original Article; Phenotype; Plant Biochemistry; Plant breeding; Plant Breeding/Biotechnology; Plant Diseases - genetics; Plant Diseases - microbiology; Plant Genetics and Genomics; Population genetics; Quantitative genetics; Quantitative Trait Loci; Seedlings; Tan spot; Triticum; Triticum - genetics; Triticum - microbiology; Variation; Wheat; Yellow leaf; Yellow spot
  • Article

    The Role of Major Facilitator Superfamily Domain-Containing 2a in the Central Nervous System

    Authors
    He, Zhidong; Zhao, Yanan; Sun, Jing
    Part of
    Cellular and molecular neurobiology, 2023-03, Vol.43 (2), p.639-647
    Abstract
    • Major facilitator superfamily-domain containing 2a (Mfsd2a) is selectively expressed in vascular endotheliocytes and plays a crucial role in maintaining the integrity of the blood‒brain barrier and the transport of docosahexaenoic acid. It is currently recognized as the only molecule that inhibits endocytosis mediated by caveolae in brain endothelial cells. Mfsd2a gene knockout leads to an increase in the permeability of the blood–brain barrier from embryonic stages to adulthood while maintaining the normal pattern of the vascular network. In Mfsd2a knockout mice, the docosahexaenoic acid content is significantly reduced and associated with neuron loss, resulting in microcephaly and cognitive impairment. Based on the role of Mfsd2a in the central nervous system, it has been preliminarily suggested as a potential therapeutic target for drug delivery to the central nervous system. This paper reviews the current progress in Mfsd2a research and summarizes the physiological functions of Mfsd2a in the central nervous system and its role in the occurrence and development of a variety of neurological diseases. Graphical Abstract
    Subjects
    Biomedical and Life Sciences; Biomedicine; Blood-brain barrier; Caveolae; Cell Biology; Central nervous system; Cognitive ability; Docosahexaenoic acid; Drug delivery; Embryos; Endocytosis; Endothelial cells; Membrane permeability; Microencephaly; Nervous system; Neurobiology; Neurological diseases; Neurosciences; Review Paper; Therapeutic targets
  • Article

    Phase 1b/2a study of galunisertib, a small molecule inhibitor of transforming growth factor-beta receptor I, in combination with standard temozolomide-based radiochemotherapy in patients with newly diagnosed malignant glioma

    Authors
    Wick, Antje; Desjardins, Annick; Suarez, Cristina; Forsyth, Peter; Gueorguieva, Ivelina; Burkholder, Tiana; Cleverly, Ann Louise; Estrem, Shawn T.; Wang, Shuaicheng; Lahn, Michael M.; Guba, Susan C.; Capper, David; Rodon, Jordi
    Part of
    Investigational new drugs, 2020-10, Vol.38 (5), p.1570-1579
    Abstract
    • Summary Purpose Galunisertib, a TGF-β inhibitor, has demonstrated antitumor effects in preclinical and radiographic responses in some patients with malignant glioma. This Phase 1b/2a trial investigated the clinical benefit of combining galunisertib with temozolomide-based radiochemotherapy (TMZ/RTX) in patients with newly diagnosed malignant glioma (NCT01220271). Methods This is an open-label, 2-arm Phase 1b/2a study ( N = 56) of galunisertib (intermittent dosing: 14 days on/14 days off per cycle of 28 days) in combination with TMZ/RTX ( n = 40), versus a control arm (TMZ/RTX, n = 16). The primary objective of Phase 1b was to determine the safe and tolerable Phase 2 dose of galunisertib. The primary objective of Phase 2a was to confirm the tolerability and pharmacodynamic profile of galunisertib with TMZ/RTX, and the secondary objectives included determining the efficacy and pharmacokinetic (PK) profile of galunisertib with TMZ/RTX in patients with glioblastoma. This study also characterized the changes in the major T-cell subsets during TMZ/RTX plus galunisertib treatment. Results In the Phase 2a study, efficacy results for patients treated with galunisertib plus TMZ/RTX or TMZ/RTX were: median overall survival (18.2 vs 17.9 months), median progression-free survival (7.6 vs 11.5 months), and disease control rate (80% [32/40] vs 56% [9/16] patients) respectively. PK profile of galunisertib plus TMZ/RTX regimen was consistent with previously published PK data of galunisertib. The overall safety profile across treatment arms was comparable. Conclusion No differences in efficacy, safety or pharmacokinetic variables were observed between the two treatment arms.
    Subjects
    Anticancer properties; Antitumor activity; Disease control; Dosage; Glioblastoma; Glioma; Growth factors; Health services; Inhibitors; Life Sciences & Biomedicine; Lymphocytes T; Medicine; Medicine & Public Health; Oncology; Patients; Pharmacodynamics; Pharmacokinetics; Pharmacology; Pharmacology & Pharmacy; Pharmacology/Toxicology; Phase II Studies; Safety; Science & Technology; Survival; Temozolomide; Transforming growth factor-b
  • Article

    Long-term safety of brazikumab in the open-label period of a randomized phase 2a study of patients with Crohn's disease

    Authors
    Danese, Silvio; Beaton, Andrew; Duncan, Elizabeth A; Mercier, Anne-Kristina; Neisen, Jessica; Seth, Henrik; Zetterstrand, Sofia; Sands, Bruce E
    Part of
    BMC gastroenterology, 2023-12, Vol.23 (1), p.451-451, Article 451
    Abstract
    • Short-term efficacy and safety of brazikumab (MEDI2070), a human monoclonal antibody and anti-p19 subunit inhibitor of interleukin-23, was demonstrated in a phase 2a trial in patients with moderate-to-severe active Crohn's disease (CD). We report brazikumab long-term safety and tolerability from the open-label period of this phase 2a study. Patients who completed the 12-week, double-blind induction period were eligible for inclusion in an open-label period where all patients received subcutaneous brazikumab (210 mg) every 4 weeks for 100 weeks. Patients had moderate-to-severe active CD and had failed or were intolerant to ≥ 1 anti-tumour necrosis factor alpha (TNFα) agent. Safety assessments included treatment-emergent adverse events (TEAEs); further assessments were pharmacokinetics and immunogenicity. Of the 104 patients who entered the open-label period, 57 (54.8%) continued to the end of the open-label period and 47 (45.2%) discontinued brazikumab. The most common reasons for discontinuation were lack of response (14.4%), patient decision (12.5%), and TEAEs (11.5%). In total, 44 (84.6%) in the group switching from placebo to brazikumab (placebo/brazikumab) and 43 (82.7%) in the group continuing brazikumab (brazikumab/brazikumab) experienced 1 or more TEAEs. Most TEAEs were mild-to-moderate in severity. Common TEAEs included nasopharyngitis and headache. Numbers of treatment-emergent serious adverse events (TESAEs) were similar between groups. Infections occurred in 40.4% of patients in the placebo/brazikumab group and 50% in the brazikumab/brazikumab group. There were 5 TESAEs of infection, none of which were opportunistic. No major adverse cardiac events, malignancies, or deaths were reported. Brazikumab was well tolerated with an acceptable safety profile over a 100-week period in patients with moderate-to-severe active CD who failed or were intolerant to 1 or more anti-TNFα agents. NCT01714726; registered October 26, 2012.
    Subjects
    Adverse events; Analysis; Antibodies, Monoclonal - adverse effects; Biologics; Crohn Disease - chemically induced; Crohn Disease - drug therapy; Crohn's disease; Crohns disease; Demographics; Dosage and administration; Double-Blind Method; Drug therapy; Gastrointestinal diseases; Headache; Humans; Immunogenicity; Infections; Inflammatory bowel disease; Interleukin 23; Interleukins; Malignancy; Monoclonal antibodies; Patient outcomes; Patients; Pharmacokinetics; Placebos; Rhinopharyngitis; Safety; Steroids; Testing; Treatment Outcome; Tumor necrosis factor; Tumor necrosis factor inhibitors; Tumor necrosis factor-α
  • Article

    "Self-cleaving" 2A peptide from porcine teschovirus-1 mediates cleavage of dual fluorescent proteins in transgenic Eimeria tenella

    Authors
    Tang, Xinming; Liu, Xianyong; Tao, Geru; Qin, Mei; Yin, Guangwen; Suo, Jingxia; Suo, Xun
    Part of
    Veterinary research (Paris), 2016-06, Vol.47 (1), p.68-68, Article 68
    Abstract
    • The "self-cleaving" 2A sequence of picornavirus, which mediates ribosome-skipping events, enables the generation of two or more separate peptide products from one mRNA containing one or more "self-cleaving" 2A sequences. In this study, we introduced a single 2A sequence of porcine teschovirus-1 (P2A) linked to two fluorescent protein genes, the enhanced yellow fluorescent protein (EYFP) gene and the red fluorescent protein (RFP) gene, in a single cassette into transgenic Eimeria tenella (EtER). As expected, we obtained two separated protein molecules rather than a fused protein, although the two molecules were translated from the same mRNA carrying a single "self-cleaving" 2A sequence. Importantly, RFP led by a secretion signal was secreted into parasitophorous vacuoles, while EYFP localized mainly to the nucleus of EtER. Our results demonstrate that the "self-cleaving" 2A sequence actively mediated cleavage of polyproteins in the apicomplexan parasite E. tenella.
    Subjects
    Animals; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Chickens - parasitology; Eimeria tenella - genetics; Eimeria tenella - metabolism; Genetic aspects; Genetic translation; Health aspects; Life Sciences; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Medical research; Organisms, Genetically Modified - genetics; Organisms, Genetically Modified - metabolism; Picornaviruses; Properties; Red Fluorescent Protein; Short Report; Teschovirus - genetics; Teschovirus - metabolism; Veterinary medicine; Viral proteins
  • Article

    Biosurfactant from endophytic Bacillus pumilus 2A: physicochemical characterization, production and optimization and potential for plant growth promotion

    Authors
    Marchut-Mikołajczyk, Olga; Drożdżyński, Piotr; Polewczyk, Arkadiusz; Smułek, Wojciech; Antczak, Tadeusz
    Part of
    Microbial cell factories, 2021-02, Vol.20 (1), p.40-11, Article 40
    Abstract
    • Microbial surfactants called biosurfactants, thanks to their high biodegradability, low toxicity and stability can be used not only in bioremediation and oil processing, but also in the food and cosmetic industries, and even in medicine. However, the high production costs of microbial surfactants and low efficiency limit their large-scale production. This requires optimization of management conditions, including the possibility of using waste as a carbon source, such as food processing by-products. This papers describes the production and characterization of the biosurfactant obtained from the endophytic bacterial strain Bacillus pumilus 2A grown on various by-products of food processing and its potential applications in supporting plant growth. Four different carbon and nitrogen sources, pH, inoculum concentration and temperature were optimized within Taguchi method. Optimization of bioprocess within Taguchi method and experimental analysis revealed that the optimal conditions for biosurfactant production were brewer's spent grain (5% w/v), ammonium nitrate (1% w/v), pH of 6, 5% of inoculum, and temperature at 30 °C, leading to 6.8 g/L of biosurfactant. Based on gas chromatography-mass spectrometry and Fourier transform infrared spectroscopy analysis produced biosurfactant was determined as glycolipid. Obtained biosurfactant has shown high and long term thermostability, surface tension of 47.7 mN/m, oil displacement of 8 cm and the emulsion index of 69.11%. The examined glycolipid, used in a concentration of 0.2% significantly enhanced growth of Phaseolus vulgaris L. (bean), Raphanus L. (radish), Beta vulgaris L. (beetroot). The endophytic Bacillus pumilus 2A produce glycolipid biosurfactant with high and long tem thermostability, what makes it useful for many purposes including food processing. The use of brewer's spent grain as the sole carbon source makes the production of biosurfactants profitable, and from an environmental point of view, it is an environmentally friendly way to remove food processing by products. Glycolipid produced by endophytic Bacillus pumilus 2A significantly improve growth of Phaseolus vulgaris L. (bean), Raphanus L. (radish), Beta vulgaris L. (beetroot). Obtained results provide new insight to the possible use of glycolipids as plant growth promoting agents.
    Subjects
    Ammonium; Ammonium nitrate; Bacillus (Bacteria); Bacillus pumilus; Bacillus pumilus 2A; Bacteria; Beans; Beta vulgaris; Biodegradability; Biodegradation; Biological products; Bioremediation; Biosurfactant; Biosurfactants; Breweries; By products; Byproducts; Carbon; Carbon sources; Chemical properties; Cooking; Design of experiments; Endophytes; Food; Food industry; Food industry wastes; Food processing; Food processing industry; Food production; Food sources; Fourier analysis; Fourier transforms; Gas chromatography; Glycolipids; Industrial microorganisms; Infrared analysis; Infrared spectroscopy; Inoculum; Lipids; Mass spectrometry; Mass spectroscopy; Microorganisms; Nitrogen sources; Optimization; pH effects; Phaseolus vulgaris; Physiological aspects; Plant growth; Plant growth promoting substances; Plant-growth promotion; Pollutants; Production costs; Production processes; Radishes; Raphanus; Sugar beets; Surface active agents; Surface tension; Surfactants; Syrups & sweeteners; Taguchi methods; Temperature; Thermal stability; Toxicity
  • Article

    Natural course of pontocerebellar hypoplasia type 2A

    Authors
    Sánchez-Albisua, Iciar; Frölich, Saskia; Barth, Peter G; Steinlin, Maja; Krägeloh-Mann, Ingeborg
    Part of
    Orphanet journal of rare diseases, 2014-05, Vol.9 (1), p.70-70
    Abstract
    • Pontocerebellar hypoplasia Type 2 (PCH2) is a rare autosomal recessive condition, defined on MRI by a small cerebellum and ventral pons. Clinical features are severe developmental delay, microcephaly and dyskinesia.Ninety percent carry a p.A307S mutation in the TSEN54-gene. Our aim was to describe the natural course including neurological and developmental features and other aspects of care in a homogeneous group of PCH2 patients all carrying the p.A307S mutation. Patients were recruited via the German patients' organizations. Inclusion criteria were imaging findings of PCH2 and a p.A307S mutation. Data were collected using medical reports and patient questionnaires discussed in a standardized telephone interview. Thirty-three patients were included. When considering survival until age 11 years, 53% of children had died Weight, length and head circumference, mostly in the normal range at birth, became abnormal, especially head circumference (-5.58 SD at age 5 yrs). Neurologic symptoms: Choreathetosis was present in 88% (62% with pyramidal signs), 12% had pure spasticity. Epileptic seizures were manifest in 82%, status epilepticus in 39%. Non-epileptic dystonic attacks occurred in 33%. General symptoms: feeding difficulties were recorded in 100%, sleep disorder in 96%, apneas in 67% and recurrent infections in 52%; gastroesophageal reflux disease was diagnosed in 73%, 67% got percutaneous endoscopic gastrostomy and 36% a Nissen-fundoplication. Neurodevelopmental data: All children made progress, but on a low level: such as fixing and following with the eyes was seen in 76%, attempting to grasp objects (76%), moderate head control (73%), social smile (70%), rolling from prone to supine (58%), and sitting without support (9%). Ten percent lost achieved abilities on follow-up. The presence of prenatal symptoms did not correlate with outcome. Phenotype of this genetically homogeneous group of PCH2 children was severe with reduced survival, but compatible with some developmental progress. Our data support the hypothesis of an early onset degeneration which thereafter stabilizes.
    Subjects
    Care and treatment; Development and progression; Endoribonucleases - genetics; Epilepsy; Gastroesophageal reflux; Genetic aspects; Health aspects; Humans; Mutation; Olivopontocerebellar Atrophies - genetics; Olivopontocerebellar Atrophies - physiopathology; Risk factors; Sleep disorders
  • Article

    Increased expression of adenosine 2A receptors in metastatic renal cell carcinoma is associated with poorer response to anti-vascular endothelial growth factor agents and anti-PD-1/Anti-CTLA4 antibodies and shorter survival

    Authors
    Kamai, Takao; Kijima, Toshiki; Tsuzuki, Toyonori; Nukui, Akinori; Abe, Hideyuki; Arai, Kyoko; Yoshida, Ken-Ichiro
    Part of
    Cancer Immunology, Immunotherapy, 2021-07, Vol.70 (7), p.2009-2021
    Abstract
    • Background Adenosine and its adenosine 2A receptors (A2AR) mediate the immunosuppressive mechanism by which tumors escape immunosurveillance and impede anti-tumor immunity within the tumor microenvironment. However, we do not know whether the adenosine pathway (CD39/CD73/A2AR) plays a role in renal cell carcinoma (RCC). Therefore, we studied the role of immunosuppression in RCC by assessing the adenosine pathway in patients with RCC treated with anti-vascular endothelial growth factor (anti-VEGF) agents or immune checkpoints inhibitors (ICIs) or both. Methods In 60 patients with metastatic RCC, we examined the expression of CD39, CD73, A2AR, and programmed cell death 1 ligand 1 (PD-L1) immunohistochemically in surgically resected tumor tissues and studied the clinicopathological characteristics of these patients. Patients were treated by cytoreductive nephrectomy with systemic therapy with anti-VEGF agent or a combination of the ICIs anti-cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) antibody and programmed cell death 1 (PD-1) antibody. Results Increased expression of A2AR in the primary tumors was associated with metastatic profiles. Patients treated with anti–PD-1 antibody in monotherapy, a combination of anti-PD-1 and anti-CTLA4 antibodies, or anti-VEGF agents showed better response and longer overall survival if the primary tumor had higher PD-L1 expression and lower A2AR expression. In Cox multivariate regression analysis, higher expression of A2AR was associated with shorter overall survival. Conclusions Our findings suggest that the expression of A2AR and PD-L1 in the primary tumors in RCC might predict the outcomes of treatment with anti-VEGF agents and ICIs and that the A2AR pathway might be a molecular target for immunotherapy.
    Subjects
    Adenosine; Adenosine receptors; Adolescent; Adult; Aged; Antibodies; Antineoplastic Combined Chemotherapy Protocols - therapeutic use; Apoptosis; Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Cancer Research; Carcinoma, Renal Cell - drug therapy; Carcinoma, Renal Cell - metabolism; Carcinoma, Renal Cell - mortality; Carcinoma, Renal Cell - pathology; Case-Control Studies; CD73 antigen; Cell death; CTLA-4 Antigen - antagonists & inhibitors; CTLA-4 protein; Cytotoxicity; Female; Follow-Up Studies; Humans; Immune Checkpoint Inhibitors - therapeutic use; Immunology; Immunosuppression; Immunosurveillance; Immunotherapy; Kidney cancer; Kidney Neoplasms - drug therapy; Kidney Neoplasms - metabolism; Kidney Neoplasms - mortality; Kidney Neoplasms - secondary; Lymphocytes T; Male; Medicine; Medicine & Public Health; Metastases; Metastasis; Middle Aged; Nephrectomy; Oncology; Original; Original Article; PD-1 protein; PD-L1 protein; Prognosis; Programmed Cell Death 1 Receptor - antagonists & inhibitors; Receptor, Adenosine A2A - genetics; Receptor, Adenosine A2A - metabolism; Renal cell carcinoma; Retrospective Studies; Survival Rate; Tumor microenvironment; Tumors; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - antagonists & inhibitors; Young Adult
  • Article

    Escherichia coli O157:H7 strains harbor at least three distinct sequence types of Shiga toxin 2a-converting phages

    Authors
    Yin, Shuang; Rusconi, Brigida; Sanjar, Fatemeh; Goswami, Kakolie; Xiaoli, Lingzi; Eppinger, Mark; Dudley, Edward G
    Part of
    BMC genomics, 2015-09, Vol.16 (1), p.733-733, Article 733
    Abstract
    • Shiga toxin-producing Escherichia coli O157:H7 is a foodborne pathogen that causes severe human diseases including hemolytic uremic syndrome (HUS). The virulence factor that mediates HUS, Shiga toxin (Stx), is encoded within the genome of a lambdoid prophage. Although draft sequences are publicly available for a large number of E. coli O157:H7 strains, the high sequence similarity of stx-converting bacteriophages with other lambdoid prophages poses challenges to accurately assess the organization and plasticity among stx-converting phages due to assembly difficulties. To further explore genome plasticity of stx-converting prophages, we enriched phage DNA from 45 ciprofloxacin-induced cultures for subsequent 454 pyrosequencing to facilitate assembly of the complete phage genomes. In total, 22 stx2a-converting phage genomes were closed. Comparison of the genomes distinguished nine distinct phage sequence types (PSTs) delineated by variation in obtained sequences, such as single nucleotide polymorphisms (SNPs) and insertion sequence element prevalence and location. These nine PSTs formed three distinct clusters, designated as PST1, PST2 and PST3. The PST2 cluster, identified in two clade 8 strains, was related to stx2a-converting phages previously identified in non-O157 Shiga-toxin producing E. coli (STEC) strains associated with a high incidence of HUS. The PST1 cluster contained phages related to those from E. coli O157:H7 strain Sakai (lineage I, clade 1), and PST3 contained a single phage that was distinct from the rest but most related to the phage from E. coli O157:H7 strain EC4115 (lineage I/II, clade 8). Five strains carried identical stx2a-converting phages (PST1-1) integrated at the same chromosomal locus, but these strains produced different levels of Stx2. The stx2a-converting phages of E. coli O157:H7 can be categorized into at least three phage types. Diversification within a phage type is mainly driven by IS629 and by a small number of SNPs. Polymorphisms between phage genomes may help explain differences in Stx2a production between strains, however our data indicates that genes encoded external to the phage affect toxin production as well.
    Subjects
    Bacteriophages - genetics; Ciprofloxacin - pharmacology; Escherichia coli O157 - genetics; Escherichia coli O157 - pathogenicity; Genes; Genetic aspects; Genome; Hemolytic-Uremic Syndrome - genetics; Hemolytic-Uremic Syndrome - microbiology; Humans; Polymorphism, Single Nucleotide; Shiga Toxin 2 - genetics
  • Article

    Norepinephrine protects against cochlear outer hair cell damage and noise-induced hearing loss via [alpha]2A-adrenergic receptor

    Authors
    Tian, Chaoyong; Yang, Yang; Wang, Renfeng; Li, Yao; Sun, Fei; Chen, Jun; Zha, Dingjun
    Part of
    BMC neuroscience, 2024-01, Vol.25 (1)
    Abstract
    • Background The cochlear sympathetic system plays a key role in auditory function and susceptibility to noise-induced hearing loss (NIHL). The formation of reactive oxygen species (ROS) is a well-documented process in NIHL. In this study, we aimed at investigating the effects of a superior cervical ganglionectomy (SCGx) on NIHL in Sprague-Dawley rats. Methods We explored the effects of unilateral and bilateral Superior Cervical Ganglion (SCG) ablation in the eight-ten weeks old Sprague-Dawley rats of both sexes on NIHL. Auditory function was evaluated by auditory brainstem response (ABR) testing and Distortion product otoacoustic emissions (DPOAEs). Outer hair cells (OHCs) counts and the expression of [alpha].sub.2A-adrenergic receptor (AR) in the rat cochlea using immunofluorescence analysis. Cells culture and treatment, CCK-8 assay, Flow cytometry staining and analysis, and western blotting were to explore the mechanisms of SCG fibers may have a protective role in NIHL. Results We found that neither bilateral nor unilateral SCGx protected the cochlea against noise exposure. In HEI-OC1 cells, H.sub.2O.sub.2-induced oxidative damage and cell death were inhibited by the application of norepinephrine (NE). NE may prevent ROS-induced oxidative stress in OHCs and NIHL through the [alpha].sub.2A-AR. Conclusion These results demonstrated that sympathetic innervation mildly affected cochlear susceptibility to acoustic trauma by reducing oxidative damage in OHCs through the [alpha].sub.2A-AR. NE may be a potential therapeutic strategy for NIHL prevention. Keywords: [alpha].sub.2A- adrenergic receptor, Noise-induced hearing loss, Outer hair cells, Sympathetic nervous system, Superior cervical ganglion
    Subjects
    Cell death; Drug therapy; Epinephrine; Hair cells (Mechanoreceptors); Health aspects; Hearing loss; Noradrenaline; Receptors; Testing
  • Article

    Association between serotonin 2A receptor (HTR2A) genetic variations and risk of hypertension in a community-based cohort study

    Authors
    Choi, Jung Ran; Jeon, Minhee; Koh, Sang Baek
    Part of
    BMC medical genetics, 2020-01, Vol.21 (1), p.5-5, Article 5
    Abstract
    • Background Hypertension is one of the risk factors for obesity-related cardiovascular diseases. We investigated whether genetic variations in serotonin 2A receptor (HTR2A) were associated with hypertension. Methods We carried out a cross-sectional study in cohorts A (Ansan-Ansung cohort, N = 6039) and B (Wonju-Pyengchang cohort, N = 7524). Several genetic variants in HTR2A including rs7330636, rs9590999, rs2183057, and rs4942595 were selected and genotyped. Results In hypertensive participants in cohort A, the baseline systolic blood pressure and body mass index were 141.80 +/- 17.20 mg/dL and 24.48 +/- 4.75 kg/m(2), respectively, which were higher than in those without hypertension (p < 0.001). rs4942595TC genotype was associated with hypertension in cohort A (OR = 0.739), after adjusting for variables. Subjects with rs4942578AA genotype had a decreased risk of hypertension after adjusting for clinical factor (OR = 0.735) in cohort B, and an elevated risk of hypertension in cohort A (OR = 1.562). The logistic regression analysis showed that participants with rs4941573TC genotype were 1.327 times more likely to have a higher blood pressure than those with TT genotype (95% CI 1.101-1.599) in cohort B. Whereas, the OR for developing hypertension in subjects with rs17069883CC genotype compared to those with AA genotype was 1.447 (95% CI 1.018-2.056; p for trend = 0.040) in cohort A. Conclusions HTR2A genetic variations were associated with hypertension risk in our study.
    Subjects
    Age; Aged; Blood pressure; Body Mass Index; Cardiovascular diseases; Cholesterol; Cohort analysis; Cross sectional study; Cross-sectional studies; Demographics; Diseases; Epidemiology; Exercise; Fasting; Female; Gender; Genes; Genetic aspects; Genetic Association Studies; Genetic diversity; Genetic Predisposition to Disease; Genetic variation; Genetics & Heredity; Genomes; Genotype; Genotype & phenotype; Genotypes; Health aspects; Hospitals; Humans; Hypertension; Hypertension - epidemiology; Hypertension - genetics; Hypertension - pathology; Life Sciences & Biomedicine; Male; Middle Aged; Obesity; Phenols (Class of compounds); Polymorphism, Single Nucleotide - genetics; Population; Receptor, Serotonin, 5-HT2A - genetics; Receptors; Regression analysis; Risk Factors; Science & Technology; Serotonin; Serotonin 2A receptor gene; Serotonin receptors; Smoking; Studies
  • Article

    Ceramide/protein phosphatase 2A axis is engaged in gap junction impairment elicited by PCB153 in liver stem-like progenitor cells

    Authors
    Squecco, Roberta; Pierucci, Federica; Idrizaj, Eglantina; Frati, Alessia; Lenci, Elena; Vicenti, Catia; Iachini, Maria Chiara; Martinesi, Maria; Garella, Rachele; Baccari, Maria Caterina; Francini, Fabio; Meacci, Elisabetta
    Part of
    Molecular and cellular biochemistry, 2021-08, Vol.476 (8), p.3111-3126
    Abstract
    • The widespread environmental pollutant 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB153) is a non-dioxin-like toxicant. It is a potential carcinogen compound able to induce gap junction (GJ) intercellular communication impairment, probably the first non-genomic event leading to tumor promotion. Although PCBs have been known for many years, the molecular mode of PCB153 action is still unclear. Recent studies from our research group have shown that the toxicant elicits a transient modulation of connexin (Cx) 43-formed GJs in hepatic stem-like WB-F344 cells involving sphingosine 1-phosphate (S1P) path. Taking into account that other strictly related bioactive sphingolipids, such as ceramide (Cer), may have different effects from S1P, here we aim to clarify the signaling paths engaged by PCB153 in the control of GJs, focusing primarily on the role of Cer. Accordingly, we have achieved a combined biomolecular and electrophysiological analysis of GJs in cultured WB-F344 cells treated with PCB153 at different time points. We have found that the toxicant elicited a time-dependent regulation of GJs formed by different Cx isoforms, through a transient modulation of Cer/Cer kinase (CerK) axis and, in turn, of protein phosphatase 2A (PP2A). Our new findings demonstrate the existence of a specific molecular mechanism downstream to Cer, which distinctly affects the voltage-dependent and -independent GJs in liver stem-like cells, and open new opportunities for the identification of additional potential targets of these environmental toxicants.
    Subjects
    Animals; Aprotinin; Biochemistry; Biomedical and Life Sciences; Carcinogens; Cardiology; Cell Communication; Cell signaling; Cells (biology); Cells, Cultured; Ceramide; Ceramides - metabolism; Contaminants; Dioxin; Dioxins; Gap Junctions - drug effects; Gap Junctions - metabolism; Gap Junctions - pathology; Hepatocytes; Hexachlorobiphenyl; Impairment; Isoforms; Kinases; Life Sciences; Liver; Liver - drug effects; Liver - metabolism; Liver - pathology; Medical Biochemistry; Membrane lipids; Modulation; Oncology; Phosphatase; Phosphatases; Phosphates; Phosphoprotein phosphatase; Pollutants; Polychlorinated Biphenyls - pharmacology; Progenitor cells; Protein phosphatase; Protein Phosphatase 2 - genetics; Protein Phosphatase 2 - metabolism; Proteins; Rats; Signal Transduction; Sphingolipids; Sphingosine; Sphingosine 1-phosphate; Stem cells; Stem Cells - drug effects; Stem Cells - metabolism; Stem Cells - pathology; Target recognition; Time dependence; Toxicants
  • Article

    Intravesical liposomal tacrolimus for hemorrhagic cystitis: a phase 2a multicenter dose-escalation study

    Authors
    Hafron, Jason; Breyer, Benjamin N.; Joshi, Shreyas; Smith, Christopher; Kaufman, Melissa R.; Okonski, Janet; Chancellor, Michael B.
    Part of
    International urology and nephrology, 2024-01, Vol.56 (1), p.87-96
    Abstract
    • Background Hemorrhagic cystitis (HC) is an inflammatory disease of the bladder with sustained hematuria for which there is currently no approved drug treatment. We evaluated a liposomal tacrolimus preparation (LP-10) in patients with refractory moderate to severe sterile HC. Methods This phase 2a dose-escalation study assessed the safety and efficacy of up to 2 intravesical instillations of LP-10 (2, 4, or 8 mg tacrolimus) in 13 patients with HC. Primary efficacy outcomes were changes from baseline in the number of bleeding sites on cystoscopy, microscopic urine analysis for red blood cells (RBCs), and hematuria on dipstick. Additional efficacy measures included urinary incontinence, frequency, and urgency on a 3-day diary and cystoscopy global response assessment (GRA). Blood samples for pharmacokinetic (PK) assessment were obtained in all patients. Results Intravesical LP-10 was well tolerated, with no treatment-related severe or serious adverse events (AEs) and only 3 drug-related AEs (artificial urinary sphincter malfunction, dysuria, and bladder spasms). LP-10 blood levels showed short durations of minimal systemic uptake. Treatment resulted in significant improvements in bleeding on cystoscopy, RBC counts in urine, hematuria on dipstick, and urinary incontinence. Bleeding on cystoscopy and urinary incontinence showed dose-dependent improvements that were more pronounced in the 4 mg and 8 mg dose groups. All dose groups showed a significant improvement in cystoscopy GRA. Conclusion LP-10 was well tolerated, with clinically relevant efficacy seen in improvements in cystoscopic bleeding, hematuria, and urinary incontinence. The benefit-risk profile supports the further clinical development of LP-10 at a tacrolimus dose of 4 mg.
    Subjects
    Administration, Intravesical; Bleeding; Blood cells; Blood levels; Cystitis; Cystitis - drug therapy; Cystitis, Hemorrhagic; Drug dosages; Erythrocytes; Hematuria; Hematuria - drug therapy; Hematuria - etiology; Hemorrhage - etiology; Hemorrhagic cystitis; Humans; Inflammatory diseases; Medicine; Medicine & Public Health; Nephrology; Patients; Pharmacokinetics; Sphincter; Tacrolimus; Tacrolimus - therapeutic use; Urinary Bladder; Urinary Incontinence; Urology; Urology - Original Paper
  • Article

    A first-in-human, open-label Phase 1b and a randomised, double-blind Phase 2a clinical trial in recent-onset type 1 diabetes with AG019 as monotherapy and in combination with teplizumab

    Authors
    Mathieu, Chantal; Wiedeman, Alice; Cerosaletti, Karen; Long, S. Alice; Serti, Elisavet; Cooney, Laura; Vermeiren, Joan; Caluwaerts, Silvia; Van Huynegem, Karolien; Steidler, Lothar; Blomme, Sven; Rottiers, Pieter; Nepom, Gerald T.; Herold, Kevan C.; AG019-T1D-101 Trial Investigators; on behalf of the AG019-T1D-101 Trial Investigators
    Part of
    Diabetologia, 2024-01, Vol.67 (1), p.27-41
    Abstract
    • Aims/hypothesis We hypothesised that islet beta cell antigen presentation in the gut along with a tolerising cytokine would lead to antigen-specific tolerance in type 1 diabetes. We evaluated this in a parallel open-label Phase 1b study using oral AG019, food-grade Lactococcus lactis bacteria genetically modified to express human proinsulin and human IL-10, as a monotherapy and in a parallel, randomised, double-blind Phase 2a study using AG019 in combination with teplizumab. Methods Adults (18–42 years) and adolescents (12–17 years) with type 1 diabetes diagnosed within 150 days were enrolled, with documented evidence of at least one autoantibody and a stimulated peak C-peptide level >0.2 nmol/l. Participants were allocated to interventions using interactive response technology. We treated 42 people aged 12–42 years with recent-onset type 1 diabetes, 24 with Phase 1b monotherapy (open-label) and 18 with Phase 2a combination therapy. In the Phase 2a study, after treatment of the first two open-label participants, all people involved were blinded to group assignment, except for the Data Safety Monitoring Board members and the unblinded statistician. The primary endpoint was safety and tolerability based on the incidence of treatment-emergent adverse events, collected up to 6 months post treatment initiation. The secondary endpoints were pharmacokinetics, based on AG019 detection in blood and faeces, and pharmacodynamic activity. Metabolic and immune endpoints included stimulated C-peptide levels during a mixed meal tolerance test, HbA 1c levels, insulin use, and antigen-specific CD4 + and CD8 + T cell responses using an activation-induced marker assay and pooled tetramers, respectively. Results Data from 24 Phase 1b participants and 18 Phase 2a participants were analysed. No serious adverse events were reported and none of the participants discontinued AG019 due to treatment-emergent adverse events. No systemic exposure to AG019 bacteria, proinsulin or human IL-10 was demonstrated. In AG019 monotherapy-treated adults, metabolic variables were stabilised up to 6 months (C-peptide, insulin use) or 12 months (HbA 1c ) post treatment initiation. In participants treated with AG019/teplizumab combination therapy, all measured metabolic variables stabilised or improved up to 12 months and CD8 + T cells with a partially exhausted phenotype were significantly increased at 6 months. Circulating preproinsulin-specific CD4 + and CD8 + T cells were detected before and after treatment, with a reduction in the frequency of preproinsulin-specific CD8 + T cells after treatment with monotherapy or combination therapy. Conclusions/interpretation Oral delivery of AG019 was well tolerated and safe as monotherapy and in combination with teplizumab. AG019 was not shown to interfere with the safety profile of teplizumab and may have additional biological effects, including changes in preproinsulin-specific T cells. These preliminary data support continuing studies with this agent alone and in combination with teplizumab or other systemic immunotherapies in type 1 diabetes. Trial registration ClinicalTrials.gov NCT03751007, EudraCT 2017-002871-24 Funding This study was funded by Precigen ActoBio Graphical Abstract
    Subjects
    Adolescent; Adult; Adverse events; Antigen presentation; Antigens; Autoantibodies; Beta cells; C-Peptide; CD4 antigen; CD8 antigen; CD8-Positive T-Lymphocytes - metabolism; Cell activation; Clinical trials; Diabetes; Diabetes mellitus (insulin dependent); Diabetes Mellitus, Type 1; Double-Blind Method; Human Physiology; Humans; Immunological tolerance; Immunotherapy; Insulin; Interleukin 10; Internal Medicine; Lymphocytes; Lymphocytes T; Medicine; Medicine & Public Health; Metabolic Diseases; Metabolism; Monoclonal antibodies; Peptides; Pharmacodynamics; Pharmacokinetics; Phenotypes; Preproinsulin; Proinsulin; Safety

Can't find what you're looking for?

Keyboard Shortcuts

Close

Available anywhere

?
Shortcut help message
s
Highlight search box
esc
Close dialog

Available in search results

n
Next page
p
Previous page
f
Toggle filters
1-5
Open nth result on page