Injúria microbiana e a qualidade microbiológica dos alimentos / Microbial injury and the microbiological quality of food

Introdução: a contaminação microbiológica dos alimentos se apresenta como um grande problema para a indústria, agências reguladoras e consumidores. Os métodos de conservação utilizados como garantia da inocuidade dos alimentos bem como de extensão da vida de prateleira garantem uma redução significativa do número de células viáveis e/esporos. Entretanto, em virtude da aplicação de técnicas como calor, frio, redução da atividade de água e adição de conservantes, parte da população microbiana pode apresentar células com danos letais bem como danos reversíveis, tornando-se injuriadas. Objetivo: realizar uma revisão bibliográfica acerca da ocorrência de injúrias microbianas decorrentes da aplicação de métodos de conservação em alimentos. Metodologia: consulta das bases de dados Pubmed e Scielo, com seleção de artigos publicados entre os anos de 2000 e 2019. Revisão de literatura: os microrganismos se tornam injuriados após sobreviverem a condições estressantes e perderem parte de suas características funcionais, o que justifica um maior período para multiplicação nos alimentos, assim como em meios de cultura tradicionais. Cabe ressaltar que, caso estes microrganismos sejam expostos à ambientes favoráveis, é possível a recuperação dos danos sofridos. Uma vez regenerados, estes agentes representam um perigo potencial, visto a capacidade de se multiplicarem novamente no alimento, oferecendo riscos aos consumidores e acelerando a deterioração de produtos alimentícios. Conclusão: diante da ocorrência de injúrias microbianas, percebe-se a necessidade de incorporação de procedimentos adequados para recuperação de danos celulares aos métodos oficiais empregados para detecção e enumeração de microrganismos, a fim de garantir a qualidade e inocuidade de alimentos.

Introduction: Microbiological contamination of food is a major problem for industry, regulatory agencies and consumers. Preservation methods used to guarantee the safety of food as well as extending the shelf life ensure a significant reduction in the number of viable cells and/or spores. However, due to the application of techniques such as heat, cold, reduction of water activity and addition of preservatives, part of the microbial population may present cells with lethal damage as well as reversible damage, becoming injured. Objective: to carry out a literature review about the occurrence of microbial injuries resulting from the application of preservation methods in food. Methodology: pubmed and Scielo databases were consulted, with a selection of articles published between 2000 and 2019. Literature review: microorganisms become injured after surviving stressful conditions and losing part of their functional characteristics, which justifies a longer period for multiplication in food, as well as in traditional culture media. It should be noted that, if these microorganisms are exposed to favourable environments, it is possible to recover from the damage suffered. Once regenerated, these agents represent a potential hazard, given their ability to multiply again in food, posing risks to consumers and accelerating the deterioration of food products. Conclusion: in view of the occurrence of microbial injuries, it is necessary to incorporate adequate cell damage recovery procedures to the official methods used for detection and enumeration of microorganisms, in order to guarantee the quality and safety of food.

Protective effect of Lycium barbanun glycopeptide on human keratinocytes against radiation-induced oxidative stress damage and its mechanism / 中国生物制品学杂志

@#Objective To investigate the protective effect of Lycium barbanun glycopeptide(LbGP)on human keratinocyte HaCaT cells against radiation-induced cytotoxicity and its mechanism. Methods HaCaT cells were exposed to radiation with linear accelerator(rate 6 Gy/min)at doses of 4,8,12,16,20,24 and 28 Gy respectively,and the cell activity was detected by CCK-8 assay. HaCaT cells were treated with LbGP(0,0. 05,0. 1,0. 5,0. 8,1. 0,1. 5 and 3 mg/mL)for 4 h,exposed to radiation of 12 Gy,and detected for the cell viability by CCK-8 assay. HaCaT cells were divided into control group(without LbGP and radiation),radiation group(radiation of 12 Gy)and LbGP + radiation group(0. 8 mg/mL LbGP for 24 h,radiation of 12 Gy). After irradiation for 1 h,the reactive oxygen species(ROS)production was measured by flow cytometry,the superoxide dismutase(SOD)activity was determined by WST-8 assay and the expression of nuclear factor-E2 related factor 2(Nrf2),p-Nrf2,NADPH quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)were detected by Western blot;The mRNA transcription levels of Nrf2,HO-1 and NQO1 were detected by qRT-PCR at 1,3 and 5 h after irradiation. Results Radiation of12 Gy induced about 50% cell death,and 0. 8 mg/mL LbGP showed the best protective effect on the activity of irradiated cells. After irradiation for 1 h,compared with the control group,the content of ROS in HaCaT cells increased significantly in radiation group(F = 2. 55,P < 0. 001),the activity of SOD decreased significantly(F = 1. 23,P < 0. 01),the contents of NQO1 and Nrf2 protein had no significant difference(F = 1. 78 and 1. 00,respectively,P > 0. 05),the content of HO-1protein increased significantly(F = 1. 37,P < 0. 05),and the content of p-Nrf2 protein decreased significantly(F = 2. 75,P < 0. 01);Compared with the radiation group,the content of ROS in HaCaT cells of LbGP + radiation group decreased significantly(F = 3. 61,P < 0. 001),the activity of SOD increased significantly(F = 1. 23,P < 0. 05),and the contents of Nrf2,p-Nrf2,HO-1 and NQO1 protein increased significantly(F = 4. 00,2. 25,6. 25 and 1. 27,respectively,P < 0. 05). In addition,the mRNA levels of Nrf2,HO-1 and NQO1 genes in LbGP + radiation group were significantly higher than those in radiation group at 1,3 and 5 h after irradiation(F = 0. 20～36. 00,P < 0. 05). Conclusion LbGP can mitigate oxidative stress damage of HaCaT cells induced by radiation and protect cell activity,which may play a role by activating Nrf2 and increasing the levels of its downstream antioxidant enzymes SOD,HO-1 and NQO1.

Shoutaiwan Regulate Lipopolysaccharide-induced Oxidative Stress and Pyroptosis in Human Extravillous Trophoblast Cells / 中国实验方剂学杂志

ObjectiveTo investigate the regulatory effect of Shoutaiwan on oxidative stress and pyroptosis in lipopolysaccharide （LPS）-induced human extravillous trophoblast （HTR-8/SVneo） cells and provide a new direction for deciphering the mechanism of action of Shoutaiwan. MethodLPS （100 μg∙L-1） was used to induce the injury of HTR-8/SVneo cells （modeling）. Five groups were designed in this study， including a blank group， a model group， a Shoutaiwan （10% Shoutaiwan-containing serum） group， an antioxidant （1 mmol·L-1 NAC） group， and NOD like receptor thermoprotein domain 3 （NLRP3） inhibitor （50 μmol·L-1 MCC950） group. Cell viability was detected by cell counting kit-8 （CCK-8） kit. Hochest 33342/PI double fluorescence staining and flow cytometry were employed to observe cell death. The levels of interleukin-18 （IL-18）， interleukin-1β （IL-1β）， malondialdehyde （MDA）， and superoxide dismutase （SOD） in cell supernatant was determined by enzyme-linked immunosorbent assay （ELISA）. DCFH-DA probe was used to measure the level of intracellular reactive oxygen species （ROS）. Western blot was employed to determine the protein levels of NLRP3， Caspase-1， gastermin D （GSDMD）， and IL-1β in cells， and Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） to measure the mRNA levels of NLRP3 and Caspase-1 in cells. ResultCompared with the blank group， the modeling decreased the cell viability （P<0.01）， elevated the levels of IL-1β， IL-18， ROS， and MDA， and weakened the activity of SOD （P<0.01）. Furthermore， it up-regulated the protein levels of NLRP3， Caspase-1， GSDMD， and IL-1β and the mRNA levels of NLRP3 and Caspase-1 （P<0.01）. Compared with the model group， Shoutaiwan， NAC， and MCC950 increased the cell viability （P<0.01）. Further， Shoutaiwan and NAC lowered the levels of MDA and ROS and increased the activity of SOD （P<0.01）. Shoutaiwan and MCC950 reduced the IL-1β and IL-18 in cell supernatant （P<0.01）， and down-regulated the protein levels of NLRP3， Caspase-1， GSDMD， and IL-1β and the mRNA levels of NLRP3， Caspase-1， and IL-1β （P<0.05，P<0.01）. ConclusionShoutaiwan can regulate oxidative stress and pyroptosis to attenuate the LPS-induced damage of HTR-8/SVneo cells， which may be the mechanism of Shoutaiwan in preventing recurrent spontaneous abortion.

Effects of elderflower extract enriched with polyphenols on antioxidant defense of salmon leukocytes

BACKGROUND: In fish farming, the plant extracts containing antioxidant compounds have been added to the diet for enhancing pathogen resistance. In vitro studies evaluating the antioxidant effect of herbal extracts on fish cell models have focused on ROS production and the respiratory burst mechanism. However, the effects on enzymatic antioxidant defense on salmon leukocytes have not been evaluated. This study aims to evaluate the enzymatic antioxidant defense and ROS-induced cell damage in Salmon Head Kidney-1 (SHK-1) cell line exposed to polyphenol-enriched extract from Sambucus nigra flowers. RESULTS: Firstly, the Total Reactive Antioxidant Power (TRAP) assay of elderflower polyphenol (EP) was evaluated, showing 459 and 489 times more active than gallic acid and butyl hydroxy toluene (BHT), respectively. The toxic effect of EP on salmon cells was not significant at concentrations below 120 mg/ mL and no hemolysis activity was observed between 20 and 400 mg/mL. The treatment of SHK-1 cell line with EP decreased both the lipid peroxidation and protein oxidation induced by H2O2, which could be associated with decreasing oxidative stress in the SHK-1 cells since the GSH/GSSG ratio increased when only EP was added. CONCLUSIONS: These results suggest that plant extracts enriched with polyphenols could improve the enzymatic antioxidant defense of salmon leukocytes and protect the cells against ROS-induced cell damage

Intrauterine infection and fetal cardiovascular system development / 国际儿科学杂志

When the pathogen infects the fetus,the pathogenic microorganism and the infection product are recognized by the corresponding receptor.The fetus innate immune system is passively activated,which produces proinflammatory cytokines,induces cascade reaction of cytokines,and releases a large number of inflammatory factors secreted by the body.Its toxic effect can cause damage to the brain,lung,small intestine and heart and other important organs of the whole body,which seriously threatens the life of the perinatal infants and their subsequent survival quality.It has been found that fetal cardiovascular system is one of the important target organs of intrauterine infection.Cytokines produced by cardiac inflammation and induced by intrauterine infection can damage myocardial cells,affect the proliferation of myocardial cells,and cause damage to cardiac function.Moreover,the persistent influence of infection on fetus leads to fetal vascular remodeling and changes in fetal cardiopulmonary hemodynamics.This article reviews the effects of pathogens of intrauterine infection and fetal cardiac inflammation,cardiac hemodynamics,cardiomyocyte development,gene program of cardiomyocyte and cardiac structure development on fetal cardiovascular system.

Effects of ethylbenzene on cell injury and mitochondrial membrane potential of CPCs / 中华劳动卫生职业病杂志

Objective@#To investigate the effects of ethylbenzene on growth morphology、proliferation ability and mitochondrial membrane potential (MMP) of cochlear progenitor cells (CPCs) , and to lay a foundation for the mechanism of hearing loss induced by ethylbenzene.@*Methods@#We can use the fluorescence microscopy to identify the original CPCs isolated from the newborn rats, and followed by the addition of different concentrations of ethylbenzene (0, 15, 30, 45 μmol/L) for 24 hours. The morphological changes of cell injury were observed by inverted optical microscope. The proliferation ability of cells was detected by MTT colorimetry, and the change of MMP was detected by fluorescent probe JC-1.@*Results@#The results of CPCs identification showed the expression of Myosin VIIa and Epsin positive; The results observed by inverted optical microscope showed all groups of CPCs morphological changes compared with the control group; MTT results showed that the decreased significantly proliferation ability of CPCs groups compared with the control group and a dose effect relationship with statistically significant difference (P<0.05) ; JC-1 test results showed the decreased significantly mitochondrial membrane potential in the treated group compared with the control group, and there was a statistically significant difference (P<0.05) .@*Conclusion@#Ethylbenzene may cause damage to CPCs, inhibition of cell proliferation and decrease of MMP in rats.

On the effects of NLRP3 inflammasome on mice pancreatic β-cell damage induced by advanced glycation end products / 中华内分泌代谢杂志

Objective To explore the role of the pyrin domain-containing 3 ( NLRP3) inflammasome in advanced glycation end products ( AGEs )-induced mice pancreatic β-cell damage. Methods AGEs were administered intraperitoneally for 6 weeks in NLRP3 knockout mice or C57BL/6J mice. Intraperitoneal glucose tolerance test and insulin releasing test were performed. Pancreatic sections were stained with haematoxylin and eosin, or with F4/80 and NLRP3 antibodies. Insulin and pancreatic tissue monocyte chemotactic protein 1 ( MCP-1) as well as interleukin-1β( IL-1β) levels were measured with ELISA kits. Expression of MCP-1 protein was determined by western blot. MIN6 cells and mouse peritoneal macrophages cells were treated with AGEs and different interventions (antioxidant NAC, adenovirus NLRP3 shRNA or NLRP3 knockout). Reactive oxygen species production, NLRP3 mRNA expression, IL-1β secretion, caspase 1 activity, apoptosis and glucose stimulated insulin release were determined. Results Injection of AGEs induced an abnormal response to glucose, enhanced the insulitis score, and increased the levels of pancreatic tissue MCP-1 and IL-1β, as well as raised the expression of NLRP3 and F4/80 in pancreatic islet. Remarkably, co-localization of NLRP3 and macrophage marker F4/80 was observed in islet. The damages were improved in NLRP3 knockout mice. After incubation with AGEs, reactive oxygen species production and cell apoptosis was enhanced, NLRP3 inflammasome activated, with glucose-stimulated insulin release impaired in MIN6 cells. NAC treatment alliviated the above damages, but NLRP3 gene silencing had no effect on ROS level, apoptosis, and insulin secretion. Finally NAC treatment and NLRP3 gene knockout inhibited activation of NLRP3 inflammasome induced by AGEs in mouse peritoneal macrophages cells. Conclusion NLRP3 knockout ameliorates the islet β-cell damage induced by AGEs. These effects were associated with AGEs-induced islets macrophage infiltrating by up-regulation of MCP-1 expression, and AGEs-induced activation of NLRP3 inflammasome in macrophage through ROS pathway, which results in the release of active IL-1βand leads to the lesions of β-cell.

Mitochondria-rich cells changes induced by nitrite exposure in tambaqui (Colossoma macropomum Cuvier, 1818)

ABSTRACT The gill mitochondria-rich cells of the juvenile Amazonian fish Colossoma macropomum were analyzed using light and scanning and transmission electron microscopy after 96 h exposure to 0.04 and 0.2 mM nitrite. Although the number of mitochondria-rich cells decreased significantly in the lamellar epithelium, no decrease was found in the interlamellar region of the gill filament. Nitrite exposure caused significant reduction on the apical surface area of individual mitochondria-rich cells (p < 0.05), with a resulting reduction of the fractional area of these cells in both the lamellar and filament epithelium. Swelling of endoplasmic reticulum cisternae, nuclear envelope and mitochondria were the main changes found in the mitochondria-rich cells. Cristae lysis and matrix vacuolization characterized the mitochondrial changes. The overall ultrastructural changes indicated cellular functional disruption caused by exposure to nitrite. The changes observed in the gill indicate that the cellular structures involved in the process of energy production become severely damaged by exposure to nitrite indicating irreversible damage conducting to cell death.

Amelioration effect and mechanisms of phenylethanoid glycosides from Pedicularis muscicola maxim on high altitude memory impairment / 中国药学杂志

OBJECTIVE: To investigate the ameliorated effect and mechanisms of phenylethanoid glycosides from Pedicularis muscicola Maxim on high altitude memory impairment. METHODS: After successfully trained in the 8-arms radial maze, fifty Wistar rats were randomly divided into normoxic control group, hypoxia group, phenylethanoid glycosides 50, 200 and 400 mg · kg-1 groups (given corresponding dose). Normoxic control and hypoxia groups were administered with distilled water for a week. When drug delivery in the fourth day, hypoxia and phenylethanoid glycosides groups rats were exposed to a simulated of 7 500 m in a specially designed animal decompression chamber. Eight arms radial maze was used to measure spatial memory, HE stained was used to observe the cell morphology in brain tissue and biochemical technique was used to detect the content of MDA and ROS and enzymatic activity of GSH and SOD in brain tissue and serum. RESULTS: Compared with the normoxic control group, for hypoxia group rats, WME, RWE and TE were respectively increased by 800%, 71%, and 127.1% (P < 0.01) and neuron damage was significantly increased, the enzymatic activity of GSH and SOD were respectively decreased by 60.9% and 18.11% (P < 0.05, P < 0.01) in brain tissue and plasma while the content of MDA was increased in brain tissueby 74.8% (P < 0.01). Compared with the hypoxia group, for phenylethanoid glycosides 200, 400 mg · kg-1 groups rats, WME, RWE, TE were respectively decreased by 68.44%, 63.11%; 33.14%, 25.34% and 43.91%, 36.72% (P < 0.05, P < 0.01) and neuron damage was significantly decreased, the enzymatic activity of GSH were respectively increased by 219.76%, 180.75% and 32 81%, 24.10% (P < 0.05, P < 0.01) and the enzymatic activity of SOD were respectively increased by 9.57%, 13.88% and 15.41%, 15.45% (P < 0.05) in brain tissue and plasma, while the content of MDA in plasma were respectively decreased by 42.73%, 42.73% (P < 0.01) and MDA and ROS in brain tissue were respectively decreased by 61.71%, 42.79% and 40.76%, 23.53% (P < 0.01); for phenylethanoid glycosides 50 mg · kg-1 group rats, the corresponding indicators had been ameliorated, but there was no significant difference. CONCLUSION: Phenylethanoid glycosides of Pedicularis muscicola Maxim can ameliorate high altitude memory impairment, which its involved mechanism may be antioxidant stress and inhibition on cell damage.

¿La exposición crónica al alcohol induce neurodegeneración en el Sistema Nervioso Central de la rata? / Does chronic alcohol exposure induce neurodegeneration in the rat Central Nervous System?

Antecedentes: La exposición crónica al alcohol se asocia con procesos neurotóxicos y neurodegenerativos relacionados con disfunciones cognitivas y de memoria. El daño inducido por alcohol depende de los patrones de consumo de etanol. La exposición prolongada al alcohol induce daño en distintas regiones cerebrales (cortezas prefrontal, perirrinal, entorrinal y parahipocampal, tálamo, hipotálamo, hipocampo y cerebelo) en pacientes alcohólicos y modelos animales de alcoholismo. Sin embargo, no se han estudiado las regiones cerebrales asociadas con el circuito de reforzamiento y recompensa de drogas de abuso.Objetivo: Investigar si la exposición crónica al alcohol induce daño neurodegenerativo en el cerebro de la rata, en particular en el sistema mesocorticolímbico y la amígdala.Método: Ratas Wistar macho fueron expuestas a etanol (10% v/v) o agua por consumo oral durante 30 días y se les privó de la droga por 0, 24 y 48h. Los animales fueron sacrificados y se les extrajo la sangre troncal y el cerebro. Para evaluar el daño neurodegenerativo, se utilizó el marcador fluorescente Fluoro-Jade B. La concentración de alcohol en sangre se determinó por espectrofotometría.Resultados: Se observó un escaso número de células positivas a Fluoro-Jade en las cortezas piriforme y frontal de asociación, el caudado-putamen y el tálamo dorsal. No se encontraron diferencias entre el tratamiento crónico o la privación de alcohol versus el grupo control.Discusión y conclusión: La exposición crónica al alcohol no indujo neurodegeneración en las condiciones utilizadas en este estudio. Probablemente, las concentraciones de alcohol en sangre alcanzadas durante el tratamiento no fueron suficientes para inducir muerte celular.

Background: Chronic alcohol exposure is associated to neurotoxic and neurodegenerative mechanisms that lead to several cognitive and memory dysfunctions. Alcohol-induced damage depends on ethanol consumption patterns. Prolonged alcohol exposure induces damage in distinct brain regions (prefrontal, perirhinal, entorhinal and parahippocampal cortices, thalamus, hypothalamus, hippocampus and cerebellum) in both alcoholic patients and animal models of alcoholism. However, brain areas of the drug reinforcement and reward circuit have not been investigated.Objective: To investigate if chronic alcohol exposure induces neurodegenerative damage in the rat brain, particularly in the mesocorticolimbic system and the amygdala.Method: Male Wistar rats were exposed to ethanol (10% v/v) or water by oral consumption during 30 days. In another set of experiments, animals similarly treated with ethanol were withdrawn from the drug for 24 and 48 h. At the end of the treatments, animals were sacrificed, whole blood samples were obtained and the brains were removed. A fluorescence marker (Fluoro-Jade B) was used to assess neurodegenerative damage in the brain. Blood alcohol concentration was evaluated by spectrophotometry.Results: We observed a low number of Fluoro-Jade B positive cells in different brain regions, including the piriform cortex, frontal cortex of association, caudate-putamen and dorsal thalamus. No differences were found between chronic alcohol or ethanol withdrawn groups versus control animals.Discussion and conclusion: Our results suggest that chronic alcohol exposure does not induce neurodegeneration under the present experimental conditions. Alcohol blood concentrations attained during treatment may not be sufficient to induce cell death.

The Effect of Human Placental Extracts in Suppressing Tumor Cell and Preventing Normal Cell Damage / 日本補完代替医療学会誌

Objectives: To analyze the effect of human placental extracts (HPEx) on hepatocellular carcinoma cells in vitro. Methods: The hepatocellular carcinoma cell lines, namely, HLE and Huh-7, were used. The cells were subjected to a growth assay using the formazan dye method; the effect of combination treatment with sorafenib and HPEx was also assessed. The preventing normal cell damage effect of HPEx was analyzed by virtual therapy where possible; the experimental protocol was constructed on the basis of pre- and post-sorafenib treatment data. Cytotoxicity was measured by lactate dehydrogenase (LDH) assay. Results: HPEx caused significant dose-dependent suppression in the growth of HLE and Huh-7 cells. These tumor cells were significantly suppressed by combination treatment with HPEx and sorafenib. In addition, HPEx potentiated sorafenib sensitivity against tumor cells, and significantly prevented sorafenib-induced cytotoxicity in primary cultured rat hepatocytes under all designed experimental conditions. Specifically, pre-treatment with HPEx had a greater effect than post-treatment with HPEx. Conclusion: HPEx suppresses tumor cell growth, potentiates sorafenib efficacy, and has a preventing normal cell damage effect; this triple functionality of HPEx makes it a useful agent for liver cancer therapy.

The effects of nonyl phenoxypolyethoxyl ethanol on cell damage pathway gene expression in SK-NSH cells

BACKGROUND/AIMS: Most pesticide formulations contain both chief and additive ingredients. But, the additives may not have been tested as thoroughly as the chief ingredients. The surfactant, nonyl phenoxypolyethoxylethanol (NP40), is an additive frequently present in pesticide formulations. We investigated the effects of NP40 and other constituents of a validamycin pesticide formulation on cell viability and on the expression of genes involved in cell damage pathways. METHODS: The effects of validamycin pesticide ingredients on cell viability and of NP40 on the mRNA expression of 80 genes involved in nine key cellular pathways were examined in the human neuroblastoma SK-N-SH cell line. RESULTS: The chemicals present in the validamycin pesticide formulation were cytotoxic to SK-N-SH cells and NP40 showed the greatest cytotoxicity. A range of gene expression changes were identified, with both up- and down-regulation of genes within the same pathway. However, all genes tested in the necrosis signaling pathway were down-regulated and all genes tested in the cell cycle checkpoint/arrest pathway were up-regulated. The median fold-change in gene expression was significantly higher in the cell cycle checkpoint/arrest pathway than in the hypoxia pathway category (p = 0.0064). The 70 kDa heat shock protein 4 gene, within the heat shock protein/unfolded protein response category, showed the highest individual increase in expression (26.1-fold). CONCLUSIONS: NP40 appeared to be particularly harmful, inducing gene expression changes that indicated genotoxicity, activation of the cell death (necrosis signaling) pathway, and induction of the 70 kDa heat shock protein 4 gene.

Role of rVvhA in inducing THP-1 cells damage / 中华微生物学和免疫学杂志

Objective To investigate the role of recombinant Vibrio vulnificus cytolysin (rVvhA) in inducing THP-1 cells damage and study the pathway of associated calcium influx .Methods Inverted mi-croscope, CCK-8 cell proliferation kit, Fluo3/AM staining and caspase activity detection were performed to analyze the damage of THP-1 cells induced by rVvhA and the pathway of calcium influx .Results rVvhA had cytotoxic effects on THP-1 cells in a dose-dependent manner .The concentrations of extracellular K +and LDH were respectively up-regulated after 1 h and 6 h of 12 μg/ml rVvhA intervention .Verapamil , Mibe-fradil and SKF-96365 could not prevent the influx of free Ca 2+induced by rVvhA .The activities of caspase-3 and caspase-9 were singanificantly enhanced by rVvhA in a time-dependant manner .Conclusion rVvhA can induce THP-1 cells damage through triggering extracellular calcium influx via porous channel on cell membrane.Moreover, rVvhA might induce THP-1 cell apoptosis through activating caspase-9/3-dependent pathway .

Relação entre biomarcadores inflamatórios, de adesão celular, de estresse oxidativo, de lesão endotelial, remodelamento tecidual e vascular e os diferentes estágios da doença venosa crônica primária (classes clínicas CEAP C0a, C2, C3, C4) / Relationship between biomarkers of inflammation, cell adhesion, oxidative stress, endothelial cell damage, vascular and tissue remodeling and the different stages of primary chronic venous disease (CEAP clinical classes C0a, C2, C3, C4)

A doença venosa crônica (DVC) é uma desordem complexa que compreende sinais e sintomas que variam das telangiectasias às úlceras ativas. A DVC é classificada de acordo com aspectos clínicos, etiológicos, anatômicos e fisiopatológicos (CEAP) em sete classes variando de C0 à C6. A principal causa da DVC é a hipertensão venosa que altera o fluxo venoso e, consequentemente, a força de cisalhamento que induz alterações fenotípicas nas células endoteliais que passam a expressar mediadores pró-inflamatórios e pró-trombóticos, que levam à adesão de leucócitos, ao aumento do estresse oxidativo, da permeabilidade vascular e do dano endotelial e ao remodelamento tecidual e vascular.Em virtude dos inúmeros mecanismos e da diversidade de moléculas envolvidas na patogênese e progressão da DVC, é essencial conhecer a interação entre elas e também saber quais são as moléculas (biomarcadores) que se correlacionam positivamente ou negativamente com a gravidade da doença. Foram avaliados os níveis de Interleucina-6 (IL-6), sL-selectina, sE-selectina, sP-selectina, molécula de adesão intercelular-1solúvel (sICAM-1), molécula de adesão das células vasculares-1 solúvel (sVCAM-1), ativador tecidual do plasminogênio (tPA), atividade do inibidor do ativador do plasminogênio-1 (PAI-1), trombomodulina solúvel (sTM), fator de von Willebrand (vWF), metaloproteinase de matriz (MMP)-2, MMP-3, MMP-9, inibidor tecidual das MMPs -1 (TIMP-1), angiopoietina-1 e -2, sTie-2 e s-Endoglina e fator de crescimento do endotélio vascular (VEGF) no sangue coletado da veia braquial de 173 mulheres com DVC primária divididas em grupos C2, C3, C4 e C4 menopausadas (C4m) e de 18 voluntárias saudáveis (grupo C0a). Foram também analisados os níveis urinários de ent-prostaglandina F2α nesses grupos. Não foram encontradas diferenças estatisticamente significativas com relação às concentrações sanguíneas e urinárias de sE-selectina, sP-selectina, sICAM-1, atividade de PAI-1, MMP-3, razão TIMP-1/MMP-3 ...

Chronic Venous Disease (CVD) is a complex disorder, which encompasses signs and symptoms that vary from telangiectasias to active ulcers. The CVD is classified according Clinical, Etiologic, Anatomical and Pathophysiological (CEAP) aspects into seven classes varying from C0 to C6. The main cause of CVD is venous hypertension, which alters venous flow and consequently, shear stress. Abnormal shear stress induces phenotypic changes in endothelial cells that start to express pro-inflammatory and pro-thrombotic mediators that lead to leukocyte adhesion, oxidative stress, increased vascular permeability and endothelial cell damage and tissue and vascular remodeling. Due to several mechanisms and the diversity of molecules involved in the pathogenesis and progression of CVD, is essential to know the interplay between them and which are the molecules (biomarkers) that correlate positively and negatively with the severity of the disease. We investigated the levels of interleukin-6 (IL-6), sL-selectin, sE-selectin, sP-selectin, soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1) activity, soluble thrombomodulin (sTM), von Willebrand factor (vWf), matrix metalloproteinase (MMP)-2, MMP-3, MMP-9, tissue inhibitor of metaloproteinases-1 (TIMP-1), angiopoietin-1 and -2, sTie-2, s-Endoglin, vascular endothelial growth factor (VEGF) in the blood taken from the brachial vein of 173 patients with primary CVD divided into C2, C3, C4 and menopaused C4 (C4m) groups and 18 healthy volunteers (C0a group).We also investigated the urinary levels of ent-prostaglandin F2α in these groups. There was no statistically significant difference between groups with respect to blood or urinary levels of sE-selectin, sP-selectin, sICAM-1, PAI-1 activity, MMP-3, TIMP-1/MMP-3 ratio, angiopoietin-2, angiopoietin-1/angiopoietin-2 ratio, s-Endoglin ...

Protective Effect of Fisetin (3,7,3',4'-Tetrahydroxyflavone) against gamma-Irradiation-Induced Oxidative Stress and Cell Damage

Ionizing radiation can induce cellular oxidative stress through the generation of reactive oxygen species, resulting in cell damage and cell death. The aim of this study was to determine whether the antioxidant effects of the flavonoid fisetin (3,7,3',4'-tetrahydroxyflavone) included the radioprotection of cells exposed to gamma-irradiation. Fisetin reduced the levels of intracellular reactive oxygen species generated by gamma-irradiation and thereby protected cells against gamma-irradiation-induced membrane lipid peroxidation, DNA damage, and protein carbonylation. In addition, fisetin maintained the viability of irradiated cells by partially inhibiting gamma-irradiation-induced apoptosis and restoring mitochondrial membrane potential. These effects suggest that the cellular protective effects of fisetin against gamma-irradiation are mainly due to its inhibition of reactive oxygen species generation.

Effect of estrogen and its receptors on glutamate-induced neural cell injury / 第二军医大学学报

It has been proven that glutamate-induced excitatory toxicity can bring damage to nerve cells. Previous studies indicate that glutamate excitotoxicity-associated neuron damage play an important role in the pathogenesis of a number of nervous system diseases, and estrogen can protect neurons from glutamate excitotoxicity-associated neuron damage, which may be mediated by estrogen receptor. This paper reviews the latest progress regarding the protective effects of estrogen and its receptors against glutamate-induced neuron damage.

Oxidative Stress of Mouse Fed with gamma -Irradiated Soybean Diet

Food irradiation has been steadily increased in many countries concomitantly with increasing international trades. Harmful contaminants naturally occurred from foods which contain high levels of unsaturated fatty acids that are easily oxidized can affect the human anti-oxidation system through the generation of free radicals. Moreover, previous studies proved that gamma-irradiation may cause production of free radicals in food. We investigated the effect of gamma-irradiated soybeans in relation to oxidative stress in mice. Oxidative index of mice was evaluated by TBARS, DNA fragmentation in various organs such as blood lymphocytes, liver and kidney. Forty male ICR mice were equally divided into 4 groups and fed control diet or gamma-irradiated diet containing 50% soybeans (5, 10, and 20 kGy, respectively )for 8 weeks. Pero-xide values of the irradiated diets were higher than that of the non-irradiated one and increased according to the storage period. There was no significant difference in weight gain as well as in TBARS value in plasma and kidney of all groups. Liver TBARS value of the group fed with irradiated diet at 20 kGy increased significantly compared with the control group (p <0.05 ). DNA oxidative damage as measured by alkaline comet assay showed that % tail DNA in the blood lymphocytes of 5 kGy and 10 kGy groups increased significantly over the control group (p <0.05 ). Also, tail moments of 5 kGy and 10 kGy groups were higher than that of the control group. Ultrastructural examination shows myeline figures and swollen mitochondria in parietal and intestinal epithelial cells of the group fed with irradiated diet. Therefore, considering unsaturated fatty acid content, consumption of soybeans gamma-irradiated with over 20 kGy or repe-atedly may decrease the body's antioxidant mechanism.

Mechanism of Bovine Coronary Artery Endothelial Cells Damage Induced by Cigarette Smoke Extract / 环境与健康杂志

Objective To study the bovine coronary artery endothelial cells(BCAEC)damage induced by cigarette abstracts and further clarify the relationship between smoking and cardiovascular diseases.Methods BCAEC were treated with nicotine, mainstream smoke extract(MSW)and sidestream smoke extract(SSW)which had the normal concentration(1.0?10~(-5),0.8?10~(-5), 0.9?10~(-5)mol/L)of nicotine in smoker.The morphological changes of BCAEC were recorded by microscope digital image system. The quantification of apoptotic BCAEC cells was performed by visualization of nuclei stained with 4,6'-diamidino-2-phenylindole and trypan blue exclusion assay was used to examine the percentage of necrotic BCAEC.The caspase activity assay was employed to discuss the mechanism of BCAEC apoptosis.Results BCAEC exposed to nicotine and MSW appeared the typical morphological alteration of apoptosis and necrotic morphological alteration were observed after BCAWC were treated with SSW. 5.89% and 11.94% apoptotic ceils were found after BCAEC were exposed to nicotine and MSW for 24 hours.The level of BCAEC necrosis after treated with SSW was 62.84%.Caspase-3 activity was induced by nicotine and MSW.Conclusion Cigarette smoke extract can induce the cell death of BCAEC.Nicotine and MSW can induce caspase-3 activity increase.Even in the presence of a non-cytotoxic concentration of nicotine and mainstream smoke solution,protease-induced apoptosis of BCAEC can be significantly increased.Sidestream smoke solution may cause BCAEC necrosis instead of apoptosis.Caspase-3 activation is probably the mechanism of BCAEC apoptosis.

Endothelial Cell Damage in Microincison Cataract Surgery and Coaxial Phacoemulsification

PURPOSE: To evaluate the damage to corneal endothelial cells following coaxial phacoemulsification and bimanual microincision cataract surgery (MICS). METHODS: We measured and compared the changes in the corneal endothelial mean cell density, cell size variation coefficient, hexagonality, and central corneal thickness in senile cataract patients who had received either coaxial phacoemulsification (Group 1, n=20), MICS using ultrasound (Group 2, n=20), and MICS using laser (Group 3, n=20). The endothelial cell parameters and corneal thickness were evaluated preoperatively and at 1 week, 1 month, and 2 months postoperatively. RESULTS: There was no significant difference among the three groups in terms of the endothelial cell parameters and corneal thickness during two months (p>0.05). CONCLUSIONS: MICS is a safe technique that does not appear to be associated with more damage to the corneal endothelium than coaxial phacoemulsification. A longer follow-up study is necessary to investigate its potential benefits for replacing conventional surgery.

A Comparison of the Efficacy of Cataract Surgery Using Aqualase(R) with Phacoemulsification Using MicroFlow(R) System

PURPOSE: To compare the outcomes after phacoemulsification performed with the AquaLase(R) and phacoemulsification in MicroFlow(R) system, including surgically induced astigmatism (SIA), corneal endothelial cell damage and postoperative recovery of visual acuity. METHODS: The cataracts of Lens Opacities Classification System, version III (LOCS III) nuclear grade below 2 were subjected in this study. Nineteen eyes underwent cataract operation using AquaLase(R) (Alcon Laboratories, Fort Worth, Texas, U.S.A.). A control group (19 eyes) used the MicroFlow(R) system (Millenium, Stortz, U.S.A.) and was selected by matching age, sex, systemic disease, corneal astigmatism and corneal endothelial cell density. All the surgeries were performed by the same operator. SIA, corneal endothelial cell loss, visual acuity, and corneal thickness were evaluated postoperatively. RESULTS: SIA in the group using AquaLase(R) was less than that of the group using MicroFlow(R) system (P=0.022) at 2 months postoperatively. Evaluation of corneal endothelial cell loss, recovery of visual acuity and corneal thickness found no statistically significant differences between the two groups. CONCLUSIONS: Cataract surgery using AquaLase(R) induces less surgically induced astigmatism in mild to moderate cataracts.