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1.
Crit Rev Eukaryot Gene Expr ; 32(2): 91-105, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35381133

RESUMO

Glucose metabolism has significant impact on cancer cell survival and proliferation. Our previous studies have shown that level of glucose in the medium affects the cell attachment to and detachment from the substratum. Control of glucose metabolism in cancer cells has potential to serve as an anti-cancer therapy. Different anti-diabetic drugs have been reported to inhibit glucose uptake at cellular level by glucose transporters. Metformin chloride is commonly used as antidiabetic drug. It is known that use of metformin reduces chances of developing cancer in diabetic patients. Here we have investigated the effect of metformin on cell adhesion proteins and other related factors in different cancer cells, both metastatic and non-metastatic. The object was to evaluate the effect of different doses of metformin on the onset of metastasis after these are detached from their primary site of origin and re-attached at the secondary site. For this purpose, we grew different cancer cells (MDA-MB231, MCF7, HCT116, and SF767) in culturing media containing different concentrations of metformin chloride. Quantitative real-time PCR was used to evaluate the expression profile of the genes involved in cell adhesion. It was observed that metformin treatment increased the expression of epithelial isoforms of cell adhesion molecules along with integrins responsible for cell-to-matrix adhesion and inducing specific morphological changes such as development of cytoskeletal structures in different cancer cell lines which normally lead to attainment of mesenchymal phenotype. The effect of metformin appeared to be different in different doses. The glioblastoma cells (SF767) were observed to be the most sensitive cells among all cancer cells under study. Our data supports the idea that the metformin prevents the cancer cells from acquiring mesenchymal phenotype and hence prevents onset of metastasis, and if the process has already started then it has potential to prevent cancer cell attachment to the secondary site.


Assuntos
Metformina , Neoplasias , Adesão Celular , Linhagem Celular Tumoral , Proliferação de Células , Cloretos/farmacologia , Humanos , Hipoglicemiantes/farmacologia , Metformina/farmacologia
2.
Org Biomol Chem ; 20(9): 1974-1981, 2022 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-35179161

RESUMO

Trehalose-based probes are useful tools that allow the detection of the mycomembrane of mycobacteria through the metabolic labeling approach. Trehalose analogues conjugated to fluorescent probes can be used, and other probes are functionalized with a bioorthogonal chemical reporter for a two-step labeling approach. The synthesis of such trehalose-based probes mainly relies on the desymmetrization of natural trehalose using a large number of regioselective protection-deprotection steps to differentiate the eight hydroxyl groups. Herein, in order to avoid these time-consuming steps, we reinvestigated our previously reported tandem protocol mediated by FeCl3·6H2O, with the aim of modifying the ratio of the products to allow the challenging desymmetrization of the C2-symmetrical disaccharide trehalose. We demonstrate the usefulness of this method in providing easy access to trehalose analogues with a bioorthogonal moiety or a fluorophore in C-2, and also present their use in a one-step and two-step labeling approach, either of which can be used to study the mycomembrane in live mycobacteria.


Assuntos
Antibacterianos/farmacologia , Membrana Celular/efeitos dos fármacos , Cloretos/farmacologia , Corynebacterium/efeitos dos fármacos , Compostos Férricos/farmacologia , Trealose/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , Cloretos/química , Compostos Férricos/química , Testes de Sensibilidade Microbiana , Trealose/síntese química , Trealose/química
3.
Life Sci ; 295: 120380, 2022 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-35143825

RESUMO

AIMS: the main purpose of this study was to identify new selective antitumor agents. MAIN METHODS: several hydrazonoyl chlorides (HCs) were synthesized and human tumor cell line viability was determined using the MTT assay. Tumor development was assessed using Ehrlich ascites carcinoma (EAC)-bearing mice. KEY FINDINGS: our results showed that 2-oxo-N-phenyl-2-(phenylamino)acetohydrazonoyl chloride (compound 4; CPD 4) and 2-oxo-2-(phenylamino)-N-(p-tolyl)acetohydrazonoyl chloride (CPD 5) were the most cytotoxic HCs to human cervical tumor HeLa (IC50: 20 and 25 µM for CPD 4 and 5 respectively), breast MCF7 (IC50: 29 and 34 µM for CPD 4 and 5 respectively) and colon HCT116 cancer cells (IC50: 26 and 25 µM for CPD 4 and 5 respectively) with the least cytotoxicity to human non-tumor CCD-18Co colon fibroblasts as well as murine splenocytes. The active compounds significantly inhibited colony formation as well as tumor development in EAC-bearing mice. We also observed that PTEN-deficient cells displayed greater sensitivity than cells expressing wild type PTEN. At the molecular level, comet and cell cycle analyses indicated that the active compounds generate DNA damage. In light of the PTEN-dependent sensitivity and genomic instability we examined the influence of HCs on the DNA repair enzyme polynucleotide kinase/phosphatase (PNKP) and the PI3K/AKT/mTOR pathway, which are each known to be synthetic lethal with PTEN. We found that both PNKP and the PI3K/AKT/mTOR pathway to be adversely affected by the HCs, which may partially account for their toxicity. SIGNIFICANCE: hydrazonoyl chlorides can be considered as hit compounds for the development of new antitumor agents.


Assuntos
Antineoplásicos/síntese química , Hidrazonas/síntese química , Hidrazonas/farmacologia , Animais , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cloretos/química , Cloretos/farmacologia , Enzimas Reparadoras do DNA/metabolismo , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Feminino , Humanos , Hidrazonas/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo
4.
J Nutr Biochem ; 100: 108901, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34748925

RESUMO

A nutrition deficiency is one of the various causes of hearing loss. Zinc is an essential element for cell proliferation, antioxidant reactions, and the maintenance of hearing ability. Our previous studies have reported that the auditory brainstem response (ABR) threshold is increased in mice fed with zinc-deficient diets. However, the molecular mechanism of zinc involved in auditory system remains to be elucidated. In the present study, we examined the detrimental effects of zinc deficiency on cell cycle progression in murine auditory cells (HEI-OC1). The treatment of HEI-OC1 cells with 0.5 µM TPEN (N,N,N',N'-Tetrakis (2-pyridylmethyl) ethylenediamine) for 24 h inhibited cell proliferation, accumulation of reactive oxygen species (ROS), and induction of apoptosis. The cell proliferation block was caused by a G1/S phase arrest. Supplementation of the cell growth medium with 5 µM ZnCl2 after exposure to TPEN attenuated ROS accumulation and the arrest caused by the zinc deficiency. The ABR threshold was elevated in mice fed with a zinc-deficient diet. Additionally, we observed an increased expression of p21 and decreased expression of cyclin E and pRb in the spiral ganglion (SG), the organ of Corti (OC), Limbus (L), and stria vascularis (SV) in the zinc-deficient mouse cochlea. These results indicated that zinc is an essential nutrient for proliferation via the cell cycle and that a dysregulation of the cell cycle may cause hearing loss.


Assuntos
Ciclo Celular , Células Ciliadas Auditivas/citologia , Células Ciliadas Auditivas/metabolismo , Zinco/deficiência , Zinco/fisiologia , Animais , Apoptose , Pontos de Checagem do Ciclo Celular , Linhagem Celular , Proliferação de Células , Sobrevivência Celular , Cloretos/farmacologia , Cóclea/metabolismo , Etilenodiaminas/farmacologia , Potenciais Evocados Auditivos do Tronco Encefálico , Audição , Homeostase , Masculino , Camundongos , Camundongos Endogâmicos CBA , Oxirredução , Espécies Reativas de Oxigênio , Compostos de Zinco/farmacologia
5.
Genes Genomics ; 44(1): 1-7, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34800260

RESUMO

BACKGROUND: Cisplatin (CP) is an effective anticancer drug broadly used for various types of cancers, but it has shown ototoxicity that results from oxidative stress. Berberine has been reported for its anti-oxidative stress suggesting its therapeutic potential for many diseases such as colitis, diabetes, and vascular dementia. OBJECTIVE: Organ of Corti of postnatal day 3 mouse cochlear explants were used to compare hair cells after the treatment with cisplatin alone or with berberine chloride (BC) followed by CP. METHODS: We investigated the potential of the anti-oxidative effect of BC against the cisplatin-induced ototoxicity. We observed a reduced aberrant bundle of stereocilia in hair cells in CP with BC pre-treated group. Caspase-3 immunofluorescence and TUNEL assay supported the hypothesis that BC attenuates the apoptotic signals induced by CP. Reactive oxygen species level in the mitochondria were investigated by MitoSOX Red staining and the mitochondrial membrane potentials were compared by JC-1 assay. RESULTS: BC decreased ROS generation with preserved mitochondrial membrane potentials in mitochondria as well as reduced DNA fragmentation in hair cells. In summary, our data indicate that BC might act as antioxidant against CP by reducing the stress in mitochondria resulting in cell survival. CONCLUSION: Our result suggests the therapeutic potential of BC for prevention of the detrimental effect of CP-induced ototoxicity.


Assuntos
Berberina/farmacologia , Cloretos/farmacologia , Cisplatino/efeitos adversos , Ototoxicidade/prevenção & controle , Animais , Antineoplásicos/efeitos adversos , Apoptose/efeitos dos fármacos , Berberina/química , Caspase 3/metabolismo , Células Cultivadas , Cloretos/química , Cóclea/citologia , Cóclea/efeitos dos fármacos , Cóclea/metabolismo , Células Ciliadas Auditivas/efeitos dos fármacos , Células Ciliadas Auditivas/metabolismo , Marcação In Situ das Extremidades Cortadas , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Órgão Espiral/citologia , Órgão Espiral/efeitos dos fármacos , Órgão Espiral/metabolismo , Ototoxicidade/etiologia , Ototoxicidade/metabolismo , Substâncias Protetoras/farmacologia , Espécies Reativas de Oxigênio/metabolismo
6.
J Mater Sci Mater Med ; 33(2): 22, 2022 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-35133505

RESUMO

Wound dressings that exert an antimicrobial effect in order to prevent and treat wound infections can be harmful to the wound healing process. Dressings with hydrophobic coatings, however, have been suggested to both reduce the microbial load and promote the healing process. Therefore, the potential effects of a dialkylcarbamoyl chloride (DACC)-coated dressing on fibroblasts and keratinocytes in wound healing were studied using mechanical scratch wounding of confluent cell layers as an in vitro model. Additionally, gene expression analysis by qRT-PCR was used to elucidate the longitudinal effects of the DACC-coated dressing on cell responses, specifically inflammation, growth factor induction and collagen synthesis. DACC promoted cell viability, did not stick to the cell layers, and supported normal wound healing progression in vitro. In contrast, cells became attached to the uncoated reference material, which inhibited scratch closure. Moreover, DACC slightly induced KGF, VEGF, and GM-CSF expression in HaCaT cells and NHDF. Physiological COL1A1 and COL3A1 gene expression by NHDF was observed under DACC treatment with no observable effect on S100A7 and RNASE7 levels in HaCaT cells. Overall, the DACC coating was found to be safe and may positively influence the wound healing outcome. Graphical abstract.


Assuntos
Bandagens , Cloretos , Cloretos/farmacologia , Fibroblastos , Queratinócitos , Cicatrização
7.
Cells ; 10(12)2021 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-34943852

RESUMO

Depression is an independent risk factor for cardiovascular disease (CVD). We have previously shown that repeated social defeat (RSD) exaggerates atherosclerosis development by enhancing neutrophil extracellular trap (NET) formation. In this study, we investigated the impact of RSD on arterial thrombosis. Eight-week-old male wild-type mice (C57BL/6J) were exposed to RSD by housing with larger CD-1 mice in a shared home cage. They were subjected to vigorous physical contact daily for 10 consecutive days. After confirming depression-like behaviors, mice underwent FeCl3-induced carotid arterial injury and were analyzed after 3 h. Although the volume of thrombi was comparable between the two groups, fibrin(ogen)-positive areas were significantly increased in defeated mice, in which Ly-6G-positive cells were appreciably co-localized with Cit-H3-positive staining. Treatment with DNase I completely diminished exaggerated fibrin-rich clot formation in defeated mice. Flow cytometric analysis showed that neutrophil CD11b expression before FeCl3 application was significantly higher in defeated mice than in control mice. In vitro NET formation induced by activated platelets was significantly augmented in defeated mice, which was substantially inhibited by anti-CD11b antibody treatment. Our findings demonstrate that RSD enhances fibrin-rich clot formation after arterial injury by enhancing NET formation, suggesting that NET can be a new therapeutic target in depression-related CVD.


Assuntos
Coagulação Sanguínea , Plaquetas/metabolismo , Comunicação Celular , Armadilhas Extracelulares/metabolismo , Fibrina/metabolismo , Neutrófilos/metabolismo , Derrota Social , Animais , Anticorpos/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Plaquetas/efeitos dos fármacos , Antígeno CD11b/metabolismo , Comunicação Celular/efeitos dos fármacos , Cloretos/farmacologia , Desoxirribonuclease I/metabolismo , Armadilhas Extracelulares/efeitos dos fármacos , Compostos Férricos/farmacologia , Masculino , Camundongos Endogâmicos C57BL , Neutrófilos/efeitos dos fármacos , Selectina-P/metabolismo , Agregação Plaquetária/efeitos dos fármacos , Trombose/patologia
8.
Biomolecules ; 11(12)2021 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-34944502

RESUMO

Metal-based drugs represent a rich source of chemical substances of potential interest for the treatment of COVID-19. To this end, we have developed a small but representative panel of nine metal compounds, including both synthesized and commercially available complexes, suitable for medical application and tested them in vitro against the SARS-CoV-2 virus. The screening revealed that three compounds from the panel, i.e., the organogold(III) compound Aubipyc, the ruthenium(III) complex KP1019, and antimony trichloride (SbCl3), are endowed with notable antiviral properties and an acceptable cytotoxicity profile. These initial findings prompted us to perform a computational study to unveil the likely molecular basis of their antiviral actions. Calculations evidenced that the metalation of nucleophile sites in SARS-CoV-2 proteins or nucleobase strands, induced by Aubipyc, SbCl3, and KP1019, is likely to occur. Remarkably, we found that only the deprotonated forms of Cys and Sec residues can react favorably with these metallodrugs. The mechanistic implications of these findings are discussed.


Assuntos
2,2'-Dipiridil/análogos & derivados , Antimônio/farmacologia , Antivirais/farmacologia , COVID-19/tratamento farmacológico , Cloretos/farmacologia , Indazóis/farmacologia , Compostos Organoáuricos/farmacologia , Compostos Organometálicos/farmacologia , Compostos de Rutênio/farmacologia , SARS-CoV-2/efeitos dos fármacos , 2,2'-Dipiridil/química , 2,2'-Dipiridil/farmacologia , Animais , Antimônio/química , Antivirais/química , Linhagem Celular , Cloretos/química , Chlorocebus aethiops , Descoberta de Drogas , Humanos , Indazóis/química , Compostos Organoáuricos/química , Compostos Organometálicos/química , Compostos de Rutênio/química , Células Vero
9.
Viruses ; 13(12)2021 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-34960706

RESUMO

Epidemic RNA viruses seem to arise year after year leading to countless infections and devastating disease. SARS-CoV-2 is the most recent of these viruses, but there will undoubtedly be more to come. While effective SARS-CoV-2 vaccines are being deployed, one approach that is still missing is effective antivirals that can be used at the onset of infections and therefore prevent pandemics. Here, we screened FDA-approved compounds against SARS-CoV-2. We found that atovaquone, a pyrimidine biosynthesis inhibitor, is able to reduce SARS-CoV-2 infection in human lung cells. In addition, we found that berberine chloride, a plant-based compound used in holistic medicine, was able to inhibit SARS-CoV-2 infection in cells through direct interaction with the virion. Taken together, these studies highlight potential avenues of antiviral development to block emerging viruses. Such proactive approaches, conducted well before the next pandemic, will be essential to have drugs ready for when the next emerging virus hits.


Assuntos
Antivirais/farmacologia , Atovaquona/farmacologia , Berberina/farmacologia , SARS-CoV-2/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Células Epiteliais Alveolares , Animais , Berberina/química , Proliferação de Células/efeitos dos fármacos , Cloretos/química , Cloretos/farmacologia , Chlorocebus aethiops , Sinergismo Farmacológico , Humanos , Proguanil/farmacologia , Células Vero , Vírion/efeitos dos fármacos
10.
Anticancer Res ; 41(11): 5453-5459, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34732414

RESUMO

BACKGROUND: Zinc is a mineral that is essential for biological molecules, such as transcription factors, and is involved in the maintenance of intestinal homeostasis. Vitamin D signaling is mediated by vitamin D receptor (VDR) activated by 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] and is also important in intestinal functions, such as calcium absorption and epithelial barrier maintenance. However, the crosstalk between vitamin D signaling and zinc signaling in intestinal cells remains poorly understood. MATERIALS AND METHODS: Colon cancer SW480 and HCT116 cells were treated with zinc chloride (ZnCl2) with/without 1,25(OH)2D3 Expression of zinc-inducible genes [metallothionein 1A (MT1A) and MT2A] and VDR target genes [cytochrome P450 family 24 subfamily A member 1 (CYP24A1), transient receptor potential cation channel subfamily V member 6 (TRPV6) and cadherin 1 (CDH1)] was examined. RESULTS: Treatment of cells with ZnCl2 effectively induced MT1A and MT2A mRNA expression, and interestingly suppressed mRNA expression of CDH1, which was induced by 1,25(OH)2D3 in both cell lines. ZnCl2 also reduced the CDH1 protein level in HCT116 cells. CONCLUSION: Zinc signaling suppresses VDR-induced expression of CDH1.


Assuntos
Antígenos CD/metabolismo , Caderinas/metabolismo , Calcitriol/farmacologia , Cloretos/farmacologia , Neoplasias do Colo/metabolismo , Receptores de Calcitriol/agonistas , Compostos de Zinco/farmacologia , Antígenos CD/genética , Caderinas/genética , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Regulação Neoplásica da Expressão Gênica , Células HCT116 , Humanos , Metalotioneína/genética , Metalotioneína/metabolismo , Receptores de Calcitriol/metabolismo , Transdução de Sinais
11.
Eur J Histochem ; 65(s1)2021 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-34755507

RESUMO

Oxaliplatin is a third-generation chemotherapy drug mainly used for colorectal cancer treatment. However, it is also known to trigger neuropathy whose underlying neurobiological mechanisms are still under investigation and currently available treatments show limited efficacy. It is now established that neurons are not the only cell type involved in chronic pain and that glial cells, mainly astrocytes and microglia, are involved in the initiation and maintenance of neuropathy. Among all the pathogenetic factors involved in neuropathic pain, an oxaliplatin-dependent oxidative stress plays a predominant role. In our study, the antioxidant properties of magnesium (Mg), manganese (Mn) and zinc (Zn) salts were evaluated in order to counteract microglial activation induced by oxaliplatin. The antioxidant efficacy of these metals was evaluated by the means of molecular and morphological assays on the BV-2 microglial cell line. Our data clearly show that Mg, Mn and Zn are able to prevent oxaliplatin-dependent microglial alterations by reducing both oxidative and endoplasmic reticulum stress.


Assuntos
Antioxidantes/farmacologia , Cloretos/farmacologia , Cloreto de Magnésio/farmacologia , Compostos de Manganês/farmacologia , Oxaliplatina/toxicidade , Compostos de Zinco/farmacologia , Animais , Antígeno B7-2/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Linhagem Celular , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Camundongos , Proteínas dos Microfilamentos/metabolismo , Microglia/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo
12.
Daru ; 29(2): 321-328, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34417987

RESUMO

PURPOSE: Mohs' paste, which is composed of zinc chloride and zinc oxide starch, is used for hemostasis of superficial malignancy in the clinical setting. We investigated the concentration of intramuscular zinc in mice after Mohs' paste application and evaluated its relationship with angiogenesis from the perspective of blood flow levels within 24 h. METHODS: Male C57BL/6JJmsSlc mice were administered single dose of Mohs' paste at 25%, 50%, and 75% after unilateral hind limb surgery, and glycerin, a viscosity modifier, was administered to the control group (0%). Hind limb blood flow levels were measured with a laser Doppler perfusion imaging system (n = 6). The amounts of intramuscular zinc and vascular endothelial growth factor-A (VEGF-A) expression were analyzed using inductively coupled plasma mass spectrometry (ICP-MS) and western blotting, respectively (n = 5 or 3). RESULTS: Blood flow levels were significantly decreased in the 50% group after 8 h, and significantly decreased in the 25% and 50% groups after 24 h. Intramuscular zinc was significantly increased in the 50% and 75% groups after 8 h. Western blotting showed that VEGF-A levels were significantly increased in the 25% and 50% groups after 8 h. Based on analytical experiments and biological investigation, we predicated the pharmacological effect of Mohs' paste and found over 50% of it is critical in the blood flow and angiogenesis suppression after more than 8 h of its application. CONCLUSIONS: The results suggest that the mechanism of blood flow suppression is independent of VEGF-A levels and might suppress future angiogenesis. Our findings support that of previous studies, in which Mohs' paste was expected to induce hemostasis and suppress angiogenesis. It is an excellent ointment that facilitates hemostasis by suppressing blood flow regardless of angiogenesis, and may be apt for situations where hemostasis is required in the clinical setting.


Assuntos
Inibidores da Angiogênese/administração & dosagem , Circulação Sanguínea/efeitos dos fármacos , Cloretos/administração & dosagem , Membro Posterior/cirurgia , Músculo Esquelético/química , Fator A de Crescimento do Endotélio Vascular/metabolismo , Compostos de Zinco/administração & dosagem , Zinco/análise , Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacologia , Animais , Cloretos/química , Cloretos/farmacologia , Relação Dose-Resposta a Droga , Glicerol/química , Membro Posterior/diagnóstico por imagem , Fluxometria por Laser-Doppler , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/diagnóstico por imagem , Imagem de Perfusão , Espectrofotometria Atômica , Compostos de Zinco/química , Compostos de Zinco/farmacologia
13.
Proc Natl Acad Sci U S A ; 118(34)2021 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-34413188

RESUMO

TMEM16A Ca2+-activated chloride channels are involved in multiple cellular functions and are proposed targets for diseases such as hypertension, stroke, and cystic fibrosis. This therapeutic endeavor, however, suffers from paucity of selective and potent modulators. Here, exploiting a synthetic small molecule with a biphasic effect on the TMEM16A channel, anthracene-9-carboxylic acid (A9C), we shed light on sites of the channel amenable for pharmacological intervention. Mutant channels with the intracellular gate constitutively open were generated. These channels were entirely insensitive to extracellular A9C when intracellular Ca2+ was omitted. However, when physiological Ca2+ levels were reestablished, the mutants regained sensitivity to A9C. Thus, intracellular Ca2+ is mandatory for the channel response to an extracellular modulator. The underlying mechanism is a conformational change in the outer pore that enables A9C to enter the pore to reach its binding site. The explanation of this structural rearrangement highlights a critical site for pharmacological intervention and reveals an aspect of Ca2+ gating in the TMEM16A channel.


Assuntos
Anoctamina-1/metabolismo , Antracenos/farmacologia , Cálcio/farmacologia , Cloretos/farmacologia , Animais , Anoctamina-1/genética , Estimulação Elétrica , Fenômenos Eletrofisiológicos , Regulação da Expressão Gênica/efeitos dos fármacos , Células HEK293 , Humanos , Camundongos , Simulação de Dinâmica Molecular , Técnicas de Patch-Clamp , Mutação Puntual
14.
Toxicol In Vitro ; 76: 105232, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34365006

RESUMO

Skin permeation and distribution of three of the most common skin sensitizers was investigated using a previously developed animal-free exposure method combined with imaging mass spectrometry. Nickel, cobalt, and chromium (III) salts were dissolved in a buffer and exposed to human skin ex vivo, to be analyzed using time of flight secondary ion mass spectrometry (ToF-SIMS). Our findings demonstrate that metal haptens mainly accumulated in the stratum corneum, however all three metal sensitizers could also be detected in the epidermis. Cobalt and chromium (III) species penetrated into the epidermis to a larger extent than nickel species. The degree of penetration into the epidermis is suggested to be affected by the sensitization potency of the metal salts, as well as their speciation, i.e. the amount of the respective metal present in the solution as bioaccessible and solubilised ions. Our method provided permeation profiles in human skin for known sensitizers, on a level of detail that is not possible to achieve by other means. The findings show that the permeation profiles are different, despite these sensitizers being all metal ions and common causes of contact allergy. Studying skin uptake by only considering penetration through the skin might therefore not give accurate results.


Assuntos
Cloretos/farmacologia , Compostos de Cromo/farmacologia , Cobalto/farmacologia , Haptenos/farmacologia , Níquel/farmacologia , Pele/metabolismo , Feminino , Humanos , Técnicas In Vitro , Espectrometria de Massas , Pele/diagnóstico por imagem , Absorção Cutânea
15.
Toxins (Basel) ; 13(7)2021 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-34208992

RESUMO

Cultures of the mussel Mytilus galloprovincialis are frequently affected by accumulation of the amnesic shellfish poisoning toxin domoic acid (DA). This species is characterized by a fast uptake and release of the toxin. In this work, the main characteristics of the uptake mechanism have been studied by incubation of digestive gland thin slices in media with different composition and DA concentration. DA uptake seems to follow Michaelis-Menten kinetics, with a very high estimated KM (1722 µg DA mL-1) and a Vmax of 71.9 µg DA g-1 h-1, which is similar to those found for other amino acids in invertebrates. Replacement of NaCl from the incubation media by Cl-choline (Na+-free medium) did not significantly reduce the uptake, but replacement by sorbitol (Na+-free and Cl--depleted medium) did. A new experiment replacing all chlorides with their equivalent gluconates (Na+- and Cl--free medium) showed an important reduction in the uptake that should be attributed to the absence of chloride, pointing to a Na+-independent, Cl- (or anion-) dependent transporter. In media with Na+ and Cl-, neither decreasing the pH nor adding cyanide (a metabolic inhibitor) had significant effect on DA uptake, suggesting that the transport mechanism is not H+- or ATP-dependent. In a chloride depleted medium, lowering pH or adding CN increased the uptake, suggesting that other anions could, at least partially, substitute chloride.


Assuntos
Trato Gastrointestinal/metabolismo , Ácido Caínico/análogos & derivados , Mytilus/metabolismo , Animais , Cloretos/farmacologia , Cianetos/farmacologia , Gluconatos/farmacologia , Concentração de Íons de Hidrogênio , Ácido Caínico/farmacologia , Água do Mar/química
16.
Glycobiology ; 31(10): 1295-1307, 2021 11 18.
Artigo em Inglês | MEDLINE | ID: mdl-34224566

RESUMO

Skeletal muscle has the intrinsic ability to self-repair through a multifactorial process, but many aspects of its cellular and molecular mechanisms are not fully understood. There is increasing evidence that some members of the mammalian ß-galactoside-binding protein family (galectins) are involved in the muscular repair process (MRP), including galectin-3 (Gal-3). However, there are many questions about the role of this protein on muscle self-repair. Here, we demonstrate that endogenous Gal-3 is required for: (i) muscle repair in vivo by using a chloride-barium myolesion mouse model and (ii) mouse primary myoblasts myogenic programming. Injured muscle from Gal-3 knockout mice (GAL3KO) showed persistent inflammation associated with compromised muscle repair and the formation of fibrotic tissue on the lesion site. In GAL3KO mice, osteopontin expression remained high even after 7 and 14 d of the myolesion, while Myoblast differentiation transcription factor (MyoD) and myogenin had decreased their expression. In GAL3KO mouse primary myoblast cell culture, Paired Box 7 (Pax7) detection seems to sustain even when cells are stimulated to differentiation and MyoD expression is drastically reduced. The detection and temporal expression levels of these transcriptional factors appear to be altered in Gal-3-deficient myoblast. Gal-3 expression in wild-type mice for GAL3KO states, both in vivo and in vitro, in sarcoplasm/cytoplasm and myonuclei; as differentiation proceeds, Gal-3 expression is drastically reduced, and its location is confined to the sarcolemma/plasma cell membrane. We also observed a change in the temporal-spatial profile of Gal-3 expression and muscle transcription factors levels during the myolesion. Overall, these results demonstrate that endogenous Gal-3 is required for the skeletal muscle repair process.


Assuntos
Galectina 3/metabolismo , Músculo Esquelético/metabolismo , Animais , Compostos de Bário/administração & dosagem , Compostos de Bário/farmacologia , Cloretos/administração & dosagem , Cloretos/farmacologia , Galectina 3/deficiência , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/patologia
17.
Int J Mol Sci ; 22(14)2021 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-34299039

RESUMO

Zinc chloride is known to be effective in combatting hepatitis A virus (HAV) infection, and zinc ions seem to be especially involved in Toll-like receptor (TLR) signaling pathways. In the present study, we examined this involvement in human hepatoma cell lines using a human TLR signaling target RT-PCR array. We also observed that zinc chloride inhibited mitogen-activated protein kinase kinase 3 (MAP2K3) expression, which could downregulate HAV replication in human hepatocytes. It is possible that zinc chloride may inhibit HAV replication in association with its inhibition of MAP2K3. In that regard, this study set out to determine whether MAP2K3 could be considered a modulating factor in the development of the HAV pathogen-associated molecular pattern (PAMP) and its triggering of interferon-ß production. Because MAP2K3 seems to play a role in antiviral immunity against HAV infection, it is a promising target for drug development. The inhibition of MAP2K3 may also prevent HAV patients from developing a severe hepatitis A infection.


Assuntos
Carcinoma Hepatocelular/virologia , Cloretos/farmacologia , Hepatite A/complicações , Hepatócitos/virologia , Neoplasias Hepáticas/virologia , MAP Quinase Quinase 3/antagonistas & inibidores , Replicação Viral , Compostos de Zinco/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/enzimologia , Hepatite A/virologia , Vírus da Hepatite A/isolamento & purificação , Hepatócitos/efeitos dos fármacos , Hepatócitos/enzimologia , Interações Hospedeiro-Patógeno , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/enzimologia , Células Tumorais Cultivadas
18.
Br J Cancer ; 125(2): 265-276, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33981016

RESUMO

BACKGROUND: Anti-microtubule agents are widely used to treat ovarian cancers, but the efficacy is often compromised by drug resistance. We investigated co-targeting the actin/tropomyosin cytoskeleton and microtubules to increase treatment efficacy in ovarian cancers and potentially overcome resistance. METHODS: The presence of tropomyosin-3.1 (Tpm3.1) was examined in clinical specimens from ovarian cancer patients using immunohistochemistry. Combinatorial effects of an anti-Tpm3.1 compound, ATM-3507, with vinorelbine and paclitaxel were evaluated in ovarian cancer cells via MTS and apoptosis assays. The mechanisms of action were established using live- and fixed-cell imaging and protein analysis. RESULTS: Tpm3.1 is overexpressed in 97% of tumour tissues (558 of 577) representing all histotypes of epithelial ovarian cancer. ATM-3507 displayed synergy with both anti-microtubule agents to reduce cell viability. Only vinorelbine synergised with ATM-3507 in causing apoptosis. ATM-3507 significantly prolonged vinorelbine-induced mitotic arrest with elevated activity of the spindle assembly checkpoint and mitotic cell death; however, ATM-3507 showed minor impact on paclitaxel-induced mitotic defects. Both combinations substantially increased post-mitotic G1 arrest with cyclin D1 and E1 downregulation and an increase of p21Cip and p27Kip. CONCLUSION: Combined targeting of Tpm3.1/actin and microtubules is a promising treatment strategy for ovarian cancer that should be further tested in clinical settings.


Assuntos
Carcinoma Epitelial do Ovário/metabolismo , Cloretos/farmacologia , Neoplasias Ovarianas/metabolismo , Paclitaxel/farmacologia , Tropomiosina/metabolismo , Regulação para Cima , Vinorelbina/farmacologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Epitelial do Ovário/tratamento farmacológico , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sinergismo Farmacológico , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/tratamento farmacológico , Tropomiosina/antagonistas & inibidores , Regulação para Cima/efeitos dos fármacos
19.
Molecules ; 26(7)2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33916013

RESUMO

Periodontal diseases like gingivitis and periodontitis are primarily caused by dental plaque. Several antiplaque and anti-microbial agents have been successfully incorporated into toothpastes and mouthwashes to control plaque biofilms and to prevent and treat gingivitis and periodontitis. The aim of this article was to review recent developments in the antiplaque, anti-gingivitis, and anti-periodontitis properties of some common compounds in toothpastes and mouthwashes by evaluating basic and clinical studies, especially the ones published in the past five years. The common active ingredients in toothpastes and mouthwashes included in this review are chlorhexidine, cetylpyridinium chloride, sodium fluoride, stannous fluoride, stannous chloride, zinc oxide, zinc chloride, and two herbs-licorice and curcumin. We believe this comprehensive review will provide useful up-to-date information for dental care professionals and the general public regarding the major oral care products on the market that are in daily use.


Assuntos
Antissépticos Bucais/análise , Antissépticos Bucais/química , Doenças Periodontais/prevenção & controle , Cremes Dentais/análise , Cremes Dentais/química , Anti-Infecciosos Locais/química , Anti-Infecciosos Locais/farmacologia , Cetilpiridínio/química , Cetilpiridínio/farmacologia , Cloretos/química , Cloretos/farmacologia , Humanos , Doenças Periodontais/etiologia , Doenças Periodontais/patologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Fluoreto de Sódio/química , Fluoreto de Sódio/farmacologia , Fluoretos de Estanho/análise , Fluoretos de Estanho/química , Fluoretos de Estanho/farmacologia , Compostos de Zinco/química , Compostos de Zinco/farmacologia
20.
J Mater Chem B ; 9(13): 3079-3087, 2021 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-33885669

RESUMO

This study demonstrates the fabrication of ambient light enabled antimicrobial functional fabrics by coating flower-like bismuth oxyhalide i.e. BiOCl0.875Br0.125, with the use of poly(vinyl alcohol) (PVA) and poly(acrylic acid) (PAA) as binders for improved coating robustness and durability. The uniformity of the microparticles was ensured with simultaneous probe sonication during the stages of crystal nucleation and growth. The polymeric binders not only strongly anchor the particle on the fabric, but also serve as an ultra-thin protective layer on the BiOClBr that mitigates bismuth leaching. The efficacy of inhibiting bacteria was investigated over the BiOClBr-coated fabrics i.e. cotton and polyester, and the results showed that the coated fabrics could effectively inhibit both Gram-positive and Gram-negative bacteria, i.e. S. aureus and E. coli. In comparison with fabrics coated with other photocatalytic materials including bismuth oxide (Bi2O3) and zinc oxide (ZnO), an exceptionally better antimicrobial efficacy was observed for BiOClBr-coated fabrics. The BiOClBr-coated cotton showed ∼5.0 and ∼6.8 times higher disinfection efficacy towards E. coli compared to that of ZnO and Bi2O3-coated cotton with the same particle weight percentage, respectively. Further elucidation of the probable mechanism by BiOClBr-coated fabrics is related to the excess amount of reactive oxygen species (ROS). Overall, BiOClBr has been shown to be a promising material to fabricate cost-effective antimicrobial functional surfaces for both environmental and biomedical applications e.g. protective laboratory and factory clothing.


Assuntos
Antibacterianos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Luz , Antibacterianos/síntese química , Antibacterianos/química , Bismuto/química , Bismuto/farmacologia , Brometos/química , Brometos/farmacologia , Cloretos/química , Cloretos/farmacologia , Testes de Sensibilidade Microbiana , Oxigênio/química , Oxigênio/farmacologia , Tamanho da Partícula
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