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1.
Molecules ; 28(6)2023 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-36985499

RESUMO

Supramolecular and biocompatible hydrogels with a tunable pH ranging from 5.5 to 7.6 lead to a wide variety of formulations useful for many different topical applications compatible with the skin pH. An in vitro viability/cytotoxicity test of the gel components demonstrated that they are non-toxic, as the cells continue to proliferate after 48 h. An analysis of the mechanical properties demonstrates that the hydrogels have moderate strength and an excellent linear viscoelastic range with the absence of a proper breaking point, confirmed with thixotropy experiments. Two cosmetic active peptides (Trifluoroacetyl tripeptide-2 and Palmitoyl tripeptide-5) were successfully added to the hydrogels and their transdermal permeation was analysed with Franz diffusion cells. The liquid chromatography-mass spectrometry (HPLC-MS) analyses of the withdrawn samples from the receiving solutions showed that Trifluoroacetyl tripeptide-2 permeated in a considerable amount while almost no transdermal permeation of Palmitoyl tripeptide-5 was observed.


Assuntos
Hidrogéis , Peptídeos , Hidrogéis/química , Peptídeos/química , Administração Cutânea , Composição de Medicamentos , Materiais Biocompatíveis/química
2.
Molecules ; 28(10)2023 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-37241793

RESUMO

Metallodrugs represent a combination of multifunctionalities that are present concomitantly and can act differently on diverse biotargets. Their efficacy is often related to the lipophilic features exhibited both by long carbo-chains and the phosphine ligands. Three Ru(II) complexes containing hydroxy stearic acids (HSAs) were successfully synthesized in order to evaluate possible synergistic effects between the known antitumor activity of HSA bio-ligands and the metal center. HSAs were reacted with [Ru(H)2CO(PPh3)3] selectively affording O,O-carboxy bidentate complexes. The organometallic species were fully characterized spectroscopically using ESI-MS, IR, UV-Vis, and NMR techniques. The structure of the compound Ru-12-HSA was also determined using single crystal X-ray diffraction. The biological potency of ruthenium complexes (Ru-7-HSA, Ru-9-HSA, and Ru-12-HSA) was studied on human primary cell lines (HT29, HeLa, and IGROV1). To obtain detailed information about anticancer properties, tests for cytotoxicity, cell proliferation, and DNA damage were performed. The results demonstrate that the new ruthenium complexes, Ru-7-HSA and Ru-9-HSA, possess biological activity. Furthermore, we observed that the Ru-9-HSA complex shows increased antitumor activity on colon cancer cells, HT29.


Assuntos
Antineoplásicos , Complexos de Coordenação , Rutênio , Humanos , Antineoplásicos/farmacologia , Antineoplásicos/química , Rutênio/química , Ligantes , Células HeLa , Proliferação de Células , Complexos de Coordenação/farmacologia , Complexos de Coordenação/química , Linhagem Celular Tumoral
3.
Molecules ; 27(13)2022 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-35807399

RESUMO

Amino acids are ubiquitous components of mammalian milk and greatly contribute to its nutritional value. The compositional analysis of free amino acids is poorly reported in the literature even though their determination in the biological fluids of livestock animals is necessary to establish possible nutritional interventions. In the present study, the free amino acid profiles in mature swine milk, colostrum and plasma were assessed using a targeted metabolomics approach. In particular, 20 amino acids were identified and quantified via two alternative and complementary reversed-phase HPLC methods, involving two stationary phases based on core-shell technology, i.e., Kinetex C18 and Kinetex F5, and two detection systems, i.e., a diode array detector (DAD) and a fluorescence detector (FLD). The sample preparation involved a de-proteinization step, followed by pre-chromatographic derivatization with 9-fluorenylmethylchloroformate (FMOC-Cl). The two optimized methods were validated for specificity, linearity, sensitivity, matrix effect, accuracy and precision and the analytical performances were compared. The analytical methods proved to be suitable for free amino acid profiling in different matrices with high sensitivity and specificity. The correlations among amino acid levels in different biological fluids can be useful for the evaluation of physio-pathological status and to monitor the effects of therapeutic or nutritional interventions in humans and animals.


Assuntos
Aminoácidos , Leite , Aminoácidos/química , Animais , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida , Colostro/química , Feminino , Mamíferos , Leite/química , Gravidez , Suínos
4.
Molecules ; 27(17)2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-36080409

RESUMO

A series of naphthoquinones, namely, 1,4-naphthoquinone, menadione, plumbagin, juglone, naphthazarin, and lawsone, were reacted with N-acetyl-L-cysteine, and except for lawsone, which did not react, the related adducts were obtained. After the tuning of the solvent and reaction conditions, the reaction products were isolated as almost pure from the complex reaction mixture via simple filtration and were fully characterized. Therefore, the aim of this work was to evaluate whether the antitumor activity of new compounds of 1,4-naphthoquinone derivatives leads to an increase in ROS in tumor cell lines of cervical carcinoma (HeLa), neuroblastoma (SH-SY5Y), and osteosarcoma (SaOS2, U2OS) and in normal dermal fibroblast (HDFa). The MTT assay was used to assay cell viability, the DCF-DA fluorescent probe to evaluate ROS induction, and cell-cycle analysis to measure the antiproliferative effect. Compounds 8, 9, and 12 showed a certain degree of cytotoxicity towards all the malignant cell lines tested, while compound 11 showed biological activity at higher IC50 values. Compounds 8 and 11 induced increases in ROS generation after 1 h of exposure, while after 48 h of treatment, only 8 induced an increase in ROS formation in HeLa cells. Cell-cycle analysis showed that compound 8 caused an increase in the number of G0/G1-phase cells in the HeLa experiment, while for the U2OS and SH-SY5Y cell lines, it led to an accumulation of S-phase cells. Therefore, these novel 1,4-naphthoquinone derivatives may be useful as antitumoral agents in the treatment of different cancers.


Assuntos
Naftoquinonas , Neuroblastoma , Acetilcisteína/farmacologia , Linhagem Celular Tumoral , Células HeLa , Humanos , Naftoquinonas/metabolismo , Naftoquinonas/farmacologia , Espécies Reativas de Oxigênio/metabolismo
5.
Molecules ; 25(19)2020 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-32992652

RESUMO

Ovarian cancer remains the leading cause of mortality among gynecological tumors. Estrogen receptor beta (ERß) expression has been suggested to act as a tumor suppressor in epithelial ovarian cancer by reducing both tumor growth and metastasis. ERß expression abnormalities represent a critical step in the development and progression of ovarian cancer: for these reasons, its re-expression by genetic engineering, as well as the use of targeted ERß therapies, still constitute an important therapeutic approach. 3-{[2-chloro-1-(4-chlorobenzyl)-5-methoxy-6-methyl-1H-indol-3-yl]methylene}-5-hydroxy-6-methyl-1,3-dihydro-2H-indol-2-one, referred to here as compound 3, has been shown to have cytostatic as well cytotoxic effects on various hormone-dependent cancer cell lines. However, the mechanism of its anti-carcinogenic activity is not well understood. Here, we offer a possible explanation of such an effect in the human ovarian cancer cell line IGROV1. Chromatin binding protein assay and liquid chromatography mass spectrometry were exploited to localize and quantify compound 3 in cells. Molecular docking was used to prove compound 3 binding to ERß. Mass spectrometry-based approaches were used to analyze histone post-translational modifications. Finally, gene expression analyses revealed a set of genes regulated by the ERß/3 complex, namely CCND1, MYC, CDKN2A, and ESR2, providing possible molecular mechanisms that underline the observed antiproliferative effects.


Assuntos
Receptor beta de Estrogênio , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Indóis , Simulação de Acoplamento Molecular , Proteínas de Neoplasias , Neoplasias Ovarianas/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Receptor beta de Estrogênio/agonistas , Receptor beta de Estrogênio/química , Receptor beta de Estrogênio/metabolismo , Feminino , Humanos , Indóis/química , Indóis/farmacologia , Proteínas de Neoplasias/agonistas , Proteínas de Neoplasias/química , Proteínas de Neoplasias/metabolismo , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/patologia
6.
Mol Pharm ; 16(3): 931-942, 2019 03 04.
Artigo em Inglês | MEDLINE | ID: mdl-30702899

RESUMO

Histone deacetylases, HDACs, have been demonstrated to play a critical role in epigenetic signaling and were found to be overexpressed in several type of cancers; therefore, they represent valuable targets for anticancer therapy. 9-Hydroxystearic acid has been shown to bind the catalytic site of HDAC1, inducing G0/G1 phase cell cycle arrest and activation of the p21WAF1 gene, thus promoting cell growth inhibition and differentiation in many cancer cells. Despite the ( R) enantiomer of 9-hydroxystearic acid (9R) displaying a promising in vitro growth-inhibitory effect on the HT29 cell line, its scarce water solubility and micromolar activity require novel solutions for improving its efficacy and bioavailability. In this work, we describe the synthesis and in vitro biological profiling of 9R keratin nanoparticles (9R@ker) obtained through an in-water drug-induced aggregation process. The anticancer activity of 9R@ker was investigated in the HT29 cell line; the results indicate an increased fluidity of cell membrane and a higher intracellular ROS formation, resulting in an unexpected S phase cell cycle arrest (25% increase as compared to the control) induced by 9R@ker with respect to free 9R and an induction of cell death.


Assuntos
Adenocarcinoma/patologia , Neoplasias do Colo/patologia , Descoberta de Drogas/métodos , Queratinas/química , Nanopartículas/química , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Ácidos Esteáricos/química , Albuminas/química , Membrana Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Química Sintética/métodos , Células HCT116 , Células HT29 , Histona Desacetilase 1/antagonistas & inibidores , Humanos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Solubilidade , Ácidos Esteáricos/farmacologia
7.
Biochim Biophys Acta Bioenerg ; 1859(3): 182-190, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29269267

RESUMO

A marked stimulation of complex II enzymatic activity was detected in cybrids bearing a homoplasmic MTCYB microdeletion causing disruption of both the activity and the assembly of complex III, but not in cybrids harbouring another MTCYB mutation affecting only the complex III activity. Moreover, complex II stimulation was associated with SDHA subunit tyrosine phosphorylation. Despite the lack of detectable hydrogen peroxide production, up-regulation of the levels of mitochondrial antioxidant defenses revealed a significant redox unbalance. This effect was also supported by the finding that treatment with N-acetylcysteine dampened the complex II stimulation, SDHA subunit tyrosine phosphorylation, and levels of antioxidant enzymes. In the absence of complex III, the cellular amount of succinate, but not fumarate, was markedly increased, indicating that enhanced activity of complex II is hampered due to the blockage of respiratory electron flow. Thus, we propose that complex II phosphorylation and stimulation of its activity represent a molecular mechanism triggered by perturbation of mitochondrial redox homeostasis due to severe dysfunction of respiratory complexes. Depending on the site and nature of the damage, complex II stimulation can either bypass the energetic deficit as an efficient compensatory mechanism, or be ineffectual, leaving cells to rely on glycolysis for survival.


Assuntos
Complexo III da Cadeia de Transporte de Elétrons/metabolismo , Complexo II de Transporte de Elétrons/metabolismo , Homeostase , Mitocôndrias/metabolismo , Acetilcisteína/farmacologia , Citocromos b/genética , Citocromos b/metabolismo , Transporte de Elétrons/efeitos dos fármacos , Complexo II de Transporte de Elétrons/genética , Complexo III da Cadeia de Transporte de Elétrons/genética , Sequestradores de Radicais Livres/farmacologia , Humanos , Células Híbridas/metabolismo , Peróxido de Hidrogênio/metabolismo , Mitocôndrias/genética , Mutação , Oxirredução , Fosforilação/efeitos dos fármacos , Succinatos/metabolismo
8.
Diabetes Metab Res Rev ; 33(8)2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28806487

RESUMO

BACKGROUND: Reactive hypoglycemia is a metabolic disorder that provokes severe hypoglycemic episodes after meals. Over recent years, the gut microbiota has been recognized as potential target for the control of metabolic diseases, and the possibility to correct gut microbiota dysbioses through diet, favouring the recovery of metabolic homeostasis, has been considered. METHODS: We investigate the impact of 2 short-term (3-day) nutritional interventions, based on the macrobiotic Ma-Pi 2 diet and a control Mediterranean diet, on the structure and functionality of the gut microbiota in 12 patients affected by reactive hypoglycemia. The gut microbiota composition was characterized by next-generation sequencing of the V3 to V4 region of the 16S rRNA gene, and the ecosystem functionality was addressed by measuring the faecal concentration of short-chain fatty acids (SCFAs). In order to measure the short-term physiological gut microbiota fluctuation, the microbiomes of 7 healthy people were characterized before and after 3 days of constant diet. RESULTS: While no convergence of the gut microbiota compositional profiles was observed, a significant increase in SCFA faecal levels was induced only in the Ma-Pi 2 diet group, suggesting the potential of this diet to support a short-term functional convergence of the gut microbiota, regardless of the individual compositional layout. CONCLUSIONS: The Ma-Pi 2 diet, with its high fibre load, was effective in increasing the production of SCFAs by the gut microbiota. Because these metabolites are known for their ability to counterbalance the metabolic deregulation in persons with glucose impairment disorders, their increased bioavailability could be of some relevance in reactive hypoglycemia.


Assuntos
Microbioma Gastrointestinal/fisiologia , Hipoglicemia/microbiologia , Adulto , Idoso , Dieta Macrobiótica , Fezes/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento
9.
Electrophoresis ; 36(21-22): 2820-2827, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26200671

RESUMO

Pharmaceutical iron sucrose is an iron (III) replacement for the treatment of iron deficiency anemia in patients with chronic kidney disease. The drug product (injection) is a colloidal solution of ferric hydroxide in complex with sucrose, containing 20 mg/mL elemental iron; according to United States pharmacopoeia (USP), the limit of iron (II) is 0.4% w/v. A selective CE method for the simultaneous determination of iron (III) and its potential impurity iron (II), was developed by applying a dual precapillary complexation. In particular, 1,10-phenanthroline and 1,2-diaminocyclohexanetetraacetic acid were used for complexation of iron (II) and iron (III), respectively. Sample preparation was optimized to achieve mineralization of pharmaceuticals using HCl 6 M, by avoiding perturbation of the oxidation status of both iron species. Simple CZE conditions, involving a 60 mM (pH 9.3) tetraborate buffer at the constant voltage of 25 KV and 25°C, allowed fast separation of iron (II) and iron (III) complexes that were detected at 265 nm. Sensitivity for iron (II) determination was found to be 4.80 µM (LOQ) corresponding to 0.15% w/w with respect to the total iron test level. The method was validated by following International Conference on Harmonization guidelines for specificity, linearity, precision, accuracy, and robustness and it was applied to real pharmaceutical samples. The obtained results suggested that the method can be a useful alternative to the official USP and British pharmacopoeia polarographic method.

10.
Sci Adv ; 10(25): eadk9117, 2024 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-38905343

RESUMO

The microbiome plays a key role in the health of all metazoans. Whether and how the microbiome favors the adaptation processes of organisms to extreme conditions, such as those of Antarctica, which are incompatible with most metazoans, is still unknown. We investigated the microbiome of three endemic and widespread species of Antarctic polychaetes: Leitoscoloplos geminus, Aphelochaeta palmeri, and Aglaophamus trissophyllus. We report here that these invertebrates contain a stable bacterial core dominated by Meiothermus and Anoxybacillus, equipped with a versatile genetic makeup and a unique portfolio of proteins useful for coping with extremely cold conditions as revealed by pangenomic and metaproteomic analyses. The close phylosymbiosis between Meiothermus and Anoxybacillus and these Antarctic polychaetes indicates a connection with their hosts that started in the past to support holobiont adaptation to the Antarctic Ocean. The wide suite of bacterial cryoprotective proteins found in Antarctic polychaetes may be useful for the development of nature-based biotechnological applications.


Assuntos
Congelamento , Microbiota , Poliquetos , Poliquetos/microbiologia , Animais , Regiões Antárticas , Filogenia , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética
11.
Food Chem ; 425: 136453, 2023 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-37271683

RESUMO

The release of hazardous chemicals into aquatic environments has long been a known problem, but its full impact has only recently been realized. This study presents a validated liquid chromatography-mass spectrometry (HPLC-MS/MS) method for detecting pharmaceutical and pesticide residues in mussels (Mytilus galloprovincialis). An innovative MS-compatible extraction method was developed and validated, demonstrating successful recovery rates for analytes at three different concentration levels (25-95%). The method detected the target analytes at ng/g concentrations with high accuracy (-7% to 11%) and low relative standard deviation (<10%) for both intra-day and inter-day analyses. After validation, the method was applied to mussel samples collected from a commercial farm near Senigallia, Adriatic Sea, detecting different contaminants in the range of 2-40 ng/g (dry weight). The study provides a valuable tool for investigating the potential threats posed by diverse contaminant classes with high annual tonnage, including analytes with known persistence and/or illegal status.


Assuntos
Poluentes Ambientais , Mytilus , Poluentes Químicos da Água , Animais , Espectrometria de Massas em Tandem , Poluentes Ambientais/análise , Poluentes Químicos da Água/análise , Mytilus/química , Substâncias Perigosas
12.
Metabolites ; 13(9)2023 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-37755260

RESUMO

Bile acids (BAs), endogenous acidic steroids synthetized from cholesterol in the liver, play a key role in the gut-liver axis physiopathology, including in hepatotoxicity, intestinal inflammatory processes, and cholesterol homeostasis. Faecal Oxo-BAs, relatively stable intermediates of oxidation/epimerization reactions of the BA hydroxyls, could be relevant to investigating the crosstalk in the liver-gut axis and the relationship between diseases and alterations in microbiota composition. A paucity of information currently exists on faecal BA profiles in dogs with and without chronic inflammatory enteropathy (CIE). Comprehensive assessment of 31 molecules among faecal BAs and related microbiota metabolites was conducted with high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Odds ratios (ORs) for associations of BAs with CIE were estimated using logistic regression. Principal component analysis was performed to find differences between the control and pathological dogs. Higher levels of primary BAs and muricholic acids, and lower levels of secondary BAs were found in pathological dogs. Higher concentrations in faecal oxo-metabolites were associated with the absence of CIE (OR < 1). This study shows a marked difference in faecal BA profiles between dogs with and without CIE. Further research will be needed to better understand the role of oxo-BAs and muricholic acids in CIE dogs.

13.
Sci Total Environ ; 887: 163948, 2023 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-37149185

RESUMO

Marine mussels, especially Mytilus galloprovincialis, are well-established sentinel species, being naturally resistant to the exposure to multiple xenobiotics of natural and anthropogenic origin. Even if the response to multiple xenobiotic exposure is well known at the host level, the role of the mussel-associated microbiome in the animal response to environmental pollution is poorly explored, despite its potential in xenobiotic detoxification and its important role in host development, protection, and adaptation. Here, we characterized the microbiome-host integrative response of M. galloprovincialis in a real-world setting, involving exposure to a complex pattern of emerging pollutants, as occurs in the Northwestern Adriatic Sea. A total of 387 mussel individuals from 3 commercial farms, spanning about 200 km along the Northwestern Adriatic coast, and in 3 different seasons, were collected. Multiresidue analysis (for quantitative xenobiotic determination), transcriptomics (for host physiological response), and metagenomics (for host-associated microbial taxonomical and functional features) analyses were performed on the digestive glands. According to our findings, M. galloprovincialis responds to the presence of the complex pattern of multiple emerging pollutants - including the antibiotics sulfamethoxazole, erythromycin, and tetracycline, the herbicides atrazine and metolachlor, and the insecticide N,N-diethyl-m-toluamide - integrating host defense mechanisms, e.g., through upregulation of transcripts involved in animal metabolic activity, and microbiome-mediated detoxification functions, including microbial functionalities involved in multidrug or tetracycline resistance. Overall, our data highlight the importance of the mussel-associated microbiome as a strategic player for the orchestration of resistance to the multixenobiotic exposure at the holobiont level, providing strategic functionalities for the detoxification of multiple xenobiotic substances, as occurring in real world exposure settings. Complementing the host with microbiome-dependent xenobiotic degradative and resistance genes, the M. galloprovincialis digestive gland associated microbiome can have an important role in the detoxification of emerging pollutants in a context of high anthropogenic pressure, supporting the relevance of mussel systems as potential animal-based bioremediation tool.


Assuntos
Microbiota , Mytilus , Praguicidas , Poluentes Químicos da Água , Animais , Mytilus/metabolismo , Estações do Ano , Praguicidas/análise , Xenobióticos/metabolismo , Poluentes Químicos da Água/análise
14.
Electrophoresis ; 33(22): 3380-6, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22961751

RESUMO

Progression of Alzheimer's disease involves aggregation of amyloid-beta (Aß) peptides, a complex process that involves the formation of several soluble intermediates and ends up with the deposition of ordered fibrillar architectures. The determination of the Aß42 self-assembly species targeted by an inhibitor is a key step in the identification of new inhibitors endowed with a suitable profile. In this context, the subtle characterization of myricetin (Myr) mode of action at a molecular level was performed by using different MS techniques, which allowed the monitoring of the modifications induced by Myr on the first occurring Aß42 self-assembly species. Results showed a persistent level of monomer and decreased formation of ordered Aß42 aggregates in the presence of Myr, further, nano-LC-nano-ESI-QTOF, MALDI-TOF, and ESI-IT highlighted the formation of a new oxidized Aß42 species, which is less prone to aggregation, in the presence of Myr. Coupling tryptic digestion and nano-LC-nano-ESI-QTOF also allowed the identification of Met(35) as the specific site of oxidation along the Aß aminoacid chain. Therefore, the detailed investigation by different MS techniques allowed better understanding of the molecular modification at the basis of the antiaggregating properties of Myr and highlighted its oxidizing action on the residue Met(35) in Aß monomers.


Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Amiloide/antagonistas & inibidores , Flavonoides/farmacologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Sequência de Aminoácidos , Amiloide/química , Peptídeos beta-Amiloides/química , Peptídeos beta-Amiloides/metabolismo , Cromatografia Líquida , Flavonoides/química , Dados de Sequência Molecular , Oxirredução , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Espectrometria de Massas por Ionização por Electrospray , Tripsina/química , Tripsina/metabolismo
16.
Sci Rep ; 12(1): 2866, 2022 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-35190565

RESUMO

There is a growing interest in the named "acidic sterolbiome" and in the genetic potential of the gut microbiome (GM) to modify bile acid (BA) structure. Indeed, the qualitative composition of BAs in feces correlates with the bowel microorganisms and their collective genetic material. GM is responsible for the production of BA metabolites, such as secondary and oxo-BAs. The specific BA profiles, as microbiome-host co-metabolic products, could be useful to investigate the GM-host interaction in animals under physiological conditions, as well as in specific diseases. In this context, we developed and validated an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry method for the simultaneous analysis of up to 21 oxo-BAs and their 9 metabolic precursors. Chromatographic separation was achieved in 7 min with adequate analytical performance in terms of selectivity, sensitivity (LOQ from 0.05 to 0.1 µg/mL), accuracy (bias% < 5%), precision (CV% < 5%) and matrix effect (ME% < 10%). A fast solvent extraction protocol has been fine-tuned, achieving recoveries > 90%. In parallel, the gut microbiota assessment in farming animals was evaluated by 16S rRNA next-generation sequencing, and the correlation with the BA composition was performed by multivariate analysis, allowing to reconstruct species-specific associations between the BA profile and specific GM components.


Assuntos
Animais Domésticos/metabolismo , Animais Domésticos/microbiologia , Ácidos e Sais Biliares/análise , Ácidos e Sais Biliares/metabolismo , Fezes/química , Microbioma Gastrointestinal , Espectrometria de Massas/métodos , Animais , Cromatografia Líquida/métodos , Microbioma Gastrointestinal/genética , Sequenciamento de Nucleotídeos em Larga Escala , Interações entre Hospedeiro e Microrganismos , RNA Bacteriano/genética , Sensibilidade e Especificidade , Especificidade da Espécie
17.
J Chromatogr A ; 1681: 463452, 2022 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-36058082

RESUMO

Glyphosate or N-(phosphonomethyl)glycine, widely used as herbicide in agriculture to control weeds and to facilitate harvesting, has been included in Group 2A pollutants (probably carcinogenic to humans) by the International Agency for Research on Cancer (IARC). In intensive agricultural areas, runoff and soil leaching are likely to drive glyphosate to surface waters, where the compound is often detected together with its main microbial metabolite, aminomethylphosphonic acid (AMPA). In the present study a method based on capillary electrophoresis coupled with light-emitting diode-induced fluorescence detection has been developed and validated for the determination of the two compounds in whole soft mass of marine mussels (Mytilus galloprovincialis). The method is based on the acidic hydrolysis of lyophilized tissue using 6 M HCl (oven at 110 °C for 22 h) to release the target analytes; their subsequent derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole, was found to be suitable for the sensitive fluorescence detection. To achieve optimum separation of the analytes from the matrix and degradation reagent interferences, the background electrolyte constituted by borate buffer (pH 9.2, 30 mM) was supplemented with 10 mM heptakis(2,6-di-O-methyl)-ß-cyclodextrin. The method was validated for linearity, precision, accuracy, robustness and sensitivity showing LOQ of 0.2 and 1.0 µg/g in fresh tissues, for AMPA and glyphosate, respectively; the recovery values ranged within 88.5 - 94.6% for glyphosate and 70.4 - 76.6% for AMPA. Experimental samples of Mediterranean mussels M. galloprovincialis treated with 100 µg/L or 500 µg/L of both glyphosate and AMPA, showed a dose dependent bioaccumulation of the compounds reaching maximum level of 77.0 µg/g and 11.3 µg/g of AMPA and glyphosate, respectively. The study demonstrates for the first time M. galloprovincialis as potential sentinel organisms for the environmental occurrence of these small amphoteric pollutants.


Assuntos
Bivalves , Herbicidas , Poluentes Químicos da Água , Animais , Bioacumulação , Boratos/análise , Eletroforese Capilar , Glicina/análogos & derivados , Herbicidas/análise , Humanos , Organofosfonatos , Compostos Organofosforados , Solo/química , Poluentes Químicos da Água/análise , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiônico/análise , Glifosato
18.
Environ Toxicol Pharmacol ; 96: 103997, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36216252

RESUMO

This study investigates the effects of glyphosate (GLY) and its metabolite AMPA on cytoprotective and detoxification mechanisms in haemocytes of Mytilus galloprovincialis. Cells were treated in vitro with 0.1 and 1.0 µg/L GLY, 0.1 µg/L, 0.1 and 1.0 µg/L AMPA, or two mixtures GLY+AMPA (0.1 µg/L GLY + 0.1 µg/L AMPA, 1.0 µg/L GLY + 1.0 µg/L AMPA). GLY and AMPA increased MXR efflux activity and modulated expression of the ABCB transcript encoding a MXR related ABC transporter P-glycoprotein. The mixtures GLY+AMPA reduced efflux activity with ABCB down-regulation (at 1 µg/L GLY/AMPA). Modulation of lysosomal and immune related transcripts generally agree with known effects of the chemicals on these physiological functions. Given their cumulative action as chemosensitizers of the MXR system, and their interactive effects on haemocyte parameters, glyphosate and AMPA at environmental concentrations should be addressed as a concern factor for the biological vulnerability of marine habitats.


Assuntos
Mytilus , Poluentes Químicos da Água , Animais , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiônico/metabolismo , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiônico/farmacologia , Glicina , Hemócitos , Poluentes Químicos da Água/metabolismo , Glifosato
19.
Front Plant Sci ; 13: 936991, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36017264

RESUMO

Increasing temperatures, heat waves, and reduction of annual precipitation are all the expressions of climate change (CC), strongly affecting bread wheat (Triticum aestivum L.) grain yield in Southern Europe. Being temperature the major driving force of plants' phenological development, these variations also have effects on wheat phenology, with possible consequences on grain quality, and gluten protein accumulation. Here, through a case study in the Bolognese Plain (North of Italy), we assessed the effects of CC in the area, the impacts on bread wheat phenological development, and the consequences on grain gluten quality. The increasing trend in mean annual air temperature in the area since 1952 was significant, with a breakpoint identified in 1989, rising from 12.7 to 14.1°C, accompanied by the signals of increasing aridity, i.e., increase in water table depth. Bread wheat phenological development was compared in two 15-year periods before and after the breakpoint, i.e., 1952-1966 (past period), and 2006-2020 (present period), the latest characterized by aridity and increased temperatures. A significant shortening of the chronological time necessary to reach the main phenological phases was observed for the present period compared to the past period, finally shortening the whole life cycle. This reduction, as well as the higher temperature regime, affected gluten accumulation during the grain-filling process, as emerged analyzing gluten composition in grain samples of the same variety harvested in the area both before and after the breakpoint in temperature. In particular, the proportion of gluten polymers (i.e., gliadins, high and low molecular weight glutenins, and their ratio) showed a strong and significant correlation with cumulative growing degree days (CGDDs) accumulated during the grain filling. Higher CGDD values during the period, typical of CC in Southern Europe, accounting for higher temperature and faster grain filling, correlated with gliadins, high molecular weight glutenins, and their proportion with low molecular weight glutenins. In summary, herein reported, data might contribute to assessing the effects of CC on wheat phenology and quality, representing a tool for both predictive purposes and decision supporting systems for farmers, as well as can guide future breeding choices for varietal innovation.

20.
Appl Environ Microbiol ; 77(19): 7072-6, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21821753

RESUMO

Human plasmin(ogen) is regarded as a component of the molecular cross talk between the probiotic species Bifidobacterium animalis subsp. lactis and the human host. However, up to now, only in vitro studies have been reported. Here, we demonstrate that the probiotic strain B. animalis subsp. lactis BI07 is capable of recruiting plasmin(ogen) present at physiological concentrations in crude extracts from human feces. Our results provide evidence that supports the significance of the B. lactis-plasmin(ogen) interaction in the human gastrointestinal tract.


Assuntos
Proteínas de Bactérias/metabolismo , Bifidobacterium/fisiologia , Proteínas de Transporte/metabolismo , Trato Gastrointestinal/microbiologia , Plasminogênio/metabolismo , Bifidobacterium/crescimento & desenvolvimento , Bifidobacterium/metabolismo , Fezes/microbiologia , Humanos , Probióticos
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