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1.
J Inorg Biochem ; 248: 112363, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37689038

RESUMO

In order to discover new anticancer drugs, novel ruthenium(III) complexes [Ru(L)Cl(H2O)], where L is tetradentate Schiff base bis(acetylacetone)ethylendiimine (acacen, 1), bis(benzoylacetone)ethylendiimine (bzacen, 2), (acetylacetone)(benzoylaceton)ethylendiimine (acacbzacen, 3), bis(acetylacetone)propylendiimine (acacpn, 4), bis(benzoylacetone)propylendiimine (bzacpn, 5) or (acetylacetone)(benzoylaceton)propylendiimine (acacbzacpn, 6), were synthesized. The complexes 1 - 6 were characterized by elemental analysis, molar conductometry, and by various spectroscopic techniques, such as UV-Vis, IR, EPR, and ESI-MS. Based on in vitro DNA/BSA experiments, complexes 2 (bzacen) and 5 (bzacpn) with two aromatic rings showed the highest DNA/BSA-activity, suggesting that the presence of the aromatic ring on the tetradentate Schiff base ligand contributes to increased activity. Moreover, these two compounds showed the highest cytotoxic effects toward human, A549 and murine LLC1 lung cancer cells. These complexes altered the ratio of anti- and pro-apoptotic molecules and induced apoptosis of A549 cells. Further, complexes 2 and 5 reduced the percentage of Mcl1 and Bcl2 expressing LLC1 cells, induced their apoptotic death and exerted an antiproliferative effect against LLC1. Finally, complex 5 reduced the volume of mouse primary heterotopic Lewis lung cancer, while complex 2 reduced the incidence and mean number of metastases per lung. Additionally, molecular docking with DNA revealed that the reduced number of aromatic rings or their absence causes lower intercalative properties of the complexes in order: 2 > 5 > 6 > 3 > 4 > 1. It was observed that conventional hydrogen bonds and hydrophobic interactions contribute to the stabilization of the structures of complex-DNA. A molecular docking study with BSA revealed a predominance of 1 - 6 in binding affinity to the active site III, a third D-shaped hydrophobic pocket within subdomain IB.


Assuntos
Neoplasias Pulmonares , Rutênio , Humanos , Animais , Camundongos , Simulação de Acoplamento Molecular , Rutênio/farmacologia , Bases de Schiff/farmacologia
2.
Food Chem ; 426: 136669, 2023 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-37352716

RESUMO

This study aimed to purify, characterise and stabilise the natural food colourant, R-phycocyanin (R-PC), from the red algae Porphyra spp. (Nori). We purified R-PC from dried Nori flakes with a high purity ratio (A618/A280 ≥ 3.4) in native form (α-helix content 53%). SAXS measurements revealed that R-PC is trimeric ((αß)3) in solution. The thermal denaturation of α-helix revealed one transition (Tm at 52 °C), while the pH stability study showed R-PC is stable in the pH range 4-8. The thermal treatment of R-PC at 60 °C has detrimental and irreversible effects on R-PC colour and antioxidant capacity (22 % of residual capacity). However, immobilisation of R-PC within calcium alginate beads completely preserves R-PC colour and mainly retains its antioxidant ability (78 % of residual capacity). Results give new insights into the stability of R-PC and preservation of its purple colour and bioactivity by encapsulation in calcium alginate beads.


Assuntos
Corantes de Alimentos , Porphyra , Ficocianina/química , Porphyra/química , Antioxidantes , Espalhamento a Baixo Ângulo , Difração de Raios X , Verduras
4.
Polymers (Basel) ; 15(6)2023 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-36987254

RESUMO

We investigate mode coupling in a multimode graded-index microstructured polymer optical fiber (GI mPOF) with a solid core by solving the time-independent power flow equation (TI PFE). Using launch beams with various radial offsets, it is possible to calculate for such an optical fiber the transients of the modal power distribution, the length Lc at which an equilibrium mode distribution (EMD) is reached, and the length zs for establishing a steady-state distribution (SSD). In contrast to the conventional GI POF, the GI mPOF explored in this study achieves the EMD at a shorter length Lc. The earlier shift to the phase of slower bandwidth decrease would result from the shorter Lc. These results are helpful for the implementation of multimode GI mPOFs as a part of communications and optical fiber sensory systems.

5.
Dalton Trans ; 52(5): 1323-1344, 2023 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-36629870

RESUMO

This paper presents the synthesis and structural characterization of a series of new ruthenium(II) complexes 1-7, with the general formula mer-[RuL3(N-N)Cl]Cl, where L is 2,2':6',2''-terpyridine (tpy) or 4'-(4-chlorophenyl)-2,2':6',2''-terpyridine (Cl-Ph-tpy) and N-N is o-benzoquinonediimine (o-bqdi), 2,3-naphthoquinonediimine (nqdi), 4,4'-dimethyl-2,2'-bipyridine (dmbpy) or 2,2'-bipyridine-4,4'-dicarboxylic acid (dcbpy). The kinetic results showed that the ligand substitution reactions of new Ru(II)-polypyridyl complexes with biomolecules were affected by different substituents and the aromaticity of meridional tridentate and bidentate spectator ligands as well as by the nature of the entering nucleophile. The reactivity of the complexes increases in the order: dmbpy < dcbipy < nqdi < o-bqdi. In addition, quantum chemical calculations were performed to support the interpretation and discussion of the experimental data. Furthermore, combining ethidium bromide (EB) and Hoechst 33258 (2-(4-hydroxyphenyl)-5-[5-(4-methylpiperazine-1-yl)benzimidazo-2-yl]-benzimidazole) fluorescence assay results implied that 1-7 might interact with calf thymus DNA through partial intercalation and/or minor groove binding. The human serum albumin (HAS)-fluorescence binding studies involving the site markers, eosin Y, as a marker for site I of subdomain IIA, and ibuprofen, as a marker for site II of subdomain IIIA, showed that Ru(II) compounds bind to both sites with moderately strong affinity (Kb = 104-106 M-1). Moreover, these DNA/HSA experimental results were confirmed by molecular docking. Complexes 2, 5 and 6 exerted good to strong and highly selective cytotoxic activity against breast adenocarcinoma (MDA-MB 231), colorectal carcinoma (HCT116) and cervix adenocarcinoma (HeLa). Depending on their structure and cell line, the complexes acted differently in terms of their influence on autophagy, the cell cycle and the engaged apoptotic pathway.


Assuntos
Adenocarcinoma , Antineoplásicos , Complexos de Coordenação , Rutênio , Humanos , Rutênio/farmacologia , Rutênio/química , Ligantes , Simulação de Acoplamento Molecular , Antineoplásicos/química , DNA/química , Quinonas , Complexos de Coordenação/química , Linhagem Celular Tumoral
6.
Int J Mol Sci ; 25(1)2023 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-38203504

RESUMO

In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52-48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19-100.00%) and sensitivity was 52.94% (95% CI, 35.13-70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67-65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.


Assuntos
COVID-19 , Proteínas do Nucleocapsídeo , Animais , Humanos , Coelhos , SARS-CoV-2 , COVID-19/diagnóstico , Anticorpos , Ensaio de Imunoadsorção Enzimática
7.
Opt Express ; 30(14): 24667-24675, 2022 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-36237015

RESUMO

By solving the time-independent power flow equation (TI PFE), we study mode coupling in a multimode W-type microstructured polymer optical fiber (mPOF) with a solid-core. The multimode W-type mPOF is created by modifying the cladding layer and reducing the core of a multimode singly clad (SC) mPOF. For such optical fiber, the angular power distributions, the length Lc at which an equilibrium mode distribution (EMD) is achieved, and the length zs for establishing a steady state distribution (SSD) are determined for various arrangements of the inner cladding's air-holes and different launch excitations. This information is useful for the implement of multimode W-type mPOFs in telecommunications and optical fiber sensors.

8.
Food Chem ; 374: 131780, 2022 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-34894468

RESUMO

This study aimed to characterise the stability of R-phycoerythrin (R-PE), a vivid natural colourant with emerging potential for application in the food industry. High-quality (A560/A280 ≥ 5), native (α-helix content 75%) R-PE was purified from commercial dried Nori (Porphyra sp.) flakes. Thermal unfolding revealed two transitions (at 56 and 72 °C), ascribed to different protein subunits. Contrary to elevated temperature, high-pressure (HP) treatment showed significant advantages: The R-PE unfolding was partly reversible and the colour bleaching was minimal. Binding of Cu2+ (6.3 × 105 M-1) and Zn2+ (1.7 × 103 M-1) influenced conformational changes in the protein tetrapyrrole chromophore without affecting R-PE structure and stability (colour). The results give new insights into the stability of R-PE suggesting its usefulness for the replacement of toxic synthetic dyes. Preservation of the red colour of R-PE could be considered in fortified food and beverages by HP processing. R-PE may act as a biosensor for Cu2+ in aquatic systems.


Assuntos
Corantes de Alimentos , Porphyra , Ficoeritrina , Subunidades Proteicas
9.
Polymers (Basel) ; 13(23)2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34883721

RESUMO

By solving the time-dependent power flow equation, we present a novel approach for evaluating the bandwidth in a multimode step-index polymer photonic crystal fiber (SI PPCF) with a solid core. The bandwidth of such fiber is determined for various layouts of air holes and widths of Gaussian launch beam distribution. We found that the lower the NA of SI PPCF, the larger the bandwidth. The smaller launch beam leads to a higher bandwidth for short fibers. The influence of the width of the launch beam distribution on bandwidth lessens as the fiber length increases. The bandwidth tends to its launch independent value at a particular fiber length. This length denotes the onset of the steady state distribution (SSD). This information is useful for multimode SI PPCF applications in telecommunications and optical fiber sensing applications.

10.
Polymers (Basel) ; 13(22)2021 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-34833273

RESUMO

The bandwidth of multimode W-type plastic optical fibers (POFs) with graded-index (GI) core distribution is investigated by solving the time-dependent power flow equation. The multimode W-type GI POF is designed from a multimode single-clad (SC) GI POF fiber upon modification of the cladding layer of the latter. Results show how the bandwidth in W-type GI POFs can be enhanced by increasing the wavelength for different widths of the intermediate layer and refractive indices of the outer cladding. These fibers are characterized according to their apparent efficiency to reduce modal dispersion and increase bandwidth.

11.
Opt Express ; 29(19): 29587-29594, 2021 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-34614700

RESUMO

A new method is proposed for investigating the bandwidth in multimode graded-index plastic optical fibers (GI POFs). By numerically solving the time-dependent power flow equation, bandwidth is reported for a varied launch conditions (radial offsets) of multimode GI POF. Our theoretical results are supported by the experimental results which show that bandwidth decreases with increasing radial offset. This decrease is more pronounced at short fiber lengths. At fiber length close to the coupling length Lc at which an equilibrium mode distribution (EMD) is achieved, this decrease becomes slower, indicating that mode coupling improves bandwidth at larger fiber lengths. With further increase of fiber length, bandwidth becomes nearly independent of the radial offset, indicating that a steady-state distribution (SSD) is achieved. Such a fiber characterization can be applied to optimize fiber performance in POF links.

12.
Dalton Trans ; 50(22): 7686-7704, 2021 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-33982702

RESUMO

We synthesized and characterized the ruthenium(iii) pincer-type complex [RuCl3(H2Lt-Bu] (H2Lt-Bu = 2,6-bis(5-tert-butyl-1H-pyrazol-3-yl)pyridine, 1) by elemental analysis, IR and UV-Vis spectroscopy, and the mass spectrometry (MS) method ESI Q-TOF. For comparison reasons, we also studied ruthenium(iii) terpyridine complexes of the general formula [Ru(N-N-N)Cl3], where N-N-N = 4'-chloro-terpyridine (Cl-tpy; 2) or 4'-chlorophenyl-terpyridine (Cl-Ph-tpy; 3). A kinetic study of the substitution reactions of 1-3 with biomolecules showed that the rate constants depend on the properties of the spectator ligand and the nature of the entering nucleophile. The DNA/HSA binding study showed that in comparison to complex 1 (bis-pyrazolylpyridine), the other two (2 and 3) terpyridine complexes had a slightly better binding affinity to calf thymus DNA (CT DNA), while in the case of human serum albumin (HSA), complex 1 exhibited the strongest quenching ability. We demonstrated that 1 possesses significant in vitro cytotoxic activity against mouse colon carcinoma CT26 cells and in vivo antitumor activity in murine heterotopic colon carcinoma. Complex 1 induced G0/G1 cell cycle arrest and apoptotic death in CT26 cells. Additionally, 1 showed antiproliferative activity, as evaluated by the detection of the expression levels of the Ki67 protein. Furthermore, the in vivo results showed that 1 reduced primary tumour growth and the number and growth of lung and liver metastases, significantly prolonging the treated mice's survival rate. This study highlighted that 1 does not show hepato- and nephrotoxicity. Our data demonstrated the considerable antitumor activity of the ruthenium(iii) pincer complex against CT26 tumour cells and implicated further investigations of its role as a potential chemotherapeutic agent for colon carcinoma.


Assuntos
Antineoplásicos , Neoplasias do Colo/tratamento farmacológico , Complexos de Coordenação , Pirazóis , Piridinas , Rutênio , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/patologia , Complexos de Coordenação/química , Complexos de Coordenação/farmacologia , Complexos de Coordenação/uso terapêutico , DNA/química , Guanosina Monofosfato/química , Histidina/química , Masculino , Metionina/química , Camundongos Endogâmicos BALB C , Simulação de Acoplamento Molecular , Pirazóis/química , Pirazóis/farmacologia , Pirazóis/uso terapêutico , Piridinas/química , Piridinas/farmacologia , Piridinas/uso terapêutico , Rutênio/química , Rutênio/farmacologia , Rutênio/uso terapêutico , Albumina Sérica Humana/química , Carga Tumoral/efeitos dos fármacos
13.
Virology ; 557: 15-22, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33582454

RESUMO

Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production. SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58-419 aa) was expressed in E. coli in soluble form, purified and characterized biochemically and immunologically. Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of ß-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower (94% and 96.5%, respectively). Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.


Assuntos
Anticorpos Antivirais/sangue , COVID-19/diagnóstico , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina G/sangue , SARS-CoV-2/imunologia , Sequência de Aminoácidos , COVID-19/sangue , COVID-19/imunologia , Teste Sorológico para COVID-19/métodos , Estudos de Casos e Controles , Clonagem Molecular , Proteínas do Nucleocapsídeo de Coronavírus/genética , Ensaio de Imunoadsorção Enzimática/normas , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Soros Imunes/química , Imunoglobulina M/sangue , Fosfoproteínas/genética , Fosfoproteínas/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , SARS-CoV-2/genética , Sensibilidade e Especificidade
14.
Molecules ; 25(20)2020 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-33066568

RESUMO

Ruthenium complexes have attracted considerable interest as potential antitumor agents. Therefore, antitumor activity and systemic toxicity of ruthenium(II) terpyridine complexes were evaluated in heterotopic mouse colon carcinoma. In the present study, cytotoxic effects of recently synthesized ruthenium(II) terpyridine complexes [Ru(Cl-tpy)(en)Cl][Cl] (en = ethylenediamine, tpy = terpyridine, Ru-1) and [Ru(Cl-tpy)(dach)Cl][Cl] (dach = 1,2-diaminocyclohexane, Ru-2) towards human and murine colon carcinoma cells were tested in vitro and in vivo and compared with oxaliplatin, the most commonly used chemotherapeutic agent against colorectal carcinoma. Ruthenium(II) complexes showed moderate cytotoxicity with IC50 values ranging between 19.1 to 167.3 µM against two human, HCT116 and SW480, and one mouse colon carcinoma cell line, CT26. Both ruthenium(II) terpyridine complexes exerted a moderate apoptotic effect in colon carcinoma cells, but induced significant necrotic death. Additionally, both complexes induced cell cycle disturbances, but these effects were specific for the cell line. Further, Ru-1 significantly reduced the growth of primary heterotopic tumor in mice, similarly to oxaliplatin. Renal damage in Ru-1 treated mice was lower in comparison with oxaliplatin treated mice, as evaluated by serum levels of urea and creatinine and histological evaluation, but Ru-1 induced higher liver damage than oxaliplatin, evaluated by the serum levels of alanine aminotransferase. Additionally, the interaction of these ruthenium(II) terpyridine complexes with the tripeptide glutathione (GSH) was investigated by proton nuclear magnetic resonance (1H NMR) spectroscopy. All reactions led to the formation of monofunctional thiolate adducts [Ru(Cl-tpy)(en)GS-S] (3) and [Ru(Cl-tpy)(dach)GS-S] (4). Our data highlight the significant cytotoxic activity of [Ru(Cl-tpy)(en)Cl][Cl] against human and mouse colon carcinoma cells, as well as in vivo antitumor activity in CT26 tumor-bearing mice similar to standard chemotherapeutic oxaliplatin, accompanied with lower nephrotoxicity in comparison with oxaliplatin.


Assuntos
Antineoplásicos/farmacologia , Neoplasias do Colo/tratamento farmacológico , Compostos de Rutênio/química , Compostos de Rutênio/farmacologia , Animais , Antineoplásicos/química , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Neoplasias do Colo/patologia , Glutationa/química , Humanos , Espectroscopia de Ressonância Magnética , Masculino , Camundongos Endogâmicos BALB C , Piridinas/química , Ensaios Antitumorais Modelo de Xenoenxerto
15.
J Inorg Biochem ; 208: 111090, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32389890

RESUMO

Interactions of three Ru(II) chlorophenyl terpyridine complexes: [Ru(Cl-Ph-tpy)(en)Cl]Cl (1), [Ru(Cl-Ph-tpy)(dach)Cl]Cl (2) and [Ru(Cl-Ph-tpy)(bpy)Cl]Cl (3) (Cl-Ph-tpy = 4'-(4-chlorophenyl)-2,2':6',2''-terpyridine, en = 1,2-diaminoethane, dach = 1,2-diaminocyclohexane, bpy = 2,2'-bipyridine) with human serum albumin (HSA), calf thymus DNA and a double-helical oligonucleotide d(CGCGAATTCGCG)2 (1BNA) were examined. Fluorescence emission studies were used to assess the interactions of complexes with HSA, which were of moderate strength for 1 and 2. Molecular docking allowed us to predict mostly π-π stacking and van der Waals interactions between the complexes and the protein. We suggest that the complexes bind to a novel site on HSA, which is different from its druggable sites I, II or III. We suggest a partial intercalation of complexes through the minor groove as a possible mode of interaction with double-helical DNA. Finally, when applied to normal extravillous cell line HTR8/SVneo and JAr choriocarcinoma cell line, complexes 1 and 2 exerted anti-adhesive properties at very low doses, whereas complex 3 had a negligible effect. The obtained results are completion of our studies of Ru(II) terpyridyl complexes that carry N-N ancillary ligands. We suggest a new research direction towards studying the cellular effects of Ru(II) polypyridyl compounds.


Assuntos
Complexos de Coordenação , DNA/química , Pirimidinas , Rutênio , Albumina Sérica Humana/química , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Simulação por Computador , Complexos de Coordenação/síntese química , Complexos de Coordenação/química , Complexos de Coordenação/farmacologia , Humanos , Pirimidinas/química , Pirimidinas/farmacologia , Rutênio/química , Rutênio/farmacologia
16.
Foods ; 8(10)2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31658590

RESUMO

Resistance to digestion by digestive proteases represents a critical property of many food allergens. Recently, a harmonized INFOGEST protocol was proposed for solid food digestion. The protocol proposes digestion conditions suitable for all kinds of solid and liquid foods. However, peanuts, as a lipid-rich food, represent a challenge for downstream analyses of the digestome. This is particularly reflected in the methodological difficulties in analyzing proteins and peptides in the presence of lipids. Therefore, the removal of the lipids seems to be a prerequisite for the downstream analysis of digestomes of lipid-rich foods. Here, we aimed to compare the digestomes of raw and thermally treated (boiled and roasted) peanuts, resulting from the INFOGEST digestion protocol for solid food, upon defatting the digests in two different manners. The most reproducible results of peanut digests were obtained in downstream analyses on TCA/acetone defatting. Unfortunately, defatting, even with an optimized TCA/acetone procedure, leads to the loss of proteins and peptides. The results of our study reveal that different thermal treatments of peanuts affect protein extraction and gastric/gastrointestinal digestion. Roasting of peanuts seems to enhance the extraction of proteins during intestinal digestion to a notable extent. The increased intestinal digestion is a consequence of the delayed extraction of thermally treated peanut proteins, which are poorly soluble in acidic gastric digestion juice but are easily extracted when the pH of the media is raised as in the subsequent intestinal phase of the digestion. Thermal processing of peanuts impaired the gastrointestinal digestion of the peanut proteins, especially in the case of roasted samples.

17.
Dalton Trans ; 47(38): 13696-13712, 2018 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-30209465

RESUMO

With the aim of assessing whether Au(iii) compounds with pincer type ligands might be utilized as potential antitumor agents, three new monofunctional Au(iii) complexes of the general formula [Au(N-N'-N)Cl]Cl2, where N-N'-N = 2,6-bis(5-tert-butyl-1H-pyrazol-3-yl)pyridine (H2LtBu, 1), 2,6-bis(5-tert-butyl-1-methyl-1H-pyrazol-3-yl)pyridine (Me2LtBu, 2) or 2,6-bis((4S,7R)-1,7,8,8-tetramethyl-4,5,6,7-tetrahydro-1H-4,7-methanoindazol-3-yl)pyridine (Me2*L, 3) were synthesized. All complexes were characterized by elemental analysis, spectroscopic techniques (IR, UV-Vis, 1D and 2D NMR) and mass spectrometry (MALDI TOF MS). The chemical behavior of the complexes under physiological conditions was studied by UV-Vis spectroscopy, which showed that all compounds were remarkably stable and that the gold center remained in the 3+ oxidation state. The kinetics and the mechanism of the reaction of complexes 1-3 with guanine derivatives (i.e. guanosine (Guo) and guanosine-5'-monophosphate (5'-GMP)) and calf thymus DNA (CT DNA) were studied by stopped-flow spectroscopy. The three complexes displayed moderately different rate constants in their reactions with Guo, 5'-GMP and CT DNA, which can be explained by the steric hindrance and σ-donicity of the methyl substituent on the bis-pyrazolylpyridine fragment in complexes 2 and 3. The measured enthalpies and entropies of activation (ΔH≠ > 0, ΔS≠ < 0) supported an associative mechanism for the substitution process. The interaction of the newly synthesized complexes 1-3 with CT DNA was investigated by UV-Vis and fluorescence spectroscopy, and also by viscosity measurements, which all indicated that complexes 1-3 bound to CT DNA with moderate binding affinity (Kb = 1.6-5.7 × 103 M-1) and stabilized the duplex of CT DNA. Molecular docking indicated that complexes 1-3 interacted with DNA via intercalation. Complex 1 reduced the cell survival of all the investigated cell lines (A549, A375, and LS-174) with IC50 values being up to 20 µM. We have shown that 1 induced perturbations of the cell cycle and led to apoptosis in human melanoma A375 cells. Complex 1 also affected the level of reactive oxygen species (ROS) in the same cells. However, pre-treatment of A375 cells with NAC (ROS scavenger) reversed the effect of 1 on their survival.


Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Complexos de Coordenação/síntese química , Complexos de Coordenação/farmacologia , DNA/metabolismo , Ouro/química , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Técnicas de Química Sintética , Complexos de Coordenação/química , Complexos de Coordenação/metabolismo , DNA/química , Humanos , Cinética , Simulação de Acoplamento Molecular , Conformação de Ácido Nucleico
18.
J Biol Inorg Chem ; 22(7): 1007-1028, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28695374

RESUMO

With the aim of assessing how the aromaticity of the inert chelating ligand can influence the activity of ruthenium(II) polypyridyl complexes, two new monofunctional ruthenium(II) complexes, [Ru(Cl-Ph-tpy)(phen)Cl]Cl (1) and [Ru(Cl-Ph-tpy)(o-bqdi)Cl]Cl (2) (where Cl-Ph-tpy = 4'-(4-chlorophenyl)-2,2':6',2″-terpyridine, phen = 1,10-phenanthroline, o-bqdi = o-benzoquinonediimine), were synthesized. All complexes were fully characterized by elemental analysis and spectroscopic techniques (IR, UV-Vis, 1D and 2D NMR, XRD). Their chemical behavior in aqueous solution was studied by UV-Vis and NMR spectroscopy showing that both compounds are relatively labile leading to the formation of the corresponding aqua species 1a and 2a. 1H NMR spectroscopy studies performed on complexes 1 and 2 demonstrated that after the hydrolysis of the Cl ligand, they are capable to interact with guanine derivatives (i.e., 9-methylguanine (9MeG) and 5'-GMP) through the N7, forming monofunctional adduct. The kinetics and the mechanism of the reaction of complexes 1 and 2 with the biologically more relevant 5'-GMP ligand were studied by UV-Vis spectroscopy. DNA/protein interactions of the complexes have been examined by photophysical studies, which demonstrated a bifunctional binding mode of the complexes with DNA and the complexes strongly quench the fluorescence intensity of bovine serum albumin (BSA) through the mechanism of both static and dynamic quenching. Complexes 1 and 2 strongly induced apoptosis of treated cancer cells with high percentages of apoptotic cells and negligible percentage of necrotic cells. In addition, both ruthenium complexes decreased Bcl-2/Bax ratio causing cytochrome c mitochondrial release, the activation of caspase-3 and induction of apoptosis.


Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Neoplasias/tratamento farmacológico , Piridinas/química , Piridinas/farmacologia , Rutênio/química , Rutênio/farmacologia , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Complexos de Coordenação/química , Complexos de Coordenação/farmacologia , DNA/metabolismo , Humanos , Modelos Moleculares , Neoplasias/metabolismo , Soroalbumina Bovina/metabolismo
19.
Appl Opt ; 51(20): 4896-901, 2012 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-22781271

RESUMO

For different depth and width of the intermediate layer, a power flow equation is used to calculate spatial transients and steady state of power distribution in W-type optical fibers (doubly clad fibers with three layers). A numerical solution has been obtained by the explicit finite difference method. Results show how the power distribution in W-type optical fibers varies with the depth of the intermediate layer for different values of intermediate layer width and coupling strength. We have found that with increasing depth of the intermediate layer, the fiber length at which the steady-state distribution is achieved increases. Such characterization of these fibers is consistent with their manifested effectiveness in reducing modal dispersion and improving bandwidth.

20.
Appl Opt ; 50(21): 4170-3, 2011 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-21772404

RESUMO

Using the power flow equation, the state of mode coupling in 100-400 µm core step-index silica optical fibers is investigated in this article. Results show the coupling length L(c) at which the equilibrium mode distribution is achieved and the length z(s) of the fiber required for achieving the steady-state mode distribution. Functional dependences of these lengths on the core radius and wavelength are also given. Results agree well with those obtained using a long-established calculation method. Since large core silica optical fibers are used at short distances (usually at lengths of up to 10 m), the light they transmit is at the stage of coupling that is far from the equilibrium and steady-state mode distributions.

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