RESUMO
In recently developed approaches for high-resolution imaging within intact tissue, molecular characterization over large volumes has been largely restricted to labeling of proteins. But volumetric nucleic acid labeling may represent a far greater scientific and clinical opportunity, enabling detection of not only diverse coding RNA variants but also non-coding RNAs. Moreover, scaling immunohistochemical detection to large tissue volumes has limitations due to high cost, limited renewability/availability, and restricted multiplexing capability of antibody labels. With the goal of versatile, high-content, and scalable molecular phenotyping of intact tissues, we developed a method using carbodiimide-based chemistry to stably retain RNAs in clarified tissue, coupled with amplification tools for multiplexed detection. The resulting technology enables robust measurement of activity-dependent transcriptional signatures, cell-identity markers, and diverse non-coding RNAs in rodent and human tissue volumes. The growing set of validated probes is deposited in an online resource for nucleating related developments from across the scientific community.
Assuntos
Química Encefálica , Hibridização In Situ/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA/análise , Transcriptoma , Adolescente , Animais , Cianatos/química , Etildimetilaminopropil Carbodi-Imida/química , Feminino , Humanos , Masculino , Maleimidas/química , Camundongos , Pessoa de Meia-Idade , Oligonucleotídeos/química , Succinimidas/químicaRESUMO
The mammalian immune system uses various pattern recognition receptors to recognize invaders and host damage and transmits this information to downstream immunometabolic signalling outcomes. Laccase domain-containing 1 (LACC1) protein is an enzyme highly expressed in inflammatory macrophages and serves a central regulatory role in multiple inflammatory diseases such as inflammatory bowel diseases, arthritis and clearance of microbial infection1-4. However, the biochemical roles required for LACC1 functions remain largely undefined. Here we elucidated a shared biochemical function of LACC1 in mice and humans, converting L-citrulline to L-ornithine (L-Orn) and isocyanic acid and serving as a bridge between proinflammatory nitric oxide synthase (NOS2) and polyamine immunometabolism. We validated the genetic and mechanistic connections among NOS2, LACC1 and ornithine decarboxylase 1 (ODC1) in mouse models and bone marrow-derived macrophages infected by Salmonella enterica Typhimurium. Strikingly, LACC1 phenotypes required upstream NOS2 and downstream ODC1, and Lacc1-/- chemical complementation with its product L-Orn significantly restored wild-type activities. Our findings illuminate a previously unidentified pathway in inflammatory macrophages, explain why its deficiency may contribute to human inflammatory diseases and suggest that L-Orn could serve as a nutraceutical to ameliorate LACC1-associated immunological dysfunctions such as arthritis or inflammatory bowel disease.
Assuntos
Inflamação , Peptídeos e Proteínas de Sinalização Intracelular , Macrófagos , Óxido Nítrico Sintase Tipo II , Animais , Artrite/imunologia , Artrite/metabolismo , Citrulina/metabolismo , Cianatos/metabolismo , Humanos , Inflamação/enzimologia , Inflamação/imunologia , Inflamação/metabolismo , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Óxido Nítrico Sintase Tipo II/metabolismo , Ornitina/metabolismo , Ornitina Descarboxilase/metabolismo , Poliaminas/metabolismo , Salmonella typhimurium/imunologiaRESUMO
Most of the currently available cytotoxic agents for tackling cancer are devoid of selectivity, thus causing severe side-effects. This situation stimulated us to develop new antiproliferative agents with enhanced affinity towards tumour cells. We focused our attention on novel chalcogen-containing compounds (thiosemicarbazones, disulfides, selenoureas, thio- and selenocyanates), and particularly on selenium derivatives, as it has been documented that this kind of compounds might act as prodrugs releasing selenium-based reactive species on tumour cells. Particularly interesting in terms of potency and selectivity was a pharmacophore comprised by a selenocyanato-alkyl fragment connected to a p-phenylenediamine residue, where the nature of the second amino moiety (free, Boc-protected, enamine-protected) provided a wide variety of antiproliferative activities, ranging from the low micromolar to the nanomolar values. The optimized structure was in turn conjugated through a peptide linkage with biotin (vitamin B7), a cellular growth promoter, whose receptor is overexpressed in numerous cancer cells; the purpose was to develop a selective vector towards malignant cells. Such biotinylated derivative behaved as a very strong antiproliferative agent, achieving GI50 values in the low nM range for most of the tested cancer cells; moreover, it was featured with an outstanding selectivity, with GI50 > 100 µM against human fibroblasts. Mechanistic studies on the mode of inhibition of the biotinylated selenocyanate revealed (Annexin-V assay) a remarkable increase in the number of apoptotic cells compared to the control experiment; moreover, depolarization of the mitochondrial membrane was detected by flow cytometry analysis, and with fluorescent microscopy, what supports the apoptotic cell death. Prior to the apoptotic events, cytostatic effects were observed against SW1573 cells using label-free cell-living imaging; therefore, tumour cell division was prevented. Multidrug resistant cell lines exhibited a reduced sensitivity towards the biotinylated selenocyanate, probably due to its P-gp-mediated efflux. Remarkably, antiproliferative levels could be restored by co-administration with tariquidar, a P-gp inhibitor; this approach can, therefore, overcome multidrug resistance mediated by the P-gp efflux system.
Assuntos
Antineoplásicos , Citostáticos , Selênio , Humanos , Citostáticos/farmacologia , Linhagem Celular Tumoral , Selênio/farmacologia , Cianatos/farmacologia , Apoptose , Proliferação de Células , Antineoplásicos/farmacologia , Antineoplásicos/química , Relação Estrutura-AtividadeRESUMO
Carbamylation is a nonenzymatic post-translational modification observed during the reaction between cyanate and amino acids and/or proteins that may occur during some pathologies such as chronic kidney disease. Evidence suggests that carbamylation may interfere with the quantification of some analytes measured using immunoturbidimetric assays. C-reactive protein (CRP) is an inflammatory response protein that is commonly quantified through immunoturbidimetry in clinical laboratories. Because the presence of modified proteins in serum can lead to impaired quantification, this study aimed to verify the impact of in vitro carbamylation on the measurement of CRP in a CRP standard solution and serum pool. The samples were incubated with 150 nM, 150 µM, or 150 mM potassium cyanate (KOCN) or 20, 100, or 500 mg/dL urea at 37 °C for 24 h. CRP concentrations were measured using an immunoturbidimetric assay. The results showed a 61%-72% decrease in the CRP detection rate after incubation with KOCN. Incubation with urea resulted in a 0.7%-8% lower CRP detection rate. The results of this study indicate that high concentrations of cyanate can lead to falsely decreased CRP levels, as measured by immunoturbidimetry.
Assuntos
Proteína C-Reativa , Carbamilação de Proteínas , Humanos , Cianatos , UreiaRESUMO
(Thio)ureas ((T)Us) and benzothiazoles (BTs) each have demonstrated to have a great variety of biological activities. When these groups come together, the 2-(thio)ureabenzothizoles [(T)UBTs] are formed, improving the physicochemical as well as the biological properties, making these compounds very interesting in medicinal chemistry. Frentizole, bentaluron and methabenzthiazuron are examples of UBTs used for treatment of rheumatoid arthritis and as wood preservatives and herbicides in winter corn crops, respectively. With this antecedent, we recently reported a bibliographic review about the synthesis of this class of compounds, from the reaction of substituted 2-aminobenzothiazoles (ABTs) with iso(thio)cyanates, (thio)phosgenes, (thio)carbamoyl chlorides, 1,1'-(thio)carbonyldiimidazoles, and carbon disulfide. Herein, we prepared a bibliographic review about those features of design, chemical synthesis, and biological activities relating to (T)UBTs as potential therapeutic agents. This review is about synthetic methodologies generated from 1968 to the present day, highlighting the focus to transform (T)UBTs to compounds containing a range substituents, as illustrated with 37 schemes and 11 figures and concluded with 148 references. In this topic, the scientists dedicated to medicinal chemistry and pharmaceutical industry will find useful information for the design and synthesis of this interesting group of compounds with the aim of repurposing these compounds.
Assuntos
Benzotiazóis , Ureia , Benzotiazóis/química , CianatosRESUMO
Isocyanic acid (HNCO) is known to be inert to strong oxidants and photolysis in the atmosphere but often appears in different forms of smoke; therefore, it is linked to various smoke-related illnesses due to tobacco usage or wildfire events. To date, the major loss pathway of HNCO is believed to be through its uptake on aerosol droplets. However, the molecular mechanisms underlying such an uptake process are still incompletely understood. Herein, we use the Born-Oppenheimer molecular dynamics (BOMD) simulations to study solvation and hydrolysis reactions of HNCO on water droplets at ambient temperature. The BOMD simulations indicate that the scavenging of HNCO by water droplets is largely attributed to the preferential adsorption of HNCO at the air-water interface, rather than inside bulk water. Specifically, the H atom of HNCO interacts with the O atom of interfacial water, leading to the formation of a hydrogen bond (H-bond) of (HNCO)H···O(H2O), which prevents HNCO from evaporating. Moreover, the interfacial water can act as H-bond acceptors/donors to promote the proton transfer during the HNCO hydrolysis reaction. Compared to the gas phase, the activation barrier is lowered from 45 to 14 kcal·mol-1 on the water surface, which facilitates the formation of the key intermediate of NH2COOH. This intermediate eventually decomposes into NH3 and CO2, consistent with the previous study [ Atmos. Chem. Phys. 2016, 16, 703-714]. The new molecular insight into HNCO solvation and reaction on the water surface improves our understanding of the uptake of HNCO on aerosols.
Assuntos
Cianatos , Água , Atmosfera/química , Cianatos/química , Hidrólise , Prótons , Água/químicaRESUMO
Biological carbon dioxide (CO2 ) reduction is an important step by which organisms form valuable energy-richer molecules required for further metabolic processes. The Mo-dependent formate dehydrogenase (FDH) from Rhodobacter capsulatus catalyzes reversible formate oxidation to CO2 at a bis-molybdopterin guanine dinucleotide (bis-MGD) cofactor. To elucidate potential substrate binding sites relevant for the mechanism, we studied herein the interaction with the inhibitory molecules azide and cyanate, which are isoelectronic to CO2 and charged as formate. We employed infrared (IR) spectroscopy in combination with density functional theory (DFT) and inhibition kinetics. One distinct inhibitory molecule was found to bind to either a non-competitive or a competitive binding site in the secondary coordination sphere of the active site. Site-directed mutagenesis of key amino acid residues in the vicinity of the bis-MGD cofactor revealed changes in both non-competitive and competitive binding, whereby the inhibitor is in case of the latter interaction presumably bound between the cofactor and the adjacent Arg587.
Assuntos
Dióxido de Carbono , Formiato Desidrogenases , Aminoácidos/metabolismo , Azidas , Sítios de Ligação , Dióxido de Carbono/química , Cianatos , Formiato Desidrogenases/química , Formiatos/química , OxirreduçãoRESUMO
Isocyanates such as 1,6-hexamethylene diisocyanate (HDI), 4,4'-methylenediphenyl diisocyanate, and toluene diisocyanate are highly reactive compounds that have a variety of commercial applications, including manufacturing polyurethane foam, elastomers, paints, adhesives, coatings, insecticides, and many other products. Their primary route of occupational exposure is through inhalation. Due to their high chemical reactivity, they are toxic and have adverse effects at the cellular and subcellular levels, leading to irritative and immunological reactions associated with lung disease. High concentrations of isocyanates are strong respiratory irritants. Bronchial sensitization and asthma are among the major adverse clinical reactions associated with low-level chronic exposure to isocyanates. Albumin adducts have been linked to the mechanism of occupational asthma caused by isocyanates. Isocyanates react in vivo with albumin, which is recognized by the immune system. Albumin adducts of isocyanates trigger immune responses and are probably the antigenic basis for isocyanate asthma. Sensitization to isocyanates is the main pathway for adverse health effects. Therefore, markers for the biologically effective dose such as albumin adducts of HDI are needed. A new isocyanate adduct of HDI with lysineâNε-[(6-amino-hexyl-amino)carbonyl]-lysine (HDI-Lys)âwas synthesized and characterized by 1H-NMR, 13C-NMR, and mass spectrometry (MS). Appropriate internal standardsâHDI-Lys-4,4'-5,5'-d4 (HDI-d4-Lys) and Nε-[(7-amino-heptyl-amino)carbonyl]-lysine (Hep-Lys)âwere synthesized to establish a LC-MS/MS method for the analysis of HDI adducts in in vitro modified albumin and in workers. The presence of HDI-Lys was found after pronase digestion of albumin and confirmed by two independent chromatographic approaches: with a C8 reversed-phase column and with a hydrophilic interaction liquid chromatography column. Quantification was performed with positive electrospray ionization (ESI)-MS. The adduct peak found in vivo was confirmed with the less sensitive negative ESI-MS. In summary, these are new compounds and methods to determine isocyanate-specific adducts with albumin in workers exposed to HDI.
Assuntos
Asma Ocupacional , Exposição Ocupacional , Humanos , Lisina/química , Cromatografia Líquida , Espectrometria de Massas em Tandem , Isocianatos/toxicidade , Isocianatos/química , Asma Ocupacional/induzido quimicamente , Exposição Ocupacional/efeitos adversos , Exposição Ocupacional/análise , Albuminas/química , CianatosRESUMO
A practical strategy for the synthesis of spiro[5.5]trienones-fused selenocyanates and spiro[4.5]trienones-fused selenocyanates through electrophilic selenocyanogen cyclization and dearomative spirocyclization is reported. This approach was conducted under mild conditions with broad substrate scope and good functional group tolerance. The utility of this procedure is exhibited in the late-stage functionalization of nature product and drug molecules.
Assuntos
Compostos de Espiro , Cianatos , Ciclização , Estrutura Molecular , Compostos de Selênio , Compostos de Espiro/químicaRESUMO
The urgent threat of new psychoactive substances worldwide promotes rapid detection and identification demand for public security. Ambient ionization mass spectrometry (AIMS) has become mainstream among various detection techniques. Still, scant successful applications have been fulfilled toward dried blood spot (DBS) or plasma spot (DPS) as an easy-to-implement sampling format in AIMS. This work bridged the gap between dielectric barrier discharge ionization mass spectrometry (DBDI-MS) and DPS/DBS samples by thermal desorption (TD) assistance. It made the impossible mission of direct DBDI-MS measurement on DPS/DBS samples containing fentanyl analogs (FTNs) possible. Guided by finite element simulations and a customized three-dimensional printing interface, we constructed a semi-covered flat-TD surface for sufficient desorption and ionization of FTNs from DPS/DBS samples without any sample pretreatment or sample separation. We successfully quantified eight FTNs in DPS samples using deuterated fentanyl as internal standard by triple quadrupole tandem mass spectrometry (MS/MS) and proved its practical applicability in the fentanyl-exposed rat plasma samples. This DBDI-TD-MS method also fits well with DBS samples, and it only took 20 s to analyze each sample. Further, based on summarized fragmentation characteristics of twenty FTNs, we established a backbone alerting ion-guided screening rule for suspect screening of FTNs in DPS samples via quadrupole time-of-flight MS/MS and built a chemometric approach for convenient mutual verification of screening "unknown" artificial samples. We hope this ideal DBDI-TD-MS method finds its valuable role in national security, doping control, and public health for routine large-scale analysis or on-site detection.
Assuntos
Teste em Amostras de Sangue Seco , Espectrometria de Massas em Tandem , Animais , Cianatos , Teste em Amostras de Sangue Seco/métodos , Fentanila , Plasma , Ratos , Espectrometria de Massas em Tandem/métodosRESUMO
Mononuclear non-heme Fe(II)- and α-ketoglutarate-dependent oxygenases (FeDOs) catalyze a site-selective C-H hydroxylation. Variants of these enzymes in which a conserved Asp/Glu residue in the Fe(II)-binding facial triad is replaced by Ala/Gly can, in some cases, bind various anionic ligands and catalyze non-native chlorination and bromination reactions. In this study, we explore the binding of different anions to an FeDO facial triad variant, SadX, and the effects of that binding on HO⢠vs X⢠rebound. We establish not only that chloride and bromide enable non-native halogenation reactions but also that all anions investigated, including azide, cyanate, formate, and fluoride, significantly accelerate and influence the site selectivity of SadX hydroxylation catalysis. Azide and cyanate also lead to the formation of products resulting from N3â¢, NCOâ¢, and OCN⢠rebound. While fluoride rebound is not observed, the rate acceleration provided by this ligand leads us to calculate barriers for HO⢠and F⢠rebound from a putative Fe(III)(OH)(F) intermediate. These calculations suggest that the lack of fluorination is due to the relative barriers of the HO⢠and F⢠rebound transition states rather than an inaccessible barrier for F⢠rebound. Together, these results improve our understanding of the FeDO facial triad variant tolerance of different anionic ligands, their ability to promote rebound involving these ligands, and inherent rebound preferences relative to HO⢠that will aid efforts to develop non-native catalysis using these enzymes.
Assuntos
Ácidos Cetoglutáricos , Oxigenases , Azidas , Cianatos , Compostos Férricos , Compostos Ferrosos/química , Fluoretos , Ácidos Cetoglutáricos/química , Ligantes , Oxigenases/metabolismoRESUMO
Isocyanic acid (HNCO) and other nitrogen-containing volatile chemicals (organic isocyanates, hydrogen cyanide, nitriles, amines, amides) were measured during the House Observation of Microbial and Environmental Chemistry (HOMEChem) campaign. The indoor HNCO mean mixing ratio was 0.14 ± 0.30 ppb (range 0.012-6.1 ppb), higher than outdoor levels (mean 0.026 ± 0.15 ppb). From the month-long study, cooking and chlorine bleach cleaning are identified as the most important human-related sources of these nitrogen-containing gases. Gas oven cooking emits more isocyanates than stovetop cooking. The emission ratios HNCO/CO (ppb/ppm) during stovetop and oven cooking (mean 0.090 and 0.30) are lower than previously reported values during biomass burning (between 0.76 and 4.6) and cigarette smoking (mean 2.7). Bleach cleaning led to an increase of the HNCO mixing ratio of a factor of 3.5 per liter of cleaning solution used; laboratory studies indicate that isocyanates arise via reaction of nitrogen-containing precursors, such as indoor dust. Partitioned in a temperature-dependent manner to indoor surface reservoirs, HNCO was present at the beginning of HOMEChem, arising from an unidentified source. HNCO levels are higher at the end of the campaign than the beginning, indicative of occupant activities such as cleaning and cooking; however the direct emissions of humans are relatively minor.
Assuntos
Poluentes Atmosféricos , Poluição do Ar em Ambientes Fechados , Compostos Orgânicos Voláteis , Poluentes Atmosféricos/análise , Poluição do Ar em Ambientes Fechados/análise , Culinária , Cianatos , Monitoramento Ambiental , Gases , Humanos , Isocianatos , NitrogênioRESUMO
New tetrahydropyrazino[2,3-c]quinolin-5(6H)-ones were prepared from 3-chloroquinoline-2,4(1H,3H)-diones and ethylene diamine. In their reaction with HNCO, an unprecedented molecular rearrangement produced new types of hydantoin derivatives. All prepared compounds were characterized on the basis of their 1H, 13C, and 15N NMR and ESI mass spectra and some were authenticated by X-ray analysis of single crystalline material. A proposed mechanism for rearrangement is discussed in this essay. The CDK and ABL inhibition activity as well as in vitro cytotoxicity of the prepared compounds was also tested.
Assuntos
Espectroscopia de Ressonância Magnética , CianatosRESUMO
The (thio)urea and benzothiazole (BT) derivatives have been shown to have a broad spectrum of biological activities. These groups, when bonded, result in the 2-(thio)ureabenzothizoles (TBT and UBT), which could favor the physicochemical and biological properties. UBTs and TBTs are compounds of great importance in medicinal chemistry. For instance, Frentizole is a UBT derivative used for the treatment of rheumatoid arthritis and systemic lupus erythematosus. The UBTs Bentaluron and Bethabenthiazuron are commercial fungicides used as wood preservatives and herbicides in winter corn crops. On these bases, we prepared this bibliography review, which covers chemical aspects of UBTs and TBTs as potential therapeutic agents as well as their studies on the mechanisms of a variety of pharmacological activities. This work covers synthetic methodologies from 1935 to nowadays, highlighting the most recent approaches to afford UBTs and TBTs with a variety of substituents as illustrated in 42 schemes and 13 figures and concluded with 187 references. In addition, this interesting review is designed on chemical reactions of 2-aminobenzothiazoles (2ABTs) with (thio)phosgenes, iso(thio)cyanates, 1,1'-(thio)carbonyldiimidazoles [(T)CDI]s, (thio)carbamoyl chlorides, and carbon disulfide. This topic will provide information of utility for medicinal chemists dedicated to the design and synthesis of this class of compounds to be tested with respect to their biological activities and be proposed as new pharmacophores.
Assuntos
Dissulfeto de Carbono , Fungicidas Industriais , Herbicidas , Benzotiazóis/farmacologia , Cloretos , Cianatos , Fungicidas Industriais/farmacologia , Herbicidas/farmacologia , UreiaRESUMO
The reactivity of thiophene in Diels-Alder reactions is investigated with different maleimide derivatives. In this paper, we have synthesized for the first time the Diels-Alder adducts of thiophene at room temperature and atmospheric pressure. Maleimido-thiophene adducts were promoted by AlCl3. The effects of solvent, time, temperature and the use of different Lewis acids were studied, showing dramatic effects for solvent and Lewis acid. Furthermore, the catalysis with AlCl3 is highly stereoselective, preferably providing the exo form of the adduct. Additionally, we also discovered the ability of AlCl3 to catalyze the arylation of maleimides to yield 3-aryl succinimides in a straightforward manner following a Friedel-Crafts-type addition. The inclusion of a selenocyanate group contributes to the cytotoxic activity of the adduct. This derivatization (from compound 7 to compound 15) results in an average GI50 value of 1.98 µM in the DTP (NCI-60) cell panel, resulting in being especially active in renal cancer cells.
Assuntos
Antineoplásicos/farmacologia , Cianatos/farmacologia , Compostos de Selênio/farmacologia , Tiofenos/química , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Cianatos/química , Reação de Cicloadição , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Ácidos de Lewis/química , Compostos de Selênio/química , Tiofenos/farmacologiaRESUMO
The electrochemical urea oxidation reaction (UOR) to N2 represents an efficient route to simultaneous nitrogen removal from N-enriched waste and production of renewable fuels at the cathode. However, the overoxidation of urea to NOx - usually dominates over its oxidation to N2 at Ni(OH)2 -based anodes. Furthermore, detailed reaction mechanisms of UOR remain unclear, hindering the rational catalyst design. We found that UOR to NOx - on Ni(OH)2 is accompanied by the formation of near stoichiometric amount of cyanate (NCO- ), which enabled the elucidation of UOR mechanisms. Based on our experimental and computational findings, we show that the formation of NOx - and N2 follows two distinct vacancy-dependent pathways. We also demonstrate that the reaction selectivity can be steered towards N2 formation by altering the composition of the catalyst, e.g., doping the catalyst with copper (Ni0.8 Cu0.2 (OH)2 ) increases the faradaic efficiency of N2 from 30 % to 55 %.
Assuntos
Níquel , Nitritos , Catálise , Cobre/química , Cianatos , Eletrólise , Níquel/química , Nitrogênio/química , UreiaRESUMO
We report the visible-light-promoted selenocyanation of cyclobutanone oxime esters using potassium selenocyanate in the presence of a fac-Ir(ppy)3 catalyst for the first time. Because of the mild conditions employed and use of readily accessible potassium selenocyanate, this method is an effective and green strategy for the synthesis of cyano and selenocyano bifunctional substituted alkanes.
Assuntos
Ésteres , Oximas , Cianatos , Potássio , Compostos de SelênioRESUMO
Three new classes of ionic organoselenium compounds containing cationic benzimidazolium and imidazolium ring systems with selenocyanates as counterions are described. The cyclization of N,N'-disubstituted benzimidazolium and imidazolium bromides having N-(CH2)2-Br and N-(CH2)3-Br groups in the presence of potassium selenocyanate (KSeCN) led to formation of the corresponding selenazolium selenocyanates (21a, 21b, 22a, and 22b) and selenazinium selenocyanates (21c, 21d, 22c, and 22d). However, the open-chain selenocyanates with additional selenocyanate counterions (21e, 21f, 22e, and 22f) were formed from the N,N'-disubstituted benzimidazolium and imidazolium bromides having N-(CH2)6-Br groups. Mechanistic studies were carried out to understand the feasibility of such cyclization processes in the presence of KSeCN. The compounds were studied further for their potencies to catalytically reduce H2O2 in the presence of thiols. Interestingly, the cyclic selenazolium (21a, 21b, 22a, and 22b) and selenazinium compounds (21c, 21d, 22c, and 22d) exhibited significantly higher antioxidant activities than the corresponding acyclic selenocyanates (21f, 22e, and 22f). Selected compounds (22d and 22e) were further evaluated for their potencies in modulating the intracellular level of reactive oxygen species (ROS) in a representative macrophage cell line (RAW 264.7). Owing to the cationic nature of compounds, they may target and scavenge mitochondrial ROS in the cellular medium.
Assuntos
Antioxidantes/farmacologia , Benzimidazóis/farmacologia , Compostos Organosselênicos/farmacologia , Animais , Antioxidantes/síntese química , Benzimidazóis/síntese química , Cianatos/síntese química , Cianatos/farmacologia , Ciclização , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Camundongos , Compostos Organosselênicos/síntese química , Oxirredução , Células RAW 264.7 , Compostos de Selênio/síntese química , Compostos de Selênio/farmacologiaRESUMO
Ammonia- and nitrite-oxidizing microorganisms are collectively responsible for the aerobic oxidation of ammonia via nitrite to nitrate and have essential roles in the global biogeochemical nitrogen cycle. The physiology of nitrifiers has been intensively studied, and urea and ammonia are the only recognized energy sources that promote the aerobic growth of ammonia-oxidizing bacteria and archaea. Here we report the aerobic growth of a pure culture of the ammonia-oxidizing thaumarchaeote Nitrososphaera gargensis using cyanate as the sole source of energy and reductant; to our knowledge, the first organism known to do so. Cyanate, a potentially important source of reduced nitrogen in aquatic and terrestrial ecosystems, is converted to ammonium and carbon dioxide in Nitrososphaera gargensis by a cyanase enzyme that is induced upon addition of this compound. Within the cyanase gene family, this cyanase is a member of a distinct clade also containing cyanases of nitrite-oxidizing bacteria of the genus Nitrospira. We demonstrate by co-culture experiments that these nitrite oxidizers supply cyanase-lacking ammonia oxidizers with ammonium from cyanate, which is fully nitrified by this microbial consortium through reciprocal feeding. By screening a comprehensive set of more than 3,000 publically available metagenomes from environmental samples, we reveal that cyanase-encoding genes clustering with the cyanases of these nitrifiers are widespread in the environment. Our results demonstrate an unexpected metabolic versatility of nitrifying microorganisms, and suggest a previously unrecognized importance of cyanate in cycling of nitrogen compounds in the environment.
Assuntos
Archaea/metabolismo , Cianatos/metabolismo , Nitrificação , Aerobiose , Amônia/metabolismo , Compostos de Amônio/metabolismo , Archaea/enzimologia , Archaea/genética , Archaea/crescimento & desenvolvimento , Dióxido de Carbono/metabolismo , Carbono-Nitrogênio Liases/genética , Carbono-Nitrogênio Liases/metabolismo , Microbiologia Ambiental , Metagenoma/genética , Nitratos/metabolismo , Nitritos/metabolismo , Nitrogênio/metabolismo , Ciclo do Nitrogênio , OxirreduçãoRESUMO
Two strains of the chlorophyte Chlamydomonas reinhardtii, a wild type (WT) and a transgenic strain (C.CYN) contained an exogenous cyanase gene (CYN), were used to investigate the growth and cyanate biosorption capability through the analysis of the adsorption equilibrium isotherm. The potential antioxidants activity of the algal strains was also investigated under cyanate concentration. The antioxidants activity of both C.CYN and WT were enhanced by the application of cyanate.Two adsorption isotherm models and the sorption kinetics were used to check the efficiency of the cyanate removal process. The results showed the biosorbent efficiency of Chlamydomonas in the removal of KCNO from aqueous solution. The C.CYN strain has great efficiency to remove cyanate as compared to the WT. The maximum percentage of cyanate removal was 83.75% for the C.CYN and 50% for the WT as treated with 0.8 mg.ml-1 KCNO. The data were adapted to the nonlinear Langmuir model on the basis of the coefficient of determination. The calculated qmax was 0.54 and 0.42 µg.mg-1 for C.CYN and WT which correlated to the experimental one (0.67 and 0.4 µg.mg-1, respectively). Our data highlight the application of the transgenic algal strain toward the removal of highly toxic materials as cyanate.Novelty statement The main objective of this work is to find out an efficient genetically-modified Chlamydomonas strain to remove the highly toxic cyanate compound from contaminated area. Moreover, to evaluate the biosorption ability of this transgenic strain with its wild one via two adsorption isotherm (the Langmuir and Freundlich) models. Also, to estimate the antioxidants activity of these strains under the cyanate toxicity through four different assays.