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Soy isoflavones are considered important sources of bioactive compounds, but they are poorly absorbable, due to their large hydrophilic structures. Some biotransformation strategies have been used to convert the glycosidic form into aglycones, making them available for absorption. This study evaluated the potential of enzymatic and/or microbial fermentation combined bioprocesses in a soymilk extract before and after gastrointestinal in vitro digestion. Commercial β-glucosidase (ET) and a mix of commercial probiotics (F) containing Lactobacillus acidophilus , Lactobacillus casei , Lactococcus lactis , Bifidobacterium bifidum , and Bifidobacterium lactis were used to biotransform the soymilk phenolic extract. An isoflavone profile was identified using HPLC-DAD, total phenolic content was identified using the Folin-Ciocalteu test, and antioxidant capacity was identified using ORAC and FRAP. Soymilk enzymatically treated (ET) followed by microbial fermentation (ET + T) resulted in better conversion of glycosylated isoflavones (6-fold lower than control for daidzin and 2-fold for genistin) to aglycones (18-fold greater than control for dadzein and genistein). The total phenolic content was increased (3.48 mg/mL for control and 4.48 mg/mL for ET + T) and the antioxidant capacity was improved with treatments of ET + T (120 mg/mL for control and 151 mg/mL with ORAC) and with FRAP (285 µL/mL for control and 317 µL/mL). After the in vitro digestion, ET + T samples resulted in a higher content of genistein (two-fold higher than control); also, increases in the total phenolic content (2.81 mg/mL for control and 4.03 mg/mL for ET + T) and antioxidant capacity measured with ORAC were greater compared to undigested samples. In addition, the isolated microbial fermentation process also resulted in positive effects, but the combination of both treatments presented a synergistic effect on soy-based products.

The aim of this study was to evaluate the effects of dose, start time, and application interval of recombinant bovine somatotropin (rbST) on performance, health, and reproduction parameters of crossbred Holstein × Gyr dairy cows (3/4 and 7/8). A 2 × 2 × 2 factorial study was designed to test the effects of application dose (250 mg rbST or 500 mg rbST), start of application (40 or 63 days in milk (DIM)), and application interval (12 or 14 days); 180 crossbred Holstein × Gyr cows were enrolled. Treatments with 500 mg rbST resulted in increased milk production, 3.5% fat-corrected milk (FCM), and energy-corrected milk (ECM). In the factorial design analysis, greater FCM, ECM, and milk solid production (kg/day) were observed when 500 mg rbST was started at 63 DIM, while 250 mg rbST resulted in greater milk production (kg/day), FCM, ECM, and milk solids (kg/day) when administered at 12-day intervals. Administration of 500 mg rbST resulted in an increase of days open and number of services per pregnancy (P = 0.01). Holstein × Gyr cows (3/4 and 7/8) presented a superior productive response to rbST when treated with 500 mg starting at 63 DIM and at 14-day intervals., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Objective:  To evaluate the seroprevalence of toxoplasmosis among puerperal women cared for at a tertiary university hospital and the level of understanding of these puerperal women about toxoplasmosis, vertical transmission, and its prophylaxis., Methods:  For this cross-sectional study, we evaluated 225 patients using presential interviews, prenatal documentation, and electronic medical records. Data were stored using Research Electronic Data Capture (REDCap) software. Prevalence rates were estimated by the presence of reactive IgG antibodies against Toxoplasma gondii . Data analysis was performed using the chi-square test and calculation of the odds ratio (OR). Seroreactivity to T. gondii and exposure variables (age, educational level, and parity) were analyzed using a confidence interval (95%CI) and a significance level of 5% (p < 0.05)., Results:  The seropositivity rate for T. gondii was 40%. There was no association between seroprevalence and age. Primiparity was a protective factor against seropositivity and low education was a risk factor., Conclusion:  Knowledge of T. gondii infection and its transmission forms was significantly limited, presenting a risk for acute maternal toxoplasmosis and vertical transmission of this protozoan. Increasing the education level regarding the risk of toxoplasmosis during pregnancy could reduce the rates of infection and vertical transmission of this parasite., Competing Interests: The authors have no conflict of interest to declare., (Federação Brasileira de Ginecologia e Obstetrícia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/).)

Introduction: Several studies demonstrated that deferoxamine, an iron chelator, can improve inflammatory alterations in adipose tissue induced by obesity. Obesity alterations in adipose tissue are also associated with tissue remodeling, and deferoxamine has anti-fibrosis action previously described in sites like the skin and liver., Methods: In this work, we analyzed deferoxamine effects on adipose tissue fibro-inflammation during obesity induced by diet in mice. in vitro approaches with fibroblasts and macrophages were also carried out to elucidate deferoxamine activity., Results: Our results demonstrated that in addition to exerting anti-inflammatory effects, reducing the cytokine production in adipose tissue of obese mice and by human monocyte differentiated in macrophage in vitro, deferoxamine can alter metalloproteinases expression and extracellular matrix production in vivo and in vitro ., Conclusion: Deferoxamine could be an alternative to control fibro-inflammation in obese adipose tissue, contributing to the metabolic improvements previously described., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Objective: to identify the rate of reactive treponemal and non-treponemal tests in pregnant women during childbirth and to analyze the factors associated with this seroreactivity., Method: this is a cross-sectional, quantitative study with secondary sources of sociodemographic and clinical data on 2,626 pregnant women treated at a public maternity hospital in the interior of São Paulo, in 2020. For statistical analysis, Fisher's exact test, Mann-Whitney test and the logistic regression model were used. A difference of p < 0.05 was considered statistically significant., Results: the rate of seropositivity for syphilis among pregnant women in this series was 2.74%. Among the groups with positive and non-reactive tests, marital status, occupation, place of residence and use of licit drugs indicated significant differences, but, in the final model, only unmarried marital status was associated with reactive tests (Odds Ratio: 0.169; Confidence Interval: 0.04-0.72; and p: 0.016)., Conclusion: in this study, unmarried marital status was the only independent factor associated with seroreactivity for syphilis. Therefore, it is necessary to create strategies aimed at women in this condition, potentially reducing the rate of congenital syphilis.

Purpose: To evaluate how the induction of liver damage by ischemia and reperfusion affects the adipose tissue of lean and obese mice., Methods: Lean and diet-induced obese mice were subjected to liver ischemia (30 min) followed by 6 h of reperfusion. The vascular stromal fraction of visceral adipose tissue was analyzed by cytometry, and gene expression was evaluated by an Array assay and by RT-qPCR. Intestinal permeability was assessed by oral administration of fluorescein isothiocyanate (FITC)-dextran and endotoxemia by serum endotoxin measurements using a limulus amebocyte lysate assay., Results: It was found that, after liver ischemia and reperfusion, there is an infiltration of neutrophils, monocytes, and lymphocytes, as well as an increase in the gene expression that encode cytokines, chemokines and their receptors in the visceral adipose tissue of lean mice. This inflammatory response was associated with the presence of endotoxemia in lean mice. However, these changes were not observed in the visceral adipose tissue of obese mice., Conclusions: Liver ischemia and reperfusion induce an acute inflammatory response in adipose tissue of lean mice characterized by an intense chemokine induction and leukocyte infiltration; however, inflammatory alterations are already present at baseline in the obese adipose tissue and liver ischemia and reperfusion do not injure further.

Women who are in the pregnancy-puerperal cycle or are lactating have been deliberately excluded from participating in COVID-19 vaccine clinical trials that aimed to evaluate either the efficacy of the vaccines in inducing the formation of neutralizing antibodies or the investigational products' safety profile. The exclusion of pregnant and lactating women from such studies certainly and inequitably denies these women access to COVID-19 vaccines, since these products have become increasingly available to nonpregnant people and even to those who are pregnant and are in high-income settings. In this clinical opinion article, we discuss some aspects of the prolonged pandemic, the emergence of viral variants, the risks of severe complications of COVID-19 in pregnant women, and the disproportionate impact of the above on low- and middle-income countries. We argue that the decision to receive the COVID-19 vaccine should be a joint decision between the pregnant or lactating women and the healthcare providers, while considering the available data on vaccine efficacy, safety, the risks of SARS-CoV-2 infection in pregnant women, and the women's individual risks for infection and serious illness. The various types of vaccines that are already in use and their safety, effectiveness, and the potential risks and benefits of their administration to pregnant or lactating women are also reviewed., (© 2021 The Authors.)

The objective of this work was to collect information on the chemical constituents that demonstrate the larvicidal activity against Aedes aegypti , as well as the antioxidant, microbiological, and cytotoxicity potential of the essential oil of Pogostemon cablin leaves. The chemical characterization was performed by gas chromatography coupled to mass spectrometer (GC-MS). The larvicidal activity was performed according to the protocol of the World Health Organization. The antioxidant activity was evaluated through the sequestering capacity of 2,2-diphenyl-1-picryl-hydrazine (DPPH). As for the microbiological evaluation, the microdilution technique was used, according to the protocol of the Clinical and Laboratory Standards Institute. The cytotoxic activity was evaluated against the larvae of Artemia salina . The species P. cablin presented the following compounds: Patchouli alcohol (33.25%), Seyshellene (6.12%), α-bulnesene (4.11%), Pogostol (6.33%), and Norpatchoulenol (5.72%), which was in synergy with the other substances may significantly potentiate the larvicidal action of the species with the LC 50 of 28.43 μg·mL -1 . There was no antioxidant activity, however, it presented antimicrobial activity against all bacteria tested with Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of 62.5 μg·mL -1 . The species demonstrated significant toxic action with LC 50 of 24.25 μg·mL -1 . Therefore, the P. cablin species showed significant larvicidal potential, antimicrobial activity, the absence of antioxidant action, and high toxicity.

Purpose: Maternal obesity can program the offspring, increasing the risk of overweight and obesity in adult life. Guarana (Paullinia cupana) is a Brazilian plant that has weight-reducing effects. Thus, this study aimed to evaluate the effects of Guarana on metabolic and inflammatory parameters in mice programmed by maternal obesity., Methods: Swiss female mice were divided into two groups: control and high fat (HF), who received a standard diet or a high-fat diet (HFD), respectively, for 8 weeks prior to mating, gestation, and lactation. After post-natal day (PN) 21, the offspring of the HF group were subdivided into three groups: HF without treatment; HF early treatment, offspring treated with Guarana (1 g/kg bodyweight) in PN25-PN30; HF late treatment, offspring treated with Guarana (1 g/kg bodyweight) in PN65-PN75. Basal energy expenditure, the lipid profile and fasting glucose levels were determined. Body composition was evaluated by dissecting adipose tissue depots. Gene expression was analyzed using real-time PCR., Results: During mating, the weight of HF females increased; after lactation, their adipose tissue depots and fasting glycemic levels also increased. The offspring of the HF group showed an increased body weight at PN21. At PN80, in the mice treated with Guarana (with both treatments), VO 2 and energy expenditure increased, adipose tissue depots decreased, and the expression of leptin, IL-6, TNF-α, and MCP-1 decreased compared with that in the HF group., Conclusions: Guarana treatment at both stages of life reversed some of the alterations developed by the offspring of HF animals in adult life.

No scientific report proves the action of the phytochemicals from the mangrove tree Rhizophora mangle in the treatment of diabetes. The aim of this work is to evaluate the effects of the acetonic extract of R. mangle barks (AERM) on type 2 diabetes. The main chemical constituents of the extract were analyzed by high-performance liquid chromatography (HPLC) and flow injection analysis electrospray-iontrap mass spectrometry (FIA-ESI-IT-MS/MS). High-fat diet (HFD)-fed mice were used as model of type 2 diabetes associated with obesity. After 4 weeks of AERM 5 or 50 mg/kg/day orally, glucose homeostasis was evaluated by insulin tolerance test (kiTT). Hepatic steatosis, triglycerides and gene expression were also evaluated. AERM consists of catechin, quercetin and chlorogenic acids derivatives. These metabolites have nutritional importance, obese mice treated with AERM (50 mg/kg) presented improvements in insulin resistance resulting in hepatic steatosis reductions associated with a strong inhibition of hepatic mRNA levels of CD36. The beneficial effects of AERM in an obesity model could be associated with its inhibitory α-amylase activity detected in vitro. Rhizophora mangle partially reverses insulin resistance and hepatic steatosis associated with obesity, supporting previous claims in traditional knowledge.

Extracellular matrix (ECM) remodeling is necessary for a health adipose tissue (AT) expansion and also has a role during weight loss. We investigate the ECM alteration during weight cycling (WC) in mice and the role of matrix metalloproteinases (MMPs) was assessed using GM6001, an MMP inhibitor, during weight loss (WL). Obesity was induced in mice by a high-fat diet. Obese mice were subject to caloric restriction for WL followed by reintroduction to high-fat diet for weight regain (WR), resulting in a WC protocol. In addition, mice were treated with GM6001 during WL period and the effects were observed after WR. Activity and expression of MMPs was intense during WL. MMP inhibition during WL results in inflammation and collagen content reduction. MMP inhibition during WL period interferes with the period of subsequent expansion of AT resulting in improvements in local inflammation and systemic metabolic alterations induced by obesity. Our results suggest that MMPs inhibition could be an interesting target to improve adipose tissue inflammation during WL and to support weight cyclers., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Chronic non-communicable diseases such as obesity are preceded by increased macrophage infiltration in adipose tissue and greater secretion of pro-inflammatory cytokines. We evaluated the anti-inflammatory potential of Biotransformed extract, and two control extracts: In Natura and Autoclaved. The assays were performed using a cellular model with RAW264.7, 3T3-L1 cells, and RAW264.7 and 3T3-L1 co-culture. The innovation of the study was the use of Biotransformed extract, a unique phenolic extract of a bioprocessed citrus residue. LPS stimulated RAW264.7 cells treated with the Biotransformed extract exhibited lower secretion of TNF-α and NO and lower protein expression of NFκB. In RAW264.7 and 3T3-L1 co-culture, treatment with 1.0mg/mL of the Biotransformed extract reduced secretion of TNF-α (30.7%) and IL-6 (43.4%). Still, the Biotransformed extract caused higher increase in adiponectin in relation to control extracts. When the co-culture received a LPS stimulus, the Autoclaved extract at 1.0mg/mL reduced IL-6 and TNF-α concentrations, and raised adiponectin. However, it was noteworthy that the Biotransformed extract was also able to significantly reduce IL-6 concentration while the Natural extract was not. The Biotransformed citrus extract evaluated in this study showed anti-inflammatory activity in macrophages and in co-culture, indicating that bioprocess of citrus residue can contribute to new product development with anti-inflammatory potential., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Adenosine receptors are expressed in adipose tissue and control physiological and pathological events such as lipolysis and inflammation. The aim of this study was to evaluate the activity of N 6 -cyclopentyladenosine (CPA), a potent and selective A 1 adenosine receptor agonist; 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxyamidoadenosine hydrochloride (CGS-21680), an A 2A adenosine receptor agonist; and 5'-N-ethylcarboxamidoadenosine (NECA), a potent non-selective adenosine receptor agonist on adipose tissue inflammatory alterations induced by obesity in mice. Swiss mice were fed with a high-fat diet for 12 weeks and agonists were administered in the last two weeks. Body weight, adiposity and glucose homeostasis were evaluated. Inflammation in adipose tissue was assessed by evaluation of adipokine production and macrophage infiltration. Adenosine receptor signaling in adipose tissue was also evaluated. Mice that received CGS21680 presented an improvement in glucose homeostasis in association with systemically reduced inflammatory markers (TNF-α, PAI-1) and in the visceral adipose tissue (TNF-α, MCP-1, macrophage infiltration). Activation of p38 signaling was found in adipose tissue of this group of mice. NECA-treated mice presented some improvements in glucose homeostasis associated with an observed weight loss. Mice that received CPA presented only a reduction in the ex vivo basal lipolysis rate measured within visceral adipose tissue. In conclusion, administration of the A 2A receptor agonist to obese mice resulted in improvements in glucose homeostasis and adipose tissue inflammation, corroborating the idea that new therapeutics to treat obesity could emerge from these compounds., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Iron is an important micronutrient, but it can also act as a dangerous element by interfering with glucose homeostasis and inflammation, two features that are already disturbed in obese subjects. In this work, we study the effects of systemic iron supplementation on metabolic and inflammatory responses in mice with hypoferremia induced by obesity to better characterize whether iron worsens the parameters that are already altered after 24 weeks of a high-fat diet (HFD). Mice were maintained on a control diet or a HFD for 24 weeks and received iron-III polymaltose (50 mg/kg/every 2 days) during the last two weeks. Glucose homeostasis (basal glucose and insulin test tolerance) and systemic and visceral adipose tissue (VAT) inflammation were assessed. Iron levels were measured in serum. The Prussian blue reaction was used in isolated macrophages to detect iron deposition. Iron supplementation resulted in an increased number of VAT macrophages that were positive for Prussian blue staining as well as increased serum iron levels. Systemic hepcidin, leptin, resistin, and monocyte chemoattractant protein-1 (MCP-1) levels were not altered by iron supplementation. Local adipose tissue inflammation was also not made worse by iron supplementation because the levels of hepcidin, MCP-1, leptin, and interleukin (IL)-6 were not altered. In contrast, iron supplementation resulted in an increased production of IL-10 by adipose tissue and VAT macrophages. Leukocytosis and VAT plasminogen activator inhibitor-1 (PAI-1) level were reduced, but insulin resistance was not altered after iron supplementation. In conclusion, systemic iron supplementation in mice with hypoferremia induced by obesity did not worsen inflammatory marker or adipose tissue inflammation or the metabolic status established by obesity. Iron deposition was observed in adipose tissue, mainly in macrophages, suggesting that these cells have mechanisms that promote iron incorporation without increasing the production of inflammatory mediators.

Despite reports that food-borne parasitic infections have been increasing worldwide, the methodologies employed to detect food contamination by helminths are still largely based on methodologies used to detect these pathogens in feces and water. This study sought to improve the diagnosis of parasitic contaminants in lettuce by standardizing a method for detecting helminth eggs and larvae and estimating their percentage of recovery. Sanitized lettuces were artificially contaminated with different amounts of Ascaris suum and hookworm eggs and larvae. To standardize the method, we tested liquid extractors, vegetable washing steps, and spontaneous sedimentation times. Higher percentages of egg and larvae recovery were obtained using 1 M glycine as the liquid extractor, manual shaking for 3 min and 2 h of sedimentation. Five different levels of artificial contamination (ten replicates each; n = 50) were tested using these standardized conditions, yielding an average recovery of 62.6 % (±20.2), 51.9 % (±20.0), and 50.0 % (±27.3) for A. suum eggs, hookworm eggs, and larvae, respectively. Tests were performed with a different matrix to evaluate the performance of the method. Furthermore, collaborative analytical studies performed by different laboratories produced satisfactory results. The method for the identification of helminth eggs and larvae proposed in this study proved to be simpler and more efficient than previously published procedures, thereby demonstrating its potential contribution to health surveillance and epidemiological studies.

Aim: To study pentoxifylline effects in liver and adipose tissue inflammation in obese mice induced by high-fat diet (HFD)., Methods: Male swiss mice (6-wk old) were fed a high-fat diet (HFD; 60% kcal from fat) or AIN-93 (control diet; 15% kcal from fat) for 12 wk and received pentoxifylline intraperitoneally (100 mg/kg per day) for the last 14 d. Glucose homeostasis was evaluated by measurements of basal glucose blood levels and insulin tolerance test two days before the end of the protocol. Final body weight was assessed. Epididymal adipose tissue was collected and weighted for adiposity evaluation. Liver and adipose tissue biopsies were homogenized in solubilization buffer and cytokines were measured in supernatant by enzyme immunoassay or multiplex kit, respectively. Hepatic histopathologic analyses were performed in sections of paraformaldehyde-fixed, paraffin-embedded liver specimens stained with hematoxylin-eosin by an independent pathologist. Steatosis (macrovesicular and microvesicular), ballooning degeneration and inflammation were histopathologically determined. Triglycerides measurements were performed after lipid extraction in liver tissue., Results: Pentoxifylline treatment reduced microsteatosis and tumor necrosis factor (TNF)-α in liver (156.3 ± 17.2 and 62.6 ± 7.6 pg/mL of TNF-α for non-treated and treated obese mice, respectively; P < 0.05). Serum aspartate aminotransferase levels were also reduced (23.2 ± 6.9 and 12.1 ± 1.6 U/L for non-treated and treated obese mice, respectively; P < 0.05) but had no effect on glucose homeostasis. In obese adipose tissue, pentoxifylline reduced TNF-α (106.1 ± 17.6 and 51.1 ± 9.6 pg/mL for non-treated and treated obese mice, respectively; P < 0.05) and interleukin-6 (340.8 ± 51.3 and 166.6 ± 22.5 pg/mL for non-treated and treated obese mice, respectively; P < 0.05) levels; however, leptin (8.1 ± 0.7 and 23.1 ± 2.9 ng/mL for non-treated and treated lean mice, respectively; P < 0.05) and plasminogen activator inhibitor-1 (600.2 ± 32.3 and 1508.6 ± 210.4 pg/mL for non-treated and treated lean mice, respectively; P < 0.05) levels increased in lean adipose tissue. TNF-α level in the liver of lean mice also increased (29.6 ± 6.6 and 75.4 ± 12.6 pg/mL for non-treated and treated lean mice, respectively; P < 0.05) while triglycerides presented a tendency to reduction., Conclusion: Pentoxifylline was beneficial in obese mice improving liver and adipose tissue inflammation. Unexpectedly, pentoxifylline increased pro-inflammatory markers in the liver and adipose tissue of lean mice.

Context: Intestinal inflammation can induce a local reduction in oxygen levels that triggers an adaptive response centered on the expression of hypoxia-inducible factors (HIFs). Nitric oxide, a well-described inflammatory mediator, may interfere with hypoxia signaling., Objectives: We aimed to evaluate the role of nitric oxide in hypoxia signaling during colonic inflammation., Methods: Colitis was induced by single (acute) or repeated (reactivated colitis) trinitrobenzenosulfonic acid administration in rats. In addition, one group of rats with reactivated colitis was also treated with Nw-Nitro-L-arginine methyl ester hydrochloride to block nitric oxide synthase. Colitis was assessed by macroscopic score and myeloperoxidase activity in the colon samples. Hypoxia was determined using the oxygen-dependent probe, pimonidazole. The expression of HIF-1α and HIF-induced factors (vascular endothelial growth factor - VEGF and apelin) was assessed using Western blotting., Results: The single or repeated administration of trinitrobenzenosulfonic acid to rats induced colitis which was characterized by a high macroscopic score and myeloperoxidase activity. Hypoxia was observed with both protocols. During acute colitis, HIF-1α expression was not increased, but VEGF and apelin were increased. HIF-1α expression was inhibited during reactivated colitis, and VEGF and apelin were not increased. Nw-Nitro-L-arginine methyl ester hydrochloride blockade during reactivated colitis restored HIF-1α, VEGF and apelin expression., Conclusions: Nitric oxide could interfere with hypoxia signaling during reactivated colitis inflammation modifying the expression of proteins regulated by HIF-1α.

Background: Dietary vitamin K intake has been considered a major factor that influences stability of oral anticoagulation (OA) with coumarins. Few studies have evaluated the relationship between amounts of dietary vitamin K intake and stability of anticoagulation., Objective: To assess whether high dietary vitamin K intake is associated to stability of International Normalized Ratio (INR) of the prothrombin time., Methods: We performed a sub-analysis of a randomized clinical trial involving outpatients from the anticoagulation clinic of a university hospital. INR and vitamin K intake were prospectively collected at baseline, 15, 30, 60 and 90 days after randomization. Patients were considered with a stable anticoagulation when their INR coefficient of variation was less than 10%. Dietary vitamin K intake was assessed by a food frequency questionnair and a score of intake was derived., Results: We studied 132 patients on chronic OA (57 ± 13 years; 55% males); 23 patients (17%) were achieved stable anticoagulation. Stable and unstable patients had no significant differences in baseline characteristics. The dietary vitamin K score over the entire follow-up for stable patients was significantly lower than that for unstable patients (p = 0.012)., Discussion: Our findings suggest that INR stability could be achieved with relatively low amounts of dietary vitamin K.

Objective: Infliximab is a monoclonal anti-TNF-α antibody that is used therapeutically to treat Crohn's disease (CD). High levels of pro-inflammatory cytokines, especially TNF-α, have been observed in the gastrointestinal tract of CD patients and were associated with alterations in the mesenteric adipose tissue, which also contributed to the high levels of adipokine release. The authors used a rat model of colitis that produces mesenteric adipose tissue alterations that are associated with intestinal inflammation to study the effects that infliximab treatment has on adipokine production, morphological alterations in adipose tissue and intestinal inflammation., Material and Methods: The ability of infliximab to neutralize rat TNF-α was evaluated in vitro using U937 cells. Colitis was induced by repeated intracolonic trinitrobenzene sulfonic acid instillations and was evaluated by macroscopic score, histopathological analysis, myeloperoxidase activity, TNF-α and IL-10 expression as well as iNOS (inducible NO synthase) expression and JNK phosphorylation in colon samples. The alterations in adipose tissue were assessed by TNF-α, IL-10, leptin, adiponectin and resistin levels as well as adipocyte size and peroxisome proliferator-activated receptor (PPAR)-γ expression., Results: Infliximab treatment controlled intestinal inflammation, which reduced lesions and neutrophil infiltration. Inflammatory markers, such as iNOS expression and JNK phosphorylation, were also reduced. In mesenteric adipose tissue, infliximab increased the production of IL-10 and resistin, which was associated with the restoration of adipocyte morphology and PPAR-γ expression., Conclusions: Our results suggest that infliximab could contribute to the control of intestinal inflammation by modifying adipokine production by mesenteric adipose tissue.

This paper reports the development and use of a Real Time PCR for detection of Babesia canis canis, B. canis rossi, and B. canis vogeli in endemic areas of Brazil. The sequences of the internal transcribed spacer (ITS) of several organisms were aligned and five primers and four probes were designed for amplification of a fragment (around 125 bp) which differentiates subspecies of B. canis. Blood samples collected from dogs living in farms in three distinct rural regions within the State of Minas Gerais (Lavras, Belo Horizonte and Nanuque) were tested. Blood samples had been collected during a dry season (Lavras, n=100; Belo Horizonte, n=50; Nanuque, n=102); the dogs were re-sampled in the subsequent rainy season (Lavras, n=71; Belo Horizonte, n=29; Nanuque, n=66). From each sample, DNA was extracted and Giemsa stained smears were microscopically examined for direct detection of Babesia parasites. B. canis vogeli was the only subspecies found, with an overall prevalence of 9.9% during the dry season and 10.8% during the rainy season. Dogs living in Nanuque and Belo Horizonte showed significantly higher prevalence rates than those living in Lavras (13.7%, 12.0% and 5.0%, respectively). The Real Time PCR developed proved to be appropriate to detect B. canis subspecies in endemic areas., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Deer are important natural reservoir hosts of Anaplasmataceae. The present study used nested PCR and nucleotide sequencing to evaluate the occurrence of Anaplasmataceae species in 23 free-living and six captive specimens of the cervids Mazama gouazoubira and Blastocerus dichotomus in Minas Gerais State, Brazil. Blood samples were tested for the presence of Ehrlichia and Anaplasma spp. using nPCR assays and sequencing of the msp4, msp1 and 16S rRNA genes. The identity of each sequence was confirmed by comparison with sequences available from GenBank using BLAST software. Of the animals investigated, 93.1% (27/29) were infected with haemoparasites including Anaplasma marginale (79.3%), Ehrlichia chaffeensis (3.4%), Anaplasma bovis (3.4%) and Anaplasma spp. (assigned to A. platys and A. phagocytophilum) (17.2%). Co-infection occurred in 20% (6/29) of the deer examined. Four (13.8%) were infected with A. marginale and Anaplasma sp., one (3.4%) was infected with A. marginale and E. chaffeensis, and one (3.4%) was infected with A. marginale and A. bovis. The results of the present study suggest that cross-protection does not occur in these deer. Immunological cross-reaction occurs when sera are tested diagnostically because these bacteria are closely related taxonomically, reinforcing the importance of molecular diagnosis followed by nucleotide sequencing., (© 2011 Blackwell Verlag GmbH.)

Purpose: To compare whether health-related quality of life (HRQoL) is altered in patients undergoing a treatment strategy guided by changes in dietary vitamin K., Methods: This study is a randomized clinical trial carried out with chronic oral anticoagulation outpatients randomized into a control group (conventional dose adjustment of oral anticoagulants) (n = 66) and an intervention group (strategy based on changes in dietary vitamin K intake) (n = 66). HRQoL was measured using the Duke Anticoagulation Satisfaction Scale (DASS) at baseline and 90 days of follow-up., Results: Patients with worse HRQoL were younger (p = 0.005) and were using a higher dose of baseline oral anticoagulants (p = 0.008), while those with better HRQoL scores had a higher level of education (p = 0.01). Both groups had significant improvements in HRQoL from baseline to 90 days in the global DASS score (p < 0.001), as well as in the negative and positive psychological impact (p < 0.001) domains. We did not observe differences in the variations of HRQoL scores in any of the DASS domains (p values > 0.05) between groups of interventions. Patients who achieved oral anticoagulation stability (n = 23) had significantly better HRQoL scores than patients who did not achieve stability (p = 0.003)., Conclusion: Patients receiving the treatment strategy based on changes in dietary vitamin K intake did not have better HRQoL scores; however, both treatment approaches to manage oral anticoagulation improved HRQoL. Patients with greater oral anticoagulation stability had better HRQoL scores.

Visceral leishmaniasis (VL), also known as kala-azar, is an important public health problem. If not treated, virtually all clinically symptomatic patients die within months. The diagnosis is based on the Montenegro skin test (MST) and anti-Leishmania titers. Nevertheless, the time required for cured individuals living in a leishmaniasis-endemic area to present a positive skin test and negative anti-Leishmania serology is known. To determine the cellular and humoral immune response profile in relation to different times post-VL cure, a cross-sectional study was conducted on subjects from a kala-azar endemic area in Paço do Lumiar, MA, Brazil, on the basis of 1995-2005 notifications reported by the National Health Foundation/Regional Coordination of Maranhão. We visited cured individuals with a history of VL within the last 10 years. Seventy-four subjects (30 females) ranging in age from 1 to 44 years were included, all of them symptom free at the time of the study. A cellular immune response was observed in 73 (98.6%) subjects, whereas no significant antibody titers were detected by indirect immunofluorescence (IIF) in the sera of 69 (93.2%) cases. Ten years post-cure, 39 (52%) subjects had a positive MST and negative IIF reaction, while in one subject the skin and anti-Leishmania serology tests were negative. Two other subjects were positive in both tests 1 year after cure. These data suggest that a cellular immune response may still be present in subjects cured of VL regardless of post-cure time, and that the parasite persists in the host after clinical cure of the disease. This would explain the persistence of significant Leishmania sp antibody titers in some subjects after treatment.

This study investigated whether a low pathogenicity isolate of Anaplasma marginale with an appendage (UFMG1) could protect calves from infection with a pathogenic A. marginale isolate (UFMG2). Two groups of five Friesian calves were each inoculated with UFMG1 by intravenous injections of either A. marginale-infected tick cell cultures (group 1) or blood stabilates (group 2); a third (control) group was injected with saline. All animals were inoculated with a blood stabilate containing a high pathogenicity A. marginale isolate (UFMG2) 75 days after the UFMG1 inoculation. After infection with UFMG2, animals in groups 1 and 2 presented low rickettsaemia, but no clinical signs and no reduction in packed cell volume (PCV). Control animals became sick, with high rickettsaemia (16% infected erythrocytes) and a reduction in PCV (71%), resulting in 60% deaths. Up to 2 weeks after the UFMG2 inoculation, msp1α UFMG1 sequences were detected in groups 1 and 2. Four weeks after UFMG2 inoculation, UFMG2 sequences were detected in these animals, along with a new msp1α genotype sequence, closely related to that of the UFMG2 isolate. Control animals had UFMG2 msp1α sequences up to 4weeks after inoculation with UFMG2 and the new msp1α genotype sequence could be detected on the sixth week. The origin of the new A. marginale genotype was unknown, but may represent the first example of MSP1a antigenic variation in infected cattle. The results confirmed the low pathogenicity of the UFMG1 isolate, which provided clinical protection against the highly pathogenic A. marginale UFMG2. Infection with UFMG1 did not prevent the establishment of a second isolate, suggesting protection without infection-exclusion among A. marginale isolates., (Published by Elsevier Ltd.)

Members of the leucine-rich repeat protein family are involved in diverse functions including protein phosphatase 2-inhibition, cell cycle regulation, gene regulation and signalling pathways. A novel Schistosoma mansoni gene, called SmLANP, presenting homology to various genes coding for proteins that belong to the super family of leucine-rich repeat proteins, was characterized here. SmLANP was 1184bp in length as determined from cDNA and genomic sequences and encoded a 296 amino acid open reading frame that spanning from 6 to 894bp. The predicted amino acid sequence had a calculated molecular weight of 32kDa. Analysis of the predicted sequence indicated the presence of 3 leucine-rich domains (LRR) located in the N-terminal region and an aspartic acid rich region in the C-terminal end. SmLANP transcript is expressed in all stages of the S. mansoni life cycle analyzed, exhibiting the highest expression level in males. The SmLANP protein was expressed in a GST expression system and antibodies raised in mice against the recombinant protein. By immunolocalization assay, using adult worms, it was shown that the protein is mainly present in the cell nucleus through the whole body and strongly expressed along the tegument cell body nuclei of adult worms. As members of this family are usually involved in protein-protein interaction, a yeast two hybrid assay was conducted to identify putative binding partners for SmLANP. Thirty-six possible partners were identified, and a protein ATP synthase subunit alpha was confirmed by pull down assays, as a binding partner of the SmLANP protein.

This epidemiological survey on canine babesiosis was carried out in three distinct rural regions (Lavras, Belo Horizonte and Nanuque) of the State of Minas Gerais, Brazil. Ticks and blood samples were collected during a dry season (Lavras, n=92; Belo Horizonte, n=50; Nanuque, n=102) and the subsequent rainy season (Lavras, n=71; Belo Horizonte, n=28; Nanuque, n=66) from dogs living on farms. Plasma samples were analyzed by the indirect fluorescent antibody test for detection of anti-Babesia canis vogeli antibodies. DNA was extracted from blood of serologically positive dogs and molecular characterization of Babesia species was performed. Rhipicephalus sanguineus, Amblyomma cajennense and Boophilus microplus were the tick species identified in all regions. In Lavras, in addition to those tick species, A. tigrinum and A. ovale were also identified. The most prevalent tick species was A. cajennense (35.3%), followed by R. sanguineus (19%) and B. microplus (4.0%). Dogs living in Nanuque region were more heavily infested with ticks than dogs living in Belo Horizonte and Lavras regions. The overall frequency of anti-B. c. vogeli antibodies in the canine population in rural areas of Minas Gerais was 28.7%, with prevalence rates of 49.0% in Nanuque, 34.0% in Belo Horizonte and 3.3% in Lavras. The age of the animals and tick infestation were associated with seroprevalence of B. c. vogeli. The sequence analysis showed that B. c. vogeli was the only Babesia species present in all three regions. This study showed different rates of prevalence and incidence of canine babesiosis among the three rural regions sampled in Minas Gerais State. The results point to the importance of canine babesiosis in rural areas and to the need for further studies related to its transmission and maintenance in nature.

In a previous work Ancylostoma caninum sex-specific genes had being identified, however without confirmation, by using an non specific RT-PCR comparing male and female cDNA. In this work more fragments had been identified and a semi-quantitative RT-PCR carried out in order to confirm the sex-specific character of the transcripts obtained. Four fragments were confirmed as sex-specific or as being expressed more in one sex than the other.

Background: Scarce data are available to predict in-hospital mortality for decompensated heart failure (HF) in South American populations., Methods and Results: We evaluated 779 consecutive HF admissions defined by the Boston criteria in a tertiary care hospital. Stepwise logistic regression was used to determine independent correlates of in-hospital mortality, derived from 83 potential predictors collected on hospital admission. A clinical score rule (HF Revised Score) was created using the regression coefficient estimates derived from multivariate modeling. During hospital stay, 77 (10%) deaths occurred and 6 clinical characteristics were independently associated with in-hospital mortality: presence of cancer (P < .001), systolic blood pressure < or =124 mm Hg (P < .001), serum creatinine >1.4 mg/dL (P = .02), blood urea nitrogen >37 mg/dL (P = .03), serum sodium <136 mEq/L (P = .03), and age >70 years old (P = .03). Both the Acute Decompensated Heart Failure National Registry stratification algorithm and the proposed HF Revised Score performed adequately to predict in-hospital mortality ("c" statistics = 0.71 and 0.76, respectively). The newly proposed score, however, discriminated a very low-risk group (101 [13%]) in whom all patients were discharged home, representing patients admitted with none of the 6 predictors of risk., Conclusion: HF risk stratification can be accurately accomplished during the first day of admission with simple and easily obtained clinical variables.

Effects of dietary energy density during late gestation and early lactation on metabolic status of periparturient cows were studied. Four weeks before expected calving, animals were fed a low (DL; 1.58 Mcal of NEL/kg) or high energy density diet (DH; 1.70 Mcal of NEL/kg). After calving, half of the cows from each prepartum treatment were assigned to a low (L; 1.57 Mcal of NEL/kg) or high energy density diet (H; 1.63 Mcal of NEL/kg) until d 20 postpartum. After d 20, all animals were fed H until d 70. Animals fed DH had a more positive energy balance during the prepartum period. Animals fed DH had higher plasma concentrations of glucose and insulin and lower concentrations of plasma nonesterified fatty acid (NEFA) on d -7 relative to calving compared with animals fed DL. No differences in blood concentrations of metabolites, insulin and liver triglycerides (TG) content were observed on d 1. Liver TG content at d 1 and 21 were more related to magnitude of change in energy intake prepartum than to energy intake in the last week of gestation. Cows fed H had higher concentrations of plasma glucose and insulin, but similar plasma NEFA during the postpartum period compared with cows fed L. Plasma concentrations of beta-hydroxybutyrate (BHBA) and liver TG content on d 21 were 46 and 30% lower, respectively, for cows fed H compared with cows fed L. Interactions between prepartum and postpartum treatments indicated that negative effects of delaying higher concentrate feeding until d 21 postpartum can be partially offset by increasing concentrate in the diet before calving. Cows fed L had a higher increase in white line hemorrhage scores between prepartum and 10 wk postpartum compared with cows fed H. Energy density of prepartum diets had a minor influence on metabolic status of cows postpartum. A more favorable metabolic profile occurs when increasing the concentrate content of the diet immediately postpartum compared with delaying the increase until d 21 postpartum.

Forty cows and twenty heifers were used to study the effects of dietary energy density during late gestation and early lactation on lactation performance and ruminal parameters. A 2 x 2 factorial arrangement of treatments was used. During prepartum (-28 d to calving), animals were fed a low energy density diet [DL; 1.58 Mcal of net energy for lactation (NE(L))/kg, 40% neutral detergent fiber (NDF) and 38% nonfiber carbohydrate (NFC)] or a high energy diet (DH; 1.70 Mcal NE(L)/kg, 32% NDF and 44% NFC). After calving, half of the cows from each prepartum treatment group were assigned to a low energy density diet (L; 1.57 Mcal NE(L)/kg, 30% NDF and 41% NFC) or a high energy density diet (H; 1.63 Mcal NE(L)/kg, 25% NDF and 47% NFC) until d 20 postpartum. After d 20, all cows were fed H until d 70. Animals fed DH had 19.8% greater dry matter intake (DMI; % of body weight) and 21.5% greater energy intake than animals fed DL prepartum and the response was greater for cows compared to heifers. Animals fed DH had lower ruminal pH compared to animals fed DL, but no major changes in volatile fatty acid concentrations were observed. Effects of dietary energy density during prepartum on postpartum production responses were dependent on parity. Primiparous cows fed DL had higher 3.5% fat-corrected milk yield and milk fat production and percentage during the first 10 wk of lactation than those fed DH. Prepartum diet did not affect lactation performance of multiparous cows. Cows fed H had higher DMI and energy intake for the first 20 d of lactation compared to cows fed L. Diets did not affect DMI after the third wk of lactation. Milk production increased faster for cows fed H compared to cows fed L. Animals fed DL-L sequence of treatments tended to have the lowest energy intake during the first 10 wk of lactation. Prepartum treatments did not affect ruminal fermentation characteristics postpartum. Cows fed H had lower ruminal pH and higher propionate concentrations than cows fed L. No prepartum x postpartum interactions were observed for ruminal fermentation parameters. The effects of DH on prepartum DMI did not carry over to the postpartum period or influence early postpartum production. Increasing concentrate content of the diet immediately postpartum instead of delaying the increase until d 21 postpartum is associated with a higher rate of increase.in milk production and higher DMI.

Objective: To report the hemodynamic and functional responses obtained with clinical optimization guided by hemodynamic parameters in patients with severe and refractory heart failure., Methods: Invasive hemodynamic monitoring using right heart catheterization aimed to reach low filling pressures and peripheral resistance. Frequent adjustments of intravenous diuretics and vasodilators were performed according to the hemodynamic measurements., Results: We assessed 19 patients (age = 48+/-12 years and ejection fraction = 21+/-5%) with severe heart failure. The intravenous use of diuretics and vasodilators reduced by 12 mm Hg (relative reduction of 43%) pulmonary artery occlusion pressure (P<0.001), with a concomitant increment of 6 mL per beat in stroke volume (relative increment of 24%, P<0.001). We observed significant associations between pulmonary artery occlusion pressure and mean pulmonary artery pressure (r=0.76; P<0.001) and central venous pressure (r=0.63; P<0.001). After clinical optimization, improvement in functional class occurred (P< 0.001), with a tendency towards improvement in ejection fraction and no impairment to renal function., Conclusion: Optimization guided by hemodynamic parameters in patients with refractory heart failure provides a significant improvement in the hemodynamic profile with concomitant improvement in functional class. This study emphasizes that adjustments in blood volume result in immediate benefits for patients with severe heart failure.

The objective of this trial was to compare the effects of increasing dietary energy density from 1.51 to 1.65 Mcal/kg of dry matter (DM) by replacing forage with concentrate or by further increasing concentrate via the substitution of corn silage and alfalfa silage by a mixture of straw, starch, and soybean meal. Our hypothesis was that the latter diet would be more glucogenic while increasing rumen fill and be potentially desirable for transition cows. Nine far-off dry cows (greater than 3 wk before parturition at the end of the trial) were fed three diets: low energy diet, [LE, 1.51 Mcal/kg of DM, 14.0% crude protein (CP) and 35% nonfiber carbohydrates (NFC)], high energy diet, (HE, 1.65 Mcal/kg of DM, 13.9% CP and 39.5% NFC) and high energy diet, where a portion of alfalfa and corn silage was replaced by straw, soybean meal, and cornstarch (HES, 1.65 Mcal/kg of DM, 13.5% CP and 40.5% NFC). The experiment was a replicated 3 x 3 Latin square design with 21-d periods. Six cows from two squares were used to examine kinetics of DM disappearance from nylon bags suspended in the rumen. Two contrasts of interest were: LE versus HE, HES (effects of energy density) and HE versus HES (method of increasing energy density). Increasing energy density increased the potentially degradable (B) and decreased the undergradable (C) DM fractions of the diets. Because HES had greater B and a faster rate of degradation of fraction B (k), effective rumen degradable DM (ERDDM) was higher in HES compared to HE. Cows fed high energy diets had greater DM intake. No differences in DM intake were observed between HE and HES. Rumen volume or DM pool sizes were not affected by treatment. High energy diets increased total ruminal fluid volatile fatty acid concentration compared with LE. Propionate concentration was higher in cows fed high energy diets compared with cows fed LE. The partial replacement of alfalfa and corn silage by straw, soybean meal, and cornstarch further increased propionate concentration. The greatest increase in serum insulin concentration following feeding was observed in cows fed HE. Cows consuming high energy diets had lower plasma nonesterified fatty acids (NEFA) before and after feeding. The HES diet was less effective in decreasing plasma NEFA concentration after feeding compared to HE. In conclusion, increasing diet energy density of far-off dry cows positively affected DMI, ruminal propionate, serum insulin, and plasma NEFA. Increasing energy density with a blend of feeds that represent extremes in rates of carbohydrate fermentation may be a strategy to provide greater amounts of glucogenic precursors. Applicability of this strategy should be examined in transition cows.

Adriamycin cardiotoxicity is associated with oxidative stress in the presence of globally depressed cardiac function. It is unknown if there is a similar profile with early diastolic changes and how it relates to baroreflex control of circulation. In this study, we evaluated baroreflex control of circulation in adriamycin-treated Wistar rats compared with controls, using invasive blood pressure recording processed by a data acquisition system (CODAS, 1 KHz). Baroreflex sensitivity was evaluated by modulating blood pressure with phenylephrine and sodium nitroprusside. Oxidative stress was quantified by chemiluminescence and by glutathione peroxidase enzyme activity. Diastolic dysfunction was characterized by increased left ventricle end-diastolic pressure in adriamycin-treated rats compared with controls with preserved ascending aortic flow. Baroreflex sensitivity in response to blood pressure elevation and reduction were similar in adriamycin (-2+/-0.27 and -3.19+/-0.56 bpm/mm Hg) and control rats (-1.35+/-0.15 and -2.52+/-0.39 bpm/mm Hg). Chemiluminescence was higher (20450+/-1286 versus 16517+/-1020 counts per second/mg protein) and glutathione peroxidase activity was lower (45.6+/-4.3 versus 76.4+/-6.9 micromol. min(-1). mg(-1) protein) in adriamycin rats compared with controls. Inverse correlations were observed between glutathione peroxidase activity and left ventricle end-diastolic pressure (r=-0.72, P=0.02), between baroreflex sensitivity to phenylephrine and left ventricle end-diastolic pressure (r=-0.77, P=0.004), and between chemiluminescence and baroreflex sensitivity to sodium nitroprusside (r=-0.75, P=0.02), whereas a positive correlation was observed between baroreflex sensitivity to sodium nitroprusside and glutathione peroxidase activity (r=0.7, P=0.04). Thus, adriamycin led to increased left ventricle end-diastolic pressure without changes in baroreflex sensitivity, and associated increased oxidative stress appeared to be related to reduction of reflex control of circulation.

Schistosoma are dioecious digenetic trematodes carrying a large (270 Mb) genome. Gaining knowledge about the genome of these parasites is of importance for the understanding of their biology, mechanisms of drug resistance and antigenic variation that determine escape from the host's immune system. This review will provide an update on the Schistosoma Gene Discovery Program, which is part of the Schistosoma Genome Project created in 1992. One of the main objectives of this program is the discovery and characterisation of new genes of Schistosoma mansoni and Schistosoma japonicum in an attempt to search for new targets for drugs and vaccine development. The success of the Schistosoma Gene Discovery Program is demonstrated by the number of catalogued genes, that now reaches 15 to 20% of the full gene complement of its genome.

With the widespread use of obstetric echography the incidence of fetal hydronephrosis has been reported more frequently. Consequently, many uropathies have been detected in asymptomatic neonates. The authors report their experience with prenatally detected primary non-refluxing megaureter. Newborns with fetal hydronephrosis were investigated by ultrasonography and micturating cystourethrogram after the beginning of chemoprophylaxis. If primary megaureter was identified, after 1 month the children underwent 99tm-DMSA, diuretic 99tm-DTPA, and intravenous urography. Eight infants with primary megaureter (bilateral in 3 cases) were identified, for a total of 11 renal units for study. All children were submitted to non-operative management. We performed ultrasonography and diuretic 99tm-DTPA during follow-up, which lasted on average 75 months. The mean cross-sectional diameter of the dilated ureter was 13.6 mm during neonatal period, and reached 8.4 mm at the end of follow-up. The renal function and the diuretic renogram remained stable throughout follow-up. Two neonates presented transitory hypertension. Our results support the notion that conservative management is safe for primary megaureter detected in asymptomatic neonates, with most cases showing spontaneous regression during a prolonged follow-up.

ESTs constitute rapid and informative tools with which to study gene-expression profiles of the diverse stages of the schistosome life cycle. Following a comprehensive EST study of adult worms, analysis has now targeted the cercaria, the parasite larval form responsible for infection of the vertebrate host. Two Schistosoma mansoni cercarial cDNA libraries were examined and partial sequence obtained from 957 randomly selected clones. On the basis of database searches, 551 (57.6%) ESTs generated had no homologs in the public databases whilst 308 (32.2%) were putatively identified, totaling 859 informative ESTs. The remaining 98 (10.2%) were uninformative ESTs (ribosomal RNA and non-coding mitochondrial sequences). By clustering analysis we have identified 453 different genes. The most common sequences in both libraries represented Sm8 calcium binding protein (8% of ESTs), fructose-1,6-bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase, cytochrome oxidase subunit 1, ATP guanidine kinase and triose phosphate isomerase. One hundred and nineteen identified genes were sorted into 11 functional categories, with genes associated with energy metabolism being the most abundant (13%) and diverse. The diversity and abundance of genes associated with the transcription/translation machinery and with regulatory/signaling functions were also marked. A paramyosin transcript was identified, indicating that this gene is not exclusively expressed in adult worms and sporocysts (as had been suggested previously). The possible physiological relevance to cercariae of the presence of transcripts with homology to calcium binding proteins of the EF-hand superfamily, Gq-coupled rhodopsin photoreceptor, rod phosphodiesterase 8 subunit and peripheral-type benzodiazepine receptor is discussed.

Primary vesicoureteric reflux (VUR) diagnosed on investigation of foetal hydronephrosis accounts for many antenatally detected uropathies. In order to study foetal VUR and its consequences, newborns with foetal hydronephrosis were investigated by ultrasound, micturating cystourethrogram and 99mTechnetium-dimercapto-succinic acid (DMSA), after beginning of chemoprophylaxis. Twenty-eight infants with VUR (bilateral in 15 cases) were identified giving a total 43 renal units for study. There was a predominance of males (86%), moderate/severe reflux (84%) and renal damage (51%). Presence of renal damage was correlated with the severity of reflux. VUR should be investigated in cases of foetal hydronephrosis and our results support that renal damage is frequently congenital and not secondary to urinary tract infection.

Continuing the Schistosoma mansoni Genome Project 363 new templates were sequenced generating 205 more ESTs corresponding to 91 genes. Seventy four of these genes (81%) had not previously been described in S. mansoni. Among the newly discovered genes there are several of significant biological interest such as synaptophysin, NIFs-like and rho-GDP dissociation inhibitor.

A comparative study of the gene expression profile in different developmental stages of Schistosoma mansoni has been initiated based on the expressed sequence tag (EST) approach. A total of 1401 ESTs were generated from seven different cDNA libraries constructed from four distinct stages of the parasite life cycle. The libraries were first evaluated for their quality for a large-scale cDNA sequencing program. Most of them were shown to have less than 20% useless clones and more than 50% new genes. The redundancy of each library was also analyzed, showing that one adult worm cDNA library was composed of a small number of highly frequent genes. When comparing ESTs from distinct libraries, we could detect that most genes were present only in a single library, but others were expressed in more than one developmental stage and may represent housekeeping genes in the parasite. When considering only once the genes present in more than one library, a total of 466 unique genes were obtained, corresponding to 427 new S. mansoni genes. From the total of unique genes, 20.2% were identified based on homology with genes from other organisms, 8.3% matched S. mansoni characterized genes and 71.5% represent unknown genes.

Many postembryonic developmental processes are regulated by an intricate interplay among hormones and growth factors. Thyroid hormone (TH) and estrogen are well known to be individually and obligatorily required for the initiation and progression of amphibian metamorphosis and vitellogenesis. However, whether or not a possible interplay between these two hormones would affect these two developmental processes is not known. Here we report on how triiodothyronine (T3) enhances the precocious activation of vitellogenin (Vit) genes by estradiol (E2) in Xenopus tadpoles during metamorphosis. Using a combination of filter hybridization, RNase protection assay and in situ hybridization, we first show that very low doses (10(-9) M) of exogenous T3 will autoinduce thyroid hormone receptor (TR) mRNA in several tissues of premetamorphic tadpoles. The same treatment enhances and accelerates the precocious activation of the silent vitellogenin genes by E2 at metamorphic climax (stages 60-64) but not before mid-metamorphosis (stages 56-58). This developmental stage dependency may be explained by our finding that, under the same experimental conditions, T3 fails to alter the autoinduction of ER mRNA at mid-metamorphosis but strongly potentiates it at metamorphic climax. Thus a developmental stage specific interplay between thyroid hormone and estrogen determines the kinetics and extent of activation of vitellogenin and estrogen receptor genes during Xenopus postembryonic development.

Although the important role of thyroid hormones in regulating metamorphosis of amphibian larvae is well known, it has not been clearly established if thyroid hormones have any function in the activities of adult amphibian tissues. We now describe a strong effect of 3,3',5-triiodothyronine (T3) on adult Xenopus liver cells. Low doses of T3 rapidly (within 6-12 h) potentiate the activation of vitellogenin (Vit) genes by estradiol-17 beta (E2) in primary cultures of adult male and female Xenopus hepatocytes. This effect is developmentally regulated and is first manifested during metamorphic climax. In an attempt to explain this potentiation, we find that T3 also upregulates thyroid hormone receptor beta, but not alpha, transcripts and rapidly enhances the autoinduction of estrogen receptor (ER) mRNA in adult Xenopus hepatocytes. In transient transfection of the Xenopus cell line XTC-2 with an estrogen response element--chloramphenicol transacetylase (ERE-CAT) construct T3 was found to potentiate the transcription by E2 from the transfected ERE, thus suggesting that it enhances the accumulation of functional ER. We conclude that T3 can function in adult amphibian tissues, and discuss the significance of thyroid hormone potentiation of responses to estrogen in reproductive processes.

Although downregulation of receptors by their respective hormonal ligands is a well-studied phenomenon, relatively less is known about autoupregulation of receptors. However, an increasing number of observations of the latter process are now being reported. Here, we discuss the phenomenon of autoinduction of nuclear receptors of the steroid/thyroid hormone gene family, and its significance in the context of the developmental and gene regulatory function of the ligands. Much of this review is illustrated by recent work from our laboratory on the autoregulation of Xenopus estrogen (ER) and thyroid hormone (TR) receptors and their transcripts, accompanying or anticipating vitellogenesis and metamorphosis, respectively. The activation by estrogen (E2) of the silent vitellogenin genes and the induction of FOSP-1 genes in primary cultures of hepatocytes from male Xenopus and oviduct cells, respectively, are tightly coupled to a substantial upregulation of ER protein and its transcript. The developmental competence to activate vitellogenin in response to E2 was found to be acquired during late metamorphosis. Since the latter process is obligatorily controlled by thyroid hormones (TH), we extended our studies to the developmental and hormonal regulation of Xenopus TR genes. Although very low levels of TR alpha and beta mRNAs are detectable in embryos and early larvae, there is a large increase in the accumulation of both transcripts before the onset of metamorphosis (stage 54 tadpoles), by which time the larval thyroid gland has first begun to secrete TH. Filter and in situ hybridization revealed that the two transcripts were differentially regulated and were not equally distributed in all regions or tissues of the tadpole. Their concentration peaks at metamorphic climax and drops to low levels in froglets and adult Xenopus. Exogenous TH given to pre-metamorphic tadpoles is known to induce metamorphosis precociously. Administration of triiodothyronine (T3) to early tadpoles (stages 50/52) caused a rapid upregulation of TR alpha and beta mRNAs which was particularly marked for the beta transcript (20- to 50-fold increase in steady-state levels). This autoinduction, which is the earliest response to T3, is mimicked to variable degrees in some Xenopus cell lines. In XTC-2 cells, in which the in vivo process is fully reproduced, it was possible to show with cycloheximide that the increase in TR mRNA requires protein synthesis. It was also possible to show by transfection of XTC-2 cells with a reporter-promoter construct of Xenopus albumin gene, which is a target for T3, that the extra TR mRNA increases functional receptor in the cell. Although the role of TH is well-known in metamorphosis, we found that TR is also autoinduced in primary culture of adult male Xenopus hepatocytes. The significance of this finding lies in the fact that T3 potentiates the autoinduction of ER and the de novo activation of vitellogenin genes by E2. Prolactin (PRL) is known to exert a "juvenilizing" action by preventing the induction of amphibian metamorphosis by TH. It is therefore highly significant that PRL prevented both the autoinduction of TR alpha and beta mRNAs in whole tadpoles and organ cultures and the activation of TR target genes, such as those encoding albumin and 63 kDa adult-type keratin. Although how PRL exerts its antimetamorphic effect is not known, these findings lead us to propose a dual threshold model for the autoinduction of TR, whereby the autoinduction of TR genes requires a very low level of TR and TH to rapidly augment the amount of functional TR. This higher amount of receptor would be required to achieve a higher threshold to activate "downstream" or target genes which specify the adult phenotype at the end of metamorphosis. Finally, a survey of recent literature shows that the phenomenon of autoinduction is not restricted to Xenopus ER and TR but is more widespread among members of the nuclear receptor family.

A pool of nuclear proteins from adult worms of Schistosoma mansoni was analyzed for amino acid composition and found to be compatible with high mobility group (HMG) proteins. One of the schistosome HMG proteins was identified as HMG 2 by one-dimensional and two-dimensional PAGE. Stage-specific differences in the HMG-like protein composition were encountered when adult worms were compared to schistosomula, the larval form. Immobilization of the adult male and female nuclear proteins onto nitrocellulose, followed by hybridization against 32P-F-10, a schistosome sex specific gene encoding a major egg shell protein, revealed distinct banding patterns. On the other hand, a synthetic oligonucleotide, derived from the 3' untranslated end of the F-10 gene and possibly containing one regulatory element of the gene, bound mainly to male low MW proteins.