Blood-based gene expression profile of oxidative stress and antioxidant genes for identifying surrogate markers of liver tissue injury in chronic hepatitis C patients.

Oxidative stress is the process by which reactive molecules and free radicals are formed in cells. In this study, we report the blood-based gene expression profile of oxidative stress and antioxidant genes for identifying surrogate markers of liver tissue in chronic hepatitis C (CHC) patients by using real-time PCR. A total of 144 untreated patients diagnosed with CHC having genotype 3a and 20 healthy controls were selected for the present study. Liver biopsy staging and grading of CHC patients were performed using the METAVIR score. Total RNA was extracted from liver tissue and blood samples, followed by cDNA synthesis and real-time PCR. The relative expression of genes was calculated using the ΔΔCt method. The expression profile of 84 genes associated with oxidative stress and antioxidants was determined in liver tissue and blood samples. In liver tissue, 46 differentially expressed genes (upregulated, 27; downregulated, 19) were identified in CHC patients compared to normal samples. In blood, 61 genes (upregulated, 51; downregulated; 10) were significantly expressed in CHC patients. A comparison of gene expression in liver and whole blood showed that 20 genes were expressed in a similar manner in the liver and blood. The expression levels of commonly expressed liver and blood-based genes were also correlated with clinical factors in CHC patients. A receiver operating curve (ROC) analysis of oxidative stress genes (ALB, CAT, DHCR24, GPX7, PRDX5, and MBL2) showed that infections in patients with CHC can be distinguished from healthy controls. In conclusion, blood-based gene expression can reflect the behavior of oxidative stress genes in liver tissue, and this blood-based gene expression study in CHC patients explores new blood-based non-invasive biomarkers that represent liver damage.

Substrate specificity of IgGs with peroxidase and oxidoreductase activities from sera of patients with systemic lupus erythematosus and multiple sclerosis.

The analysis of IgGs to protect humans from oxidative stress through oxidation of harmful compounds was carried out. We have compared here for the first time peroxidase (in the presence of H2 O2 ) and oxidoreductase (in the absence of H2 O2 ) activities of IgGs from sera of healthy humans and patients with systemic lupus erythematosus (SLE) and multiple sclerosis (MS). In addition, substrate specificity of SLE and MS IgG preparations in the oxidation of different compounds was analyzed: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 3,3'-diaminobenzidine (DAB), homovanillic acid (HVA), o-phenylenediamine (OPD), α-naphthol, 3-amino-9-ethylcarbazole (AEC), p-hydroquinone (pHQ), and adrenaline. IgGs of healthy humans and SLE and MS patients oxidized DAB, ABTS, and OPD due to their peroxidase and oxidoreductase activities, while other compounds were substrates of IgGs only in the presence of H2 O2 : adrenaline was not oxidized by both activities of IgGs. The average SLE IgGs peroxidase activity increased statistically significant in comparison with abzymes from healthy humans in the order (-fold): OPD (1.2) <  DAB (1.7) < α-naphtol (2.2) ≤ AEC (2.4) < ABTS (4.5) < 5-ASA (10.6), while with oxidoreductase activity: OPD (1.8) ≤ DAB (2.1-fold) < ABTS (5.0). Only HVA was oxidized by IgGs with peroxidase activity of healthy donors faster than by SLE (1.3-fold) and MS abzymes (2.4-fold). In the oxidation of several substrates, only three IgGs of MS patients were used. The data speak of a tendency to increase the peroxidase and oxidoreductase activities of MS IgGs in comparison with healthy donors, but to a lesser extent: OPD (1.1 to 1.2-fold) ≤ ABTS (1.2 to 1.8-fold). It was shown that development of SLE and MS leads to increase in peroxidase and oxidoreductase activities of IgGs toward most of classical substrates. Thus, abzymes can serve as an additional factor of reactive oxygen species detoxification protecting of patients with SLE and MS from some harmful compounds somewhat better than healthy peoples.

Dietary tryptophan deficiency and its supplementation compromises inflammatory mechanisms and disease resistance in a teleost fish.

Tryptophan participates on several physiological mechanisms of the neuroendocrine-immune network and plays a critical role in macrophages and lymphocytes function. This study intended to evaluate the modulatory effects of dietary tryptophan on the European seabass (Dicentrarchus labrax) immune status, inflammatory response and disease resistance to Photobacterium damselae piscicida. A tryptophan deficient diet (NTRP); a control diet (CTRL); and two other diets supplemented with tryptophan at 0.13% (TRP13) and 0.17% (TRP17) of feed weight were formulated. Fish were sampled at 2 and 4 weeks of feeding and the remaining were i.p. injected with Phdp (3 × 106 cfu/fish) at 4 weeks and the inflammatory response (at 4, 24, 48 and 72 hours post-infection) as well as survival were evaluated. Results suggest that fish immune status was not altered in a tryptophan deficient scenario whereas in response to an inflammatory insult, plasma cortisol levels increased and the immune cell response was compromised, which translated in a lower disease resistance. When dietary tryptophan was offered 30% above its requirement level, plasma cortisol increased and, in response to bacterial infection, a decrease in lymphocytes, monocytes/macrophages and several immune-related genes was observed, also compromising at some degree fish disease resistance.

Effects of orange peels derived pectin on innate immune response, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus) cultured under indoor biofloc system.

The present study investigates the effects of orange peels derived pectin (OPDP) on skin mucus and serum immune parameters, disease resistance and growth performance of O. niloticus cultured under indoor biofloc system. Six hundred Nile tilapia (average weight 9.09 ±â€¯0.05 g) were distributed into 15 fiber tanks (300 L per tank) assigned to five treatments repeated in triplicate. Fish were fed experimental diets contain different levels OPDP as follows: 0 (control in clear water), 0 (control in biofloc system), 5, 10, and 20 g kg-1 OPDP for 8 weeks. At weeks 4 and 8 post feeding, skin mucus lysozyme (SMLA), peroxidase activities (SMPA), serum lysozyme (SL), serum peroxidase (SP), alternative complement (ACH50), phagocytosis (PI), and respiratory burst activities (RB) as well specific growth rate (SGR), weight gain (WG), final weight (FW), and feed conversion ratio (FCR) were measured. Also, resistance against Streptococcus agalactiae was assessed after 8 weeks post-feeding. Nile tilapia fed OPDP supplemented diets had significantly higher SMLA and SMPA compared to the controls (P < 0.05). The maximum values were observed in tilapia fed 10 g kg-1 OPDP followed by 5 and 20 g kg-1 OPDP. Nevertheless, no significant differences were observed between these two supplemented diets and between the control groups (P > 0.05). Regarding the serum immunological parameters, dietary inclusion of 10 g kg-1 OPDP showed significant higher SL and PI than other supplemented groups and control groups (P < 0.05). However, no significant differences were observed in SL and PI of fish fed 5 and 20 g kg-1 OPDP (P > 0.05). Dietary administration of OPDP significantly increased SP and ACH50 compared to the controls (P < 0.05), regardless of inclusion level. Additionally, non-significant change was found in RB of OPDP fed fish when compared with the controls (P > 0.05). The challenge test revealed that relative percent of survival (RPS) in OPDP treatments were 45.45%, 81.82%, 50%, respectively. The highest RPS was noticed in fish fed 10 g kg-1 OPDP. Furthermore, dietary administration of OPDP significantly improved SGR, WG, FW, and FCR (P < 0.05). Overall, the present findings suggested that OPDP can be taken into account as functional feed additives for O. niloticus.

Vascular peroxide 1 promotes ox-LDL-induced programmed necrosis in endothelial cells through a mechanism involving ß-catenin signaling.

BACKGROUND AND AIMS: Vascular peroxidase 1 (VPO1) plays a key role in mediation of cardiovascular oxidative injury. This study aims to determine whether VPO1 can promote programmed necrosis of endothelial cells and the underlying mechanisms. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were incubated with oxidized low-density lipoprotein (ox-LDL, 100 µg/mL) for 48 h to induce cell injury, which showed an elevation in cell necrosis (reflected by the increased propidium iodide (PI) positive-staining cells, LDH release and decreased cell viability), concomitant with an increase in programmed necrosis-relevant proteins including receptor-interacting protein kinase 1/3 (RIPK1/3), p-RIPK3 and mixed lineage kinase domain like (MLKL); these phenomena were attenuated by necrostatin-1(Nec-1) and RIPK3 siRNA. Meanwhile, VPO1 was up-regulated in ox-LDL-treated endothelial cells accompanied by a decrease in GSK-3ß activity and p-ß-catenin levels, and an elevation of ß-catenin levels; these phenomena were reversed in the presence of VPO1 siRNA or hypochlorous acid (HOCl) inhibitor; replacement of ox-LDL with HOCl could also induce endothelial programmed necrosis and activate the ß-catenin signaling; ß-catenin inhibitor could also suppress ox-LDL-induced RIPK-dependent necrosis. In hyperlipidemic patients, the plasma level of VPO1 was obviously increased concomitant with an elevation in plasma levels of RIPK1, RIPK3 and MLKL, and they were positively correlated. CONCLUSIONS: VPO1 plays an important role in promotion of endothelial programmed necrosis under hyperlipidemic conditions through activation of ß-catenin signaling. It may serve as a novel therapeutic target for prevention of endothelial dysfunction in hyperlipidemia.

[Effect of the treatment acceptance on the perinatal outcomes in women with subclinical hypothyroidism, positive thyroid gland peroxidase antibody in early pregnancy].

OBJECTIVE: To investigate if women with subclinical hypothyroidism (SCH), positive thyroid gland peroxidase antibody (TPOAb) in early pregnancy accepted treatment or not had effect on perinatal outcomes. METHODS: 15 000 pregnant women who delivered in Women and Infants Hospital of Zhengzhou from January 1, 2013 to June 30, 2014 were recruited retrospectively. Among them, 2 042 women had SCH in early pregnancy. The diagnostic standard of SCH was serum free thyroxine (FT4) between 12.91-22.35 pmol/L and TSH level between 5.22-10.00 mU/L. TPOAb level ≥ 34 U/L was defined as positive result. The 2 042 patients with SCH were divided into the treated group (1 236 cases) and the untreated group (806 cases), according to whether or not women accepted the levothyroxine treatment. Meanwhile, the 2 042 patients with SCH were divided into the TPOAb (+) treated group (1 021 cases), the TPOAb (+) untreated group (201 cases), the TPOAb (-) treated group (215 cases) and the TPOAb (-) untreated group (605 cases), according to the TPOAb result and acceptance the levothyroxine treatment. 2 000 pregnant women with normal thyroid function who delivered in the same period were selected as the control group. Perinatal outcomes were analyzed. RESULTS: (1) The incidence of SCH in early pregnancy was 13.61% (2 042/15 000). 60.53% (1 236/2 042) accepted levothyroxine treatment and 39.47% (806/2 042) did not. (2) The incidence of abortion (5.71%, 46/806), premature delivery (6.20%, 50/806), gestational hypertension disease (13.90%, 112/806), gestational diabetes mellitus (GDM; 6.58%, 53/806), fetal growth restriction (FGR; 12.28%, 99/806) and low birth weight infants (10.17%, 82/806) in the untreated group were higher than those in the treated group [3.96% (49/1 236), 4.21% (52/1 236), 10.76% (133/1 236), 4.13% (51/1 236), 8.90% (110/1 236), 7.52% (93/1 236), respectively] and the control group [3.60% (72/2 000), 4.00% (80/2 000), 10.70% (214/2 000), 3.80% (76/2 000), 9.60% (192/2 000), 7.50% (150/2 000), respectively]. The differences were statistically significant (P < 0.05). While there was no statistically significant difference in the incidence of placental abruption, anemia in pregnant women, or fetal distress among the three groups (P > 0.05). (3)The incidences of abortion (11.44%, 23/201), premature delivery (12.44%, 25/201), gestational hypertension disease (22.89%, 46/201), GDM (8.46%, 17/201), FGR (19.90%, 40/201) and low birth weight infants (16.42%, 33/201) in the TPOAb (+) untreated group were higher than those in TPOAb (+) treated group [4.02% (41/1 021), 4.21% (43/1 021), 10.77% (110/1 021), 4.11% (42/1 021), 8.72% (89/1 021), 7.35% (75/1 021), respectively] and the control group, with statistically significant differences (P < 0.05). The incidence of the pregnancy complications in the TPOAb (+) treated group was higher than those in the control group, but the differences were not statistically significant (P > 0.05). (4) There were no statistically significant difference (P > 0.05) in the incidence of abortion (3.72%, 8/215), premature delivery (4.19%, 9/215), gestational hypertension disease (10.70%, 23/215), GDM (4.19%, 9/215), FGR (9.77%, 21/215) or low birth weight infants (8.37%, 18/215) among the TPOAb (-) treated group, the TPOAb (-) untreated group [3.80% (23/605), 4.13% (25/605), 10.91% (66/605), 5.95% (36/605), 9.75% (59/605), 8.10% (49/605), respectively] and the control group. CONCLUSIONS: (1) The incidence of abortion, premature delivery, gestational hypertension disease, GDM, FGR and low birth weight infants could be increased in women with SCH in early pregnancy. (2) Thyroxine treatment could reduce the incidence of pregnancy complications in women with SCH in early pregnancy.

Oxidative stress and antioxidant parameters in patients with major depressive disorder compared to healthy controls before and after antidepressant treatment: results from a meta-analysis.

OBJECTIVE: To investigate the role of oxidative stress and antioxidants in depression. DATA SOURCES: We searched the literature without language restrictions through MEDLINE/PubMed, Cochrane Library, Fisterra, and Galenicom from database inception until December 31, 2013, supplemented by a hand search of relevant articles. Search terms included (1) oxidative stress, antioxidant*, nitrosative stress, nitrative stress, nitro-oxidative stress, free radical*, and names of individual oxidative stress markers/antioxidants and (2) depression and related disorders and antidepressant. STUDY SELECTION: Included were studies in patients with depression comparing antioxidant or oxidative stress markers with those in healthy controls before and after antidepressant treatment. DATA EXTRACTION: Two authors independently extracted the data for antioxidant or oxidative stress markers. Standardized mean differences (SMDs) ± 95% confidence intervals (CIs) for results from ≥ 3 studies were calculated. DATA SYNTHESIS: Altogether, 29 studies (N = 3,961; patients with depression = 2,477, healthy controls = 1,484) reported on the oxidative stress marker malondialdehyde (MDA) and total nitrites, the antioxidants uric acid and zinc, or the antioxidant-enhancing enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX). When patients with depression were compared with healthy controls, depression was associated with higher oxidative stress MDA levels (8 studies; n = 916; SMD = 1.34; 95% CI, 0.57 to 2.11; P < .001), lower antioxidant uric acid (4 studies; n = 512; SMD = -0.64; 95% CI, -1.22 to -0.06; P = .030) and zinc levels (13 studies; n = 2,002; SMD = -0.66; 95% CI, -0.98 to -0.34; P < .0001), and higher antioxidant-enhancing enzyme SOD levels (11 studies; n = 902; SMD = 0.62; 95% CI, 0.07 to 1.17; P = .028), while differences in total nitrites and CAT and GPX were nonsignificant. Antidepressant treatment, which significantly reduced Hamilton Depression Rating Scale scores (24.6 ± 0.7 to 16.2 ± 1.6; SMD = 2.65; 95% CI, 1.13 to 4.15; P = .00065), reduced MDA (4 studies; n = 194; SMD = -1.45; 95% CI, -2.43 to -0.47; P = .004) and increased uric acid (3 studies; n = 212; SMD = 0.76; 95% CI, 0.03 to 1.49; P = .040) and zinc levels (3 studies; n = 65; SMD = 1.22; 95% CI, 0.40 to 2.04, P = .004), without differences in MDA (P = .60), uric acid (P = .10), and zinc (P = .163) levels compared to healthy controls. CONCLUSIONS: Results suggest that oxidative stress plays a role in depression and that antidepressant activity may be mediated via improving oxidative stress/antioxidant function.

Study on the effect of black cumin (Nigella sativa Linn.) on experimental renal ischemia-reperfusion injury in rats.

PURPOSE: To evaluate the effect of Black cumin (Nigella sativa Linn.) pre-treatment on renal ischemia/reperfusion (I/R) induced injury in the rats. METHODS: A total of 40 male Wistar rats were randomly allocated into five equal groups including Sham, I/R model and three I/R+ Black cumin (0.5, 1 and 2%)-treated groups. I/R groups' kidneys were subjected to 60 min of global ischemia at 37°C followed by 24 h of reperfusion. At the end of reperfusion period, the rats were euthanized. Superoxide dismutase, catalase and glutathione peroxidase activities as well as reduced glutathione and renal malondialdehyde contents were determined in renal tissues. Kidney function tests and histopathological examination were also performed. RESULTS: High serum creatinine, blood urea nitrogen and uric acid as well as malondialhehyde (MDA) levels, and low antioxidant enzyme activities were observed in I/R rats compared to the sham rats. Pre-treatment with Black cumin for three weeks prior to IR operation improved renal function and reduced I/R induced renal inflammation and oxidative injury. These biochemical observations were supported by histopathological test of kidney sections. CONCLUSION: Black cumin significantly prevented renal ischemia/reperfusion induced functional and histological injuries.

Study on the effect of black cumin (Nigella sativa Linn.) on experimental renal ischemia-reperfusion injury in rats

PURPOSE: To evaluate the effect of Black cumin (Nigella sativa Linn.) pre-treatment on renal ischemia/reperfusion (I/R) induced injury in the rats.METHODS: A total of 40 male Wistar rats were randomly allocated into five equal groups including Sham, I/R model and three I/R+ Black cumin (0.5, 1 and 2%)-treated groups. I/R groups' kidneys were subjected to 60 min of global ischemia at 37°C followed by 24 h of reperfusion. At the end of reperfusion period, the rats were euthanized. Superoxide dismutase, catalase and glutathione peroxidase activities as well as reduced glutathione and renal malondialdehyde contents were determined in renal tissues. Kidney function tests and histopathological examination were also performed.RESULTS: High serum creatinine, blood urea nitrogen and uric acid as well as malondialhehyde (MDA) levels, and low antioxidant enzyme activities were observed in I/R rats compared to the sham rats. Pre-treatment with Black cumin for three weeks prior to IR operation improved renal function and reduced I/R induced renal inflammation and oxidative injury. These biochemical observations were supported by histopathological test of kidney sections.CONCLUSION:Black cumin significantly prevented renal ischemia/reperfusion induced functional and histological injuries.

Modulatory effects of deltamethrin-exposure on the immune status, metabolism and oxidative stress in gilthead seabream (Sparus aurata L.).

Deltamethrin, a sintetic pyrethroid, is the insecticide that has been replacing recently to others like organochlorines, organophosphates and carbamates which are less toxic for birds and mammals, although, unfortunately, all of them are highly toxic to various non-targeted aquatic organisms including fish. In the present study, the consequences of the exposition of gilthead seabream (Sparus aurata L.) specimens to sublethal bath dose of deltamethrin (0.1 ppb) on organo-somatic indexes, immunity, seric metabolic parameters, oxidative stress and liver histology were determined after 1, 3, 7 and 14 days of exposure. Deltamethrin alters gilthead seabream immune status, the hepato-somatic index and various seric metabolic parameters since the first exposure day while important progressive deleterious morphological changes in liver were also observed. However, no statistically significant deviation was detected in the expression of oxidative stress-related genes whilst the expression of cytochrome P450 gene was up-regulated in head-kidney and liver of exposed fish. Overall, the present results indicate severe immunotoxicological and metabolic effects of deltamethrin in gilthead seabream, the species with the highest rate of production in Mediterranean aquaculture. In general, the values obtained for the tested parameters during the trial seem to indicate that specimens try to adapt to this adverse situation although the continuous presence of the toxic impede the hypothetic recovery of homoeostasis. The use of deltamethrin in the proximities of seabream farms should be carefully considered.

A study on the antioxidant effect of Coriolus versicolor polysaccharide in rat brain tissues.

The objective of the study was to investigate the antioxidant effect of Chinese medicine Coriolus versicolor polysaccharide on brain tissue and its mechanism in rats. SOD, MDA and GSH-Px levels in rat brain tissues were determined with SD rats as the animal model. The results showed that Coriolus versicolor polysaccharide can reduce the lipid peroxidation level in brain tissues during exhaustive exercise in rats, and can accelerate the removal of free radicals. The study concluded that its antioxidant effect is relatively apparent.

Physiological adaptations during endurance training below anaerobic threshold in rats.

To assess the effects of continuous exercise training at intensities corresponding to 80 and 90 % of the lactate minimum test (LM), we evaluated antioxidant activity, hormone concentration, biochemical analyses and aerobic and anaerobic performance, as well as glycogen stores, during 12 weeks of swimming training in rats. One-hundred rats were separated into three groups: control (CG, n = 40), exercise at 80 (EG80, n = 30) and 90% (EG90, n = 30) of LM. The training lasted 12 weeks, with sessions of 60 min/day, 6 days/week. The intensity was based at 80 and 90% of the LM. The volume did not differ between training groups (X of EG80 = 52 ± 4 min; X of EG90 = 56 ± 2 min). The glycogen concentration (mg/100 mg) in the gastrocnemius increased after the training in EG80 (0.788 ± 0.118) and EG90 (0.795 ± 0.157) in comparison to the control (0.390 ± 0.132). The glycogen stores in the soleus enhanced after the training in EG90 (0.677 ± 0.230) in comparison to the control (0.343 ± 0.142). The aerobic performance increased by 43 and 34% for EG80 and EG90, respectively, in relation to baseline. The antioxidant enzymes remain unchanged during the training. Creatine kinase (U/L) increased after 8 weeks in both groups (EG80 = 427.2 ± 97.4; EG90 = 641.1 ± 90.2) in relation to the control (246.9 ± 66.8), and corticosterone (ng/mL) increased after 12 weeks in EG90 (539 ± 54) in comparison to the control (362 ± 44). The continuous exercise at 80 and 90% of the LM has a marked aerobic impact on endurance performance without significantly biomarkers changes compared to control.

Clinical evaluation with self-sequential longitudinal reference intervals: pregnancy outcome and neonatal thyroid stimulating hormone level associated with maternal thyroid diseases / Evaluación clínica con intervalos de referencia longitudinal auto-secuencial: evolución clínica del embarazo y niveles de la hormona estimulante de la tiroides en neonatos asociados con enfermedades tiroideas maternas

OBJECTIVE: We attempted to evaluate maternal thyroid function in a new self-sequential longitudinal reference interval (SLRI) which we established recently. By this method, we analysed the correlation between pregnancy outcome, neonatal thyroid stimulating hormone (TSH) level and maternal thyroid diseases. METHODS: A total of 1744 pregnant women participated in the study and 1747 babies were born from those women (three bore twins). The levels of TSH, free thyroxine (FT4) and thyroid peroxidase antibodies (TPO-Ab) of mothers were quantified by electrochemistry immunoassay (ECL). The levels of neonatal blood TSH were detected by time-resolved fluorescence immunoassay (TRFIA). All data were collected and statistically analysed by SPSS 13.0 software. RESULTS: With our new SLRI method, we found that 0.11%~3.84% pregnant women would get thyroid diseases. Subclinical hypothyroidism was the most common maternal thyroid disorder. Being positive for thyroid peroxidase antibodies was a significant risk factor of subclinical hypothyroidism during pregnancy. The median, P2.5~P97.5, and interquartile range (IQR) of neonatal TSH (N-TSH) of 1747 babies were 2.72 mIU/L, 0.10~8.01 mIU/L and 2.62 mIU/L, respectively; 28.6% of pregnant women with thyroid diseases developed pregnancy complications. The prevalence was significantly higher than in the normal thyroid function group (p< 0.001). The levels of N-TSH were low correlated with maternal TSH levels (p < 0.05), but there were no significant correlations between N-TSH and maternal FT4 and maternal TPO-Ab (p > 0.05). CONCLUSIONS: Thyroid disorders, especially subclinical hypothyroidism, are common in pregnant women. These disorders are associated with pregnancy and fetal outcome. Routine maternal thyroid function screening is important and should be recommended.

OBJETIVO: Intentamos evaluar la función tiroidea materna en un nuevo intervalo de referencia longitudinal auto-secuencial (SLRI) que establecimos recientemente. Por este método, analizamos la correlación entre el resultado del embarazo, el nivel de la hormona estimulante de la tiroides (TSH) en neonatos, y las enfermedades tiroideas maternas MÉTODOS: Un total de 1744 mujeres embarazadas participó en el estudio y 1747 bebés nacieron de esas mujeres (tres de ellas tuvieron gemelos). Los niveles de TSH, la tiroxina libre (FT4), y los anticuerpos de la peroxidasa tiroidea (TPO-Ab) de las madres, fueron cuantificados mediante inmunoensayo electroquímico (ECL). Los niveles de TSH en la sangre de los neonatos, fueron determinados mediante inmunoensayo por fluorescencia resuelto en el tiempo (TRFIA). Todos los datos fueron recogidos y analizados estadísticamente usando el software SPSS 13.0 RESULTADOS: Con nuestro nuevo método SLRI, encontramos que 0.11%~3.84% de las mujeres embarazadas contraerán enfermedades tiroideas. El hipotiroidismo subclínico fue el trastorno de la tiroides materna más común. Ser positivo a los anticuerpos de la peroxidasa tiroidea fue un factor de riesgo significativo del hipotiroidismo subclínico durante el embarazo. La mediana, P2.5~P97.5, y el rango intercuartil (IQR) de la TSH (N-TSH) neonatal de los 1747 bebés fueron 2.72 mIU/L, 0.10~8.01 mIU/L y 2.62 mIU/L respectivamente. El 28.6% de las mujeres embarazadas que tenían enfermedades tiroideas, desarrollaron complicaciones del embarazo. La prevalencia fue significativamente más alta que en el grupo con función tiroidea normal (p < 0.001). Los niveles de N-TSH fueron bajos en correlación con los niveles de TSH maternos (p < 0.05), pero no hubo ninguna correlación significativa entre la N-TSH y la FT4 materna, y la TPO-Ab materna (p > 0.05). CONCLUSIÓNS: Los trastornos tiroideos, especialmente el hipotiroidismo, son comunes en las mujeres embarazadas.Estos trastornos se hallan asociados con el resultado del embarazo y el resultado fetal. El tamizaje de rutina de la función tiroidea materna es importante y debe recomendarse.

Pituitary antibodies in women with Hashimoto's thyroiditis: prevalence in diagnostic and prediagnostic sera.

BACKGROUND: Pituitary antibodies have been reported with greater frequency in patients with Hashimoto's thyroiditis than in healthy controls, although there is significant variability in the strength of the association and the methodologies used. METHODS: We designed a nested case-control study to characterize the prevalence of pituitary antibodies at the time of the clinical diagnosis of Hashimoto's thyroiditis, as well as at 2, 5, and 7 years before diagnosis. Active component female service member cases (n=87) and matched female controls (n=107) were selected using the Defense Medical Surveillance System database (DMSSD) between January 1998 and December 2007. Pituitary antibodies were measured by immunofluorescence using human pituitary glands collected at autopsy as the substrate. RESULTS: At diagnosis, pituitary antibodies were present in 9% of cases with Hashimoto's (8 of 87) and 3% of controls (3 of 107). When the data were analyzed using a conditional logistic regression model, which takes into account the matching on age and work status, pituitary antibodies increased the odds of having Hashimoto's thyroiditis by sevenfold (95% confidence interval from 1.3 to 40.1, p=0.028), after adjusting for components of the DMSSD-category-termed race and for thyroperoxidase antibodies. Before diagnosis, pituitary antibodies were positive in 3 of the 11 subjects (2 cases and 1 control) at the -2-year time point, and negative in all 11 subjects at the -5- and -7-year time points. CONCLUSIONS: In summary, using a nested case-control design, we confirm that pituitary antibodies are more common in Hashimoto's thyroiditis and suggest that they appear late during its natural history.

Structural and functional analysis of native peroxiredoxin 2 in human red blood cells.

Peroxiredoxin 2, a typical 2-Cys peroxiredoxin, is the third most abundant protein in erythrocytes. It is understood that the physiologically functional state of peroxiredoxin 2 is the monomer, and that its role in scavenging low levels of H(2)O(2) results in the formation of disulfide-linked dimers, which are reversibly reduced to monomers by the thioredoxin-thioredoxin reductase system. Additionally, peroxiredoxins are highly susceptible to sulfinic acid formation through reactions with various peroxides. This overoxidized form, which is thought to convert peroxiredoxins into molecular chaperones and to be accompanied by a transition to polymeric forms, can be reversed by sulfiredoxins. However, physiological conformational changes and the antioxidant role of erythrocyte peroxiredoxin 2 are still unclear because there is low sulfiredoxin and thioredoxin-thioredoxin reductase activity in erythrocytes. In this study, we examined the structural and redox states of peroxiredoxin 2 in fresh hemolysates and estimated the activities of native and overoxidized peroxiredoxin 2 purified from red blood cells to clear the physiological roles of peroxiredoxin 2 in erythrocyte. Our findings demonstrate that native peroxiredoxin 2 exists as high molecular weight (>160 kDa) oligomers and that decamers or higher order molecular weight oligomers (260-460 kDa) have peroxidase activity. We further showed that peroxiredoxin 2 oligomers, which were predominantly composed of monomers in the reduced form, exert a chaperone activity equal to that of overoxidized peroxiredoxin 2 polymers. These results provide the novel insight that redox-active peroxiredoxin 2 functions in human red blood cells as high molecular weight oligomers that possess peroxidase and chaperone activities.

[Catalytic activity in rat blood plasma and erythrocytes that eliminates homocysteine with hydrogen peroxide].

The activity utilizing the free SH-form of homocysteine with H2O2 has been found in rat blood plasma and erythrocyte lysates with the use of the glutathione peroxidase assay with hydrogen peroxide and 5,5'-dithiobis-(2-nitrobenzoic acid) and with the substitution of glutathione by homocysteine. The presence of rat plasma or erythrocyte lysate in a reaction mixture containing D,L-homocysteine, and H2O2 resulted in a marked acceleration of homocysteine concentration decrease (the decrease of homocysteine concentration was initiated by addition of hydrogen peroxide). The data obtained suggest the presence of homocysteine peroxidase activity in plasma and erythrocytes. The observed activity may be attributed to some known thiol-dependent enzymes. In the rat brain tissue, the level of the activity is extremely low (at the detection limit). The increase of the activity in blood components during post-embryonic ontogeny has also been shown. Probably, this activity contributes to low concentrations of life SH-form of homocysteine in the blood components.

Vascular peroxidase-1 is rapidly secreted, circulates in plasma, and supports dityrosine cross-linking reactions.

Members of the peroxidase-cyclooxygenase superfamily catalyze biochemical reactions essential to a broad spectrum of biological processes, including host defense, thyroid hormone biosynthesis, and modification of extracellular matrix, as well as contributing to the pathogenesis of chronic inflammatory diseases. We recently identified a novel member of this family, vascular peroxidase-1 (VPO1), that is highly expressed in the human cardiovascular system. Its biosynthesis and enzymatic properties are largely unknown. Here, we report that VPO1 was rapidly and efficiently secreted into the extracellular space when the gene was stably expressed in human embryonic kidney (HEK) cells. Secreted VPO1 is a monomer with complex N-linked oligosaccharides and exhibits peroxidase activity. Biosynthesis of endogenous VPO1 by cultured human umbilical vein endothelial cells (HUVECs) shares features exhibited by heterologous expression of recombinant VPO1 (rVPO1) in HEK cells. The proinflammatory agents lipopolysaccharide and tumor necrosis factor-α induce expression of VPO1 mRNA and protein in HUVECs. Furthermore, murine and bovine sera and human plasma contain enzymatically active VPO1. rVPO1 exhibits spectral and enzymatic properties characteristic of the peroxidase-cyclooxygenase family, except with regard to its heat stability. rVPO1 catalyzes tyrosyl radical formation and promotes dityrosine cross-linking. Taken together, these data demonstrate that VPO1 is a glycosylated heme peroxidase that is actively secreted into circulating plasma by vascular endothelial cells and shares several features with other members of the peroxidase-cyclooxygenase family, including the catalysis of dityrosine formation.

Unusual peroxidase activity of polynitroxylated pegylated hemoglobin: Elimination of H(2)O(2) coupled with intramolecular oxidation of nitroxides.

Polynitroxylated hemoglobin (Hb(AcTPO)(12)) has been developed as a hemoglobin-based oxygen carrier. While Hb(AcTPO)(12) has been shown to exert beneficial effects in a number of models of oxidative injury, its peroxidase activity has not been characterized thus far. In the blood stream, Hb(AcTPO)(12) undergoes reduction by ascorbate to its hydroxylamine form Hb(AcTPOH)(12). Here we report that Hb(AcTPOH)(12) exhibits peroxidase activity where H(2)O(2) is utilized for intramolecular oxidation of its TPOH residues to TPO. This represents an unusual redox-catalytic mechanism whereby reduction of H(2)O(2) is achieved at the expense of reducing equivalents of ascorbate converted into those of Hb(AcTPOH)(12), a new propensity that cannot be directly associated with ascorbate.

Comprehensive peroxidase-based hematologic profiling for the prediction of 1-year myocardial infarction and death.

BACKGROUND: Recognition of biological patterns holds promise for improved identification of patients at risk for myocardial infarction (MI) and death. We hypothesized that identifying high- and low-risk patterns from a broad spectrum of hematologic phenotypic data related to leukocyte peroxidase-, erythrocyte- and platelet-related parameters may better predict future cardiovascular risk in stable cardiac patients than traditional risk factors alone. METHODS AND RESULTS: Stable patients (n=7369) undergoing elective cardiac evaluation at a tertiary care center were enrolled. A model (PEROX) that predicts incident 1-year death and MI was derived from standard clinical data combined with information captured by a high-throughput peroxidase-based hematology analyzer during performance of a complete blood count with differential. The PEROX model was developed using a random sampling of subjects in a derivation cohort (n=5895) and then independently validated in a nonoverlapping validation cohort (n=1474). Twenty-three high-risk (observed in > or =10% of subjects with events) and 24 low-risk (observed in > or =10% of subjects without events) patterns were identified in the derivation cohort. Erythrocyte- and leukocyte (peroxidase)-derived parameters dominated the variables predicting risk of death, whereas variables in MI risk patterns included traditional cardiac risk factors and elements from all blood cell lineages. Within the validation cohort, the PEROX model demonstrated superior prognostic accuracy (78%) for 1-year risk of death or MI compared with traditional risk factors alone (67%). Furthermore, the PEROX model reclassified 23.5% (P<0.001) of patients to different risk categories for death/MI when added to traditional risk factors. CONCLUSIONS: Comprehensive pattern recognition of high- and low-risk clusters of clinical, biochemical, and hematologic parameters provided incremental prognostic value in stable patients having elective diagnostic cardiac catheterization for 1-year risks of death and MI.