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1.
Molecules ; 29(8)2024 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-38675528

RESUMO

Glioblastoma (GBM), the most frequent and lethal brain cancer in adults, is characterized by short survival times and high mortality rates. Due to the resistance of GBM cells to conventional therapeutic treatments, scientific interest is focusing on the search for alternative and efficient adjuvant treatments. S-Adenosylmethionine (AdoMet), the well-studied physiological methyl donor, has emerged as a promising anticancer compound and a modulator of multiple cancer-related signaling pathways. We report here for the first time that AdoMet selectively inhibited the viability and proliferation of U87MG, U343MG, and U251MG GBM cells. In these cell lines, AdoMet induced S and G2/M cell cycle arrest and apoptosis and downregulated the expression and activation of proteins involved in homologous recombination DNA repair, including RAD51, BRCA1, and Chk1. Furthermore, AdoMet was able to maintain DNA in a damaged state, as indicated by the increased γH2AX/H2AX ratio. AdoMet promoted mitotic catastrophe through inhibiting Aurora B kinase expression, phosphorylation, and localization causing GBM cells to undergo mitotic catastrophe-induced death. Finally, AdoMet inhibited DNA repair and induced cell cycle arrest, apoptosis, and mitotic catastrophe in patient-derived GBM cells. In light of these results, AdoMet could be considered a potential adjuvant in GBM therapy.


Assuntos
Antineoplásicos , Apoptose , Proliferação de Células , Glioblastoma , S-Adenosilmetionina , Humanos , Glioblastoma/tratamento farmacológico , Glioblastoma/metabolismo , Glioblastoma/patologia , S-Adenosilmetionina/farmacologia , Linhagem Celular Tumoral , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Antineoplásicos/farmacologia , Antineoplásicos/química , Sobrevivência Celular/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Aurora Quinase B/metabolismo , Aurora Quinase B/antagonistas & inibidores , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Rad51 Recombinase/metabolismo , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Mitose/efeitos dos fármacos
2.
Int J Mol Sci ; 24(3)2023 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-36768835

RESUMO

Cannabis sativa-derived compounds, such as delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), and components of the endocannabinoids system, such as N-arachidonoylethanolamide (anandamide, AEA) and 2-arachidonoylglycerol (2-AG), are extensively studied to investigate their numerous biological effects, including powerful antioxidant effects. Indeed, a series of recent studies have indicated that many disorders are characterized by alterations in the intracellular antioxidant system, which lead to biological macromolecule damage. These pathological conditions are characterized by an unbalanced, and most often increased, reactive oxygen species (ROS) production. For this study, it was of interest to investigate and recapitulate the antioxidant properties of these natural compounds, for the most part CBD and THC, on the production of ROS and the modulation of the intracellular redox state, with an emphasis on their use in various pathological conditions in which the reduction of ROS can be clinically useful, such as neurodegenerative disorders, inflammatory conditions, autoimmunity, and cancers. The further development of ROS-based fundamental research focused on cannabis sativa-derived compounds could be beneficial for future clinical applications.


Assuntos
Canabidiol , Canabinoides , Cannabis , Canabinoides/farmacologia , Antioxidantes , Espécies Reativas de Oxigênio , Canabidiol/farmacologia , Oxirredução , Estresse Oxidativo , Dronabinol
3.
Int J Mol Sci ; 24(5)2023 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-36902485

RESUMO

Pollutants consist of several components, known as direct or indirect mutagens, that can be associated with the risk of tumorigenesis. The increased incidence of brain tumors, observed more frequently in industrialized countries, has generated a deeper interest in examining different pollutants that could be found in food, air, or water supply. These compounds, due to their chemical nature, alter the activity of biological molecules naturally found in the body. The bioaccumulation leads to harmful effects for humans, increasing the risk of the onset of several pathologies, including cancer. Environmental components often combine with other risk factors, such as the individual genetic component, which increases the chance of developing cancer. The objective of this review is to discuss the impact of environmental carcinogens on modulating the risk of brain tumorigenesis, focusing our attention on certain categories of pollutants and their sources.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , Neoplasias Encefálicas , Humanos , Poluentes Atmosféricos/análise , Poluição Ambiental , Poluição do Ar/análise , Monitoramento Ambiental , Carcinogênese , Transformação Celular Neoplásica , Encéfalo , Exposição Ambiental
4.
Int J Mol Sci ; 23(6)2022 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-35328765

RESUMO

Medical case reports suggest that cannabinoids extracted from Cannabis sativa have therapeutic effects; however, the therapeutic employment is limited due to the psychotropic effect of its major component, Δ9-tetrahydrocannabinol (THC). The new scientific discoveries related to the endocannabinoid system, including new receptors, ligands, and mediators, allowed the development of new therapeutic targets for the treatment of several pathological disorders minimizing the undesirable psychotropic effects of some constituents of this plant. Today, FDA-approved drugs, such as nabiximols (a mixture of THC and non-psychoactive cannabidiol (CBD)), are employed in alleviating pain and spasticity in multiple sclerosis. Dronabinol and nabilone are used for the treatment of chemotherapy-induced nausea and vomiting in cancer patients. Dronabinol was approved for the treatment of anorexia in patients with AIDS (acquired immune deficiency syndrome). In this review, we highlighted the potential therapeutic efficacy of natural and synthetic cannabinoids and their clinical relevance in cancer, neurodegenerative and dermatological diseases, and viral infections.


Assuntos
Canabidiol , Canabinoides , Cannabis , Neoplasias , Canabidiol/farmacologia , Canabidiol/uso terapêutico , Canabinoides/efeitos adversos , Dronabinol/farmacologia , Dronabinol/uso terapêutico , Endocanabinoides , Humanos , Neoplasias/induzido quimicamente , Neoplasias/tratamento farmacológico
5.
Int J Mol Sci ; 22(7)2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33916164

RESUMO

Cannabinoids are a family of heterogeneous compounds that mostly interact with receptors eliciting several physiological effects both in the central and peripheral nervous systems and in peripheral organs. They exert anticancer action by modulating signaling pathways involved in cancer progression; furthermore, the effects induced by their use depend on both the type of tumor and their action on the components of the endocannabinoid system. This review will explore the mechanism of action of the cannabinoids in signaling pathways involved in cancer proliferation, neovascularisation, migration, invasion, metastasis, and tumor angiogenesis.


Assuntos
Canabinoides/farmacologia , Endocanabinoides/metabolismo , Neoplasias/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Animais , Canabinoides/uso terapêutico , Progressão da Doença , Humanos , Ligantes , Neoplasias/metabolismo
6.
Int J Mol Sci ; 22(19)2021 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-34638872

RESUMO

BACKGROUND: Vasculogenic mimicry (VM) is a functional microcirculation pattern formed by aggressive tumor cells. Thus far, no effective drugs have been developed to target VM. Glioblastoma (GBM) is the most malignant form of brain cancer and is a highly vascularized tumor. Vasculogenic mimicry represents a means whereby GBM can escape anti-angiogenic therapies. METHODS: Here, using an in vitro tube formation assay on Matrigel, we evaluated the ability of N6-isopentenyladenosine (iPA) to interfere with vasculogenic mimicry (VM). RhoA activity was assessed using a pull-down assay, while the modulation of the adherens junctions proteins was analyzed by Western blot analysis. RESULTS: We found that iPA at sublethal doses inhibited the formation of capillary-like structures suppressing cell migration and invasion of U87MG, U343MG, and U251MG cells, of patient-derived human GBM cells and GBM stem cells. iPA reduces the vascular endothelial cadherin (VE-cadherin) expression levels in a dose-dependent manner, impairs the vasculogenic mimicry network by modulation of the Src/p120-catenin pathway and inhibition of RhoA-GTPase activity. CONCLUSIONS: Taken together, our results revealed iPA as a promising novel anti-VM drug in GBM clinical therapeutics.


Assuntos
Cateninas/metabolismo , Glioblastoma/tratamento farmacológico , Isopenteniladenosina/farmacologia , Neovascularização Patológica/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Proteína rhoA de Ligação ao GTP/metabolismo , Quinases da Família src/metabolismo , Antígenos CD/genética , Antígenos CD/metabolismo , Caderinas/genética , Caderinas/metabolismo , Cateninas/genética , Linhagem Celular Tumoral , Glioblastoma/irrigação sanguínea , Glioblastoma/genética , Glioblastoma/metabolismo , Humanos , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Proteína rhoA de Ligação ao GTP/genética , Quinases da Família src/genética
7.
FEBS Open Bio ; 14(5): 843-854, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38514913

RESUMO

Glioblastoma (GBM) is a primary tumor in the central nervous system with poor prognosis. It exhibits elevated glucose uptake and lactate production. This metabolic state of aerobic glycolysis is known as the Warburg effect. N6-isopentenyladenosine (iPA), a natural cytokine modified with an isopentenyl moiety derived from the mevalonate pathway, has well-established anti-tumor activity. It inhibits cell proliferation in glioma cells, inducing cell death by apoptosis and/or necroptosis. In the present study, we found that iPA inhibits aerobic glycolysis in unmodified U87MG cells and in the same cell line engineered to over-express wild-type epidermal growth factor receptor (EGFR) or EGFR variant III (vIII), as well as in a primary GBM4 patient-derived cell line. The detection of glycolysis showed that iPA treatment suppressed ATP and lactate production. We also evaluated the response of iPA treatment in normal human astrocyte primary cells, healthy counterpart cells of the brain. Aerobic glycolysis in treated normal human astrocyte cells did not show significant changes compared to GBM cells. To determine the mechanism of iPA action on aerobic glycolysis, we investigated the expression of certain enzymes involved in this metabolic pathway. We observed that iPA reduced the expression of pyruvate kinase M2 (PKM2), which plays a key role in the regulation of aerobic glycolysis, promoting tumor cell proliferation. The reduction of PKM2 expression is a result of the inhibition of the inhibitor of nuclear factor kappa-B kinase subunit, beta/nuclear factor-kappa B pathway upon iPA treatment. In conclusion, these experimental results show that iPA may inhibit aerobic glycolysis of GBM in stabilized cell lines and primary GBM cells by targeting the expression and activity of PKM2.


Assuntos
Glioblastoma , Glicólise , Isopenteniladenosina , Piruvato Quinase , Humanos , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Glioblastoma/metabolismo , Glioblastoma/patologia , Glioblastoma/tratamento farmacológico , Glioblastoma/genética , Glicólise/efeitos dos fármacos , Isopenteniladenosina/farmacologia , Isopenteniladenosina/metabolismo , Piruvato Quinase/efeitos dos fármacos , Piruvato Quinase/metabolismo
8.
Front Immunol ; 15: 1373435, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38601151

RESUMO

Introduction: The involvement of endocannabinoid system (ECS) in the inflammatory cascade, and the ability of phytocannabinoids, endocannabinoids and their synthetic analogues to modulate it has become an interesting research area for new therapeutic approaches in inflammatory skin diseases. Cannabidiol (CBD) appears to be the most promising among phytocannabinoids, due to the lack of psychotropic effects and low toxicity profile. Its anti-inflammatory action has been highlighted in different preclinical models, ranging from experimental colitis to arthritis and neuroinflammation. Our aim was to evaluate CBD immune-modulatory effects in peripheral blood mononuclear cells (PBMC) of psoriasis individuals with particular attention to both innate and adaptative immune arms. Methods: We performed in vitro immune functional experiments to analyze CBD action on various immune cells active in psoriatic lesions. Results: The results showed that CBD produced a shift from Th1 to Th2 response, while boosting cytotoxic activity of Natural Killer (NK) cells. Furthermore, it also exerted a potent action on monocyte differentiation as, after CBD treatment, monocytes from psoriatic individuals were unable to migrate in response to inflammatory stimuli and to fully differentiate into mature dendritic cells. Finally, a M2 skewing of monocyte-derived macrophages by CBD also contributed to the fine tuning of the magnitude of immune responses. Conclusions: These data uncover new potential immunomodulatory properties of this cannabinoid suggesting a possible therapeutic action in the treatment of multiple inflammatory skin diseases.


Assuntos
Canabidiol , Canabinoides , Psoríase , Humanos , Canabidiol/farmacologia , Canabidiol/uso terapêutico , Leucócitos Mononucleares , Psoríase/tratamento farmacológico , Endocanabinoides
9.
Cell Death Dis ; 14(9): 638, 2023 09 27.
Artigo em Inglês | MEDLINE | ID: mdl-37758718

RESUMO

Despite intense research efforts, glioblastoma remains an incurable brain tumor with a dismal median survival time of 15 months. Thus, identifying new therapeutic targets is an urgent need. Here, we show that the lysine methyltransferase SETD8 is overexpressed in 50% of high-grade gliomas. The small molecule SETD8 inhibitor UNC0379, as well as siRNA-mediated inhibition of SETD8, blocked glioblastoma cell proliferation, by inducing DNA damage and activating cell cycle checkpoints. Specifically, in p53-proficient glioblastoma cells, SETD8 inhibition and DNA damage induced p21 accumulation and G1/S arrest whereas, in p53-deficient glioblastoma cells, DNA damage induced by SETD8 inhibition resulted in G2/M arrest mediated by Chk1 activation. Checkpoint abrogation, by the Wee1 kinase inhibitor adavosertib, induced glioblastoma cell lines and primary cells, DNA-damaged by UNC0379, to progress to mitosis where they died by mitotic catastrophe. Finally, UNC0379 and adavosertib synergized in restraining glioblastoma growth in a murine xenograft model, providing a strong rationale to further explore this novel pharmacological approach for adjuvant glioblastoma treatment.


Assuntos
Glioblastoma , Doenças do Recém-Nascido , Humanos , Animais , Camundongos , Recém-Nascido , Glioblastoma/tratamento farmacológico , Glioblastoma/genética , Apoptose , Proteína Supressora de Tumor p53 , Linhagem Celular Tumoral , Pontos de Checagem da Fase G2 do Ciclo Celular
10.
Cell Death Discov ; 8(1): 173, 2022 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-35393392

RESUMO

Targeting necroptosis is considered a promising therapeutic strategy in cancer, including Glioblastoma Multiforme (GBM), one of the most lethal brain tumors. Necroptosis is a mechanism of programmed cell death overcoming the apoptosis resistance mechanism underlying GBM tumorigenesis and malignant progression. N6-isopentenyladenosine (iPA), adenosine modified with isoprenoid derivative, displays antitumor activity in different cancer models. In previous studies, we demonstrated that iPA interferes with EGFR signaling reducing glioma cell viability. Here, we show that iPA induces necroptosis in glioblastoma cell lines and in primary cells established from tumor explants, without affecting the viability of non-cancerous brain cell lines, (Normal Human Astrocyte). The activation of RIP1, RIP3, and MLKL and the upregulation of necrosome formation were increased upon iPA treatment while caspase-3, caspase-8, and PARP were not activated in GBM cells. Co-treatment with specific necroptosis inhibitor necrostatin-1 (Nec-1) or Necrosulfonamide (NSA) prevented cell death caused by iPA treatment while the general caspase inhibitor Z-VAD-fluoromethylketone (z-VAD-fmk) did not elicit any effect, suggesting that this molecule induces caspase-independent necroptosis. These results suggest that iPA treatment can be able to bypass the apoptosis resistance mechanism in glioblastoma thereby offering higher therapeutic efficacy.

11.
Cancers (Basel) ; 14(24)2022 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-36551529

RESUMO

Glioblastoma multiforme (GBM) is the most aggressive malignant brain tumor and is poorly susceptible to cytotoxic therapies. Amplification of the epidermal growth factor receptor (EGFR) and deletion of exons 2 to 7, which generates EGFR variant III (vIII), are the most common molecular alterations of GBMs that contribute to the aggressiveness of the disease. Recently, it has been shown that EGFR/EGFRvIII-targeted inhibitors enhance mitochondrial translocation by causing mitochondrial accumulation of these receptors, promoting the tumor drug resistance; moreover, they negatively modulate intrinsic mitochondria-mediated apoptosis by sequestering PUMA, leading to impaired apoptotic response in GBM cells. N6-isopentenyladenosine (i6A or iPA), a cytokinin consisting of an adenosine linked to an isopentenyl group deriving from the mevalonate pathway, has antiproliferative effects on numerous tumor cells, including GBM cells, by inducing cell death in vitro and in vivo. Here, we observed that iPA inhibits the mitochondrial respiration in GBM cells by preventing the translocation of EGFR/EGFRvIII to the mitochondria and allowing PUMA to interact with them by promoting changes in mitochondrial activity, thus playing a critical role in cell death. Our findings clearly demonstrate that iPA interferes with mitochondrial bioenergetic capacity, providing a rationale for an effective strategy for treating GBM.

12.
J Environ Sci Health B ; 46(6): 461-8, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21726142

RESUMO

A field experiment was performed with the aim to assess the amount of penconazole losses during field application and the spatial variability of penconazole concentration in a vineyard soil, under two different management techniques (tilled and grass covered). The field dissipation of penconazole under the two techniques was also followed for 114 days, highlighting the effects of spatial variability of the initial concentration and of the grass covering in terms of the different soil metabolic activity. Data found show that a high percentage of penconazole, from 42.5 % to 67.43 %, can reach the soil during the treatments, despite the fact that penconazole is applied to the foliage. The high values of the coefficient of variation for data within rows show considerable variability in all applications, ranging from 30 to 65 in the first application, from 35 to 79 in the second and from 36 to 75 in the third. Since the applications of penconazole occurred under almost same climatic conditions in terms of wind speed and direction, the high variability of concentration of penconazole found within mid-rows was attributed to the uneven slope of the vineyard causing an irregular speed of the sprayer equipment. However, least significant differences of the mean values did not end in a significant difference of penconazole concentration among mid-rows for all applications, indicating that the variability between rows does not contribute to the overall variability. The calculated half-life values for penconazole in tilled soil were 62.4 days for tilled and 33.0 for grassed soil, highlighting the tendency of penconazole to faster dissipate in grassed than in tilled soil. Grassed soil was characterised by a higher metabolic activity in terms of microbial biomass carbon content, basal respiration and total hydrolytic activity, than tilled soil. A significant correlation between penconazole dissipation and total hydrolytic activity, was found. The faster dissipation of penconazole in grassed soil was attributed to the higher amount of the total hydrolytic activity which, in grassed soil, was 1.5 fold-higher than in tilled soil.


Assuntos
Agricultura/métodos , Herbicidas/análise , Resíduos de Praguicidas/análise , Poluentes do Solo/análise , Solo/análise , Triazóis/análise , Biomassa , Carbono , Clima , Meia-Vida , Vento
13.
J Environ Sci Health B ; 44(7): 724-9, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20183083

RESUMO

A laboratory experiment was carried out to test the efficiency of a bio-mixture made up of pruning residues at two (PR2) and five (PR5) years of composting and wheat straw (STW) in the biological cleaning of water contaminated by different mixtures of fungicides usually employed in vineyards. The experiment was conducted and reproduced at a scale of 1:100 of operating field conditions. Commercial formulates of penconazole (PC), (RS)-1-[2-(2,4-dichlorophenyl)pentyl]-1H-1,2,4-triazole), dimetomorph (DM), (EZ)-4-[3-(4-chlorophenyl)-3-(3,4-dimethoxyphenyl)acryloyl]morpholine), azoxystrobin (AZ), (methyl (E)-2-{2-[6-(2-cyanophenoxy)pyrimidin-4-yloxy]phenyl}-3-methoxyacrylate), iprovalicarb (IP), (isopropyl 2-methyl-1-[(RS)-1-p-tolylethyl]carbamoyl-(S)-propylcarbamate), metalaxyl (MX), (methyl N-(methoxyacetyl)-N-(2,6-xylyl)-DL-alaninate), fludioxonil (FL), (4-(2,2-difluoro-1,3-benzodioxol-4-yl)-1H-pyrrole-3-carbonitrile) and cyprodinil (CY), (4-cyclopropyl-6-methyl-N-phenylpyrimidin-2-amine) were mixed in water and discharged into the bio-mixture following the time schedule of the treatments carried out in the grapevine in real field conditions. At each treatment, contaminated water with fungicides was circulated repeatedly through the bio-mixture to enhance the sorption of fungicides. In fact, it retained them between 98-100% with the exception of MX of which it was able to retain only 90.5%. The adsorption/desorption experiment showed that repeated circulation of water, instead of enhancing MX retention, can easily remove about 30% of MX already adsorbed by the bio-mixture. This finding suggests that water contaminated by very mobile pesticides should be discharged at the end of field treatments without re-circulating the water in order to avoid the release of pesticides weakly adsorbed on the bio-mixture.


Assuntos
Fungicidas Industriais/análise , Resíduos de Praguicidas/análise , Solo/análise , Poluentes Químicos da Água/análise , Purificação da Água , Monitoramento Ambiental , Poluentes do Solo/análise , Desintoxicação por Sorção , Vitis/química
14.
J Agric Food Chem ; 55(2): 396-401, 2007 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-17227070

RESUMO

Biobeds developed in Sweden bind and degrade pesticides from point sources. The objective of this work was to adapt the biobed to Italian operating conditions, for example, to identify organic materials as effective as those in the original Swedish composition. The capacity of urban and garden composts alone or mixed with citrus peel or straw to degrade chlorpyrifos and its metabolite TCP was compared to the typical Swedish biomix consisting of straw, peat, and soil. A tendency for higher 14C-chlorpyrifos mineralization and lower TCP levels was observed in the biomixes with garden compost alone or amended with straw. In a second trial, a high correlation of lower TCP with increasing levels of straw in typical Swedish biomixes was observed. Straw stimulates production of lignin-degrading enzymes such as manganese peroxidase (MnP), and further trials with pure MnP showed that this enzyme degrades TCP. Materials with an active lignin-degrading microflora are a prerequisite for effective dissipation of chlorpyrifos and non-accumulation of TCP. Thus, lignocellulosic materials as straw and garden composts should be present in biomixes to be used under Italian conditions.


Assuntos
Clorpirifos/química , Inseticidas/química , Solo/análise , Clorpirifos/metabolismo , Citrus , Europa (Continente) , Peroxidases/metabolismo
15.
Bioanalysis ; 9(22): 1807-1825, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29148835

RESUMO

The 2017 11th Workshop on Recent Issues in Bioanalysis (11th WRIB) took place in Los Angeles/Universal City, California from 3 April 2017 to 7 April 2017 with participation of close to 750 professionals from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. WRIB was once again a 5-day, weeklong event - A Full Immersion Week of Bioanalysis, Biomarkers and Immunogenicity. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small and large molecule analysis involving LCMS, hybrid LBA/LCMS and ligand-binding assay (LBA) approaches. This 2017 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2017 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 1) covers the recommendations for Small Molecules, Peptides and Small Molecule Biomarkers using LCMS. Part 2 (Biotherapeutics, Biomarkers and Immunogenicity Assays using Hybrid LBA/LCMS and Regulatory Agencies' Inputs) and Part 3 (LBA: Immunogenicity, Biomarkers and PK Assays) are published in volume 9 of Bioanalysis, issues 23 and 24 (2017), respectively.


Assuntos
Biomarcadores/análise , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Peptídeos/análise , Bibliotecas de Moléculas Pequenas/análise , Conferências de Consenso como Assunto , Guias como Assunto , Ligantes , Bibliotecas de Moléculas Pequenas/química
16.
Bioanalysis ; 8(22): 2363-2378, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27712081

RESUMO

The 2016 10th Workshop on Recent Issues in Bioanalysis (10th WRIB) took place in Orlando, Florida with participation of close to 700 professionals from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations, and regulatory agencies worldwide. WRIB was once again a 5-day, weeklong event - A Full Immersion Week of Bioanalysis including Biomarkers and Immunogenicity. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small and large molecule analysis involving LCMS, hybrid LBA/LCMS, and LBA approaches, with the focus on biomarkers and immunogenicity. This 2016 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. This white paper is published in 3 parts due to length. This part (Part 1) discusses the recommendations for small molecules, peptides and small molecule biomarkers by LCMS. Part 2 (Hybrid LBA/LCMS and regulatory inputs from major global health authorities) and Part 3 (large molecule bioanalysis using LBA, biomarkers and immunogenicity) will be published in the Bioanalysis journal, issue 23.

17.
Bioanalysis ; 7(22): 2913-25, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26573485

RESUMO

The 2015 9th Workshop on Recent Issues in Bioanalysis (9th WRIB) took place in Miami, Florida with participation of over 600 professionals from pharmaceutical and biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. It is once again a 5-day week long event - a full immersion bioanalytical week - specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest in bioanalysis. The topics covered included both small and large molecules, and involved LCMS, hybrid LBA/LCMS, LBA approaches including the focus on biomarkers and immunogenicity. This 2015 White Paper encompasses recommendations that emerged from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to advance scientific excellence, improve quality and deliver better regulatory compliance. Due to its length, the 2015 edition of this comprehensive White Paper has been divided into three parts. Part 1 covers the recommendations for small molecule bioanalysis using LCMS. Part 2 (hybrid LBA/LCMS and regulatory agencies' inputs) and Part 3 (large molecule bioanalysis using LBA, biomarkers and immunogenicity) will also be published in volume 7 of Bioanalysis, issues 23 and 24, respectively.


Assuntos
Biomarcadores/análise , Cromatografia Líquida/normas , Espectrometria de Massas/normas , Bibliotecas de Moléculas Pequenas/análise , Humanos
18.
Environ Sci Pollut Res Int ; 20(4): 2546-55, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22965543

RESUMO

Biopurification systems (BPS) have been introduced to minimise the risk for point source contamination of natural water resources by pesticides. Their depuration efficiency relies mostly on the high biodegradation of their packing substrate (biomixture). Despite that, little is known regarding the interactions between biomixture microflora and pesticides, especially fungicides which are expected to have a higher impact on the microbial community. This study reports the dissipation of the fungicides azoxystrobin (AZX), fludioxonil (FL) and penconazole (PC), commonly used in vineyards, in a biomixture composed of pruning residues and straw used in vineyard BPS. The impact of fungicides on the microbial community was also studied via microbial biomass carbon, basal respiration and phospholipid fatty acid analysis. AZX dissipated faster (t1/2 = 30.1 days) than PC (t1/2 = 99.0 days) and FL (t1/2 = 115.5 days). Fungicides differently affected the microbial community. PC showed the highest adverse effect on both the size and the activity of the biomixture microflora. A significant change in the structure of the microbial community was noted for PC and FL, and it was attributed to a rapid inhibition of the fungal fraction while bacteria showed a delayed response which was attributed to indirect effects by the late proliferation of fungi. All effects observed were transitory and a full recovery of microbial indices was observed 60 days post-application. Overall, no clear link between pesticide persistence and microbial responses was observed stressing the complex nature of interactions between pesticides in microflora in BPS.


Assuntos
Recuperação e Remediação Ambiental/métodos , Fungicidas Industriais/metabolismo , Consórcios Microbianos/efeitos dos fármacos , Componentes Aéreos da Planta/microbiologia , Vitis/microbiologia , Análise de Variância , Biodegradação Ambiental , Cromatografia Líquida de Alta Pressão , Dioxóis/metabolismo , Dioxóis/farmacologia , Fungicidas Industriais/farmacologia , Cromatografia Gasosa-Espectrometria de Massas , Meia-Vida , Metacrilatos/metabolismo , Metacrilatos/farmacologia , Consórcios Microbianos/fisiologia , Análise Multivariada , Componentes Aéreos da Planta/química , Análise de Componente Principal , Pirimidinas/metabolismo , Pirimidinas/farmacologia , Pirróis/metabolismo , Pirróis/farmacologia , Extração em Fase Sólida , Estrobilurinas , Triazóis/metabolismo , Triazóis/farmacologia , Vitis/química
19.
Pest Manag Sci ; 67(1): 107-13, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21162150

RESUMO

BACKGROUND: The composition and properties of a biomixture used in a biobed are decisive for pesticide sorption and degradation. This study was performed to investigate the capability of compost-based substrates in mixtures with citrus peel and vine branch straw and peat-based substrates in mixtures with soil and vine branch straw at different levels in order to degrade isoproturon and bentazone. RESULTS: Dissipation and mineralisation rates of both pesticides were determined, and metabolic activity was followed as respiration. Compost-based substrates showed faster pesticide dissipation in the presence of lignocellulosic materials, as in garden compost and vine branch straw. The increasing content of vine branch straw in peat-based substrates does not seem to affect dissipation of the parent compounds. Low mineralisation rate was observed in all treatments. CONCLUSION: Higher pesticide degradation was observed in the lignocellulosic substrates, probably because of the development of lignin-degrading microorganisms which have shown to be robust and are able to degrade recalcitrant pesticides. Copyright © 2010 Society of Chemical Industry.


Assuntos
Benzotiadiazinas/química , Recuperação e Remediação Ambiental , Herbicidas/química , Compostos de Fenilureia/química , Poluentes do Solo/química , Solo/química , Biodegradação Ambiental , Microbiologia do Solo , Fatores de Tempo
20.
N Biotechnol ; 29(1): 99-106, 2011 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-21439413

RESUMO

Biological systems are being developed all over EU countries to protect water-bodies from pesticide contamination at farm level. A laboratory experiment was carried out to test the efficiency of a mixture of compost and straw in bio-degrading different mixtures of fungicides usually applied in vineyards. At the same time the effects of fungicide applications on microbial community of biomixture were also evaluated. Results showed that the biomixture had a good capability of degrading pesticides. Indeed, at the end of the experiment (112 days), the concentration of most of the pesticides was close to complete degradation. Denaturing gradient gel electrophoresis (DGGE) analysis showed an evident modification of microbial diversity after the addition of fungicides. However, at the end of degradation process, no significant changes in the composition of microbial community were seen. In this specific substrate used in the biomixture, yeast flora and ascomycete filamentous fungi seemed to be involved in the degradation activity.


Assuntos
Fungicidas Industriais/metabolismo , Praguicidas/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Agricultura/métodos , Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Biodegradação Ambiental , Análise por Conglomerados , Eletroforese/métodos , Fungos/efeitos dos fármacos , Fungos/metabolismo , Fungicidas Industriais/farmacologia , Praguicidas/farmacologia
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