Egg yolk immunoglobulin Y as a promising tool to detect immune complexes in neurocysticercosis serum samples.

BACKGROUND: Neurocysticercosis (NCC) is a neglected tropical disease and its diagnosis is still a challenge due to non-specific manifestations. Neuroimaging techniques are used in the diagnosis of NCC, however, due to the high cost of these methods and the advantages presented in the use of immunological tests, such as ease of performance and satisfactory results, immunoassays are commonly used to detect antibodies against Taenia sp. antigens. The aim of the present study was to produce, characterize and apply specific polyclonal immunoglobulin Y (IgY) anti-Taenia crassiceps extracted from egg yolk of hens immunized with T. crassiceps metacestodes. METHODS: Indirect enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence tests were performed for characterization of IgY antibodies. Diagnostic performance was verified by ELISA for immune complex detection testing 90 serum samples. RESULTS: Values of sensitivity, specificity, positive and negative likelihood ratios (LR+/LR-) and area under the curve (AUC) were calculated and presented the following results: sensitivity 83.3%, specificity 96.7%, AUC 0.966, LR+ 25.0 and LR- 0.17. CONCLUSIONS: Results of this pioneering and innovative study demonstrate that anti-T. crassiceps IgY antibodies present potential applicability and can be used as an efficient tool in human NCC serodiagnosis.

IgY antibody and human neurocysticercosis: a novel approach on immunodiagnosis using Taenia crassiceps hydrophobic antigens.

Human neurocysticercosis (NCC) is a worldwide neglected disease caused by Taenia solium metacestode and responsible for various complications and neurological disorders. This study aimed to evaluate the use of specific immunoglobulin Y (IgY) produced by laying hens immunized with a hydrophobic fraction of Taenia crassiceps metacestodes (hFTc) in NCC diagnosis. Egg yolk IgY antibodies were fractionated, purified and characterized. Enzyme-linked immunosorbent assay (ELISA) was carried out to evaluate the production kinetics and avidity maturation of anti-hFTc IgY antibodies throughout the IgY obtention process. Antigen recognition tests were carried out by Western blotting and immunofluorescence antibody test using purified and specific anti-hFTc IgY antibodies for detection of parasitic antigens of T. crassiceps and T. solium metacestodes. Sandwich ELISA was performed to detect circulating immune complexes formed by IgG and parasitic antigens in human sera. The results showed high diagnostic values (93.2% sensitivity and 94.3% specificity) for immune complexes detection in human sera with confirmed NCC. In conclusion, specific IgY antibodies produced from immunized hens with hFTc antigens were efficient to detect T. solium immune complexes in human sera, being an innovative and potential tool for NCC immunodiagnosis.

Highly specific and sensitive anti-Strongyloides venezuelensis IgY antibodies applied to the human strongyloidiasis immunodiagnosis.

Due to the epidemiological problem of the neglected condition of human strongyloidiasis, rapid and effective diagnosis is extremely important, with the development of new diagnostic tools being essential to reduce infections and chronic cases. Avian immunoglobulin Y (IgY) technology is an alternative for antibody production that has high specificity and profitability. This study aimed to produce and fractionate IgY antibodies from the egg yolks of hens that were immunized with the total antigenic extracts of Strongyloides venezuelensis infectious filariform larvae (iL3) and parthenogenetic females (pF). IgY antibodies were then evaluated by their recognition of antigenic proteins, evolutive helminth forms, and serological diagnosis of human strongyloidiasis by the detection of immune complexes in serum samples. Egg yolks were fractionated to obtain IgY antibodies by thiophilic interaction chromatography. Immune complex detection in serum samples showed diagnostic values for anti-iL3 IgY and anti-pF IgY antibodies at 95.56% and 88.89% sensitivity and 95.56% and 91.11% specificity, respectively. Therefore, IgY technology is a promising tool for the detection of blood circulating Strongyloides antigens, with possible application as a serological diagnostic method.

Structure-Based Discovery of Thiosemicarbazone Metalloproteinase Inhibitors for Hemorrhage Treatment in Snakebites.

The venoms of snakes are composed by many toxins, which are responsible for various toxic effects including intense pain, bleeding disorders, and local tissue damage caused by hemorrhage and necrosis. The snake venom metalloproteinases (SVMPs) are proteolytic zinc-dependent enzymes acting in different hemostatic mechanisms. In this work, a structure-based molecular modeling strategy was used for the rational design, by means of a homology 3D model of an SVMP isolated from Bothrops pauloensis venom (BpMP-I), followed by synthesis and in vitro evaluation of new thiosemicarbazones as the first inhibitors of the B. pauloensis SVMP. Besides being effective for the SVMP inhibition, two molecules were shown to be effective also in vivo, inhibiting hemorrhage caused by the B. pauloensis whole venom. Docking studies on metalloproteinases from other snake species suggest that the thiosemicarbazones activity is not confined to BpMP-I, but seems to be a common feature of metzincins.

Produção e purificação de imunoglobulinas Y policlonais anti-Leptospira spp / Production and purification of polyclonal anti-Leptospira immunoglobulin Y

Objetivou-se verificar se galinhas imunizadas com uma solução de Leptospira interrogans inativadas e proteínas de membrana externa do sorovar Hardjo, poderiam produzir anticorpos policlonais específicos anti-leptospiras, detectáveis em testes ELISA. Foram imunizados oito galinhas com 25 semanas de idade, da raça White Leghorn, sendo três imunizadas com uma suspensão de leptospiras inativadas, três com uma solução de proteínas de membrana externa extraída do sorovar Hardjo e duas controle. Coletas de sangue foram realizadas quinzenalmente e de ovos diariamente. A IgY foi purificada a partir da gema dos ovos utilizando para a delipidação o método de diluição em água ácida e a precipitação com sulfato de amônio. Nos testes ELISA realizados para verificar a especificidade da IgY, foi demonstrada a produção de anticorpos anti-Leptospira, tanto no soro quanto nas gemas purificadas. O pico de produção de anticorpos específicos ocorreu na 5º semana após a primeira imunização. Ficou demonstrada a possibilidade da indução da produção de anticorpos específicos em galinhas imunizadas com leptospiras do sorovar Hardjo inativadas, bem como, com proteínas de membrana externa (PME) extraidas desse sorovar. As galinhas imunizadas com uma suspensão de leptospiras inativadas ou com PME de Leptospira interrogans do sorovar Hardjo produziram anticorpos reativos a PME Hardjo detectáves por teste ELISA.

The aim was to determine whether hens immunized with an inactivated suspension of Leptospira and a solution of outer membrane proteins extracted from the serovar Hardjo, could produce specific polyclonal antibodies to Leptospira, detected in ELISA assay. Eight hens White Leghorn race with 25-weeks-old were immunized, three with an inactivated suspension of Leptospira, three with a solution of outer membrane proteins (OMP) extracted from the serovar Hardjo and two controls immunized with saline. Blood samples were collected fortnightly and eggs daily. The IgY was purified from the egg yolk using the method for the delipidation of dilution with water acidic and ammonium sulfate precipitation. The ELISA assay was performed to verify the specificity of the IgY, these was possible to observe the production of specific antibody to Leptospira both in serum and purified egg yolk. The specific antibody titers peaked in the fifth week post immunization. The production of polyclonal IgY was effective for producing high titers of specific antibodies.

A cutoff point for peak oxygen consumption in the prognosis of heart failure patients with ß-blocker therapy.

PURPOSE: Beta-blockers (BB) have shown to improve outcomes among heart failure patients (HF). Adequate risk stratification is still a major concern for HF. The prognostic indexes have been detected, but only few parameters maintain consistently high power in predicting progression of disease and mortality. Peak oxygen consumption (VO(2) peak, ml kg(-1) min(-1)) is traditionally used for risk stratification in HF, however, there is limited evidence regarding predictive value of VO(2) peak in patients taking BB. METHODS: Two hundred twenty nine patients, aged 49 ± 13 years with diagnosed HF for more than 6 months due to ischemic (n=73), idiopathic dilated (n=149) and Chagas disease (n=7) underwent a cardiopulmonary exercise test (CPX). The ejection fraction was 38 ± 10%; clinical stability was defined as no change in the NYHA class or absence of hospitalization for heart failure and stable medical treatment during 3 months prior to CPX. Subjects were tracked for cardiac-related mortality after CPX. RESULTS: The mean follow-up period was 2.5 ± 1.1 years and means value for VO(2) peak was 16.3 ± 4. Current BB therapy included carvedilol (83.4%), metoprolol (7.8%), bisoprolol (3.9%) and others (4.8%). The area under the ROC curve for VO(2) peak was 0.80 (95% CI: 0.69-0.90, optimal threshold: 12.5 and 82% sensitivity/26% specificity, p<0.001). Kaplan-Meier analysis that revealed event-free survival for subjects in < and >12.5 was 28% and 2.8%, respectively (long-rank 34.8; p<0.001). CONCLUSIONS: VO(2) peak seems to maintain prognostic value in HF patients BB therapy. The present study also provides new evidence that optimal threshold value for VO(2) peak in the BB era is 12.5 ml kg(-1) min(-1).

Acute effects of continuous and interval aerobic exercise on 24-h ambulatory blood pressure in long-term treated hypertensive patients.

BACKGROUND: Despite antihypertensive therapy, it is difficult to maintain optimal systemic blood pressure (BP) values in hypertensive patients (HPT). Exercise may reduce BP in untreated HPT. However, evidence regarding its effect in long-term antihypertensive therapy is lacking. Our purpose was to evaluate the acute effects of 40-minute continuous (CE) or interval exercise (IE) using cycle ergometers on BP in long-term treated HPT. METHODS: Fifty-two treated HPT were randomized to CE (n=26) or IE (n=26) protocols. CE was performed at 60% of reserve heart rate (HR). IE alternated consecutively 2 min at 50% reserve HR with 1 min at 80%. Two 24-h ambulatory BP monitoring were made after exercise (postexercise) or a nonexercise control period (control) in random order. RESULTS: CE reduced mean 24-h systolic (S) BP (2.6+/-6.6 mm Hg, p=0.05) and diastolic (D) BP (2.3+/-4.6, p=0.01), and nighttime SBP (4.8+/-6.4, p<0.001) and DBP (4.6+/-5.2 mm Hg, p=0.001). IE reduced 24-h SBP (2.8+/-6.5, p=0.03) and nighttime SBP (3.4+/-7.2, p=0.02), and tended to reduce nighttime DBP (p=0.06). Greater reductions occurred in higher BP levels. Percentage of normal ambulatory BP values increased after CE (24-h: 42% to 54%; daytime: 42% to 61%; nighttime: 61% to 69%) and IE (24-h: 31% to 46%; daytime: 54% to 61%; nighttime: 46% to 69%). CONCLUSION: CE and IE reduced ambulatory BP in treated HPT, increasing the number of patients reaching normal ambulatory BP values. These effects suggest that continuous and interval aerobic exercise may have a role in BP management in treated HPT.

Acute aerobic exercise reduces 24-h ambulatory blood pressure levels in long-term-treated hypertensive patients.

BACKGROUND: Even with anti-hypertensive therapy, it is difficult to maintain optimal systemic blood pressure values in hypertensive patients. Exercise may reduce blood pressure in untreated hypertensive, but its effect when combined with long-term anti-hypertensive therapy remains unclear. Our purpose was to evaluate the acute effects of a single session of aerobic exercise on the blood pressure of long-term-treated hypertensive patients. METHODS: Fifty treated hypertensive patients (18/32 male/female; 46.5+/-8.2 years; Body mass index: 27.8+/-4.7 kg/m(2)) were monitored for 24 h with respect to ambulatory (A) blood pressure after an aerobic exercise session (post-exercise) and a control period (control) in random order. Aerobic exercise consisted of 40 minutes on a cycle-ergometer, with the mean exercise intensity at 60% of the patient's reserve heart rate. RESULTS: Post-exercise ambulatory blood pressure was reduced for 24 h systolic (126+/-8.6 vs. 123.1+/-8.7 mmHg, p=0.004) and diastolic blood pressure (81.9+/-8 vs. 79.8+/-8.5 mmHg, p=0.004), daytime diastolic blood pressure (85.5+/-8.5 vs. 83.9+/-8.8 mmHg, p=0.04), and nighttime S (116.8+/-9.9 vs. 112.5+/-9.2 mmHg, p<0.001) and diastolic blood pressure (73.5+/-8.8 vs. 70.1+/-8.4 mmHg, p<0.001). Post-exercise daytime systolic blood pressure also tended to be reduced (129.8+/-9.3 vs. 127.8+/-9.4 mmHg, p=0.06). These post-exercise decreases in ambulatory blood pressure increased the percentage of patients displaying normal 24h systolic blood pressure (58% vs. 76%, p=0.007), daytime systolic blood pressure (68% vs. 82%, p=0.02), and nighttime diastolic blood pressure (56% vs. 72%, p=0.02). Nighttime systolic blood pressure also tended to increase (58% vs. 80%, p=0.058). CONCLUSION: A single bout of aerobic exercise reduced 24h ambulatory blood pressure levels in long-term-treated hypertensive patients and increased the percentage of patients reaching normal ambulatory blood pressure values. These effects suggest that aerobic exercise may have a potential role in blood pressure management of long-term-treated hypertensive.

Acute aerobic exercise reduces 24-h ambulatory blood pressure levels in long-term-treated hypertensive patients

BACKGROUND: Even with anti-hypertensive therapy, it is difficult to maintain optimal systemic blood pressure values in hypertensive patients. Exercise may reduce blood pressure in untreated hypertensive, but its effect when combined with long-term anti-hypertensive therapy remains unclear. Our purpose was to evaluate the acute effects of a single session of aerobic exercise on the blood pressure of long-term-treated hypertensive patients. METHODS: Fifty treated hypertensive patients (18/32 male/female; 46.5±8.2 years; Body mass index: 27.8±4.7 kg/m²) were monitored for 24 h with respect to ambulatory (A) blood pressure after an aerobic exercise session (post-exercise) and a control period (control) in random order. Aerobic exercise consisted of 40 minutes on a cycle-ergometer, with the mean exercise intensity at 60 percent of the patient's reserve heart rate. RESULTS: Post-exercise ambulatory blood pressure was reduced for 24 h systolic (126±8.6 vs. 123.1±8.7 mmHg, p=0.004) and diastolic blood pressure (81.9±8 vs. 79.8±8.5 mmHg, p=0.004), daytime diastolic blood pressure (85.5±8.5 vs. 83.9±8.8 mmHg, p=0.04), and nighttime S (116.8±9.9 vs. 112.5±9.2 mmHg, p<0.001) and diastolic blood pressure (73.5±8.8 vs. 70.1±8.4 mmHg, p<0.001). Post-exercise daytime systolic blood pressure also tended to be reduced (129.8±9.3 vs. 127.8±9.4 mmHg, p=0.06). These post-exercise decreases in ambulatory blood pressure increased the percentage of patients displaying normal 24h systolic blood pressure (58 percent vs. 76 percent, p=0.007), daytime systolic blood pressure (68 percent vs. 82 percent, p=0.02), and nighttime diastolic blood pressure (56 percent vs. 72 percent, p=0.02). Nighttime systolic blood pressure also tended to increase (58 percent vs. 80 percent, p=0.058). CONCLUSION: A single bout of aerobic exercise reduced 24h ambulatory blood pressure levels in long-term-treated hypertensive patients and increased the percentage of patients reaching normal...

Normalization of right ventricular performance and remodeling evaluated by magnetic resonance imaging at late follow-up of heart transplantation: relationship between function, exercise capacity and pulmonary vascular resistance.

BACKGROUND: Right ventricular (RV) dysfunction remains one of the most prominent complications during the period immediately after heart transplantation (HT); however, late adaptation of the RV has not been well described. The aim of our study was to evaluate RV function and remodeling using magnetic resonance imaging (MRI) and to correlate it with exercise capacity and also with hemodynamic data obtained before HT. METHODS: We prospectively evaluated RV function of 25 heart-transplanted patients, without cardiac allograft vasculopathy, who were documented by negative dobutamine stress echocardiography during late follow-up (Group 1, 6 +/- 4.3 years) using MRI. We then compared Group 1 with a control group consisting of 10 patients, who were < or =1 year post-HT (Group 2), hemodynamically stable, and with the same pre-operative hemodynamic features as Group 1. Their pulmonary arterial systolic blood pressure (PSBP) varied from 17 to 67 mm Hg (43.2 +/- 15.3) and pulmonary vascular resistance (PVR) from 1.0 to 5.4 Wood units (2.5 +/- 1.12). The following parameters were studied: RV end-diastolic volume (EDV) and systolic volume (ESV); stroke volume (SV); ejection fraction (EF); and mass (M). We also evaluated the VO2 peak and slope VE/VCO2 values during a treadmill test. Data were analyzed and correlated with the hemodynamic values of PVR and PSBP obtained pre-HT. RESULTS: In Group 1, treadmill evaluation data showed exercise VO2 peak (19.9 +/- 3.19 ml/kg/min) and slope VE/VCO2 (36.9 +/- 4.5) values comparable to those of sedentary individuals; RV variables according to MRI were within normal ranges, with the following mean values for Groups 1 and 2, respectively: RVEDV, 99.6 +/- 4.0 ml vs 127 +/- 16 ml (p = 0.03); RVESV, 42 +/- 2 ml vs 58.5 +/- 9 ml (p = 0.01); RVSV, 57 +/- 3 ml vs 71 +/- 10 ml (p = 0.1); RVEF, 58 +/- 1.4% vs 54 +/- 3.8% (p = 0.29); and RVM, 43.4 +/- 1.9 g vs 74 +/- 8.8 g (p = 0.001). There was no correlation between hemodynamic pulmonary values before HT or any other index of late RV performance, including RV remodeling and hypertrophy, in our study population (p = not significant). CONCLUSIONS: In contrast to what we would expect for heart transplant patients at late follow-up, the RV may adapt to pulmonary pressure and resistance, with reverse remodeling characterized by volume and mass reduction, leading to normalization of RV function despite abnormal hemodynamic pulmonary values being measured before HT. There was no influence on the low exercise capacity observed in these patients, in the absence of cardiac allograft vasculopathy.