Heme peroxidases are responsible for the dehydrogenation and oxidation metabolism of harmaline into harmine / 中国天然药物

Harmaline and harmine are β-carboline alkaloids with effective pharmacological effects. Harmaline can be transformed into harmine after oral administration. However, enzymes involved in the metabolic pathway remain unclear. In this study, harmaline was incubated with rat liver microsomes (RLM), rat brain microsomes (RBM), blood, plasma, broken blood cells, and heme peroxidases including horseradish peroxidase (HRP), lactoperoxidase (LPO), and myeloperoxidase (MPO). The production of harmine was determined by a validated UPLC-ESI-MS/MS method. Results showed that heme peroxidases catalyzed the oxidative dehydrogenation of harmaline. All the reactions were in accordance with the Hill equation. The reaction was inhibited by ascorbic acid and excess H2O2. The transformation of harmaline to harmine was confirmed after incubation with blood, plasma, and broken blood cells, rather than RLM and RBM. Harmaline was incubated with blood, plasma, and broken cells liquid for 3 h, and the formation of harmine became stable. Results indicated an integrated metabolic pathway of harmaline, which will lay foundation for the oxidation reaction of dihydro-β-carboline. Moreover, the metabolic stability of harmaline in blood should not be ignored when the pharmacokinetics study of harmaline is carried out.

Expression of Glutathione Peroxidases and Its Effect on Clinical Prognosis in Glioma Patients / 中国医学科学院学报

Objective To investigate the relationship between the expression of glutathione peroxidase(GPX)genes and the clinical prognosis in glioma patients,and to construct and evaluate the model for predicting the prognosis of glioma. Methods The clinical information and GPX expression of 663 patients,including 153 patients of glioblastoma(GBM)and 510 patients of low-grade glioma(LGG),were obtained from The Cancer Genome Atlas(TCGA)database.The relationship between GPX expression and patient survival was analyzed.The key GPX affecting the prognosis of glioma was screened out by single- and multi-factor Cox's proportional-hazards regression models and validated by least absolute shrinkage and selection operator(Lasso)regression.Finally,we constructed the model for predicting the prognosis of glioma with the screening results and then used concordance index and calibration curve respectively to evaluate the discrimination and calibration of model. Results Compared with those in the control group,the expression levels of GPX1,GPX3,GPX4,GPX7,and GPX8 were up-regulated in glioma patients(all P<0.001).Moreover,the expression levels of other GPX except GPX3 were higher in GBM patients than in LGG patients(all P<0.001).The Kaplan-Meier curves showed that the progression-free survival of GBM with high expression of GPX1(P=0.013)and GPX4(P=0.040),as well as the overall survival,disease-specific survival,and progression-free survival of LGG with high expression of GPX1,GPX7,and GPX8,was shortened(all P<0.001).GPX7 and GPX8 were screened out as the key factors affecting the prognosis of LGG.The results were further used to construct a nomogram model,which suggested GPX7 was the most important variable.The concordance index of the model was 0.843(95%CI=0.809-0.853),and the calibration curve showed that the predicted and actual results had good consistency. Conclusion GPX7 is an independent risk factor affecting the prognosis of LGG,and the nomogram model constructed with it can be used to predict the survival rate of LGG.

Relationship between glutathione peroxidases family and survival prognosis in patients with colorectal cancer / 国际肿瘤学杂志

Objective:To investigate the relationship between glutathione peroxidases (GPXs) gene expression in colorectal cancer tissues and survival prognosis, and to construct and evaluate a nomogram prediction model of GPXs for survival prognosis of colorectal cancer patients.Methods:The GPXs gene expresion data and other clinical data of 620 patients with colorectal cancer (455 cases of colon cancer and 165 cases of rectal cancer) were downloaded from The Cancer Genome Atlas (TCGA) database, and the GPXs gene expression data of 820 normal people were downloaded as controls, preprocessed by R language, and the gene expression data were analyzed for differential expression. Spearman rank correlation was used to analyze the correlation between GPXs gene expression and tumor mutation burden (TMB) in colorectal cancer tissues. Cox risk regression model was used to analyze the influencing factors of survival and prognosis of colorectal cancer patients. Nomogram models were constructed to predict overall survival (OS) of colon cancer and rectal cancer patients, and its predictive performance was evaluated by calibration curve.Results:In the GPXs family, there were statistically significant differences in the mRNA expressions of GPX1, GPX2, GPX3, GPX4, GPX5, GPX7 and GPX8 between colon cancer patients and normal population, and the mRNA expressions of GPX1, GPX2, GPX4 and GPX8 in colon cancer patients were higher than those in normal population (all P<0.05) . There were statistically significant differences in the mRNA expressions of GPX1, GPX2, GPX3, GPX4, GPX7 and GPX8 between rectal cancer patients and normal population, and the mRNA expressions of GPX1, GPX2, GPX4, GPX7 and GPX8 in rectal cancer patients were higher than those in normal population (all P<0.05) . Spearman rank correlation analysis showed that GPX2 ( r s=-0.27, P<0.001) and GPX7 ( r s=-0.11, P=0.043) expressions were negatively correlated with TMB in colon cancer. There were no significant correlations between GPXs genes expressions and TMB in rectal cancer tissues (all P>0.05) . In colon cancer, univariate analysis showed that GPX3 ( HR=1.22, 95% CI: 1.05-1.43, P=0.012) , GPX4 ( HR=1.39, 95% CI: 1.01-1.92, P=0.045) , age ( HR=1.02, 95% CI: 1.01-1.04, P=0.010) and pTNM-stage ( HR=1.78, 95% CI: 1.43-2.21, P<0.001) were the influencing factors of OS. Multivariate analysis showed that GPX4 ( HR=1.96, 95% CI: 1.09-3.51, P=0.024) , age ( HR=1.02, 95% CI: 1.00-1.04, P=0.042) and pTNM-stage ( HR=1.61, 95% CI: 1.21-2.15, P=0.001) were the independent risk factors of OS. In rectal cancer, univariate analysis showed that age ( HR=1.08, 95% CI: 1.04-1.13, P<0.001) was the influencing factor of OS. Multivariate analysis showed that GPX7 ( HR=0.44, 95% CI: 0.22-0.88, P=0.020) , GPX8 ( HR=3.17, 95% CI: 1.63-6.17, P=0.001) and age ( HR=1.10, 95% CI: 1.04-1.16, P=0.001) were the independent influencing factors of OS. The consistency index (C-index) of the nomogram model for predicting OS in patients with colon cancer and rectal cancer were 0.71 (95% CI: 0.63-0.79) and 0.88 (95% CI: 0.82-0.94) respectively. The calibration curve showed that the prediction curve of the two models had a good fit with the real curve. Conclusion:GPX4 is an independent risk factor affecting the prognosis of colon cancer patients. Patients with high GPX4 expression have a poor prognosis. GPX7 and GPX8 are independent prognostic factors for rectal cancer patients, and the rectal cancer patients with low GPX7 expression and high GPX8 expression have poor prognosis. The nomogram constructed based on the above factors can better predict the prognosis of patients with colon cancer and rectal cancer.

Expression of glutathione peroxidases 4 in colon adenocarcinoma tissues and its relationship with clinicopathological features and prognosis of patients / 肿瘤研究与临床

Objective:To investigate the expression of glutathione peroxidases 4 (GPX4) in colon adenocarcinoma and its relationship with clinicopathological features and prognosis of patients.Methods:The data set of colon adenocarcinoma was obtained from The Cancer Genome Atlas (TCGA) database to analyze the expression of GPX4 in colon adenocarcinoma tissues and its predictive value for overall survival (OS). A total of 93 colon adenocarcinoma tissues and 87 adjacent mucosa tissues after operation from November 2009 to May 2010 provided by the National Human Genetic Resources Sharing Service Platform were selected. The expression of GPX4 protein was detected by using tissue chip immunohistochemistry. The relations between the expression of GPX4 protein and the clinicopathological features and OS of colon adenocarcinoma patients were analyzed. Cox proportional hazards regression model was used to analyze the factors affecting the prognosis. The nomogram for predicting OS rate was established and drawn.Results:The analysis of data from TCGA database showed that in 380 cases of colon adenocarcinoma, the expression of GPX4 in colon adenocarcinoma tissues were higher than that in the normal colonic mucosa tissues [the value of fragments per kilobase of exon per million fragments mapped (FPKM): 85.654 (20.351-356.237) vs. 56.230 (48.783-63.931)], and the difference was statistically significant ( Z = -6.150, P<0.05). The OS in GPX4 high-expression group (FPKM ≥83.614) were poorer than that in GPX4 low-expression group (FPKM < 83.614) (median OS time: 84.40 months vs. 94.03 months, 5-year OS rate: 58.6% vs. 72.7%), and the difference was statistically significant ( P<0.05). Tissue chip immunohistochemical staining results show that the high-expression rate of GPX4 protein in colon adenocarcinoma tissues was higher than that in adjacent normal tissues [38.0% (35/92) vs. 7.3% (6/82)], and the difference was statistically significant ( χ2 = 22.727, P<0.01); the high-expression rate of GPX4 protein in left colon adenocarcinoma tissues was higher than that in right colon adenocarcinoma tissues [47.2% (25/53) vs. 25.6% (10/39), and the difference was statistically significant ( χ2 = 4.42, P = 0.036); the 5-year OS rate of patients in GPX4 high-expression group was lower than that in GPX4 low-expression group (25.7% vs. 57.9%), and the difference was statistically significant ( χ2 = 9.051, P<0.05). Multivariate Cox proportional hazards regression model analysis showed that lymph node metastasis (stage N 1-N 3) ( HR = 2.241, 95% CI 1.242-4.046, P = 0.007) and high expression of GPX4 ( HR = 2.783, 95% CI 1.598-4.848, P<0.01) were independent factors affecting the poor prognosis of colon adenocarcinoma patients. The above factors were used to establish a nomogram for predicting the prognosis of patients with colon adenocarcinoma, the C index was 0.739, indicating that the nomogram had good predictive performance. Conclusion:The expression of GPX4 is up-regulated in colon adenocarcinoma tissues, and its high expression is related to the malignant biological behavior of the tumor and poor prognosis.

Influence of root-knot nematode infestation on antioxidant enzymes, chlorophyll content and growth in Pogostemon cablin (Blanco) Benth.

Plants adapt themselves to overcome adverse environmental conditions, and this involves a plethora of concurrent cellular activities. Physiological experiments or metabolic profiling can quantify this response. Among several diseases of Pogostemon cablin (Blanco) Benth. (Patchouli), root-knot nematode infection caused by Meloidogyne incognita (Kofoid and White) Chitwood causes severe damage to the plant and hence, the oil production. In the present study, we identified M. incognita morphologically and at molecular level using sequenced characterized amplified region marker (SCAR). M. incognita was artificially inoculated at different levels of second stage juveniles (J2) to examine the effect on Patchouli plant growth parameters. Peroxidase and polyphenol oxidase enzyme activity and changes in the total phenol and chlorophyll contents in M. incognita was also evaluated in response to infection. The results have demonstrated that nematode infestation leads to increased peroxidase activities in the leaves of the patchouli plants and thereby, increase in phenolic content as a means of defence against nematode infestation. Chlorophyll content was also found decreased but no changes in polyphenol oxidase enzyme activity.

Mannitol-induced drought stress on calli of Trigonella foenum-graecum L. Var. RMt-303.

Different explants of fenugreek, T. foenum-graecum L. (Var. RMt-303), were compared for their callus induction and subsequent shoot regeneration capabilities on Murashige and Skoog media supplemented with different phytohormones in varying concentration. The highest percentage of callus induction frequency was observed in 1ppm benzylaminopurine (BAP). Maximum shoots were induced on media supplemented with 0.5ppm BAP using leaf and stem tissues as explants. However, root tissues showed only callusing with no subsequent shooting. Cotyledonary node responded better than hypocotyls in terms of shoot induction on media supplemented with thidiazuron (0.1ppm). The callus was subjected to drought stress as simulated by reduced water potential of growth media due to addition of mannitol. Calli could withstand -2 MPa water potential till 30 days indicating that the drought stress tolerance mechanisms are functional in this variety. Chlorophyll a and b and total chlorophyll, proline and total phenolic contents, total peroxidase and catalase activities increased under stress conditions suggesting the tolerance of callus to drought stress. However, ascorbate peroxidase, guaiacol peroxidase activities were found to decrease slightly. Malondialdehyde and H2O2 contents were found to decrease while only a slight disturbance was found in membrane stability index. These results underline the mechanisms that are crucial for drought stress tolerance in fenugreek.

Descoloração de azul de bromofenol utilizando peroxidase imobilizada em pó de sabugo de milho altamente ativado / Bromofhenol blue discoloration using peroxidase immobilized on highly activated corncob powder

The aim of the present study was to evaluate the efficacy of peroxidase immobilized on corncob powder for the discoloration of dye. Peroxidase was extracted from soybean seed coat, followed by amination of the surface of the tertiary structure. The aminated peroxidase was immobilized on highly activated corncob powder and employed for the discoloration of bromophenol blue. Amination was performed with 10 or 50 mmol.L-1 carbodiimide and 1 mol.L-1 ethylenediamine. The amount of protein in the extract was 0.235 ± 0.011 mg.mL-1 and specific peroxidase activity was 86.06 ± 1.52 μmol min-1. mg -1 , using 1 mmol.L-1 ABTS as substrate. Ten mmol.L-1 and 50 mmol.L-1 aminated peroxidase retained 88 and 100% of the initial activity. Following covalent immobilization on a corncob powder-glyoxyl support, 10 and 50 mmol.L-1 aminated peroxidase retained 74 and 86% of activity, respectively. Derivatives were used for the discoloration of 0.02 mmol.L-1 bromophenol blue solution. After 30 min, 93 and 89% discoloration was achieved with the 10 mmol.L-1 and 50 mmol.L-1 derivatives, respectively. Moreover, these derivatives retained 60% of the catalytic properties when used three times. Peroxidase extracted from soybean seed coat immobilized on a low-cost corncob powder support exhibited improved thermal stability.

Nesta pesquisa a enzima peroxidase foi extraída do tegumento de sementes de soja, e a superfície da estrutura terciária foi aminada. A peroxidase aminada foi imobilizada em suporte pó de sabugo de milho altamente ativado e utilizado na descoloração de azul de bromofenol. A aminação da peroxidase foi realizada com carbodiimida em concentrações de 10 e 50 mmol.L-1 , e 1 mol.L-1 de etilenodiamina. A quantidade de proteínas no extrato foi de 0,235 ± 0,011 mg.mL-1 , e a atividade específica da peroxidase foi 86,06 ± 1,52 μmol min-1 .mg-1, usando 1 mmol.L-1 de ABTS como substrato. A peroxidase aminada a 10 mmol.L-1 reteve 88% e a aminada a 50 mmol.L-1 reteve 100% da atividade inicial. As peroxidases aminadas a 10 ou 50 mmol.L-1 foram covalentemente imobilizadas em suporte glioxil-pó de sabugo de milho com atividade recuperada de 74% e 86%, respectivamente. Os derivados obtidos foram utilizados na descoloração de solução de azul de bromofenol 0,02 mmol.L-1 . Após 30 min 93% de descoloração foram alcançados com o derivado glioxil-pó de sabugo de milho com a peroxidase aminada 10 mmol.L-1 e 89% com a aminada 50 mmol.L-1 . Estes derivados mantiveram 60% das propriedades catalíticas, quando utilizado por três vezes. A peroxidase extraída do tegumento da semente de soja imobilizada em suporte de baixo custo pó de sabugo de milho apresentou melhoria na estabilidade térmica da enzima.

Influence of pH on the growth, laccase activity and RBBR decolorization by tropical basidiomycetes

The basidiomycete fungi Lentinus crinitus and Psilocybe castanella are being evaluated in a bioremediation process of soils contaminated with organochlorine industrial residues in the Baixada Santista, São Paulo. The aim of the present study was to determine the influence of pH on the fungal growth, in vitro decolorization of anthraquinonic dye Remazol Brilliant Blue R (RBBR) and laccase activity. The pH of the culture medium influenced the growth of L. crinitus and P. castanella, which presented less growth at pH 5.9 and pH 2.7, respectively. The fungi were able to modify the pH of the culture medium, adjusting it to the optimum pH for growth which was close to 4.5. Decolorization of the RBBR was maximal at a pH of 2.5 to 3.5. Higher laccase activity was observed at pH 3.5 and pH 4.5 for L. crinitus and P. castanella, respectively. pH was found to be an important parameter for both the growth of these fungi and the enzymatic system involved in RBBR decolorization.

Os fungos basidiomicetos Lentinus crinitus e Psilocybe castanella estão sendo avaliados em processo de biorremediação de solos contaminados com resíduos industriais organoclorados, na Baixada Santista, SP. O presente estudo avaliou a influência do pH no crescimento, na descoloração in vitro do corante Azul Brilhante de Remazol R (RBBR) e na atividade de lacase durante cultivo destes fungos, de forma a subsidiar a otimização do processo. O pH do meio influenciou o crescimento de L. crinitus e de P. castanella, com menor biomassa em pH 5,9 e pH 2,7, respectivamente. Os fungos foram capazes de modificar o pH inicial do meio de cultura, de modo a ajustá-lo ao valor ótimo de crescimento, próximo a 4,5. Descoloração in vitro do RBBR foi máxima em pH 2,5 e 3,5. Maiores atividades de lacase foram obtidas em pH 3,5 e em pH 4,5 para L. crinitus e P. castanella, respectivamente. Evidenciou-se que o pH é um parâmetro importante para o crescimento destes fungos, atividade de lacase e descoloração in vitro do RBBR.

Efeito do L-Name na expressão de iNOS, MPO e na perda óssea alveolar em ratos diabéticos com periodontite experimental / Effect of L-NAME on iNOS, MPO expression and alveolar bone loss in diabetic rats with experimental periodontitis

Mediadores inflamatórios como mieloperoxidase (MPO) e óxido nítrico (NO) participam do processo inflamatório da doença periodontal e na associação ao diabetes. Inibidores da Óxido Nítrico Sintase (NOS), como o L-NAME têm sido administrados na tentativa de minimizar danos teciduais decorrentes da inflamação. O objetivo deste estudo foi avaliar o efeito do L-NAME sobre os níveis da isoforma induzível de óxido nítrico (iNOS), sobre a perda óssea alveolar e sobre os níveis de mieloperoxidase (MPO) em ratos diabéticos com periodontite induzida. Foram utilizados 192 ratos divididos em grupos de 24 animais cada: grupo C: Controle com ingestão de água; grupo C-L: Controle com ingestão de LNAME; grupo D: Ratos diabéticos com ingestão de água; grupo D-L: Ratos diabéticos com ingestão de L-NAME; grupo P: Ratos com periodontite experimental e ingestão de água; grupo P-L: Ratos com periodontite experimental e ingestão de L-NAME; grupo DP: Ratos diabéticos com periodontite experimental e ingestão de água; e grupo DP-L: Ratos diabéticos com periodontite experimental e ingestão de L-NAME. O sacrifício dos animais foram realizados aos 3,7,15,e 30 dias após a indução da periodontite experimental. Foram realizadas análises da taxa glicêmica, da perda óssea alveolar, da atividade de mieloperoxidase e da expressão de iNOS. Os grupos com D, P e DP mostraram níveis de iNOS estatisticamente mais altos quando comparados aos grupos D-L, P-L e DP-L que ingeriram L-NAME em todos os períodos. (p < 0.05). Os grupos que ingeriram L-NAME mostraram perda óssea estatisticamente menor quando comparados aos grupos que ingeriram água (p < 0.05). A alta expressão de MPO foi observada nos grupos com periodontite P e DP que ingeriram água. Foi verificado um pico nos níveis de MPO aos 7 dias em todos os grupos experimentais que ingeriram água...

Inflammatory mediators such as myeloperoxidase (MPO) and nitric oxide (NO) play a role in inflammatory processes related to periodontal disease and diabetes. NO-Synthase (NOS) inhibitors as L-NAME have been administered in attempts to reduce tissue damage resulting from such inflammation. The aim of this study was to evaluate the effects of L-NAME on alveolar bone loss, NO and MPO levels in diabetic rats with experimental periodontitis. A hundred ninety-two rats were divided into one of the following groups with 24 animals each: group C-W: control group with water intake; group C-L: control group with L-NAME intake; group D-W: diabetic rats with water intake; group D-L: diabetic rats with LNAME intake; group P-W: chronic periodontitis rats with water intake; group PL: chronic periodontitis rats with L-NAME intake; group DP-W: diabetic chronic periodontitis rats with water intake, and group DP-L: diabetic chronic periodontitis rats with L-NAME intake. The killing was performed at 3, 7, 15 and 30 days after ligature-periodontitis induction to obtain gingival specimens and to evaluate MPO and NO activity and radiographic bone loss. Groups with D-W or P-W and DP-W showed statistically higher iNOS expression compared to LNAME groups in all periods (p < 0.05). L-NAME treatment (L) statistically decreased iNOS expression in all groups with inflammatory pathological conditions, such as D, P and DP. In general, groups with L-NAME intake showed lower bone loss compared to water intake groups. In addition, P-L or DP-L rats demonstrated statistically lower bone loss compared to diabetics and controls at 30 days (p < 0.05). The highest MPO expression was verified in periodontitis groups with water intake (P-W e DP-W). There was a peak in MPO levels at 7 days in all experimental groups with water intake during experimental evolution. The L-NAME statistically decreased MPO levels from all groups...

Defense mechanisms of Solanum tuberosum L. in response to attack by plant-pathogenic bacteria

The natural resistance of plants to disease is based not only on preformed mechanisms, but also on induced mechanisms. The defense mechanisms present in resistant plants may also be found in susceptible ones. This study attempted to analyze the metabolic alterations in plants of the potato Solanum tuberosum L. cv. Agata that were inoculated with the incompatible plant-pathogenic bacteria X. axonopodis and R. solanacearum, and the compatible bacterium E. carotovora. Levels of total phenolic compounds, including the flavonoid group, and the activities of polyphenol oxidase (PPO) and peroxidase (POX) were evaluated. Bacteria compatibility was evaluated by means of infiltration of tubers. The defense response was evaluated in the leaves of the potato plants. Leaves were inoculated depending on their number and location on the stem. Multiple-leaf inoculation was carried out on basal, intermediate, and apical leaves, and single inoculations on intermediate leaves. Leaves inoculated with X. axonopodis and with R. solanacearum showed hypersensitive responses within 24 hours post-inoculation, whereas leaves inoculated with E. carotovora showed disease symptoms. Therefore, the R. solanacearum isolate used in the experiments did not exhibit virulence to this potato cultivar. Regardless of the bacterial treatments, the basal leaves showed higher PPO and POX activities and lower levels of total phenolic compounds and flavonoids, compared to the apical leaves. However, basal and intermediate leaves inoculated with R. solanacearum and X. axonopodis showed increases in total phenolic compounds and flavonoid levels. In general, multiple-leaf inoculation showed the highest levels of total phenolics and flavonoids, whereas the single inoculations resulted in the highest increase in PPO activity. The POX activity showed no significant difference between single- and multiple-leaf inoculations. Plants inoculated with E. carotovora showed no significant increase ...

Efeito do tratamento periodontal não-cirúrgico em pacientes portadores de diabetes mellitus tipo 2 com doença periodontal: análises clínica, enzimática e microbiológica / Effect of the non-surgical periodontal treatment in type 2 diabetes patients with periodontal disease: clinical, enzymatic and microbiological analysis

Avaliar o efeito do tratamento periodontal não-cirúrgico sobre parâmetros clínicos, microbiota subgengival e a atividade enzimática na saliva e no fluido sulcular gengival em pacientes portadores de diabetes e em indivíduos sistemicamente sadios, ambos com periodontite crônica. Material e método: a amostra populacional foi composta por 40 indivíduos divididos em dois grupos: 20 portadores de Diabetes mellitus tipo 2 com inadequado controle metabólico (grupo DM2) e 20 indivíduos sistemicamente sadios (grupo controle), ambos portadores de periodontite crônica. Clinicamente os indivíduos foram avaliados quanto: ao índice de placa visível (IPV), ao índice de sangramento marginal (ISM), à profundidade de sondagem (PS), ao nível clínico de inserção (NI), à presença de sangramento à sondagem (SS) e à presença de supuração (SUP). Foram realizadas coletas de saliva e de fluido sulcular gengival para determinação da atividade peroxidásica total salivar (PTS) e da mieloperoxidase (MPO), por meio de espectrofotometria. Amostras de placa bacteriana subgengival de 4 sítios rasos e 4 profundos com doença periodontal foram avaliadas pela técnica de Checkerboard DNA-DNA hybridization. Toda a amostra recebeu terapia periodontal não-cirúrgica com posterior fase de acompanhamento para realização de controle de placa profissional a cada quinze dias por um período de três meses. Todas as avaliações foram realizadas no baseline e aos três meses após o término da terapia periodontal. Resultados: No baseline, o grupo DM2 apresentou maior percentual de sítios com IPV e SS (p< 0.01) sem diferença estatisticamente significante entre grupos quanto aos demais parâmetros clínicos. O tratamento periodontal não-cirúrgico resultou em melhora significativa dos parâmetros clínicos avaliados nos 2 grupos. Aos três meses pós-tratamento foram observados maiores...

To evaluate the effect of periodontal treatment on clinical parameters as well as on subgingival microbiota and enzimatic activity in the saliva and the gingival crevicular fluid of diabetes patients and systemically healthy individuals with chronic periodontitis. Material and Methods: The sample was constituted by 40 individuals divided in two groups: 20 type 2 diabetes mellitus patients with inadequate metabolic control (DM2 group) and 20 systemically healthy individuals (control group), both groups with chronic periodontitis. Clinical assessments included visible plaque index (VPI), bleeding on probing (BOP), gingival bleeding index (GBI), suppuration (SUP), probing depth (PD) and clinical attachment level (CAL). Saliva and gingival crevicular fluid samples were collected for determination of the total salivary peroxidase (TPS) activity and myeloperoxidase (MPO) activity respectively, by spectrophotometric assays. Subgingival plaque samples from 4 shallow and 4 deep sites with periodontal disease were evaluated by the Checkerboard DNA-DNA hybridization technique. All the patients received non-surgical periodontal therapy followed of the period for accomplishment of professional plaque control performed twice a month during three months. All the evaluations were assessed at baseline and 3 months after periodontal therapy. Results: At baseline, the DM2 group presented a significantly higher percentage of sites with VPI and BOP (p<0.01). No significant differences between groups were observed for the other clinical parameters. Nonsurgical periodontal treatment resulted in a significant improvement of clinical parameters. At 3 months post-treatment, the DM2 group presented significantly higher percentage of sites with VPI, GBI, BOP and mean PD (p<0.05). Regarding the microbiological analysis, at baseline, only C. rectus and S. anginosus were founded significantly higher...

Predictive value of myeloperoxidase to identify high risk patients admitted to the hospital with acute chest pain / Valor preditivo da mieloperoxidase na identificação de pacientes de alto risco admitidos por dor torácica aguda

FUNDAMENTO: A mieloperoxidase (MPO) é uma enzima intensamente expressa diante da ativação leucocitária, com múltiplas ações aterogênicas, incluindo a oxidação do colesterol (LDL), e relacionada à instabilização da placa aterosclerótica. É preditora de eventos adversos em indivíduos sadios, coronariopatas ou em investigação de dor torácica. OBJETIVO: Analisar a contribuição da MPO na identificação de pacientes com dor torácica aguda, eletrocardiograma (ECG) sem elevação de segmento ST e com alto risco para eventos adversos intra-hospitalares. MÉTODOS: O nível sérico da MPO foi mensurado na admissão de pacientes com dor torácica aguda, ECG sem elevação de segmento ST e submetidos a protocolo estruturado de investigação. RESULTADOS: De uma coorte de 140 pacientes, 49 (35 por cento) receberam o diagnóstico de síndrome coronariana aguda, tendo sido estabelecido diagnóstico de infarto agudo do miocárdio (troponina I > 1,0 ng/ml) sem elevação de ST em 13 pacientes (9,3 por cento). O melhor ponto de discriminação da MPO para infarto agudo do miocárdio foi identificado em > 100 pM pela curva ROC (AUC = 0,662; IC 95 por cento = 0,532-0,793), que demonstrou elevada sensibilidade (92,3 por cento) e elevado valor preditivo negativo (98,1 por cento), embora com baixa especificidade (40,2 por cento). Na análise multivariada, a MPO mostrou-se a única variável independente para o diagnóstico de infarto agudo do miocárdio em evolução, com razão de chance de 8,04 (p = 0,048). CONCLUSÃO: Em pacientes com dor torácica aguda e sem elevação de ST, a MPO admissional elevada é importante ferramenta preditiva de eventos adversos intra-hospitalares, com razão de chance de oito vezes para o diagnóstico de infarto agudo do miocárdio.

BACKGROUND: Myeloperoxidase (MPO) is a highly expressed enzyme due to leukocyte activation, with multiple atherogenic actions, including LDL cholesterol oxidation, and is related to the instability of atherosclerotic plaque. It is a predictor of adverse events in healthy individuals, patients with heart disease or those undergoing chest pain investigations. OBJECTIVE: To analyze the contribution of MPO to identify patients with acute chest pain, non-ST elevation ECG and at high risk for in-hospital adverse events. METHODS: Patients presenting acute chest pain and a non-ST elevation ECG, were admitted to the hospital and submitted to serum MPO level measurements and a structured examination protocol. RESULTS: From a cohort of 140 patients, 49 (35 percent) were diagnosed with acute coronary syndrome, of which 13 patients (9.3 percent) were diagnosed with non-ST elevation acute myocardial infarction (AMI) (troponin I >1.0 ng/mL). The best MPO cut-off point for AMI was identified as >100 pM using the ROC curve (AUC=0.662; CI 95 percent=0.532-0.793) revealing elevated sensitivity (92.3 percent) and negative predictive value (98.1 percent), however with low specificity (40.2 percent). In the multivariate analysis, MPO proved to be the only independent variable to diagnose AMI in evolution, with an odds ratio of 8.04 (p=0.048). CONCLUSION: In patients with acute chest pain and no ST elevation, high MPO levels upon admission to the hospital are an important tool to predict in-hospital adverse events, with an odds ratio of eight for the diagnosis of AMI.

Application value of serum thyroid peroxidase in screening thyroid dysfunction in pregnancy / 国际检验医学杂志

Objective To research the clinical value of serum thyroid peroxidase (TPOAb) in screening thyroid dysfunction in pregnancy.Methods The concentration of serum TPOAb and TSH(thyroid-stimulating hormone) was detected by MEIA and CMIA in 75 cases of pregnant women with thyroid disfunction,and 145 cases of pregnant women without tyhroid dysfunction (as control group).Results There were significant differences in both TSH and TPOAb levels between pregnant women with and without thyroid dysfunction(P0.05). So the normal serum TSH could not completely exclude the thyroid dysfunction in pregnant women.Conclusion TPOAb can be used as a prenatal screening marker for early diagnosis of thyroid dysfunction in pregnancy. From the diagnostic point, TSH can not completely substitute for TPOAb and the abnormal TPOAb can indicate the possibility and risk for autoimmune thyroid disease in pregnant women.

Preparación de un conjugado peroxidasa-anti IgG humana (cadena ) en conejo / Preparation of a peroxidase-anti human IgG conjugate (chain g) in rabbit

Se preparó un conjugado con peroxidasa a partir de los anticuerpos específicos aislados de un suero de conejo anti cadenas g de la IgG humana. Los anticuerpos específicos se aislaron por cromatografía de afinidad, y el conjugado se preparó por el método de oxidación con peryodato. El conjugado obtenido presentó una relación molar IgG/peroxidasa de 1,07, un valor de RZ de 0,33 y resultó evaluado satisfactoriamente en cuanto a su especificidad y reactividad en los ensayos inmunoenzimáticos realizados.

A peroxidase conjugate was prepared starting from the specific antibodies isolated from an anti-chain rabbit serum and from human IgG. The specific antibodies were isolated by affinity chromatography and the conjugate was prepared by the method of oxidation with periodate. The conjugate obtained presented a molar IgG/peroxidase relation of 1.07, a RZ value of 0.33 and it was satisfactorily evaluated as regards its specificity and reactivity in the immunoenzimatic assays carried out.

An Experimental Study of Pathological Changes of Thyroid after Severe Burns and Their Mechanisms in Rats / 第三军医大学学报

Thirty percent TBSA full thickness burns due to napalm was inflicted to the rats and the pathological changes of the thyroid were studied under optical and electron microscopy and with peroxidase histochemistry for 15 days. The alterations of serum T3 and T4 levels accompaniying the pathological changes were also observed dynamically.It was found that the pathological changes of the thyroid could be distinguished into three phases as follows.1. The phase of injury inhibition. This phase existed from the 2nd to the 24th hour postburn and was characterized by marked damage on the follicular epithelium.2. The phase of structural and functional recovery. This phase entended from the 3rd to the 6th day postburn. In this phase, the damages on the thyroid became abated and its damaged structure showed a tendency to recover.3. The phase of active function. It extended from the 10th to the 15th day postburn. The organelles to synthesize and release thyroid hormone were fonud well developed.The alteration of T4 run parallelly to the pathological changes of the thyroid in all the stages, while T3 level was continously inhibited because of the lower serum concentration of T4, the metabolic disturbance of T4 deiodination, and the inhibition of thyroid secretion in the early postburn stage and the metabolic disturbance of T4 deiodination in the middle and late postburn stages. In the early postburn stage, the inhibition of thyroid peroxidase was an important factor of functional inhibition of the thyroid.