Genome-wide analysis of metallothionein gene family in maize to reveal its role in development and stress resistance to heavy metal

BACKGROUND: Maize (Zea mays L.) is a widely cultivated cereal and has been used as an optimum heavy metal phytoremediation crop. Metallothionein (MT) proteins are small, cysteine-rich, proteins that play important roles in plant growth and development, and the regulation of stress response to heavy metals. However, the MT genes for maize have not been fully analyzed so far. METHODS: The putative ZmMT genes were identified by HMMER. The heat map of ZmMT genes spatial expression analysis was generated by using R with the log2 (FPKM + 1). The expression profiles of ZmMT genes under three kinds of heavy metal stresses were quantified by using qRT-PCR. The metallothionein proteins was aligned using MAFFT and phylogenetic analysis were constructed by ClustalX 2.1. The protein theoretical molecular weight and pI, subcellular localization, TFs binding sites, were predicted using ProtParam, PSORT, PlantTFDB, respectively. RESULTS: A total of 9 ZmMT genes were identified in the whole genome of maize. The results showed that eight of the nine ZmMT proteins contained one highly conserved metallothio_2 domain, while ZmMT4 contained a Metallothio_PEC domain. All the ZmMT proteins could be classified into three major groups and located on five chromosomes. The ZmMT promoters contain a large number of hormone regulatory elements and hormone-related transcription factor binding sites. The ZmMT genes exhibited spatiotemporal specific expression patterns in 23 tissues of maize development stages and showed the different expression patterns in response to Cu, Cd, and Pb heavy metal stresses. CONCLUSIONS: We identified the 9 ZmMT genes, and explored their conserved motif, tissue expression patterns, evolutionary relationship. The expression profiles of ZmMT genes under three kinds of heavy metal stresses (Cu, Cd, Pb) were analyzed. In summary, the expression of ZmMTs have poteintial to be regulated by hormones. The specific expression of ZmMTs in different tissues of maize and the response to different heavy metal stresses are revealed that the role of MT in plant growth and development, and stress resistance to heavy metals.

Efeito da exposição a produtos fumígenos sobre a expressão de metalotioneínas durante a artrite experimental e sobre linfócitos T / Effect of exposure to tobacco-related products on metallothionein expression in the experimental arthritis and on T lymphocytes functions

Produtos liberados pela queima do cigarro convencional (CC) estão relacionados com a progressão clínica da artrite reumatoide (AR). Produtos fumígenos não combustíveis surgiram com a premissa de apresentarem menor toxicidade que o CC, dentre os quais está o tabaco aquecido (heat-not-burn tobacco; HNBT). Neste projeto investigamos os efeitos do HNBT sobre eventos envolvidos na AR, focando na sintomatologia, expressão de metalotioneínas (MTs), e na biologia de linfócitos T CD4+ primários e da linhagem Jurkat. Exposições in vivo ao ar, CC ou HNBT foram realizadas 2 vezes ao dia, 1 hora cada (12 CC ou 24 HNBT/hora), nos dias 14-21 da indução da artrite induzida por antígeno (AIA) em camundongos C57Bl/6. Foram realizadas análises dos parâmetros clínico da doenças, histopatologia e imunohistoquímica; quantificação de nicotina e cotinina séricas por cromatografia líquida acoplada a espectrometria de massas (MS). Os efeitos das exposições in vitro sobre linfócitos T foram mensurados por citometria de fluxo e ELISA. A concentração de metais emitidas pelo CC ou HNBT durante as exposições foram mensurados por MS com plasma acoplado. Camundongos expostos ao CC apresentaram intensa inflamação pulmonar, expressões acentuadas de MTs hepáticas e pulmonares e exacerbação dos parâmetros de AIA quando comparados ao grupo expostos ao HNBT. Animais expostos ao CC ou ao HNBT apresentaram redução na celularidade de órgãos linfoides. Somente a exposição in vitro ao CC causou estresse oxidativo e secreção de citocinas inflamatórias, ativação do receptor de hidrocarbonetos arila (AhR) e polarização de células Th17. Diferentemente, exposição ao CC ou ao HNBT provocaram redução da secreção de IL-2 e proliferação de células Jurkat. A exposição de células Jurkat à nicotina mimetizou os efeitos inibitórios da exposição ao HNBT sobre a secreção de IL-2 e proliferação de linfócitos T. O CC liberou maiores concentrações de metais nas câmaras de exposição. Associados, nossos resultados mostram que embora exposições ao HNBT não exacerbem parâmetros inflamatórios de AIA e nem em funções linfócitos T, ambos produtos prejudicam a celularidade de órgãos linfoides e a proliferação e secreção de IL-2 por linfócitos T

Products released by burning conventional cigarettes (CC) are related to the worsening of rheumatoid arthritis (RA). Non-combustible smoking products appeared with the premise of presenting less toxicity than the CC, among which is the heated tobacco (heat-not-burn tobacco; HNBT). Here, we investigate the effects of HNBT on events involved in RA, focusing on symptoms, expression of metallothioneins (MTs), and on the biology of primary CD4+ T lymphocytes and the Jurkat T cell lineage. In vivo exposures to air, CC or HNBT were performed twice a day, 1 hour each (12 CC or 24 HNBT / hour), on days 14-21 of the induction of antigen-induced arthritis (AIA) in C57Bl / 6 mice. Analyzes of the clinical parameters of the AIA, histopathology, and immunohistochemistry were performed; quantification of nicotine and cotinine by liquid chromatography coupled to mass spectrometry (MS). The in vitro effects of exposures on T lymphocytes were measured by flow cytometry and ELISA. The concentration of metals released by the CC or HNBT during the exposures was measured by MS with coupled plasma. Mice exposed to CC showed intense pulmonary inflammation, marked expressions of hepatic and pulmonary MTs, and exacerbation of AIA parameters when compared to the group exposed to HNBT. Animals exposed to CC or HNBT showed a reduction in the cellularity of lymphoid organs. Only in vitro exposure to CC caused oxidative stress and secretion of inflammatory cytokines, activation of the aryl hydrocarbon receptor (AhR), and polarization of Th17 cells. However, exposure to CC or HNBT led to reduced secretion of IL-2 and proliferation of Jurkat cells. The exposure of Jurkat T cells to nicotine mimicked the inhibitory effects of exposure to HNBT on IL-2 secretion and T lymphocyte proliferation. The CC released higher concentrations of metals in the exposure chambers. In association, our results show that although exposures to HNBT do not exacerbate inflammatory parameters of AIA or T lymphocyte functions, both products impair lymphoid organ cell function and the proliferation and secretion of IL-2 by T lymphocytes

Metallothionein-2a (rs1610216&rs28366003) gene polymorphisms and the risk of stomach adenocarcinoma / Polimorfismos do gene metalotioneína-2A (rs1610216& rs28366003) e o risco de adenocarcinoma de estômago

ABSTRACT BACKGROUND: Gastric cancer is the fourth most common cause of worldwide cancer. Also in contrast to the huge advances in curing, the chance of living is very low even in surgery cases. Having a genetic predisposition plays an important role in cancer development. The association between Metallothionein-2A gene polymorphisms and the risk of adenocarcinoma has been widely studied, yet there is only one study on stomach diseases. OBJECTIVE: In this study, we aimed to investigate the association between 2 (MT-2A) polymorphisms and adenocarcinoma. METHODS: This cross-sectional case control study was performed between Mach 2014 and January 2015 at the Tuba Hospital of Sari, Iran. Peripheral blood samples were collected in EDTA tube. DNA extraction was performed using the spin column procedure. The MT-2A polymorphisms MT-2A (rs1610216), (rs28366003) were determined by polymerase chain reaction-restriction fragment length polymorphism analysis in 95 a topic adenocarcinoma patients and 90 healthy individuals from Iranian population. RESULTS: The MT-2A rs1610216 polymorphism increased the risk of adeno carcinoma in our Iranian population [OR: 3.8533; 95%CI, 1.3155-11.2869; P=0.0139] and rs28366003 [OR: 4.0978; 95%CI, 1.2521-13.4108; P=0.0197]. CONCLUSION: The MT-2A gene polymorphism was associated with the risk of adenocarcinoma in the Iranian population.

RESUMO CONTEXTO: O câncer gástrico é a quarta causa mais comum de câncer em todo o mundo. Também em contraste com os enormes avanços na cura, a chance de viver é muito baixa, mesmo em casos de cirurgia. Ter uma predisposição genética desempenha um papel importante no desenvolvimento do câncer. A associação entre polimorfismos do gene metalotioneína-2A e o risco de adenocarcinoma tem sido amplamente estudada, mas há apenas um estudo sobre doenças estomacais. OBJETIVO: Neste estudo, objetivou-se investigar a associação entre 2 (MT-2A) polimorfismos e adenocarcinoma. MÉTODOS: Um estudo de controle de caso transversal foi realizado entre março de 2014 e janeiro de 2015 no hospital Tuba, Sari, Irã. Amostras de sangue periférico foram coletadas em tubo EDTA. A extração do ADN foi executada usando o procedimento da coluna da rotação. Os polimorfismos MT-2a MT-2A (rs1610216), (rs28366003) foram determinados pela análise do polimorfismo do comprimento do fragmento da reação-limitação de cadeia da polimerase em 95 pacientes com adenocarcinoma tópico e em 90 indivíduos saudáveis da população iraniana. RESULTADOS: O polimorfismo MT-2A rs1610216 aumentou o risco de adenocarcinoma de em nossa população iraniana. [OR: 3,8533; 95%CI, 1,3155-11,2869; P=0,0139] e rs28366003 [OR: 4,0978; 95%CI, 1,2521-13,4108; P=0,0197]. CONCLUSÃO: O polimorfismo do gene MT-2A foi associado ao risco de adenocarcinoma na população iraniana.

Increased expression levels of metalloprotease, tissue inhibitor of metalloprotease, metallothionein, and p63 in ectopic endometrium: an animal experimental study / Aumento nos níveis de expressão de metaloprotease, inibidor tecidual das metaloproteases, metalotioneina e p63 em endométrio ectópico: um estudo experimental animal

Abstract Objective To characterize the patterns of cell differentiation, proliferation, and tissue invasion in eutopic and ectopic endometrium of rabbits with induced endometriotic lesions via a well- known experimental model, 4 and 8 weeks after the endometrial implantation procedure. Methods Twenty-nine female New Zealand rabbits underwent laparotomy for endometriosis induction through the resection of one uterine horn, isolation of the endometrium, and fixation of tissue segment to the pelvic peritoneum. Two groups of animals (one with 14 animals, and the other with15) were sacrificed 4 and 8 weeks after endometriosis induction. The lesion was excised along with the opposite uterine horn for endometrial gland and stroma determination. Immunohistochemical reactions were performed in eutopic and ectopic endometrial tissues for analysis of the following markers: metalloprotease (MMP-9) and tissue inhibitor of metalloprotease (TIMP-2), which are involved in the invasive capacity of the endometrial tissue; and metallothionein (MT) and p63, which are involved in cell differentiation and proliferation. Results The intensity of the immunostaining for MMP9, TIMP-2, MT, and p63 was higher in ectopic endometria than in eutopic endometria. However, when the ectopic lesions were compared at 4 and 8 weeks, no significant difference was observed, with the exception of the marker p63, which was more evident after 8 weeks of evolution of the ectopic endometrial tissue. Conclusion Ectopic endometrial lesions seem to express greater power for cell differentiation and tissue invasion, compared with eutopic endometria, demonstrating a potentially invasive, progressive, and heterogeneous presentation of endometriosis.

Resumo Objetivo Caracterizar o padrão de diferenciação celular, proliferação e invasão tecidual em endométrio eutópico e ectópico de coelhas com lesões de endometriose induzidas por um modelo experimental 4 e 8 semanas após o procedimento de implantação endometrial. Métodos Vinte e nove coelhas fêmeas Nova Zelândia foram submetidas a laparotomia para indução de endometriose através da ressecção de um dos cornos uterinos, isolamento do endométrio e fixação do tecido no peritônio pélvico. Dois grupos de animais (14 animais em um grupo e 15 animais no outro) foram sacrificados 4 e 8 semanas após a indução da endometriose. A lesão foi excisada junto com o corno uterino contralateral para determinação da presença de glândulas e de estroma endometrial. Reações de imunohistoquímica foram realizadas no tecido endometrial eutópico e ectópico para análise dos seguintes marcadores: metaloprotease (MMP9) e inibidor tecidual da metaloprotease 2 (TIMP-2), os quais estão envolvidos na capacidade de invasão do tecido endometrial; e metalotioneina (MT) e p63, os quais estão envolvidos na diferenciação e proliferação celular. Resultados A intensidade da imunomarcação para MMP9, TIMP-2, MT e p63 foi mais alta nos endométrios ectópicos do que nos endométrios eutópicos. Contudo, quando as lesões foram comparadas entre 4 e 8 semanas, nenhuma diferença foi observada, com exceção do marcador p63, o qual foi mais evidente depois de 8 semanas de evolução do tecido endometrial ectópico. Conclusão Lesões endometriais ectópicas parecem expressar maior poder de diferenciação celular e de invasão tecidual comparadas com endométrios eutópicos, demonstrando o potencial de invasão, de progressão e de apresentação heterogênea da endometriose.

Analysis of BMSCs-intervened viscoelasticity of sciatic nerve in rats with chronic alcoholic intoxication

Abstract Purpose: To investigate the impact of bone mesenchymal stem cells (BMSCs) intervention on the viscoelasticity of sciatic nerve in rats with chronic alcohol intoxication (CAI). Methods: The CAI rat models were prepared, divided into model groups, and treated with either BMSCs or basic fibroblast growth factor (bFGF). Then the rats underwent electrophysiological test and the serum levels of malondialdehyde (MDA), superoxide dismutase (SOD), and metallothionein (MT) were measured. Histological observation, stress relaxation test, and creep test were performed for the sciatic nerve of the CAI model in each group. Results: The MDA level of group BMSC was significantly lower (p<0.05) than that of groups MOD (the CIA model) and bFGF. The SOD and MT levels were higher in group BMSC than in groups MOD and bFGF (p<0.05). The motor nerve conduction velocity and amplitude were higher in group BMSC than in groups MOD and bFGF (p<0.05). The amounts of 7200s stress reduction and 7200 s strain increase of the sciatic nerve in group BMSC were greater than those in groups bFGF and MOD (p<0.05). Conclusion: Bone mesenchymal stem cells can improve the metabolism of free radicals, restore the tissue morphology and viscoelasticity of the chronic alcohol intoxication animal model, and positively affect the repairing of the injured sciatic nerve.

Nuclear metallothionein in oral squamous cell carcinoma: clinicopathological parameters and patient survival

Abstract The aim of this study was to identify the immunoexpression of methallothionein in oral squamous cell carcinoma as well as to address the correlation with clinical features, histological grade and patient survival. Samples were collected from 93 patients with tongue squamous cell carcinoma who presented for follow-up. Immunohistochemical expression of methallothionein in all groups was performed. The scoring system has previously been published by Tsurutani in 2005, which is based on intensity and distribution of staining. We used Kappa index to evaluate the degree of observers' agreement under metallothionein immunostaining and histological grade. Associations between methallothionein expression and clinical parameters (age, gender, smoking, tumor size, lymph node metastasis and disease stage) were examined for statistical significance using the chi-squared test. The overall survival rates were estimated by the Kaplan-Meier method and the relationship between protein expression and survival was compared using the log-rank test (p < 0.05). Our results showed no statistically significant association between methallothionein immunostaining and the selected clinicopathological variables. Immunohistochemistry results showed positive nuclear immunostaining for metallothionein in 62,37% (58/93) and negative for metallothionein 37,63% (35/93). The degree of examiners agreement by Kappa varied from substantial to perfect and both metallothionein immunostaining and histological grade were explored. The present study suggests that positive methallothionein expression found in tongue squamous cell carcinoma may not help to predict survival in the analyzed samples, as well as no relation between the protein and histological grade and clinical features was observed. In conclusion, the present study suggests that metallothionein is not associated with tongue squamous cell carcinoma clinicopathological characteristics and aggressiveness.

Heterologous expression of a rice metallothionein isoform (OsMTI-1b) in Saccharomyces cerevisiae enhances cadmium, hydrogen peroxide and ethanol tolerance

Abstract Metallothioneins are a superfamily of low-molecular-weight, cysteine (Cys)-rich proteins that are believed to play important roles in protection against metal toxicity and oxidative stress. The main purpose of this study was to investigate the effect of heterologous expression of a rice metallothionein isoform (OsMTI-1b) on the tolerance of Saccharomyces cerevisiae to Cd2+, H2O2 and ethanol stress. The gene encoding OsMTI-1b was cloned into p426GPD as a yeast expression vector. The new construct was transformed to competent cells of S. cerevisiae. After verification of heterologous expression of OsMTI-1b, the new strain and control were grown under stress conditions. In comparison to control strain, the transformed S. cerevisiae cells expressing OsMTI-1b showed more tolerance to Cd2+ and accumulated more Cd2+ ions when they were grown in the medium containing CdCl2. In addition, the heterologous expression of GST-OsMTI-1b conferred H2O2 and ethanol tolerance to S. cerevisiae cells. The results indicate that heterologous expression of plant MT isoforms can enhance the tolerance of S. cerevisiae to multiple stresses.

Time-dependent Expression of MT1A mRNA and MT2A mRNA in the Contused Skeletal Muscle of Rats / 法医学杂志

OBJECTIVES@#To investigate the time-dependent expression of metallothionein （MT） 1A mRNA and MT2A mRNA in contused skeletal muscle of rats.@*METHODS@#A total of 54 Sprague-Dawley rats were used in this study. The rats were divided into two parts: control group （n=6） and contusion groups （0.5, 1, 6, 12, 18, 24, 30, and 36 h after contusion, n=6）. Total RNA was extracted from skeletal muscle. The expression levels of MT1A mRNA and MT2A mRNA were detected by SYBR Green I real-time PCR.@*RESULTS@#The expression trends of the two potential marker genes were related to wound age. In addition to 0.5 h, there were significant contrasts between the control group and contused group （P<0.05）, about the expression levels of MT1A mRNA and MT2A mRNA in different phases. As the extension of wound age, the relative expression of MT1A mRNA and MT2A mRNA at 1 h, 6 h, 12 h and 18 h after contusion demonstrated upgrade tendency until its expression levels in 18 h peak with 239.41±15.20 and 717.42±50.76, respectively. When time extends to 24 h after injury, the expression of above two marks decreased, respectively. The MT1A mRNA and MT2A mRNA expression levels increased at 30 h and then decreased.@*CONCLUSIONS@#Determination of MT1A mRNA and MT2A mRNA levels by real-time PCR may be useful for the estimation of wound age.

The Role of Nuclear Factor-E2-Related Factor 1 in the Oxidative Stress Response in MC3T3-E1 Osteoblastic Cells

BACKGROUND: Reactive oxygen species (ROS) and antioxidants are associated with maintenance of cellular function and metabolism. Nuclear factor-E2-related factor 1 (NFE2L1, Nrf1) is known to regulate the expression of a number of genes involved in oxidative stress and inflammation. The purpose of this study was to examine the effects of NFE2L1 on the response to oxidative stress in osteoblastic MC3T3-E1 cells. METHODS: The murine calvaria-derived MC3T3-E1 cell line was exposed to lipopolysaccharide (LPS) for oxidative stress induction. NFE2L1 effects were evaluated using small interfering RNA (siRNA) for NFE2L1 mRNA. ROS generation and the levels of known antioxidant enzyme genes were assayed. RESULTS: NFE2L1 expression was significantly increased 2.4-fold compared to the control group at 10 µg/mL LPS in MC3T3-E1 cells (P<0.05). LPS increased formation of intracellular ROS in MC3T3-E1 cells. NFE2L1 knockdown led to an additional increase of ROS (20%) in the group transfected with NFE2L1 siRNA compared with the control group under LPS stimulation (P<0.05). RNA interference of NFE2L1 suppressed the expression of antioxidant genes including metallothionein 2, glutamatecysteine ligase catalytic subunit, and glutathione peroxidase 1 in LPS-treated MC3T3-E1 cells. CONCLUSION: Our results suggest that NFE2L1 may have a distinct role in the regulation of antioxidant enzymes under inflammation-induced oxidative stress in MC3T3-E1 osteoblastic cells.

Propylthiouracil, Perchlorate, and Thyroid-Stimulating Hormone Modulate High Concentrations of Iodide Instigated Mitochondrial Superoxide Production in the Thyroids of Metallothionein I/II Knockout Mice

BACKGROUND: Increased oxidative stress has been suggested as one of the underlying mechanisms in iodide excess-induced thyroid disease. Metallothioneins (MTs) are regarded as scavengers of reactive oxygen species (ROS) in oxidative stress. Our aim is to investigate the effects of propylthiouracil (PTU), a thyroid peroxidase inhibitor, perchlorate (KClO4), a competitive inhibitor of iodide transport, and thyroid stimulating hormone (TSH) on mitochondrial superoxide production instigated by high concentrations of iodide in the thyroids of MT-I/II knockout (MT-I/II KO) mice. METHODS: Eight-week-old 129S7/SvEvBrd-Mt1(tm1Bri) Mt2(tm1Bri)/J (MT-I/II KO) mice and background-matched wild type (WT) mice were used. RESULTS: By using a mitochondrial superoxide indicator (MitoSOX Red), lactate dehydrogenase (LDH) release, and methyl thiazolyl tetrazolium (MTT) assay, we demonstrated that the decreased relative viability and increased LDH release and mitochondrial superoxide production induced by potassium iodide (100 µM) can be relieved by 300 µM PTU, 30 µM KClO4, or 10 U/L TSH in the thyroid cell suspensions of both MT-I/II KO and WT mice (P<0.05). Compared to the WT mice, a significant decrease in the relative viability along with a significant increase in LDH release and mitochondrial superoxide production were detected in MT-I/II KO mice(P<0.05). CONCLUSION: We concluded that PTU, KClO4, or TSH relieved the mitochondrial oxidative stress induced by high concentrations of iodide in the thyroids of both MT-I/II KO and WT mice. MT-I/II showed antioxidant effects against high concentrations of iodide-induced mitochondrial superoxide production in the thyroid.

Metallothinein 1E Enhances Glioma Invasion through Modulation Matrix Metalloproteinases-2 and 9 in U87MG Mouse Brain Tumor Model

Malignant glioma cells invading surrounding normal brain are inoperable and resistant to radio- and chemotherapy, and eventually lead to tumor regrowth. Identification of genes related to motility is important for understanding the molecular biological behavior of invasive gliomas. According to our previous studies, Metallothionein 1E (MT1E) was identified to enhance migration of human malignant glioma cells. The purpose of this study was to confirm that MT1E could modulate glioma invasion in vivo. Firstly we established 2 cell lines; MTS23, overexpressed by MT1E complementary DNA construct and pV12 as control. The expression of matrix metalloproteinases (MMP)-2, -9 and a disintegrin and metalloproteinase 17 were increased in MTS23 compared with pV12. Furthermore it was confirmed that MT1E could modulate MMPs secretion and translocation of NFkB p50 and B-cell lymphoma-3 through small interfering ribonucleic acid knocked U87MG cells. Then MTS23 and pV12 were injected into intracranial region of 5 week old male nude mouse. After 4 weeks, for brain tissues of these two groups, histological analysis, and immunohistochemical stain of MMP-2, 9 and Nestin were performed. As results, the group injected with MTS23 showed irregular margin and tumor cells infiltrating the surrounding normal brain, while that of pV12 (control) had round and clear margin. And regrowth of tumor cells in MTS23 group was observed in another site apart from tumor cell inoculation. MT1E could enhance tumor proliferation and invasion of malignant glioma through regulation of activation and expression of MMPs.

Effect of A One-Week Balanced Diet on Expression of Genes Related to Zinc Metabolism and Inflammation in Type 2 Diabetic Patients

To evaluate the effect of diet on metabolic control and zinc metabolism in patients with type 2 diabetes mellitus (T2DM). One-week balanced diet was provided to 10 Brazilians patients with T2DM. Nutritional assessment, laboratorial parameters and expression of zinc transporter and inflammatory genes in peripheral blood mononuclear cells (PBMC) were performed. Healthy non-diabetic subjects of the same demographic were recruited to provide baseline data. Diabetic patients had higher body mass index and greater fasting plasma glucose, plasma tumor necrosis factor alpha (TNFalpha) and plasma interleukin 6 (IL6) levels compared with healthy subjects. In addition, the expression of transporters 4 (ZnT4) mRNA was lower and IL6 mRNA was higher in PBMC of these diabetic patients than in healthy subject. One week after a balanced diet was provided, fasting plasma glucose decreased significantly as did TNFalpha, IL6 and Metallothionein 1 (MT1) mRNAs. No change was observed in zinc transporter expression in PBMC after the dietary intervention. A healthy eating pattern maintained for one week was able to improve metabolic control of diabetic patients by lowering fasting plasma glucose. This metabolic control may be related to down-regulation of zinc-related transcripts from PBMCs, as TNFalpha, IL6 and MT1 mRNA.

Metallothionein and lipid peroxidation as markers to assess health status of chronically oiled Magellanic penguins in Argentina / Metalotioneínas y peroxidación lipídica como marcadores para determinar el estado de salud del Pingüino Magallánico crónicamente empetrolado en Argentina

Chronic oiling affects seabirds, with short (individual survival) and long-term (dynamics of population) effects. Magellanic penguin is the most affected species in Argentina. The aim was to evaluate the use of blood biochemical-molecular biomarkers (MT and TBARs) to assess health status of Magellanic penguin exposed to chronic environmental oiling, in addition with routine hematological analyses (differential WBC count and PCV). Oiled specimens presented significant higher RBC metallothionein and TBARs concentrations than those from shelter; while routine hematological parameters were within normal levels for penguins and did not show any difference between both groups. These results demonstrated the higher sensibility of both biomarkers, evidencing effects on health status by chronic oiling; furthermore, they are the first report in blood metallothionein and oxidative stress in S. magellanicus. These studies will be useful for a better assessment of the health condition in rehabilitation centers (short term); and, as early warning signals in natural colonies (long term).

El empetrolamiento crónico afecta a las aves marinas, con efectos a corto (supervivencia del individuo) y largo plazo (dinámica de la población). El pingüino de Magallanes es la especie más afectada en Argentina. El objetivo fue evaluar el uso de biomarcadores bioquímicos-moleculares (MT y TBARS) en sangre para evaluar el estado de salud del pingüino de Magallanes expuesto a empetrolamiento crónico ambiental, además de los análisis hematológicos de rutina (recuento diferencial de leucocitos y hematocrito). Los ejemplares empetrolados presentaron concentraciones significativamente mayores de metalotioneínas en fracción celular y peroxidación lipídica que aquellos de albergue; por el contrario, los parámetros hematológicos de rutina estuvieron dentro de los niveles normales para los pingüinos y no mostraron ninguna diferencia entre los grupos. Estos resultados demuestran la mayor sensibilidad de los dos biomarcadores, evidenciando efectos del empetrolamiento crónico sobre el estado de salud; además, este es el primer reporte de concentraciones de metalotioneínas en sangre y el estrés oxidativo en S. magellanicus. Estos estudios son de utilidad para una mejor evaluación del estado de salud en los centros de rehabilitación (corto plazo); y, como señales de alerta temprana en las colonias naturales (largo plazo).

The Critical Roles of Zinc: Beyond Impact on Myocardial Signaling

Zinc has been considered as a vital constituent of proteins, including enzymes. Mobile reactive zinc (Zn2+) is the key form of zinc involved in signal transductions, which are mainly driven by its binding to proteins or the release of zinc from proteins, possibly via a redox switch. There has been growing evidence of zinc's critical role in cell signaling, due to its flexible coordination geometry and rapid shifts in protein conformation to perform biological reactions. The importance and complexity of Zn2+ activity has been presumed to parallel the degree of calcium's participation in cellular processes. Whole body and cellular Zn2+ levels are largely regulated by metallothioneins (MTs), Zn2+ importers (ZIPs), and Zn2+ transporters (ZnTs). Numerous proteins involved in signaling pathways, mitochondrial metabolism, and ion channels that play a pivotal role in controlling cardiac contractility are common targets of Zn2+. However, these regulatory actions of Zn2+ are not limited to the function of the heart, but also extend to numerous other organ systems, such as the central nervous system, immune system, cardiovascular tissue, and secretory glands, such as the pancreas, prostate, and mammary glands. In this review, the regulation of cellular Zn2+ levels, Zn2+-mediated signal transduction, impacts of Zn2+ on ion channels and mitochondrial metabolism, and finally, the implications of Zn2+ in health and disease development were outlined to help widen the current understanding of the versatile and complex roles of Zn2+.

The role of protein phosphatase 2A B56β holoenzyme in the regulation of heavy metal CdCl2 induced cytotoxicity / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the role of holoenzyme containing Protein Phosphatase 2A B56β in regulating CdCl2 induced cytotoxicity.</p><p><b>METHOD</b>CdCl2-induced cytotoxicity in normal human cell line L-02, AFB1-transformed hepatic cell line L-02 RT-AFB1 and tumor cell line Bel7402 was measured by modified MTT assay. Stable cell lines L-02 SHAKT, L-02 SHB56β, L-02 RT-AFB1-B56β and Bel7402-B56β were generated by infecting L-02 cells or Bel7402 cells with retroviral vectors encoding lentiviral AKT shRNA, lentiviral B56β shRNA and B56β. The relative cell viability was measured in normal human cell line AFB1-transformed hepatic cell line and tumor cell line when treated by CdCl2 (0, 20, 40, 80, 160 µmol/L). After treated by wortmannin (2.5, 5.0 µmol/L) combined with 40 µmol/L CdCl2, Western blot was applied to measure the expression of associated protein in L-02.Western blot was applied to measure the expression of B56β, MT (metallothionein), AKT, and p-AKT in these cell lines treated by CdCl2.</p><p><b>RESULTS</b>The levels of MT were 0.12 ± 0.02, 0.06 ± 0.06 in L-02 RT-AFB1 and Bel7402, which were lower than L02 (0.92 ± 0.14) (F = 1 148.16, P < 0.001) when treated by 40 µmol/L CdCl2. When treated by 40 µmol/L CdCl2, the expression of p-AKT in L-02 SHAKT-1 and L-02 SHAKT-2 were 0.08 ± 0.02, 0.08 ± 0.05, which levels were lower than L-02 SHGFP (0.18 ± 0.15) (F = 724.70, P < 0.001); and the expression of MT were both 0.62 ± 0.16 in L-02 SHAKT-1 and L-02 SHAKT-2, which levels were higher than L-02 SHGFP (0.22 ± 0.14) (F = 94.73, P < 0.001). After treated by wortmannin (2.5, 5.0 µmol/L) combined with 40 µmol/L CdCl2, the expression of p-AKT in L-02 were 0.28 ± 0.07, 0.15 ± 0.11, which levels were lower than wortmannin untreated cells (0.52 ± 0.11) (F = 578.57, P < 0.001); and the expreesion of MT were 1.62 ± 0.80, 1.08 ± 0.15, which levels were higher than wortmannin untreated cells (0.69 ± 0.18) (F = 12.34, P < 0.001). When treated by 40 µmol/L CdCl2, the levels of p-AKT in L-02 SHB56β-1 and L-02 SHB56β-2 were 0.57 ± 0.13, 0.59 ± 0.02, which were higher than L-02 SHGFP (0.32 ± 0.02) (F = 87.16, P < 0.001); and the levels of MT were 0.35 ± 0.07, 0.20 ± 0.03 in L-02 SHB56β-1 and L-02 SHB56β-2, which were lower than L-02 SHGFP (1.51 ± 0.13) (F = 2 457.10, P < 0.001). After treated by 40 µmol/L CdCl2, the expression of p-AKT in L-02 RT-AFB1-B56β and Bel7402-B56β were 0.10 ± 0.11, 0.09 ± 0.01, which were lower than L-02 RT-AFB1 (0.36 ± 0.01) and Bel7402 (0.43 ± 0.11) (F = 877.62, P < 0.001); and the levels of MT were 0.92 ± 0.13, 0.95 ± 0.08 in L-02 RT-AFB1-B56β and Bel7402-B56β,which were higher than L-02 RT-AFB1 (0.44 ± 0.12) and Bel7402 (0.77 ± 0.06) (F = 51.97, P < 0.001).</p><p><b>CONCLUSION</b>Protein phosphatase 2A complexes containing B56β participated in the regulation of MT expression through direct dephosphorylation of AKT, finally affected the cytotoxicity responding to CdCl2. Our study revealed a key signaling pathways of PP2A involved in heavy metals induced cytotoxicity.</p>

Nível de dependência e qualidade de vida da população idosa / Level of dependency and quality of life of elderly / Nivel de dependencia y calidad de vida de los ancianos

A qualidade de vida (QV) nos idosos é determinada em grande parte pelo seu estado funcional e condições de saúde. Com o objectivo de avaliar o nível de QV, os factores que a influenciam e identificar o grau de dependência dos idosos foi realizado um estudo observacional transversal do tipo exploratório-descritivo, englobando 93 idosos. Na recolha de dados utilizouse o índice de Barthel e MOS-SF 36. Na identificação dos níveis de dependência os resultados indicam-nos que 40,0% são independentes e 18,0% são dependentes mínimos, sendo 12,0% dependentes totais. No que diz respeito à QV 88,0% dos sujeitos refere uma pontuação inferior a 50,0%, em média reportam uma QV de 39±10,0%. Verificou-se que existe uma correlação positiva entre o grau de dependência e o índice de QV, sobretudo na componente física. Assim, importa promover um envelhecimento saudável procurando-se privilegiar a preservação da autonomia e capacidade funcional dos idosos.

The quality of life (QoL) in older adults is largely determined by their functional status and health conditions. With the purpose of investigate the QoL and the factors affecting it, and identify the degree of dependency of the elderly was carried out an observational cross-sectional exploratory and descriptive, involving 93 elderly. In collecting data we used the Barthel Index and MOS SF-36. In the identification of levels of dependency results indicate us that 40.0% are independent and 18.0% are dependents, minimum being 12.0% total-dependent. The results show us that, 88.0% of the subjects reported a score below 50.0% on average reported a QoL of 39±10.0%. Checking that are a positive correlation between the degree of dependence and the index of QoL, especially in the physical component. It is therefore important to promote healthy aging in an attempt to favor the preservation of autonomy and functional capacity of the elderly.

La calidad de vida (CV) en los adultos mayores es en gran parte determinado por su estado funcional y las condiciones de salud. Con el fin de evaluar el nivel de CV y los factores que influyen en ella y determinar el grado de dependencia de los ancianos se llevó a cabo un estudio observacional transversal, exploratorio y descriptivo, que involucró a 93 personas mayores. En la recopilación de datos se utilizó el Índice de Barthel y el MOS SF-36. En la identificación de los nivele de dependiencia los resultados nos indican que 40,0% son independientes, 18,0% são dependentes mínimos y 12,0% dependientes totales. En lo que respeicta a la CV, 88,0% de los sujetos reportaron una puntuación inferior a 50,0% en promedio reportó una CV de 39±10,0%. Tomando nota de que existe una correlación positiva entre el grado de dependencia y el índice de calidad de vida, especialmente en el componente físico. Por tanto, es importante promover un envejecimiento saludable, en un intento de favorecer la preservación de la autonomía y la capacidad funcional de los ancianos.

Instrumento para avaliação da qualidade do Time de Resposta Rápida em um hospital universitário público / Instrument for assessing the quality of the Rapid Response Team at a university public hospital / Instrumento para la evaluación de la calidad del Equipo de Respuesta Rápida en un hospital universitario público

O artigo descreve a construção de um questionário para avaliação da qualidade do atendimento de um Time de Resposta Rápida, em Hospital Universitário de Londrina-PR, fundamentado no modelo conceitual de Donabedian (estrutura-processoresultado). A coleta de dados ocorreu no mês de março de 2012 e o processo de adequação do questionário foi desenvolvido com a aplicação da Técnica Delphi com a participação de 15 especialistas. Ao término do estudo obteve-se um questionário com 37 enunciados, sendo alcançado índice de concordância final na pesquisa com valores superiores a 80% para todos os conceitos. Espera-se que as contribuições do grupo de especialistas tornem o instrumento confiável e seja aplicado em outros serviços semelhantes. Aplicações futuras deste instrumento poderão trazer subsídios para melhor avaliação da qualidade dos serviços de equipes de Resposta Rápida.

The paper describes the construction of a questionnaire to assess the quality of care of a Rapid Response Team at the University Hospital of Londrina, based on the conceptual model of Donabedian (structure-process-outcome). Data collection occurred in March 2012 and the process of adjusting the questionnaire was developed with the application of the Delphi technique involving 15 experts. At the end of the study the questionnaire contained 37 statements, achieving final compliance level higher than 80% in all concepts. It is hoped that the contributions of the expert group produce a more reliable questionnaire to be applied in other similar services. Future applications of this instrument may provide information to better assess the quality of teams of Rapid Response services.

El artículo describe la construcción de un cuestionario para la evaluación de la calidad de la atención de un Equipo de Respuesta Rápida en un Hospital Universitario de Londrina, Paraná, basado en el modelo conceptual de Donabedian (estructura-procesoresultado). La recolección de datos ocurrió durante el mes de marzo de 2012 y el proceso de ajuste del cuestionario fue desarrollado por medio de la Técnica Delphi con la participación de 15 especialistas. Al término del estudio se obtuvo un cuestionario con 37 enunciados, alcanzándose un índice de concordancia final en la investigación con valores superiores al 80% para todos los conceptos. Se espera que las contribuciones del grupo de especialistas afiancen la confiabilidad del instrumento y que el cuestionario sea utilizado en otros servicios semejantes. Las aplicaciones futuras podrán traer subsidios para evaluar mejor la calidad de los servicios de los equipos de Respuesta Rápida.

Role of complementary therapies in the understanding of primary healthcare professionals: a systematic review / Papel de prácticas complementarias en la comprensión de los profesionales de atención primaria: una revisión sistemática / Papel das práticas complementares na compreensão dos profissionais da Atenção Básica: uma revisão sistemática

Objective To identify the understanding of the healthcare professionals in relation to the role of complementary therapies in primary health care. Method Systematic review by way of the following information sources: PubMed, CINAHL, PeriEnf, AMED, EMBASE, Web of Science, Psicoinfo and Psicodoc, using the keyword Primary Health Care alone, and associated with the following keywords: Medicinal Plants, Herbal Medicine, Homeopathy, Traditional Chinese Medicine, Acupuncture, Anthroposophical Medicine. Results Twenty-two studies from 1986 to 2011 were included. We identified three styles of practice: conventional medicine, complementary therapies and integrative medicine. Positioning professional practices within these three styles may facilitate discussion of concepts of health care, enhancing the health care provided as a result. Conclusion The work process in primary care presents difficulties for conducting integrative and holistic health care, but this practice has been introduced over time by professionals who integrate conventional medicine and complementary therapies, concerned with the care and well-being of patients.
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Objetivo Identificar la comprensión de los profesionales sobre el papel de las prácticas complementarias en la atención primaria. Método Revisión sistemática. Fuentes de datos: PubMed, CINAHL, PeriEnf, AMED, EMBASE, Web of Science, Psicodoc y PysicoInfo. Descriptor Atención Primaria se asoció solo a los siguientes descriptores: plantas medicinales, fitoterapia, homeopatía, acupuntura, medicina tradicional china, medicina antroposófica. Resultados Se incluyeron 22 pesquisas entre 1986-2011. Tres estilos de práctica se identificaron: medicina convencional, medicina integrativa y Terapias Complementarias. Identificar la práctica profesional dentro de estos tres estilos puede facilitar la discusión de los conceptos de salud y la atención, mejorar la atención. Conclusión El proceso de trabajo en atención primaria presenta dificultades para realización de integración y atención integral, pero esta práctica se ha introducido con profesionales que integran medicina convencional y complementaria, ocupados con la atención y bienestar del paciente.


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Objetivo Identificar a compreensão dos profissionais de saúde quanto ao papel das práticas complementares na Atenção Básica. Método Revisão sistemática cujas fontes de informação foram: PubMed, CINAHL, PeriEnf, AMED, EMBASE, Web of Science, PysicoInfo e PsicoDoc, utilizando o descritor Atenção Básica associado, isoladamente, aos seguintes descritores: Plantas Medicinais, Fitoterapia, Homeopatia, Medicina Tradicional Chinesa, Acupuntura, Medicina Antroposófica. Resultados Incluíram-se 22 estudos entre 1986-2011. Identificaram-se três estilos de prática: medicina convencional, práticas integrativas e medicina integrativa. Posicionar a prática profissional dentro desses três estilos pode facilitar a discussão de concepções de saúde e cuidado, ampliando o cuidado. Conclusão O processo de trabalho na Atenção Básica apresenta dificuldades para a realização de cuidado integrativo e holístico, mas essa prática vem sendo introduzida com profissionais que integram medicina convencional e práticas complementares, preocupados com o cuidado e o bem-estar do paciente.

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Metallothionein-1 and nitric oxide expression are inversely correlated in a murine model of Chagas disease

Chagas disease, caused by Trypanosoma cruzi, represents an endemic among Latin America countries. The participation of free radicals, especially nitric oxide (NO), has been demonstrated in the pathophysiology of seropositive individuals with T. cruzi. In Chagas disease, increased NO contributes to the development of cardiomyopathy and megacolon. Metallothioneins (MTs) are efficient free radicals scavengers of NO in vitro and in vivo. Here, we developed a murine model of the chronic phase of Chagas disease using endemic T. cruzi RyCH1 in BALB/c mice, which were divided into four groups: infected non-treated (Inf), infected N-monomethyl-L-arginine treated (Inf L-NAME), non-infected L-NAME treated and non-infected vehicle-treated. We determined blood parasitaemia and NO levels, the extent of parasite nests in tissues and liver MT-I expression levels. It was observed that NO levels were increasing in Inf mice in a time-dependent manner. Inf L-NAME mice had fewer T. cruzi nests in cardiac and skeletal muscle with decreased blood NO levels at day 135 post infection. This affect was negatively correlated with an increase of MT-I expression (r = -0.8462, p < 0.0001). In conclusion, we determined that in Chagas disease, an unknown inhibitory mechanism reduces MT-I expression, allowing augmented NO levels.

Cadmio: efectos sobre la salud. Respuesta celular y molecular / Cadmium: effects on health. Cellular and molecular response

El cadmio (Cd) es un metal que se encuentra principalmente en la corteza terrestre y siempre se presenta en combinación con el zinc. Es ampliamente utilizado en la industria. Se considera un contaminante y es liberado al ambiente como subproducto de la extracción de cobre, hierro y zinc. La exposición al Cd puede producir una variedad de efectos adversos tanto en el humano como en los animales. Una vez absorbido se acumula en el organismo por tiempos largos. Dependiendo de la dosis, fuente y tipo de exposición puede dañar varios órganos como el hígado, riñón, pulmón, hueso, testículos y placenta. Los seres humanos están expuestos al Cd principalmente a través de la ingesta de alimentos, del humo del cigarro, así como del agua y aire contaminados con el metal. La entrada de Cd a las células no es uniforme en todos los sistemas y puede ser mediada por transporte pasivo o activo, o por canales de calcio. Se considera que uno de los mecanismos de toxicidad de este metal es debido, en parte, a las especies reactivas de oxígeno, las cuales pueden actuar como segundos mensajeros y por tanto alterar diferentes vías de señalización. Por todo lo expuesto el objetivo de esta revisión es analizar los efectos del Cd sobre la salud, así como sobre la respuesta celular y molecular.

Cadmium (Cd) is a metal found in the earth´s crust, always as part of several, mainly zinc-rich, ores. Cd is considered as an environmental pollutant, it is widely used in the industry. It coexists with other metals and its release into the environment is carried out in parallel with the release of copper, iron and zinc. Cd is known to have numerous undesirable effects on health in both humans and animals. Once absorbed, it is effciently retained in the body, where it accumulates throughout life. Depending on the dose, source and type of exposure it could damage several organs as the liver, kidney, lung, bones, testes and placenta. Impor-tant sources of human intoxication are food, cigarette smoke as well as contaminated water and air. Cd cell uptake is not uniform across all systems. This could be mediated by passive or active transport, or via calcium channels. It is known that the toxicity produced by this metal is due, in part to reactive oxygen species, which could act as second messengers that may alter different signaling cascades. The aim of this review is to analyze the effects of Cd on health, as well as on cellular and molecular response.