RESUMEN
Selenium is an important element in nutrition, showing great potential in the udder health of dairy goats and in the control of subclinical mastitis. However, there are few studies that evaluated the influence of selenium supplementation on subclinical mastitis in goats. The aim of this study was to evaluate the incidence of subclinical mastitis in dairy goats supplemented with organic selenium (Se yeast) in a semi-arid region. Sixteen Saanen × Toggenburg crossbred lactating goats were allocated randomly into two treatments: with and without addition of organic selenium (Se) to the concentrate. Milk samples were collected every 20 days from each udder half to determine the somatic cell count (SSC), chloride content, pH, electrical conductivity, microbiological isolation, composition, and selenium contents. The highest serum selenium concentrations in the blood of these goats occurred at 42 days of supplementation (P < 0.001). Goats which received organic selenium supplementation had higher serum selenium concentrations (P < 0.05). The milk composition variables did not differ (P > 0.05) between the tested treatments, teats, and collections. After 60 days of supplementation, a difference was observed (P < 0.05) between treatments for SSC, chloride content, and pH. Addition of organic selenium to the diet of dairy goats after 60 days of supplementation was promising in reducing the somatic cell count, consequently improving milk quality.
Asunto(s)
Enfermedades de las Cabras , Mastitis , Selenio , Animales , Femenino , Recuento de Células/veterinaria , Cloruros/análisis , Cloruros/farmacología , Dieta/veterinaria , Enfermedades de las Cabras/microbiología , Cabras , Lactancia , Mastitis/veterinaria , Leche/química , Saccharomyces cerevisiae , Selenio/farmacologíaRESUMEN
This study evaluated the effect of a mouthwash containing 0.075% cetylpyridinium chloride and 0.28% zinc lactate (CPC + Zn) in a multispecies biofilm model. A 7-days 33-species biofilm, formed on Calgary device, was 1-min treated with: 0.12% chlorhexidine (CHX), culture medium (negative control), 0.075% cetylpyridinium chloride (CPC) or CPC + Zn, 2x/day, from day 3 until day 6. The metabolic activity and the microbial composition were evaluated by colorimetric method and checkerboard DNA-DNA hybridization, respectively. The three antimicrobials (CPC, CPC + Zn and CHX) reduced metabolic activity, total biofilm count and several species counts, including Porphyromonas gingivalis, Fusobacterium nucleatum, Parvimonas micra, Campylobacter gracilis and Streptococcus mutans. However, only CPC + Zn reduced counts of the pathogen Prevotella intermedia and did not interfere with the levels of some beneficial species in relation to the negative control. The treatment of multispecies subgingival biofilm with CPC + Zn was effective in controlling periodontal pathogens and favored the colonization of health-associated bacterial species.
Asunto(s)
Cetilpiridinio , Antisépticos Bucales , Cetilpiridinio/farmacología , Antisépticos Bucales/farmacología , Zinc/farmacología , Cloruros/farmacología , Biopelículas , Clorhexidina/farmacología , Porphyromonas gingivalis , ADNRESUMEN
Mercury is considered a risk factor for the development of hypertension and other cardiovascular diseases. We investigated whether the effects of mercury exposure on haemodynamic parameters of young Wistar rats and prehypertensive SHRs might alter the time course of hypertension development. Young (4 weeks) male Wistar rats and SHRs were randomly assigned to four groups: untreated Wistar rats (Wistar Ct), Wistar rats exposed to mercury chloride for 30 days (Wistar Hg), untreated SHRs (SHR Ct) and SHRs exposed to mercury chloride (SHR Hg) for 30 days. Non-invasive and invasive arterial pressures were measured to investigate pressure reactivity; nitrite/nitrate levels, ACE activity, and lipid peroxidation were measured in plasma. The systolic blood pressure (SBP) of the Wistar rat groups did not change but increased in the SHRs from the second week to the last week. Hg exposure accelerated the increase in the SBP of SHRs. L-NAME administration increased SBP and diastolic blood pressure (DBP) in all groups, but this increase was smaller in SHRs exposed to Hg. A decrease in plasma nitrite and nitrate levels in the SHR Hg group suggested that mercury reduced NO bioavailability. Tempol-reduced blood pressure suggesting that the superoxide anion played a role in the marked increase in this parameter. These findings provide evidence that Hg exposure might activate mechanisms to accelerate hypertension development, including a reduction in NO bioavailability. Therefore, predisposed individuals under mercury exposure are at greater risk from an enhanced development of hypertension.
Asunto(s)
Hipertensión , Mercurio , Animales , Masculino , Ratas , Presión Sanguínea , Cloruros/farmacología , Hipertensión/inducido químicamente , Mercurio/farmacología , Nitratos , Nitritos , Estrés Oxidativo , Ratas Endogámicas SHR , Ratas WistarRESUMEN
Herein a systematic series of four [AuLL']n+ n = 0, +1 complexes, where L = 1,3-bis(mesityl)imidazole-2-ylidene (IMes), or triphenylphosphine (PPh3), and L' = chloride, or 4-dimethylaminopyridine (DMAP), had their in vitro antiviral activity assessed against Chikungunya virus (CHIKV). The PPh3 derivatives inhibited viral replication by 99%, whereas the IMes derivatives about 50%. The lipophilicity of the PPh3 derivatives is higher than the IMes-bearing compounds, which can be related to their more prominent antiviral activities. The dissociation of DMAP is faster than chloride in solution for both IMes and PPh3 derivatives; however, it does not significantly affect their in vitro activities, showing a higher dependence on the nature of L rather than L' towards their antiviral effects. All complexes bind to N-acetyl-L-cysteine, with the Ph3P-bearing complexes coordinating at a faster rate to this amino acid. The binding constants to bovine serum albumin are in the order of 104, slightly higher for the DMAP complexes in both PPh3 and IMes derivatives. Mechanistic investigations of the PPh3 complexes showed a ubiquitous protective effect of the compounds in the pretreatment, early stages, and post-entry assays. The most significant inhibition was observed in post-entry activity, in which the complexes blocked viral replication in 99%, followed by up to 95% inhibition of the early stages of infection. Pretreatment assays showed a 92% and 80% replication decrease for the chloride and DMAP derivatives, respectively. dsRNA binding assays showed a significant interaction of the compounds with dsRNA, an essential biomolecule to viral replication.
Asunto(s)
Virus Chikungunya , Antivirales/farmacología , Virus Chikungunya/genética , Cloruros/farmacología , Oro/farmacología , Compuestos OrganofosforadosRESUMEN
Chagas disease is a disease that is emerging in North America and Europe countries. Benznidazole is the main drug available, but it has high toxicity and low efficacy in the chronic phase. In this way, researching new antichagasic agents is necessary. Thus, the aim of this study is to evaluate the effect of novel chalcones and the influence of chlorine substitutions on Trypanosoma cruzi and host cells. Unsubstituted (1), 4-chlorine substituted (2) and 2,4-chlorine substituted (3) chalcones were synthesized by Claisen-Schmidt condensation, characterized, and electrical distribution was assessed by Density Fuctional Theory (DFT). The host cells toxicity (LLC-MK2) was performed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) reduction assay. The effect on epimastigote (24, 48 and 72h), trypomastigote (24h) and amastigotes (24 h) was evaluated. Flow cytometry assays were performed with 7-Aminoactinomycin D (7-AAD) and Annexin-PE, Dichlorofluorescein diaceteate (DCFH-DA) and Rhodamine123 (Rho123). Finally, molecular docking predicted interactions between chalcones and cruzain (TcCr) and trypanothione reductase (TcTR). The toxicity on host cells was reduced almost twenty times on chlorine substituted molecules. On epimastigote and trypomastigote forms, all substances presented similar effects. After treatment with molecule 3, it was observed a decrease in infected cells and intracellular amastigotes. Their effect is related to necrotic events, increase of cytoplasmic Reactive Oxygen Species (ROS) and mitochondrial dysfunction. Also, this effect might be associated with involvement of TcCr and TcTR enzymes. Therefore, the results showed that chlorine substitution on chalcones reduces the host cell's toxicity without compromising the effect on Trypanosoma cruzi Y strain forms, and it occurs over membrane damage, oxidative stress and possible interactions with TcCr and TcTR.
Asunto(s)
Enfermedad de Chagas , Chalcona , Chalconas , Tripanocidas , Trypanosoma cruzi , Enfermedad de Chagas/tratamiento farmacológico , Chalcona/farmacología , Chalconas/farmacología , Chalconas/uso terapéutico , Cloruros/farmacología , Cloro , Humanos , Simulación del Acoplamiento Molecular , Tripanocidas/farmacologíaRESUMEN
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.
Asunto(s)
Galectina 3/metabolismo , Músculo Esquelético/metabolismo , Animales , Compuestos de Bario/administración & dosificación , Compuestos de Bario/farmacología , Cloruros/administración & dosificación , Cloruros/farmacología , Galectina 3/deficiencia , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/patologíaRESUMEN
OBJECTIVES: To evaluate the in vitro photodynamic activity of aluminum phthalocyanine tetrasulfonate chloride (AlPcClS4) on promastigotes and amastigotes of Leishmania (Viannia) peruviana and Leishmania (Viannia) braziliensis. MATERIALS AND METHODS: The activity of photodynamic therapy using AlPcClS4 on Leishmania promastigote and amastigotes was determined by the Methyl Thiazole Tetrazolium (MTT) colorimetric method and quantitative PCR, respectively. RESULTS: Photodynamic treatment showed an inhibitory effect on promastigotes, particularly on Leishmania (V.) peruviana, to a lesser extent on Leishmania (V.) braziliensis and also on intracellular forms of both species. At 24 hours post-radiation, using concentrations of 200 µM and 350 µM, the inhibitory effect on Leishmania (V.) peruviana was 72.9% and 73.9% respectively; at 96 hours the inhibitory effect was of 78.8% and 80.6%, respectively. Regarding intracellular forms, the inhibitory effect on Leishmania (V.) peruviana amastigotes was 57.8% at 72 hours post-treatment, using a concentration of 200 µM. The IC50 was 56.5, 50, 44 and 39.7 µM, at 24, 48, 72 and 96 hours post-radiation, respectively. CONCLUSIONS: Photodynamic therapy using AlPcClS4 against Leishmania species showed encouraging results, mainly on Leishmania (V.) peruviana, suggesting a potential use as an alternative or complement to the usual treatment of tegumentary leishmaniasis. However, new trials are still required to determine the selectivity index for the intracellular form of the parasite, and to develop methods to facilitate the efficient entry of the molecule into the host cell and the parasite.
OBJETIVOS: Evaluar la actividad fotodinámica in vitro de la ftalocianina de aluminio tetrasulfonada clorada (AlPcClS4) sobre promastigotes y amastigotes de Leishmania (Viannia) peruviana y Leishmania (Viannia) braziliensis. MATERIALES Y MÉTODOS: La actividad del tratamiento fotodinámico empleando AlPcClS4 sobre promastigotes y amastigotes de Leishmania fue determinada mediante el método colorimétrico Metil Tiazol Tetrazolium (MTT) y PCR cuantitativo, respectivamente. RESULTADOS: El tratamiento fotodinámico presentó un efecto inhibitorio sobre promastigotes, principalmente sobre Leishmania (V.) peruviana, en menor proporción sobre Leishmania (V.) braziliensis y sobre las formas intracelulares de ambas especies. En Leishmania (V.) peruviana, a las 24 horas posirradiación a 200 µM y 350 µM el efecto inhibitorio fue del 72,9% y 73,9%, respectivamente y a las 96 horas fue del 78,8% y 80,6%, respectivamente. En las formas intracelulares, empleando 200 µM y evaluado a las 72 horas postratamiento, se observó una inhibición del 57,8% de amastigotes de Leishmania (V.) peruviana. El IC50 fue del 56,5; 50; 44; y 39,7 µM, que corresponde a las 24, 48, 72 y 96 horas posirradiación, respectivamente. CONCLUSIONES: El tratamiento fotodinámico empleando AlPcClS4 frente a las especies de Leishmania presentó resultados alentadores principalmente sobre Leishmania (V.) peruviana, lo cual sugiere su potencial uso como alternativa o complemento del tratamiento convencional de la leishmaniasis tegumentaria. Sin embargo, aún se requiere continuar con nuevos ensayos para determinar el índice de selectividad sobre el parásito en su forma intracelular, y desarrollar estrategias que faciliten el ingreso eficiente de la molécula hacia la célula hospedera y al parásito.
Asunto(s)
Cloruros , Indoles , Leishmania , Compuestos Organometálicos , Cloruros/farmacología , Indoles/farmacología , Leishmania/clasificación , Leishmania/efectos de los fármacos , Leishmania braziliensis/efectos de los fármacos , Compuestos Organometálicos/farmacologíaRESUMEN
PURPOSE: To develop a new wound dressing composed of alginate and Aloe vera gel and cross-linked with zinc ions. METHODS: The aloe-alginate film was characterized using scanning electron microscopy (SEM), swelling profile, mechanical properties, polysaccharide content and X-ray diffraction (XRD). Thirty Wistar rats were divided in two groups a) treated with aloe-alginate film and b) control (treated with sterile gauze). Wound contraction measurements and hystological analysis were performed on 7th, 14th and 21st days after wound surgery. RESULTS: The aloe-alginate film presented adequated mechanical resistance and malleability for application as wound dressing. There was no statistical difference in wound contraction between two groups. Histological assay demonstrated that aloe-alginate film presented anti-inflammatory activity, stimulated angiogenesis on proliferative phase and a more significant increased in collagen type I fibers and decreased type III fibers which promoted a mature scar formation when compared to control. CONCLUSIONS: The aloe-alginate film showed adequate physicochemical characteristics for wound dressing applications. The in vivo assay demonstrated that aloe-alginate film enhanced the healing process of incisional skin wounds.
Asunto(s)
Alginatos , Aloe , Cloruros , Preparaciones de Plantas , Cicatrización de Heridas , Compuestos de Zinc , Alginatos/farmacología , Animales , Cloruros/química , Cloruros/farmacología , Preparaciones de Plantas/farmacología , Ratas , Ratas Wistar , Cicatrización de Heridas/efectos de los fármacos , Compuestos de Zinc/química , Compuestos de Zinc/farmacologíaRESUMEN
Synthesis of four compounds belonging to mesoionic class, (E)-3-phenyl-5-(phenylamino)-2-styryl-1,3,4-thiadiazol-3-ium chloride derivatives (5a-d) and their biological evaluation against MT2 and C92 cell lines infected with human T-cell lymphotropic virus type-1 (HTLV-1), which causes adult T-cell leukemia/lymphoma (ATLL), and non-infected cell lines (Jurkat) are reported. The compounds were obtained by convergent synthesis under microwave irradiation and the cytotoxicity was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Results showed IC50 values of all compounds in the range of 1.51-7.70 M in HTLV-1-infected and non-infected cells. Furthermore, it was observed that 5b could induce necrosis after 24 h for Jurkat and MT2 cell lines. The experimental (fluorimetric method) and theoretical (molecular docking) results suggested that the mechanism of action for 5b could be related to its capacity to intercalate into DNA. Moreover, the preliminary pharmacokinetic profile of the studied compounds (5a-d) was obtained through human serum albumin (HSA) binding affinity using multiple spectroscopic techniques (circular dichroism, steady-state and time-resolved fluorescence), zeta potential and molecular docking calculations. The interaction HSA:5a-d is spontaneous and moderate (Ka ~ 104 M-1) via a ground-state association, without significantly perturbing both the secondary and surface structures of the albumin in the subdomain IIA (site I), indicating feasible biodistribution in the human bloodstream.
Asunto(s)
Antineoplásicos/farmacología , Cloruros/farmacología , Leucemia-Linfoma de Células T del Adulto/tratamiento farmacológico , Leucemia-Linfoma de Células T del Adulto/virología , Sitios de Unión , Línea Celular Tumoral , Supervivencia Celular , Dicroismo Circular , Ensayos de Selección de Medicamentos Antitumorales , Virus Linfotrópico T Tipo 1 Humano , Humanos , Concentración 50 Inhibidora , Células Jurkat , Espectroscopía de Resonancia Magnética , Microondas , Simulación del Acoplamiento Molecular , Unión Proteica , Especies Reactivas de Oxígeno/metabolismo , Albúmina Sérica Humana/química , Distribución TisularRESUMEN
We previously showed that microwave assisted synthesis is the best method for the synthesis of naphthoquinone amino acid and chloride-naphthoquinone amino acid derivatives by a complete evaluation of reaction conditions such as stoichiometry, bases, and pH influence. Following the same strategy, we synthesized chloride and non-chloride tyrosine, valine, and tryptophan-naphthoquinones achieving 85-95%, 80-92%, and 91-95% yields, respectively. The cyclic voltammetry profiles showed that both series of naphthoquinone amino acid derivatives mainly display one redox reaction process. Overall, chloride naphthoquinone amino acid derivatives exhibited redox potential values (E1/2) more positive than non-chloride compounds. The six newly synthesized compounds were tested in HPV positive and negative as well as in immortal and tumorigenic cell lines to observe the effects in different cellular context simulating precancerous and cancerous status. A dose-response was achieved to determine the IC50 of six newly synthesized compounds in SiHa (Tumorigenic and HPV16 positive), CaLo (Tumorigenic and HPV18 positive), C33-A (Tumorigenic and HPV negative) and HaCaT (Keratinocytes immortal HPV negative) cell lines. Non-chloride tryptophan-naphthoquinone (3c) and chloride tyrosine-naphthoquine (4a) effects were more potent in tumorigenic SiHa, CaLo, and C33-A cells with respect to non-tumorigenic HaCaT cells. Interestingly, there seems to be a differential effect in non-chloride and chloride naphthoquinone amino acid derivatives in tumorigenic versus non tumorigenic cells. Considering all naphthoquinone amino acid derivatives that our group synthesized, it seems that hydrophobic and aromatic amino acids have the greatest effect on cell proliferation inhibition. These results show promising compounds for cervical cancer treatment.
Asunto(s)
Antineoplásicos/síntesis química , Proliferación Celular/efectos de los fármacos , Naftoquinonas/química , Triptófano/química , Tirosina/química , Neoplasias del Cuello Uterino/patología , Valina/química , Antineoplásicos/farmacología , Carcinogénesis , Línea Celular Tumoral , Cloruros/química , Cloruros/farmacología , Diseño de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Células HaCaT , Humanos , Concentración 50 Inhibidora , Microondas , Oxidación-Reducción , Infecciones por Papillomavirus/complicaciones , Neoplasias del Cuello Uterino/tratamiento farmacológicoRESUMEN
Abstract Purpose To develop a new wound dressing composed of alginate and Aloe vera gel and cross-linked with zinc ions. Methods The aloe-alginate film was characterized using scanning electron microscopy (SEM), swelling profile, mechanical properties, polysaccharide content and X-ray diffraction (XRD). Thirty Wistar rats were divided in two groups a) treated with aloe-alginate film and b) control (treated with sterile gauze). Wound contraction measurements and hystological analysis were performed on 7th, 14th and 21st days after wound surgery. Results The aloe-alginate film presented adequated mechanical resistance and malleability for application as wound dressing. There was no statistical difference in wound contraction between two groups. Histological assay demonstrated that aloe-alginate film presented anti-inflammatory activity, stimulated angiogenesis on proliferative phase and a more significant increased in collagen type I fibers and decreased type III fibers which promoted a mature scar formation when compared to control. Conclusions The aloe-alginate film showed adequate physicochemical characteristics for wound dressing applications. The in vivo assay demonstrated that aloe-alginate film enhanced the healing process of incisional skin wounds.
Asunto(s)
Animales , Ratas , Cicatrización de Heridas/efectos de los fármacos , Cloruros/farmacología , Cloruros/química , Compuestos de Zinc/farmacología , Compuestos de Zinc/química , Preparaciones de Plantas/farmacología , Alginatos/farmacología , Aloe , Ratas WistarRESUMEN
Trypanosoma cruzi is an obligate intracellular parasite transmitted to vertebrate hosts by blood-sucking insects. Molecules present in parasites and mammalian cells allow the recognition and parasite internalization. Metallic ions play an essential role in the establishment and maintenance of host-parasite interaction. However, little is known about how parasites handle with essential and nonessential metal quotas. This study aimed to investigate the influence of metal ions on the biological processes of T. cruzi infected cells. Infected cells were incubated with ZnCl2, CdCl2, and HgCl2 for 12 h and labeled with different specific dyes to investigate the cellular events related to intracellular parasite death and elimination. Infected host cells and parasite's mitochondria underwent functional and structural disorders, in addition to parasite's DNA condensation and pH decrease on host cells, which led to parasite death. Further investigations suggested that lysosomes were involved in pH decrease and the double membrane of the endoplasmic reticulum formed vacuoles surrounding damaged parasites, which indicate the occurrence of autophagy for parasite elimination. In conclusion, low concentrations of nonessential and essential metals cause a series of damage to Trypanosoma cruzi organelles, leading to its loss of viability, death, and elimination, with no removal of the host cells.
Asunto(s)
Autofagia/efectos de los fármacos , Cloruro de Cadmio/farmacología , Cloruros/farmacología , Compuestos de Mercurio/farmacología , Trypanosoma cruzi/efectos de los fármacos , Compuestos de Zinc/farmacología , Lisosomas/efectos de los fármacos , Lisosomas/metabolismo , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Trypanosoma cruzi/citología , Trypanosoma cruzi/metabolismo , Vacuolas/efectos de los fármacos , Vacuolas/metabolismoRESUMEN
Finding new potential antagonists of potassium channels is a continuing task. TASK potassium channels operate over a large physiological range of membrane voltages, why they are thought to contribute to the excitability and resting potential of mammalian membrane potentials. Additionally, they are regulated by extracellular stimuli like changes in pH and K+ concentrations. TASK malfunctions are associated with diseases, which makes them popular targets for the search of new antagonists. Identification of channel inhibitors can be a time-consuming and expensive project. Here, we present an easy-to-use and inexpensive yeast system for the expression of the two-pore domain K+ channel TASK-3, and for the characterization of TASK-3 antagonists. The Saccharomyces cerevisiae strain BYT45 was used to express guinea pig TASK-3. The system allowed the expression and characterization of TASK-3 at variable pH values and K+ concentrations. Three known TASK-3 antagonists have been tested in the BYT45 yeast system: PK-THPP, ZnCl2 and Bupivacaine. Their inhibitory effect on TASK-3 was tested in solid and liquid media assays, and half maximal inhibitory concentrations were estimated. Although the system is less sensitive than more refined systems, the antagonistic activity could be confirmed for all three inhibitors.
Asunto(s)
Bloqueadores de los Canales de Potasio/farmacología , Canales de Potasio de Dominio Poro en Tándem/antagonistas & inhibidores , Canales de Potasio/efectos de los fármacos , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , ATPasa Intercambiadora de Sodio-Potasio/genética , Animales , Bupivacaína/farmacología , Cationes , Cloruros/farmacología , Cobayas , Concentración de Iones de Hidrógeno , Concentración 50 Inhibidora , Canales de Potasio de Dominio Poro en Tándem/genética , Saccharomyces cerevisiae/crecimiento & desarrollo , Compuestos de Zinc/farmacologíaRESUMEN
Toxoplasma gondii, the etiological agent of toxoplasmosis, infects nucleated cells and then resides and multiplies within a parasitophorous vacuole. For this purpose, the parasite secretes many virulence factors for the purpose of invading and subverting the host microbicidal defenses in order to facilitate its survival in the intracellular milieu. Essential metals are structural components of proteins and enzymes or cofactors of enzymatic reactions responsible for these parasitic survival mechanisms. However, an excess of non-essential or essential metals can lead to parasite death. Thus, infected host cells were incubated with 20 µM ZnCl2 in conjunction with 3 µM CdCl2 or HgCl2 for 12 h in order to investigate cellular events and organelle damage related to intracellular parasite death and elimination. In the presence of these metals, the tachyzoites undergo lipid uptake and transport impairment, functional and structural mitochondrial disorders, DNA condensation, and acidification of the parasitophorous vacuole, thus leading to parasite death. Additional research has suggested that lysosome-vacuole fusion was involved in parasite elimination since acid phosphatases were found inside the parasitophorous vacuole, and vacuoles containing parasites were also positive for autophagy. In conclusion, low concentrations of CdCl2, HgCl2, and ZnCl2 can cause damage to Toxoplasma gondii organelles, leading to loss of viability, organelle death, and elimination without causing toxic effects to host cells.
Asunto(s)
Cloruro de Cadmio/farmacología , Cloruros/farmacología , Cloruro de Mercurio/farmacología , Toxoplasma/efectos de los fármacos , Vacuolas/efectos de los fármacos , Compuestos de Zinc/farmacología , Animales , Autofagia , Transporte Biológico/efectos de los fármacos , Línea Celular , Interacciones Huésped-Parásitos/efectos de los fármacos , Lisosomas , Macaca mulatta , Mitocondrias/patología , Desnaturalización de Ácido Nucleico/efectos de los fármacos , Toxoplasma/patogenicidad , Vacuolas/parasitología , Vacuolas/patología , Factores de VirulenciaRESUMEN
This study investigated the effects of BaCl2 adsorbed to polyethylene glycol (PEG) microspheres on human blood mononuclear cells (MN) co-cultured with breast cancer cell lines (MCF-7). The MCF-7 cells were obtained from the American Type Culture Collection and the blood mononuclear (MN) cells from volunteer donors. MN cells, MCF-7 cells and their co-culture (MN and MCF-7 cells) were pre-incubated for 24 h with or without 25 and 1000 pg L-1 BaCl2 (Ba25 and Ba1000), PEG microspheres or 25 and 1000 pg L-1 BaCl2 adsorbed to PEG microspheres (PEG-Ba25 and PEG-Ba1000). Rheological parameters and apoptosis were determined. Fluorescence microscopy and flow cytometry analyses revealed that BaCl2 was able to adsorb the PEG microspheres. The blood flow and viscosity curves were similar among the treatments. In general, apoptosis rates increased in co-cultured cells, co-cultured cells incubated with Ba25 and with PEG-Ba25, but the highest rates were observed in co-cultured cells incubated with PEG-Ba1000. In conclusion, BaCl2 adsorbed to PEG microspheres exhibited dose-dependent antitumor effects against human MCF-7 breast cancer cells co-cultured with MN cells, thereby offering a possible therapeutic alternative for treating this disease provided they are administered at very low concentrations.
Asunto(s)
Antineoplásicos , Compuestos de Bario , Neoplasias de la Mama , Cloruros , Leucocitos Mononucleares/metabolismo , Microesferas , Polietilenglicoles , Antineoplásicos/química , Antineoplásicos/farmacología , Compuestos de Bario/química , Compuestos de Bario/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Cloruros/química , Cloruros/farmacología , Femenino , Humanos , Leucocitos Mononucleares/patología , Células MCF-7 , Masculino , Polietilenglicoles/química , Polietilenglicoles/farmacologíaRESUMEN
ATP and adenosine, the main signaling molecules of purinergic system, are involved in toxicological effects induced by metals. The manganese (Mn) exposure induces several cellular changes, which could interfere with signaling pathways, such as the purinergic system. In this study, we evaluated the effects of exposure to manganese(II) chloride (MnCl2) during 96 h on nucleoside triphosphate diphosphohydrolase (NTPDase), ecto-5'-nucleotidase, and adenosine deaminase (ADA) activities, followed by analyzing the gene expression patterns of NTPDases (entpd1, entpd2a.1, entpd2a.2, entpd2-like, entpd3) and ADA (ADA 1 , ADA 2.1 , ADA 2.2 , ADAasi, ADAL) families in zebrafish brain. In addition, the brain metabolism of nucleotides and nucleosides was evaluated after MnCl2 exposure. The results showed that MnCl2 exposure during 96 h inhibited the NTPDase (1.0 and 1.5 mM) and ecto-ADA (0.5, 1.0, and 1.5 mM) activities, further decreasing ADA2.1 expression at all MnCl2 concentrations analyzed. Purine metabolism was also altered by the action of MnCl2. An increased amount of ADP appeared at all MnCl2 concentrations analyzed; however, AMP and adenosine levels are decreased at the concentrations of 1.0 and 1.5 mM MnCl2, whereas decreased inosine (INO) levels were observed at all concentrations tested. The findings of this study demonstrated that MnCl2 may inhibit NTPDase and ecto-ADA activities, consequently modulating nucleotide and nucleoside levels, which may contribute for the toxicological effects induced by this metal.
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Envejecimiento/metabolismo , Encéfalo/metabolismo , Cloruros/farmacología , Compuestos de Manganeso/farmacología , Nucleósidos/metabolismo , Nucleótidos/metabolismo , Pez Cebra/metabolismo , Adenosina Desaminasa/metabolismo , Animales , Antígenos CD , Apirasa , Femenino , MasculinoRESUMEN
Trypanosoma cruzi has many molecules that need metallic elements to work, allowing cell invasion and the establishment of infection, causing Chagas disease. Nonetheless, knowledge regarding how the parasites address metals and maintain homeostasis is lacking. To study this relationship, zinc, cadmium and mercury were chosen. Epimastigote, trypomastigote and intracellular forms of T. cruzi were incubated with these metals for different times and at different concentrations. In general, epimastigotes were the most sensitive and trypomastigotes the most resistant to metals. ZnCl2 induced low toxic effects to all parasite forms. Although the parasites were very sensitive to the toxic effects of CdCl2 and HgCl2, pretreatment with ZnCl2 decreased the death rate. The trypomastigotes pretreated with CdCl2 were unable to infect the host cells, and the treated intracellular forms were damaged after 2 h of incubation, when the toxic effects were poorly reverted. New insights on metal toxicity mechanisms are provided, helping to understand how metallic ions influence the parasite's biochemical and physiological processes.
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Metales/farmacología , Trypanosoma cruzi/efectos de los fármacos , Trypanosoma cruzi/fisiología , Animales , Cloruro de Cadmio/farmacología , Cloruros/farmacología , Chlorocebus aethiops , Mercurio/farmacología , Trypanosoma cruzi/patogenicidad , Trypanosoma cruzi/ultraestructura , Células Vero , Compuestos de Zinc/farmacologíaRESUMEN
Organoselenium compounds, such as diphenyl diselenide (PhSe)2 and phenylselenium zinc chloride (PhSeZnCl), show protective activities related to their thiol peroxidase activity. However, depending on experimental conditions, organoselenium compounds can cause toxicity by oxidising thiol groups of proteins and induce the production of reactive oxygen species (ROS). Here, we analysed the toxicity of (PhSe)2 and PhSeZnCl in yeast Saccharomyces cerevisiae. Cell growth of S. cerevisiae after 1, 2, 3, 4, 6, and 16 h of treatment with 2, 4, 6, and 10 µM of (PhSe)2 was evaluated. For comparative purpose, PhSeZnCl was analysed only at 16 h of incubation at equivalent concentrations of selenium (i.e. 4, 8, 12, and 20 µM). ROS production (DCFH-DA), size, granularity, and cell membrane permeability (propidium iodide) were determined by flow cytometry. (PhSe)2 inhibited cell growth at 2 h (10 µM) of incubation, followed by increase in cell size. The increase of cell membrane permeability and granularity (10 µM) was observed after 3 h of incubation, however, ROS production occurs only at 16 h of incubation (10 µM) with (PhSe)2, indicating that ROS overproduction is a more likely consequence of (PhSe)2 toxicity and not its determinant. All tested parameters showed that only concentration of 20 µM induced toxicity in samples incubated with PhSeZnCl. In summary, the results suggest that (PhSe)2 toxicity in S. cerevisiae is time and concentration dependent, presenting more toxicity when compared with PhSeZnCl.
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Derivados del Benceno/farmacología , Permeabilidad de la Membrana Celular/efectos de los fármacos , Compuestos de Organoselenio/farmacología , Especies Reactivas de Oxígeno/metabolismo , Saccharomyces cerevisiae/efectos de los fármacos , Cloruros/farmacología , Oxidación-Reducción , Compuestos de Selenio/farmacología , Compuestos de Sulfhidrilo/farmacología , Compuestos de Zinc/farmacologíaRESUMEN
BACKGROUND: Considering that heme oxygenase-1 (HO-1) and the brain-derived neurotrophic factor (BDNF)-mediated pathway are involved in the pathophysiology of depression and that zinc has been shown to exert beneficial effects in the management of depression, this study investigated the influence of these targets on the antidepressant-like effect of zinc. METHODS: Mice were treated with sub-effective or effective doses of zinc chloride (ZnCl2, 10mg/kg, po), and 45min later, they received intracerebroventricular (icv) injections of sub-effective doses of either zinc protoporphyrin IX (ZnPP, 10µg/mouse, HO-1 inhibitor), cobalt protoporphyrin IX (CoPP, 0.01µg/mouse, HO-1 inducer) or K-252a (1µg/mouse, TrkB receptor antagonist). Immobility time and locomotor activity were evaluated through the tail suspension test (TST) and open-field test (OFT), respectively. HO-1 immunocontents were evaluated in the prefrontal cortex and hippocampus 60min after ZnCl2 (10mg/kg, po) treatment. RESULTS: The antidepressant-like effect of ZnCl2 was prevented by the treatment with ZnPP and K-252a. Furthermore, sub-effective doses of CoPP and ZnCl2 produced a synergistic antidepressant-like effect in the TST. None of the treatments altered locomotor activity. ZnCl2 administration increased HO-1 immunocontents only in the prefrontal cortex. CONCLUSIONS: The results indicate that the antidepressant-like effect of ZnCl2 in the TST may depend on the induction of HO-1, and activation of TrkB receptor.
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Antidepresivos/farmacología , Cloruros/farmacología , Depresión/tratamiento farmacológico , Hemo-Oxigenasa 1/metabolismo , Compuestos de Zinc/farmacología , Animales , Antidepresivos/administración & dosificación , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Carbazoles/farmacología , Cloruros/administración & dosificación , Depresión/fisiopatología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Femenino , Suspensión Trasera , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Alcaloides Indólicos/farmacología , Inyecciones Intraventriculares , Ratones , Actividad Motora/efectos de los fármacos , Corteza Prefrontal/efectos de los fármacos , Corteza Prefrontal/metabolismo , Protoporfirinas/farmacología , Compuestos de Zinc/administración & dosificaciónRESUMEN
Cystic fibrosis (CF) is caused by mutations in the CFTR gene, which encodes a cAMP-regulated chloride channel. Several cellular functions are altered in CF cells. However, it is not clear how the CFTR failure induces those alterations. We have found previously several genes differentially expressed in CF cells, including c-Src, MUC1, MTND4, and CISD1 (CFTR-dependent genes). Recently, we also reported the existence of several chloride-dependent genes, among them GLRX5 and RPS27. Here, varying the intracellular chloride concentration [Cl- ]i of IB3-1 CF bronchial epithelial cells, we show that IL-1ß mRNA expression and secretion are also under Cl- modulation. The response to Cl- is biphasic, with maximal effects at 75 mM Cl- . The regulation of the IL-1ß mRNA expression involves an IL-1ß autocrine effect, since in the presence of the IL-1ß receptor antagonist IL1RN or anti-IL-1ß blocking antibody, the mRNA response to Cl- disappeared. Similar effects were obtained with the JNK inhibitor SP600125, the c-Src inhibitor PP2 and the IKK inhibitor III (BMS-345541). On the other hand, the IL-1ß secretion is still modulated by Cl- in the presence of IL-1RN, IL-1ß blocking antibody, or cycloheximide, suggesting that Cl- is affecting the IL-1ß maturation/secretion, which in turn starts an autocrine positive feedback loop. In conclusion, the Cl- anion acts as a second messenger for CFTR, modulating the IL-1ß maturation/secretion. The results also imply that, depending on its intracellular concentration, Cl- could be a pro-inflammatory mediator. J. Cell. Biochem. 118: 2131-2140, 2017. © 2016 Wiley Periodicals, Inc.