RESUMO
A series of new coordination compounds of cobalt(II), copper(II) and zinc(II) with heterocyclic ester derivatives (ethyl 4-methyl-5-imidazole-carboxylate (emizco), 1-(2-(phenylsulphonyl)ethyl)-4-imidazole carboxylate (semizco)) and methyl 5-(propylthio)-2-benzimidazolecarbamate (albendazole, abz) were synthesized. They were fully characterized by different techniques such as IR, UV-Vis-NIR, elemental analysis, molar conductivity and magnetic susceptibility. Additionally, X-ray crystal structures of semizco and its [Co(semizco)2Cl2]·2CH3CN 10, [Co(smmizco)2Br2]·2CH3CN 11 and [Cu(semizco)2Br2] 15 coordination compounds are analyzed. These compounds present lone pair SOâ¯π interactions between the sulfone and the imidazolic ring. These ligands showed three coordination modes: monodentate, through an imidazolic nitrogen atom, or a bidentate chelating mode by a nitrogen and an oxygen atom from the ester group. The different coordination modes and the number of coordinated ligands gave rise to tetrahedral and octahedral compounds, or for [Cu(semizco)(µ-Br)Br]n·0.5H2O 7 a square base pyramidal geometry. A cytotoxic study was carried out with the free ligands and their copper(II) and zinc(II) halide coordination compounds on HeLa (cervix-uterine), MCF-7 (breast), HCT-15 (colon), PC3 (prostate) human carcinoma cell lines and L929 mouse fibroblast (healthy cells). A TUNEL assay (terminal deoxynucleotidyl transferase dUTP nick end labeling) was performed with the most active copper(II) compounds to determine if cell death was by apoptosis.
Assuntos
Proliferação de Células/efeitos dos fármacos , Cobalto/química , Complexos de Coordenação/farmacologia , Cobre/química , Ésteres/química , Zinco/química , Animais , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Carbamatos/química , Linhagem Celular Tumoral , Quelantes/farmacologia , Complexos de Coordenação/química , Cristalografia por Raios X/métodos , Humanos , Imidazóis/química , Marcação In Situ das Extremidades Cortadas/métodos , Ligantes , Camundongos , Estrutura MolecularRESUMO
PURPOSE: To investigate whether the ability of human spermatozoa to decondense in vitro in the presence of heparin (Hep) and glutathione (GSH) is related to assisted reproduction (ART) success. METHODS: Cross-sectional pilot study involving male partners of 129 infertile couples undergoing ICSI with (45) or without (84) donor oocytes at two infertility clinics in CABA, Argentina, between October 2012 and December 2013. In vitro decondensation kinetics with Hep and GSH and DNA fragmentation (TUNEL) were determined on the same sample used for ICSI. The possible relationship of decondensation parameters (maximum decondensation and decondensation velocity) and TUNEL values with ART success was evaluated. RESULTS: Embryo quality correlated positively with decondensation velocity (D60/D30) (Spearman's correlation, p < 0.05). According to D60/D30 values, patients were classified as slow decondensers (SlowD) (n = 68) or fast decondensers (FastD) (n = 61). Embryo quality was better in FastD (unpaired t test, p < 0.05). FastD and SlowD were subdivided according to use of donor oocytes. Among SlowD, biochemical and clinical pregnancy rates per transfer were significantly higher in donor (n = 19) vs. in non-donor (n = 31) cycles (Fisher's exact test, p < 0.05). TUNEL values were not related to embryo quality, but no clinical pregnancies or live births were achieved in TUNEL+ SlowD (n = 7). CONCLUSION: Decondensation kinetics of human spermatozoa in vitro with Hep and GSH could be related to embryo quality and ART success.
Assuntos
Embrião de Mamíferos/fisiologia , Espermatozoides/fisiologia , Argentina , Estudos Transversais , Fragmentação do DNA , Feminino , Fertilização in vitro/métodos , Humanos , Marcação In Situ das Extremidades Cortadas/métodos , Infertilidade/terapia , Nascido Vivo , Masculino , Oócitos/fisiologia , Projetos Piloto , Gravidez , Taxa de Gravidez , Injeções de Esperma Intracitoplásmicas/métodosRESUMO
The selective serotonin reuptake inhibitor fluoxetine has been used for the treatment of depression. Although sexual disorders have been reported in male patients, few studies have demonstrated the fluoxetine effect on the reproductive histophysiology, and the target of this antidepressant in testes is unknown. We evaluated the impact of short-term treatment with fluoxetine on the adult rat testes, focusing on steroidogenesis by Leydig cells (LC) and androgen-dependent testicular parameters, including Sertoli cells (SC) and peritubular myoid cells (PMC). Since UCHL1 (ubiquitincarboxyl-terminal hydrolase L1) seems to control spermatogenesis, the immunoexpression of this hydrolase was also analyzed. Adult male rats received 20 mg/kg BW of fluoxetine (FG) or saline (CG) for eleven days. In historesin-embedded testis sections, the seminiferous tubule (ST) and epithelial (Ep) areas, and the LC nuclear diameter (LCnu) were measured. The number of abnormal ST, androgen-dependent ST, SC and PMC was quantified. Testicular ß-tubulin levels and peritubular actin immunofluorescence were evaluated. Serum testosterone levels (STL) and steroidogenesis by 17ß-HSD6 immunofluorescence were analyzed, and either UCHL1-immunolabeled or TUNEL-positive germ cells were quantified. In FG, abnormal ST frequency increased whereas ST and Ep areas, androgen-dependent ST number, LCnu, 17ß-HSD6 activity and STL reduced significantly. TUNEL-positive PMC and SC was related to decreased number of these cells and reduction in peritubular actin and ß-tubulin levels. In FG, uncommon UCHL1-immunoexpression was found in spermatocytes and spermatids, and the number of UCHL1-immunolabeled and TUNEL-positive germ cells increased in this group. These findings indicate that LC may be a fluoxetine target in testes, impairing PMC-SC integrity and disturbing spermatogenesis. The increase of UCHL1 in the damaged tubules associated with high incidence of cell death confirms that this hydrolase regulates germ cell death and may be controlled by androgens. The fertility in association with the androgenic status of patients treated with fluoxetine should be carefully evaluated.
Assuntos
Androgênios/metabolismo , Morte Celular/efeitos dos fármacos , Fluoxetina/farmacologia , Células Germinativas/efeitos dos fármacos , Túbulos Seminíferos/efeitos dos fármacos , Ubiquitina Tiolesterase/metabolismo , Animais , Células Germinativas/metabolismo , Hidrolases/efeitos dos fármacos , Hidrolases/metabolismo , Marcação In Situ das Extremidades Cortadas/métodos , Células Intersticiais do Testículo/efeitos dos fármacos , Células Intersticiais do Testículo/metabolismo , Masculino , Ratos , Túbulos Seminíferos/metabolismo , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/metabolismo , Espermatogênese/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testículo/metabolismo , Ubiquitinas/metabolismoRESUMO
Estrogen maintains osteocyte viability, whereas its deficiency induces osteocyte apoptosis. As autophagy is important for osteocyte viability, we hypothesized whether the anti-apoptotic effect of estrogen is related to autophagy in osteocytes. Thirty adult female rats were sham-operated (SHAM) or ovariectomized (OVX). After three weeks, twelve rats of SHAM and OVX groups were killed before treatment (basal period), whereas the remaining rats received estrogen (OVXE) or vehicle (OVX) for 45 days. Fragments of maxilla containing alveolar process of the first molars were embedded in paraffin or Araldite. Paraffin-sections were stained with hematoxylin/eosin for histomorphometry, or subjected to the silver impregnation method for morphological analysis of osteocyte cytoplasmic processes. Autophagy was analyzed by immunohistochemical detections of beclin-1, MAP-LC3α and p62, whereas apoptosis was evaluated by immunohistochemical detections of cleaved caspase-3 and BAX, TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) method and by ultrastructural analysis. Araldite-semithin sections were subjected to the Sudan-black method for detection of lipids. OVX-basal group showed high frequency of caspase-3-, TUNEL- and p62-positive osteocytes accompanied with low frequency of beclin-1- and MAP-LC3α-positive osteocytes. At 45 days, OVXE group exhibited higher number of osteocytes, higher frequency of beclin-1- and MAP-LC3α-positive osteocytes, and lower frequency of caspase-3, BAX-, TUNEL- and p62-positive osteocytes than OVX group. Significant reduction in bone area was observed in the OVX compared to OVXE and SHAM groups. The highest frequency of Sudan-Black-positive osteocytes and osteocytes with scarce cytoplasmic processes, or showing apoptotic features were mainly observed in OVX groups. Our results indicate that estrogen deficiency decreases autophagy and increases apoptosis, whereas estrogen replacement enhances osteocyte viability by inhibiting apoptosis and maintaining autophagy in alveolar process osteocytes. These results suggest that the anti-apoptotic effect of estrogen may be, at least in part, related to autophagy regulation in osteocytes.
Assuntos
Autofagia/fisiologia , Sobrevivência Celular/fisiologia , Estrogênios/metabolismo , Osteócitos/metabolismo , Processo Alveolar/metabolismo , Animais , Apoptose/fisiologia , Proteínas Reguladoras de Apoptose/metabolismo , Caspase 3/metabolismo , Feminino , Marcação In Situ das Extremidades Cortadas/métodos , Ovariectomia/métodos , Ratos , Ratos WistarRESUMO
En los últimos años se han producido grandes avances en el diagnóstico y tratamiento del Liquen Plano Oral (LPO). Sin embargo, sigue siendo una entidad con muchos interrogantes para la estomatología, sobre todo referidos a su proceso de aparición y a su tratamiento. El propósito de este trabajo es realizar una revisión bibliográfica actualizada del LPO y su relación con la apoptosis, tema de gran interés para la investigación científica. La apoptosis por su parte adquiere gran relevancia debido al rol que cumple este mecanismo: regulación en la morfogénesis, homeostasis de las poblaciones celulares y carcinogénesis en relación a la tendencia y potencial capacidad de transformación maligna de algunas variantes de LPO en sus formas atípicas.
In recent years, there have been major advances in the diagnosis and treatment of Oral Lichen Planus (OLP). However, it remains an entity with many questions for stomatology, especially referred to the process of occurrence and treatment. The purpose of this work is to conduct a literature review updated LPO and its relationship with apoptosis, topic of great interest for scientific research. The apoptosis meanwhile acquires great importance because of the role that this mechanism regulating morphogenesis, homeostasis of cell populations and carcinogenesis in relation to the trend and potential ability of malignant transformation of some variants of LPO in their atypical forms.
Assuntos
Humanos , Apoptose/fisiologia , Transformação Celular Neoplásica , Líquen Plano Bucal/etiologia , Líquen Plano Bucal/fisiopatologia , Marcação In Situ das Extremidades Cortadas/métodos , Imuno-Histoquímica/métodos , Lesões Pré-Cancerosas/classificação , Lesões Pré-Cancerosas/complicaçõesRESUMO
PURPOSE: To determine whether Toll-like receptor 7 (TLR7) is the potential targets of prevention or progression in the renal ischemia/reperfusion (I/R) injury of STZ-induced diabetic rats. METHODS: Thirty six Sprague-Dawley rats were randomly arranged to the nondiabetic (ND) or diabetic group (DM), with each group further divided into sham (no I/R injury), I/R (ischemia-reperfusion) and CD (given by Chloroquine) group. Preoperatively, Chloroquine (40 mg/kg, intraperitoneal injection.) was administrated 6 days for treatment group. I/R animals were subjected to 25 min of bilateral renal ischemia. Renal function, histology, apoptosis, cytokines, expression of TLR7, MyD88 and NF-κB were detected. RESULTS: The serum levels of blood urea nitrogen, creatinine, IL-6 and TNF-α, apoptotic tubular epithelial cells, expression of TLR7, MyD88 and NF-κB were significantly increased in DM+I/R group, compared with ND+I/R group (p<0.05). All these changes were further improved by TLR7 inhibition Chloroquine except Paller scores (p<0.05). CONCLUSION: Toll-like receptor 7 inhibition attenuates the acute renal ischemia/reperfusion injury of STZ-induced diabetic in SD rats.
Assuntos
Injúria Renal Aguda/metabolismo , Diabetes Mellitus Experimental/metabolismo , Rim/metabolismo , Traumatismo por Reperfusão/metabolismo , Receptor 7 Toll-Like/metabolismo , Injúria Renal Aguda/patologia , Animais , Apoptose , Diabetes Mellitus Experimental/complicações , Modelos Animais de Doenças , Marcação In Situ das Extremidades Cortadas/métodos , Rim/patologia , Masculino , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Traumatismo por Reperfusão/complicações , Receptor 7 Toll-Like/sangueRESUMO
OBJECTIVE: to evaluate the influence of Tumor Necrosis Factor alpha (TNF-α) and apoptosis in rats with DM treated with chamomile extract or triamcinolone. MATERIAL AND METHODS: Wistar male rats (210.0±4.2 g) were divided into five groups: negative control group (NCG) without diabetes; positive control group (PCG) with DM (alloxan, 45 mg/kg); and groups treated with chamomile extract (normoglycemic= NCG group and diabetic= DCG group) and with triamcinolone (TG). Traumatic ulcers were performed on all animals that received topical triamcinolone, chamomile extract or saline 12/12 hours for ten days. RESULTS: On days five and ten the animals were euthanized and the ulcers were analyzed by light microscopy, TUNEL assay, and immunohistochemically (TNF-α). The NCG (p=0.0062), PCG (p=0.0285), NCG (p=0.0041), and DCG (p<0.0001) groups were completely healed on the 10th day, however, there was no healing on the TG (p=0.5127) group. The TNF-α expression showed a significant reduction from the 5th to the 10th day in NCG (p=0.0266) and DCG (p=0.0062). In connective tissue, the TUNEL assay showed a significant reduction in the number of positive cells in NCG (p=0.0273) and CNG (p=0.0469) and in the epithelium only in CDG (p=0.0320). CONCLUSIONS: Chamomile extract can optimize the healing of traumatic oral ulcers in diabetic rats through the reduction of apoptosis in the epithelium and TNF-α expression.
Assuntos
Anti-Inflamatórios/farmacologia , Colágeno/análise , Diabetes Mellitus Experimental/fisiopatologia , Matricaria/química , Úlceras Orais/tratamento farmacológico , Triancinolona/farmacologia , Fator de Necrose Tumoral alfa/análise , Aloxano , Animais , Colágeno/efeitos dos fármacos , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas/métodos , Masculino , Úlceras Orais/patologia , Extratos Vegetais/farmacologia , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do Tratamento , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Cicatrização/efeitos dos fármacosRESUMO
BACKGROUND: Pemphigus is part of a group of blistering diseases that affect the skin and mucous membranes. Based on its autoimmune origin, autoantibodies develop in pemphigus that are directed toward cell surface components of keratinocytes. However, some data cannot be explained, such as the lack of a relationship between autoantibody levels and the severity of clinical manifestations, treatment resistance, the presence of inflammatory infiltrates and the potential occurrence of apoptosis as determinants of vesicle formation. OBJECTIVE: To examine the presence of apoptosis in pemphigus vulgaris by TUNEL technique. METHODS: In this cross-sectional study, we selected 15 paraffin-embedded tissues from subjects who were diagnosed with pemphigus vulgaris by hematoxylin and eosin staining. The samples were subjected to TUNEL assay and examined under an Olympus BX61 fluorescence microscope. Positivity was categorized dichotomously, and the statistical analysis was performed using the X2 test. RESULTS: Positivity was observed in basal layer cells in 14 (93.3%) cases. In 13 (86.7%) of the positive cases, we noted espinosum and granular layers that formed the blister roof, and in 12 cases (80%), positive acantholytic cells were observed. CONCLUSIONS: TUNEL positivity was observed in pemphigus vulgaris, implicating apoptosis in the pathophysiology of this condition, which can help guide the development of apoptotic blockers as therapeutics.
Assuntos
Apoptose/fisiologia , Marcação In Situ das Extremidades Cortadas/métodos , Pênfigo/fisiopatologia , Acantólise/fisiopatologia , Adulto , Vesícula/fisiopatologia , Estudos Transversais , Humanos , Pênfigo/patologia , Pele/fisiopatologiaRESUMO
ABSTRACT PURPOSE: To determine whether Toll-like receptor 7 (TLR7) is the potential targets of prevention or progression in the renal ischemia/reperfusion (I/R) injury of STZ-induced diabetic rats. METHODS: Thirty six Sprague-Dawley rats were randomly arranged to the nondiabetic (ND) or diabetic group (DM), with each group further divided into sham (no I/R injury), I/R (ischemia-reperfusion) and CD (given by Chloroquine) group. Preoperatively, Chloroquine (40 mg/kg, intraperitoneal injection.) was administrated 6 days for treatment group. I/R animals were subjected to 25 min of bilateral renal ischemia. Renal function, histology, apoptosis, cytokines, expression of TLR7, MyD88 and NF-κB were detected. RESULTS: The serum levels of blood urea nitrogen, creatinine, IL-6 and TNF-α, apoptotic tubular epithelial cells, expression of TLR7, MyD88 and NF-κB were significantly increased in DM+I/R group, compared with ND+I/R group (p<0.05). All these changes were further improved by TLR7 inhibition Chloroquine except Paller scores (p<0.05). CONCLUSION: Toll-like receptor 7 inhibition attenuates the acute renal ischemia/reperfusion injury of STZ-induced diabetic in SD rats.
Assuntos
Animais , Masculino , Traumatismo por Reperfusão/metabolismo , Diabetes Mellitus Experimental/metabolismo , Receptor 7 Toll-Like/metabolismo , Injúria Renal Aguda/metabolismo , Rim/metabolismo , Traumatismo por Reperfusão/complicações , Distribuição Aleatória , NF-kappa B/metabolismo , Ratos Sprague-Dawley , Apoptose , Marcação In Situ das Extremidades Cortadas/métodos , Diabetes Mellitus Experimental/complicações , Modelos Animais de Doenças , Receptor 7 Toll-Like/sangue , Fator 88 de Diferenciação Mieloide/metabolismo , Injúria Renal Aguda/patologia , Rim/patologiaRESUMO
Abstract: Background: Pemphigus is part of a group of blistering diseases that affect the skin and mucous membranes. Based on its autoimmune origin, autoantibodies develop in pemphigus that are directed toward cell surface components of keratinocytes. However, some data cannot be explained, such as the lack of a relationship between autoantibody levels and the severity of clinical manifestations, treatment resistance, the presence of inflammatory infiltrates and the potential occurrence of apoptosis as determinants of vesicle formation. Objective: To examine the presence of apoptosis in pemphigus vulgaris by TUNEL technique. Methods: In this cross-sectional study, we selected 15 paraffin-embedded tissues from subjects who were diagnosed with pemphigus vulgaris by hematoxylin and eosin staining. The samples were subjected to TUNEL assay and examined under an Olympus BX61 fluorescence microscope. Positivity was categorized dichotomously, and the statistical analysis was performed using the X2 test. Results: Positivity was observed in basal layer cells in 14 (93.3%) cases. In 13 (86.7%) of the positive cases, we noted espinosum and granular layers that formed the blister roof, and in 12 cases (80%), positive acantholytic cells were observed. Conclusions: TUNEL positivity was observed in pemphigus vulgaris, implicating apoptosis in the pathophysiology of this condition, which can help guide the development of apoptotic blockers as therapeutics.
Assuntos
Humanos , Adulto , Pênfigo/fisiopatologia , Apoptose/fisiologia , Marcação In Situ das Extremidades Cortadas/métodos , Pele/fisiopatologia , Estudos Transversais , Acantólise/fisiopatologia , Vesícula/fisiopatologia , Pênfigo/patologiaRESUMO
ABSTRACT Diabetes mellitus (DM) is a disease associated with delayed wound healing of oral ulcers by increased expression of proinflammatory cytokines and cellular apoptosis. Objective to evaluate the influence of Tumor Necrosis Factor alpha (TNF-α) and apoptosis in rats with DM treated with chamomile extract or triamcinolone. Material and Methods Wistar male rats (210.0±4.2 g) were divided into five groups: negative control group (NCG) without diabetes; positive control group (PCG) with DM (alloxan, 45 mg/kg); and groups treated with chamomile extract (normoglycemic= NCG group and diabetic= DCG group) and with triamcinolone (TG). Traumatic ulcers were performed on all animals that received topical triamcinolone, chamomile extract or saline 12/12 hours for ten days. Results On days five and ten the animals were euthanized and the ulcers were analyzed by light microscopy, TUNEL assay, and immunohistochemically (TNF-α). The NCG (p=0.0062), PCG (p=0.0285), NCG (p=0.0041), and DCG (p<0.0001) groups were completely healed on the 10th day, however, there was no healing on the TG (p=0.5127) group. The TNF-α expression showed a significant reduction from the 5th to the 10th day in NCG (p=0.0266) and DCG (p=0.0062). In connective tissue, the TUNEL assay showed a significant reduction in the number of positive cells in NCG (p=0.0273) and CNG (p=0.0469) and in the epithelium only in CDG (p=0.0320). Conclusions Chamomile extract can optimize the healing of traumatic oral ulcers in diabetic rats through the reduction of apoptosis in the epithelium and TNF-α expression.
Assuntos
Animais , Masculino , Triancinolona/farmacologia , Colágeno/análise , Fator de Necrose Tumoral alfa/análise , Úlceras Orais/tratamento farmacológico , Matricaria/química , Diabetes Mellitus Experimental/fisiopatologia , Anti-Inflamatórios/farmacologia , Fatores de Tempo , Cicatrização/efeitos dos fármacos , Imuno-Histoquímica , Extratos Vegetais/farmacologia , Distribuição Aleatória , Reprodutibilidade dos Testes , Colágeno/efeitos dos fármacos , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Resultado do Tratamento , Ratos Wistar , Úlceras Orais/patologia , Marcação In Situ das Extremidades Cortadas/métodos , AloxanoRESUMO
MTCH2 has been described in liver as a protein involved in the intrinsic apoptotic pathway, although new evidence also associates this protein with cellular metabolism. In this work, the expression of MTCH2 in testis (an organ in which high levels of apoptosis normally take place as part of the spermatogenic process) is analyzed in rat, both at the mRNA and at the protein levels. Our results showed that MTCH2 was highly expressed in testis compared with other tissues and was differentially expressed according to developmental stage and testicular cell type. Protein expression was initially detected during the first spermatogenic wave at the time of meiosis onset and its levels increased in adulthood, with the highest expression levels being detected in meiotic prophase I. Specific differences in MTCH2 expression levels at the various stages of the adult seminiferous epithelium were also observed. Co-staining with TUNEL revealed a differential MTCH2 staining pattern in TUNEL-positive cells, mainly in dying primary spermatocytes, i.e., meiotic prophase I cells. Furthermore, upon mild hyperthermia (treatment shown to increase apoptosis in testis), MTCH2 levels rose concomitantly with a massive appearance of TUNEL-positive cells within the seminiferous tubules; these cells exhibited a differential MTCH2 distribution. Thus, MTCH2 is related to testicular apoptosis, especially during meiotic prophase.
Assuntos
Apoptose/fisiologia , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Túbulos Seminíferos/metabolismo , Espermatócitos/metabolismo , Testículo/metabolismo , Animais , Marcação In Situ das Extremidades Cortadas/métodos , Masculino , Meiose/fisiologia , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Espermatogênese/fisiologiaRESUMO
Environmentally relevant doses of inhaled diesel particles elicit pulmonary inflammation and impair lung mechanics. Eugenol, a methoxyphenol component of clove oil, presents in vitro and in vivo anti-inflammatory and antioxidant properties. Our aim was to examine a possible protective role of eugenol against lung injuries induced by diesel particles. Male BALB/c mice were divided into four groups. Mice received saline (10 µl in; CTRL group) or 15 µg of diesel particles DEP (15 µg in; DIE and DEUG groups). After 1 h, mice received saline (10 µl; CTRL and DIE groups) or eugenol (164 mg/kg; EUG and DEUG group) by gavage. Twenty-four hours after gavage, pulmonary resistive (ΔP1), viscoelastic (ΔP2) and total (ΔPtot) pressures, static elastance (Est), and viscoelastic component of elastance (ΔE) were measured. We also determined the fraction areas of normal and collapsed alveoli, amounts of polymorpho- (PMN) and mononuclear cells in lung parenchyma, apoptosis, and oxidative stress. Est, ΔP2, ΔPtot, and ΔE were significantly higher in the DIE than in the other groups. DIE also showed significantly more PMN, airspace collapse, and apoptosis than the other groups. However, no beneficial effect on lipid peroxidation was observed in DEUG group. In conclusion, eugenol avoided changes in lung mechanics, pulmonary inflammation, and alveolar collapse elicited by diesel particles. It attenuated the activation signal of caspase-3 by DEP, but apoptosis evaluated by TUNEL was avoided. Finally, it could not avoid oxidative stress as indicated by malondialdehyde.
Assuntos
Eugenol/farmacologia , Pulmão/efeitos dos fármacos , Pneumonia/induzido quimicamente , Pneumonia/tratamento farmacológico , Alvéolos Pulmonares/efeitos dos fármacos , Emissões de Veículos/toxicidade , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Marcação In Situ das Extremidades Cortadas/métodos , Peroxidação de Lipídeos/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Tamanho da Partícula , Pneumonia/metabolismo , Pneumonia/patologia , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/patologia , Mecânica Respiratória/efeitos dos fármacosRESUMO
Los esfingolípidos, como la ceramida (Cer), la ceramida-1-fosfato (C-1-P), la esfingosina (Sph) y la esfingosina-1-fosfato (S-1P) estan relacionados con la señalización intracelular en procesos como crecimiento celular, movilización intracelular de Ca+2 y apoptósis. En este trabajo se evaluó el efecto de estos esfingolípidos en la homeostasis de Ca+2 intracelular y en la apoptósis en células de cáncer de mama MCF-7. Se utilizaron fluoróforos específicos para el Ca+2 y microscopía confocal. Se demostró que en estas células, la Sph (20 uM), la Cer (10uM), la S-P (2uM) y la C--P (uM) aumentaron la concentración intracelular ce Ca+2, induciendo su liberación desde el retículo endoplasmático (RE). Además, se observo que la esfingosina abrioun canal de Ca2+ en la membrana plasmática. También se demostró que la Cer inhibe parcialmente la actividad de la Ca2+-ATPasa del RE (SERCA), de forma dosis dependiente, mientras que la ceramina, su análogo no hidrolisable la inhibe totalmente. La Sph también inhibe completamente la actividad de la SERCA, a la misma concentración que induce la liberación del Ca+2 del RE. Asimismo, se evaluó el efecto de estos esfingolípidos sobre la inducción de la apotósis en células MCF-7 evidenciando que el tratamiento con la Cer, la ceramida, la Sph inducen toxicidad. También se observo que mientras la ceramida activo la caspasa 7 y la caspasa 8, el esfingolipido natural, la Cer no tuvo ningún efecto. Por su parte, la Sph activa la caspasa 8 sin modificar la activdad de la caspasa 7. Tanto la Cer, como la ceramida y la Sph, disminuyeron la expresión de la proteína Bcl-2 amti-apoptótica, y también indujeron la fragmentación de ADN, visualizada mediante la técnica de TUNEL, demostrando que estos esfingolípidos inducen apoptósis en MCF-7. La agelasina B, toxina purificada a partir de la esponja marina Agelas clathrodes tiene un efecto citotóxico un orden de magnitud mayor en MCF-7, en comparación con fibroplastos humanos. La agelasina B induce la liberación del Ca+2 almacenado en el RE en celulas MCF-7, ademas de inhibir la actividad de la SERCA en un 100%. También se demostró que esta toxina induce apoptosis, ya que disminuye el potencial de membrana mitocondrial, activa la caspasa 8, disminuye la expresion de la proteina Bcl-2 e induce fragmentación del ADN de las células MCF-7. Este mecanismo es similar al efecto de la tapsigargina.
Assuntos
Humanos , Animais , Esfingolipídeos/farmacologia , Neoplasias da Mama/metabolismo , Transdução de Sinais/efeitos dos fármacos , Cálcio/metabolismo , Apoptose/efeitos dos fármacos , Agelas/química , Purinas/uso terapêutico , Purinas/farmacologia , Esfingolipídeos/toxicidade , Esfingolipídeos/uso terapêutico , Neoplasias da Mama/patologia , Neoplasias da Mama/tratamento farmacológico , Ceramidas/toxicidade , ATPases Transportadoras de Cálcio/efeitos adversos , Marcação In Situ das Extremidades Cortadas/métodos , Receptores de Detecção de Cálcio/uso terapêutico , Células MCF-7 , Naftalenos/uso terapêutico , Naftalenos/farmacologia , Antineoplásicos/uso terapêutico , Antineoplásicos/farmacologiaRESUMO
BACKGROUND: There is an association between aging ana an increased number of sperms with alterations in nuclear DNA. AIM: To study the association between age and fragmentation of sperm DNA. MATERIAL AND METHODS: Sixty two volunteers provided semen for analysis. These were separated in a group aged less than forty years and a second group aged more than forty years. Sperm DNA fragmentation was studied by TUNEL (terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end-labeling) and SCD (sperm chromatin dispersion test) assays. RESULTS: Compared with their younger counterparts, patients aged more than 40 years had a higher proportion of sperms with DNA fragmentation by TUNEL (20 ± 1.3 and 24 ± 1.9% respectively, p < 0.05) and SCD (22 ± 1.4 and 26 ± 1.6% respectively, p < 0.05). The results of both assays had a correlation coefficient of O.8. No differences between groups were observed for other seminal parameters. CONCLUSIONS: Sperm DNA fragmentation increases with age in males.
Assuntos
Cromatina/química , Fragmentação do DNA , Marcação In Situ das Extremidades Cortadas/métodos , Análise do Sêmen/métodos , Espermatozoides/ultraestrutura , Adolescente , Adulto , Fatores Etários , Humanos , Masculino , Pessoa de Meia-Idade , Contagem de Espermatozoides , Motilidade dos Espermatozoides , Espermatozoides/fisiologia , Adulto JovemRESUMO
Background: There is an association between aging ana an increased number ofsperms with alterations in nuclear DNA. Aim: To study the association between age and fragmentation of sperm DNA. Material andMethods: Sixty two volunteers provided semen for analysis. These were separated in a group aged less than forty years and a second group aged more than forty years. Sperm DNA fragmentation was studied by TÚNEL (terminal deoxynucleotidyl transferase-mediated2'-deoxyuridine 5'-triphosphate nick end-labeling) and SCD (sperm chromatin dispersión test) assays. Results: Compared with theiryounger counterparts, patients aged more than 40years had a higher proportion ofsperms with DNA fragmentation by TÚNEL (20 ±1.3 and24 ± 1.9 percent respectively, p < 0.05) and SCD (22 ± 1.4 and26 ± 1.6 percent respectively, p < 0.05). The results ofboth assays had a correlation coefficientofO.8. No differences between groups were observed for other seminal parameters. Conclusions: Sperm DNA fragmentation increases with age in males.
Assuntos
Adolescente , Adulto , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Cromatina/química , Fragmentação do DNA , Marcação In Situ das Extremidades Cortadas/métodos , Análise do Sêmen/métodos , Espermatozoides/ultraestrutura , Fatores Etários , Contagem de Espermatozoides , Motilidade dos Espermatozoides , Espermatozoides/fisiologiaRESUMO
The nervous system of echinoderms has long been considered too unique to be directly comparable to the nervous system of other Deuterostomia. Using two novel monoclonal antibodies in combination with epifluorescence, confocal, and electron microscopy, we demonstrate here that the central nervous system of the sea cucumber Holothuria glaberrima possesses a major non-neuronal cell type, which shares striking similarities with the radial glia of chordates. The basic features in common include (a) an elongated shape, (b) long radial processes, (c) short lateral protrusions branching off the main processes and penetrating into the surrounding neuropile, (d) prominent orderly oriented bundles of intermediate filaments, and (e) ability to produce Reissner's substance. Radial glia account for the majority of glia cells in echinoderms and constitutes more than half of the total cell population in the radial nerve cord and about 45% in the circumoral nerve ring. The difference in glia cell number between those regions is significant, suggesting structural specialization within the seemingly simple echinoderm nervous system. Both cell death and proliferation are seen under normal physiological conditions. Although both glia and neurons undergo apoptosis, most of the mitotic cells are identified as radial glia, indicating a key role of this cell type in cell turnover in the nervous system. A hypothesis is proposed that the radial glia could be an ancestral feature of the deuterostome nervous system, and the origin of this cell type might have predated the diversification of the Chordata and Ambulacraria lineages.
Assuntos
Sistema Nervoso Central/citologia , Neuroglia/fisiologia , Pepinos-do-Mar/anatomia & histologia , Animais , Bromodesoxiuridina/metabolismo , Contagem de Células/métodos , Morte Celular , Proliferação de Células , Marcação In Situ das Extremidades Cortadas/métodos , Microscopia Imunoeletrônica/métodos , Proteínas do Tecido Nervoso/metabolismo , Neuroglia/metabolismo , Neuroglia/ultraestruturaRESUMO
Apoptose e seus mecanismos reguladores são eventos fisiológicos cruciais para a manutenção da homeostase placentária, e o desequilíbrio desses processos pode comprometer a função placentária e, consequentemente, o sucesso da gravidez. Neste estudo, investigou-se a apoptose utilizando-se histomorfometria em lâminas coradas em HE e submetidas à reação de TUNEL. Além disso, avaliou-se a expressão de Bcl-2 e das caspases 8 e 3, pela reação de polimerase em cadeia em tempo real, em placentas saudáveis em diferentes estádios de gestação. Amostras de placentônios de vacas com quatro, seis e nove meses de gestação foram colhidas e processadas. O índice apoptótico aumentou progressivamente com o avanço da gestação. Tanto o Bcl-2 quanto as caspases 3 e 8 foram expressas nos três períodos estudados, sendo a expressão de Bcl-2 menor que a de caspase 8, que é menor que a de caspase 3. Estes resultados indicam que essas moléculas estão envolvidas na via apoptótica ativada na maturação placentária, exibindo um padrão de expressão ao longo da gestação e contribuindo para o equilíbrio fisiológico da celularidade e renovação celular na placenta bovina.
Apoptosis and its regulating mechanisms are crucial physiological events for the maintenance of the placental homostasis; and disequilibrium of these processes may compromise placental function and the success of the pregnancy. In this study, apoptosis was investigated by histomorphometry using slides stained with HE and TUNEL reaction. Besides that, Bcl-2 and caspases 8 and 3 expression were evaluated by real time polymerase chain reaction in healthy placentas under different gestacional ages. Samples of placentones of cows at 4th, 6th, and 9th months of gestation were harvested and processed. The apoptotic index gradually increased with the advance of the gestation. Bcl-2 and caspases 3 and 8 were expressed in all the studied periods, being the expression of Bcl-2 lower than that of caspase 8, which was lower than caspase 3. These results indicate that these molecules are involved in the activated apoptotic way in the placental maturation, showing a standard expression throughout the gestation and contributing for the physiological balance of the cellularity and cellular turn over in bovine placenta.
Assuntos
Animais , Bovinos , Caspases/análise , Caspases/efeitos adversos , Caspases/isolamento & purificação , Fator de Indução de Apoptose/análise , Fator de Indução de Apoptose/deficiência , Placenta/anatomia & histologia , Placenta/embriologia , Bovinos/anatomia & histologia , Bovinos/anormalidades , Bovinos/cirurgia , Homeostase , Marcação In Situ das Extremidades Cortadas/métodos , Marcação In Situ das Extremidades Cortadas/veterinária , PrenhezRESUMO
Apoptose e seus mecanismos reguladores são eventos fisiológicos cruciais para a manutenção da homeostase placentária, e o desequilíbrio desses processos pode comprometer a função placentária e, consequentemente, o sucesso da gravidez. Neste estudo, investigou-se a apoptose utilizando-se histomorfometria em lâminas coradas em HE e submetidas à reação de TUNEL. Além disso, avaliou-se a expressão de Bcl-2 e das caspases 8 e 3, pela reação de polimerase em cadeia em tempo real, em placentas saudáveis em diferentes estádios de gestação. Amostras de placentônios de vacas com quatro, seis e nove meses de gestação foram colhidas e processadas. O índice apoptótico aumentou progressivamente com o avanço da gestação. Tanto o Bcl-2 quanto as caspases 3 e 8 foram expressas nos três períodos estudados, sendo a expressão de Bcl-2 menor que a de caspase 8, que é menor que a de caspase 3. Estes resultados indicam que essas moléculas estão envolvidas na via apoptótica ativada na maturação placentária, exibindo um padrão de expressão ao longo da gestação e contribuindo para o equilíbrio fisiológico da celularidade e renovação celular na placenta bovina.(AU)
Apoptosis and its regulating mechanisms are crucial physiological events for the maintenance of the placental homostasis; and disequilibrium of these processes may compromise placental function and the success of the pregnancy. In this study, apoptosis was investigated by histomorphometry using slides stained with HE and TUNEL reaction. Besides that, Bcl-2 and caspases 8 and 3 expression were evaluated by real time polymerase chain reaction in healthy placentas under different gestacional ages. Samples of placentones of cows at 4th, 6th, and 9th months of gestation were harvested and processed. The apoptotic index gradually increased with the advance of the gestation. Bcl-2 and caspases 3 and 8 were expressed in all the studied periods, being the expression of Bcl-2 lower than that of caspase 8, which was lower than caspase 3. These results indicate that these molecules are involved in the activated apoptotic way in the placental maturation, showing a standard expression throughout the gestation and contributing for the physiological balance of the cellularity and cellular turn over in bovine placenta.(AU)
Assuntos
Animais , Bovinos , Placenta/anatomia & histologia , Placenta/embriologia , Fator de Indução de Apoptose/análise , Fator de Indução de Apoptose/deficiência , Caspases/efeitos adversos , Caspases/análise , Caspases/isolamento & purificação , Bovinos/anormalidades , Bovinos/anatomia & histologia , Bovinos/cirurgia , Prenhez , Homeostase , Marcação In Situ das Extremidades Cortadas/métodos , Marcação In Situ das Extremidades Cortadas/veterináriaRESUMO
Altered glial function may contribute to the initiation or progression of neuronal death in neurodegenerative diseases. Thus, modulation of astrocyte death may be essential for preventing pathological processes in the CNS. In recent years, metabotropic glutamate receptor (mGluR) activation has emerged as a key target for neuroprotection. We investigated the effect of subtype 3 mGluR (mGluR3) activation on nitric oxide (NO)-induced astroglial death. A mGluR3 selective agonist, LY379268, reduced inducible NO synthase expression and NO release induced by bacterial lipopolysaccharide and interferon-gamma in cultured rat astrocytes. In turn, a NO donor (diethylenetriamine/NO) induced apoptotic-like death in cultured astrocytes, which showed apoptotic morphology and DNA fragmentation, but no caspase 3 activation. LY379268 prevented astrocyte death induced by NO exposure, which correlates with a reduction in: phosphatidylserine externalization, p53 and Bax activation and mitochondrial permeability. The reported effects of LY379268 were prevented by the mGluR3 antagonist (s)-alpha-ethylglutamic acid. All together, these findings show the protective effect of mGluR3 activation on astroglial death and provide further evidence of a role of these receptors in preventing CNS injury triggered by several inflammatory processes associated with dysregulated NO production.