Characterization of microplastics in skim-milk powders.

The diffusion of microplastics in the food supply chain is prompting public concern as their impact on human health is still largely unknown. The aim of this study was to qualitatively and quantitatively characterize microplastics in skim-milk powder samples (n = 16) from different European countries (n = 8) through Fourier-transform infrared micro-spectroscopy in attenuated total reflectance mode analysis. The present study highlights that the use of hot alkaline digestion has enabled the efficacious identification of microplastics in skim-milk powders used for cheese-making across European countries. The adopted protocol allowed detection of 29 different types of polymeric matrices for a total of 536 plastic particles. The most abundant microplastics were polypropylene, polyethylene, polystyrene, and polyethylene terephthalate. Microplastics were found in skim-milk powders in 3 different shapes (fiber, sphere, and irregular fragments) and 6 different colors (black, blue, brown, fuchsia, green, and gray). Results demonstrate the presence of microplastics in all skim-milk powder samples, suggesting a general contamination. Results of the present study will help to evaluate the impact of microplastics intake on human health.

Specifying requirements for collection and analysis of online user feedback.

According to data-driven Requirements Engineering (RE), explicit and implicit user feedback can be considered a relevant source of requirements, thus supporting requirements elicitation. However, limited attention has been paid so far to the role of online feedback in RE tasks, such as requirements validation, and on how to specify what online feedback to collect and analyse. We performed an action research study, together with a company that developed a platform for online training. This paper presents the design and execution of the study, and a discussion of its results. This study provides evidence about the need of practitioners to follow a simple but systematic approach for specifying requirements for data collection and analysis, at design time. Another outcome of this study is a method to tackle this task that leverages goal-oriented requirements modelling combined with Goal-Question-Metric. The applicability of the method has been explored on two industrial evaluations, while the perceived effectiveness, efficiency and acceptance have been assessed with practitioners through a dedicated survey.

Radiofrequency catheter ablation of accessory pathways in the dog: the Italian experience (2008-2016).

INTRODUCTION: Accessory pathways (APs) in dogs are mostly right-sided, display nondecremental conduction, and mediate atrioventricular reciprocating tachycardias (AVRTs). Radiofrequency catheter ablation (RFCA) is considered the first-line therapy in human patients to abolish electrical conduction along APs. ANIMALS: Seventy-six consecutive client-owned dogs. MATERIAL AND METHODS: Retrospective study to describe the precise anatomical distribution and the electrophysiologic characteristics of APs in a large population of dogs and to evaluate long-term success and complication rates of RFCA. RESULTS: Eighty-three APs were identified in 76 dogs (92.1% with single APs and 7.9% with multiple APs); 96.4% were right-sided, 3.6% left-sided. Conduction along the APs was unidirectional and retrograde in 68.7% of the cases and bidirectional in 31.3%. Accessory pathways presented retrograde decremental properties in 6.5% of the cases. They mediated orthodromic AVRT in 92.1% of the cases and permanent junctional reciprocating tachycardia in 6.5%. In one case, no AVRT could be induced. In 97.4% of dogs, RFCA was attempted with an acute success rate of 100%. In 7.7% of cases, recurrence of the tachycardia occurred within 18 months, followed by a second definitively successful ablation. A major complication requiring pacemaker implantation was identified in 2.6% of dogs. DISCUSSION: Accessory pathway distribution and electrophysiologic properties in these 76 dogs were similar to previous report. Long-term success and complication rates of RFCA in dogs appeared very similar to results of humans. CONCLUSION: Radiofrequency catheter ablation of APs can be performed with a high success rate and low incidence of complications.

Short-term, supra-physiological rhGH administration induces transient DNA damage in peripheral lymphocytes of healthy women.

PURPOSE: While a good safety for recombinant human growth hormone (rhGH) therapy at replacement doses is recognized, a possible link between high concentration of the GH-IGF-I axis hormones and side negative effect has been reported. The aim of this pilot study was to assess whether a short-term exposure to supra-physiological doses of rhGH may affect DNA integrity in human lymphocytes (PBL). METHODS: Eighteen healthy Caucasian female (24.2 ± 3.5 years) were randomly included in a Control (n = 9) and rhGH administration group (n = 9, 3-week treatment). DNA damage (comet assay), chromosomal breaks, and mitotic index in phytohemagglutinin-stimulated PBL were evaluated before (PRE), immediately (POST), and 30 days (POST30) after the last rhGH administration (0.029 mg kg- 1 BW; 6 days/week), together with serum IGF-1 and IGFBP-3 concentrations. RESULTS: rhGH administration increased IGF-I, without evidence of persisting IGF-I and IGFBP-3 changes 30 days after withdrawal. Total DNA breakage (% DNA in tails) was not significantly different in subjects treated with rhGH in comparison with controls, although the rhGH-treated subjects showed an higher percentage of heavily damaged nuclei immediately after the treatment (POST30 vs. PRE: p = 0.003), with a lower mitogenic potential of lymphocytes, detectable up to the POST30 (PRE vs. POST: p = 0.02; PRE vs. POST30: p = 0.007). CONCLUSIONS: This pilot study showed that 3 weeks of short-term supra-physiological rhGH administration in healthy women induce a transient DNA damage and mitogenic impairment in PBL. The analysis of DNA damage should be explored as useful tool in monitoring the mid to long-term effects of high rhGH treatment or abuse.

[Treatment of eating disorders during hospitalization: presentation of a hospital intensive care program in pediatric age]. / Gestione dei disturbi del comportamento alimentare durante il ricovero: presentazione di un programma di terapia intensiva ospedaliera in eta pediatrica.

AIM: The aim of this study was to present a hospital intensive care program for patients affected by a severe eating disorders, with a significant loss of weight (BMI<15 and weight less than 25° centile), severe medical complications (orthostatic hypotension, bradycardia <40 bpm or tachycardia >110 bpm or inability to sustain core body temperature), abnormal laboratory data, especially electrolyte imbalance and refusal to take food and fluids. METHODS: In our study we reported 2 year follow-up of 16 patients treated with the hospital intensive care program between 2007 and 2008 in our department. RESULT: The proposed program was proved an efficient method in a critical phase of the alimentary behavior disorders. It was possible for all the patients to avoid alternative feeding techniques (enteral or parenteral) and to obtain a correct alimentation with a satisfactory improvement of clinical conditions. Eight patients (50%) fully recovered. 5 patients (31.25%) had a significant improvement reaching a BMI>18.5 and one of them had a regular menstrual cycle, too. However in this group of patients a strict modality to alimentation and concern about weight and physical appearance remain. In 3 patients (18.5%) the BMI is still low and amenorrhea persists. CONCLUSIONS: The hospital intensive care program, inspired by the cognitive-behavioral model, through a food rehabilitation and a psychotherapeutic and psychoeducational help, lets the patients and their family understand and modify the dysfunctional patterns, experimenting a right modality to approach alimentation, with a satisfactory improvement in clinical conditions.

Flavonone treatment reverses airway inflammation and remodelling in an asthma murine model.

BACKGROUND AND PURPOSE: Asthma is an inflammatory disease that involves airway hyperresponsiveness and remodelling. Flavonoids have been associated to anti-inflammatory and antioxidant activities and may represent a potential therapeutic treatment of asthma. Our aim was to evaluate the effects of the sakuranetin treatment in several aspects of experimental asthma model in mice. EXPERIMENTAL APPROACH: Male BALB/c mice received ovalbumin (i.p.) on days 0 and 14, and were challenged with aerolized ovalbumin 1% on days 24, 26 and 28. Ovalbumin-sensitized animals received vehicle (saline and dimethyl sulfoxide, DMSO), sakuranetin (20 mg kg(-1) per mice) or dexamethasone (5 mg kg(-1) per mice) daily beginning from 24th to 29th day. Control group received saline inhalation and nasal drop vehicle. On day 29, we determined the airway hyperresponsiveness, inflammation and remodelling as well as specific IgE antibody. RANTES, IL-5, IL-4, Eotaxin, IL-10, TNF-α, IFN-γ and GMC-SF content in lung homogenate was performed by Bioplex assay, and 8-isoprostane and NF-kB activations were visualized in inflammatory cells by immunohistochemistry. KEY RESULTS: We have demonstrated that sakuranetin treatment attenuated airway hyperresponsiveness, inflammation and remodelling; and these effects could be attributed to Th2 pro-inflammatory cytokines and oxidative stress reduction as well as control of NF-kB activation. CONCLUSIONS AND IMPLICATIONS: These results highlighted the importance of counteracting oxidative stress by flavonoids in this asthma model and suggest sakuranetin as a potential candidate for studies of treatment of asthma.

Assessment of swim-up and discontinuous density gradient in sperm sex preselection for bovine embryo production / Avaliação do swim-up e do gradiente descontinuo de densidade na pré-seleção do sexo de espermatozoides na produção de embriões bovinos

The purpose of this work was to associate the modified swim-up method with centrifugation in density gradient for the separation of X-bearing spermatozoa. Sperm viability and integrity were evaluated through the Trypan Blue/Giemsa staining method. Quality control of centrifuged spermatozoa was performed in in vitro produced embryos. The results were validated by the sex ratio of in vitro produced embryos using PCR by Y- specific sequences present in bovine male genomic DNA. After determining genetic sex of in vitro produced embryos, the results showed difference (P<0.05) in deviation of sex ratio when comparing the control group (45.2% females) with the other spermatozoa selection procedures (60.6% females) (P<0.05). The sperm selection methods are capable of selecting X-bearing spermatozoa without compromising the spermatozoa fertility (cleavage and blastocyst rates, 70% and 26%, respectively) and were considered relevant methods to be introduced in bovine in vitro produced embryo programs.

O objetivo do presente trabalho foi associar o método de swim-up modificado à centrifugação em gradiente de densidade para a separação de espermatozoides portadores do cromossomo X. A viabilidade e a integridade espermática foram avaliadas pelo método de coloração Azul de Tripan e Giemsa. O controle de qualidade dos espermatozoides centrifugados foi realizado por meio da produção in vitro de embriões bovinos. Os resultados foram validados pela técnica de PCR para verificar a proporção sexual dos embriões produzidos in vitro, com o uso de sequências Y especificas presente no DNA genômico de machos bovinos. Após determinar o sexo genético dos embriões produzidos in vitro, os resultados não mostraram diferença (P<0,05) no desvio da proporção do sexo quando comparou o grupo controle (45,2% de fêmeas) com os outros processos de seleção de espermatozoides (60,6% de fêmeas) (P<0,05). Os métodos de seleção de espermatozoides são capazes de selecionar espermatozoides portadores do cromossomo X sem comprometer a fertilidade, medida pelas taxas de clivagem e blastocisto de 70% e 26%, respectivamente, e foram considerados métodos de relevância para serem introduzidos nos programas de produção in vitro de embriões bovinos.

Exercise inhibits allergic lung inflammation.

Aerobic conditioning (AC) performed either during or after sensitization reduces allergic inflammation in mice; however, the effects of AC performed before and during allergic sensitization on airway inflammation are unknown. Mice were divided into Control, AC, OVA, and AC + OVA groups. Mice were trained in a treadmill followed by either ovalbumin (OVA) sensitization or saline administration. Peribronchial inflammation, OVA-specific IgE and IgG1 titers, the expression of Th1 and Th2 cytokines, and airway remodeling were evaluated, as well as the expression of Eotaxin, RANTES, ICAM-1, VCAM-1, TGF-ß and VEGF. Aerobic conditioning performed before and during allergic sensitization displayed an inhibitory effect on the OVA-induced migration of eosinophils and lymphocytes to the airways, a reduction of IgE and IgG1 titers and an inhibition of the expression of Th2 cytokines. The AC + OVA group also demonstrated reduced expression of ICAM-1, VCAM-1, RANTES, TGF-ß and VEGF, as well as decreased airway remodeling (p<0.05). The effects of AC before and during the sensitization process inhibit allergic airway inflammation and reduce the production of Th2 cytokines and allergen-specific IgE and IgG1.

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association.

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Validação comparativa utilizando PCR quantitativo em tempo real (qPCR) e PCR convencional de sêmen bovino centrifugado em gradiente de densidade contínuo / Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient

The objective of the present study was to determine the sperm enrichment with X-bearing spermatozoa, after one centrifugation in a Percoll or OptiPrep continuous density gradient, using quantitative real-time polymerase chain reaction (qPCR) of sperm DNA and resultant in vitro-produced bovine embryos by PCR. Frozen/thawed sperm was layered on density gradients and the tubes were centrifuged. Supernatants were gently aspirated and the sperm recovered from the bottom of the tubes. Cleavage and blastocyst rates were determined through in vitro production of embryos and PCR was performed to identify the embryos' genetic sex. A difference in blastocyst rate was found in the Percoll treatment compared to OptiPrep (P<0.05). The percentage of female embryos in the Percoll and OptiPrep groups was 62.0 percent and 47.1 percent, respectively. These results were confirmed by qPCR of spermatozoa DNA and underestimation was seen only in the Percoll group. It was possible to sexing sperm using simple approach.

O objetivo do presente estudo foi determinar o enriquecimento de espermatozoides portadores do cromossomo X após a centrifugação em gradiente de densidade contínuo de Percoll ou OptiPrep, utilizando reação em cadeia da polimerase quantitativa em tempo real (qPCR) do DNA do espermatozoide e dos embriões bovinos produzidos in vitro resultantes pela PCR convencional. Espermatozoides descongelado foram depositados em gradientes de densidade e os tubos foram centrifugados. Os sobrenadantes foram gentilmente aspirados e os espermatozoides recuperados do fundo dos tubos. As taxas de clivagem e de blastocisto foram determinadas pela produção in vitro de embriões e a PCR foi realizada para a identificação genética do sexo dos embriões. Verificou-se diferença na taxa de blastocistos entre os grupos Percoll e OptiPrep (P<0,05). A porcentagem de embriões de fêmeas nos grupos Percoll e OptiPrep foi de 62,0 por cento e 47,1 por cento, respectivamente. Estes resultados foram confirmados pela qPCR do DNA de espermatozoides e uma subestimação foi observada no grupo do gradiente de densidade de Percoll. Foi possível a sexagem de espermatozoides utilizando uma metodologia simples.

PAS-1, an Ascaris suum protein, modulates allergic airway inflammation via CD8+γδTCR+ and CD4+CD25+FoxP3+ T cells.

We have previously demonstrated that PAS-1, a 200 kDa protein from Ascaris suum, has a potent immunomodulatory effect on humoral and cell-mediated responses induced by APAS-3 (an allergenic protein from A. suum) or unrelated antigens. In this study, we investigated the mechanisms by which PAS-1 is able to induce this effect on an allergic airway inflammation induced by OVA in mice. C57BL/6 mice were adoptively transferred on day 0 with seven different PAS-1-primed cell populations: PAS-1-primed CD19(+) or B220(+) or CD3(+) or CD4(+) or CD8(+) or CD4(+) CD25⁻ or CD4(+) CD25(+) lymphocytes. These mice were immunized twice with OVA and alum by intraperitoneal route (days 0 and 7) and challenged twice by intranasal route (days 14 and 21). Two days after the last challenge, the airway inflammation was evaluated by antibody levels, cellular migration, eosinophil peroxidase levels, cytokine and eotaxin production, and pulmonary mechanical parameters. Among the adoptively transferred primed lymphocytes, only CD4(+) CD25(+) , CD8(+) or the combination of both T cells impaired the production of total IgE and OVA-specific IgE and IgG1 antibodies, eosinophilic airway inflammation, Th2-type cytokines (IL-4, IL-5 and IL-13), eotaxin release and airway hyperreactivity. Moreover, airway recruited cells from CD4(+) CD25(+) and CD8(+) T-cell recipient secreted more IL-10/TGF-ß and IFN-γ, respectively. Moreover, we found that PAS-1 expands significantly the number of CD4(+) CD25(+) FoxP3(+) and CD8(+) γδTCR(+) cells. In conclusion, these findings demonstrate that the immunomodulatory effect of PAS-1 is mediated by these T-cell subsets.

Use of transcutaneous external pacing during transvenous pacemaker implantation in dogs.

The aim of this retrospective study was to evaluate the utility of transcutaneous external pacing (TEP) during transvenous pacemaker implantation in dogs. Eighty-two pacemakers were implanted in 77 dogs because of third-degree atrioventricular block (AVB) (58 cases; 70.7 per cent), sinus node dysfunction (SND) (nine cases; 11.0 per cent), high-grade second-degree AVB (six cases; 7.3 per cent), persistent atrial standstill (PAS) (four cases; 4.9 per cent), post-radiofrequency catheter ablation of the bundle of His (four cases; 4.9 per cent) and vasovagal syncope with atrial fibrillation (one case; 1.2 per cent). TEP was initiated during general anaesthesia after the onset of asystole or profound bradycardia, and stopped when permanent pacing was started. The use of TEP was necessary in 27 cases: 19 cases of third-degree AVB, five of SND, two of PAS and one of vasovagal syncope. External pacing was successful in all but two dogs.

Induction of the acrosome reaction test to in vitro estimate embryo production in Nelore cattle / Teste de indução da reação acrossomal na estimativa de produção in vitro de embriões Nelore

The effectiveness of induction of the acrosome reaction (AR) test as a parameter to in vitro estimate embryo production (IVP) in Nelore breed and the AR pattern by the Trypan Blue/Giemsa (TB) stain were evaluated. Frozen semen samples from ten Nelore bulls were submitted to AR induction and were also evaluated for cleavage and blastocyst rates. The treatments utilized for AR induction were: control (TALP medium), TH (TALP medium + 10μg heparin), TL (TALP medium + 100μg lysophosphatidylcholine) and THL (TALP medium + 10μg heparin + 100μg lysophosphatidylcholine). Sperm acrosomal status and viability were evaluated by TB staining at 0 and after 4h incubation at 38ºC. The results obtained for AR presented a significant difference (P<0.05) in the percentage of acrosome reacted live sperm after 4h of incubation in the treatments that received heparin. The cleavage and blastocyst rates were 60 percent and 38 percent respectively and a significant difference was observed among bulls (P<0.05). It was founded a satisfactory model to estimate the cleavage and blastocyst rates by AR induction test. Therefore, it can be concluded that the induction of the AR test is a valuable tool to predict the IVP in Nelore breed.

Avaliou-se a eficiência da técnica de indução da reação acrossomal (RA) como parâmetro para estimar a produção in vitro (PIV) de embriões Nelore e analisou-se o padrão de RA pela técnica de coloração Azul de Tripan/Giemsa (TB). Amostras de sêmen congelado de dez touros foram submetidas à indução da RA e avaliadas quanto a taxa de clivagem e blastocisto. Os tratamentos utilizados para indução da RA foram: controle (meio TALP), TH (meio TALP + 10μg heparina), TL (meio TALP + 100μg lisofosfatidilcolina) e THL (meio TALP + 10μg heparina + 100μg lisofosfatidilcolina). Avaliou-se viabilidade espermática e acrossomal pela coloração TB a zero e após 4h de incubação a 38ºC. Os resultados obtidos para RA mostram uma diferença significativa (P<0,05) na porcentagem de espermatozoides vivos com acrossoma reagindo após 4h de incubação nos tratamentos que receberam heparina. As taxas de clivagem e blastocisto obtidas foram 60 por cento e 38 por cento respectivamente e observou-se uma diferença significativa entre touros (P<0,05). Delineou-se um modelo satisfatório para estimar as taxas de clivagem e blastocisto. Desta forma, conclui-se que o teste de indução da RA é uma ferramenta valiosa para predizer a PIV na raça Nelore.

Electrophysiologic characteristics and topographic distribution of focal atrial tachycardias in dogs.

BACKGROUND: Focal atrial tachycardia (FAT) is a common supraventricular tachycardia in dogs. OBJECTIVE: To evaluate electrophysiologic characteristics and topographic distribution of FAT. ANIMALS: Sixteen dogs with symptomatic FAT. METHODS: Retrospective case series. Electrophysiological studies were performed to test the inducibility of documented and no documented arrhythmias. Once induced for each dog, FAT was analyzed for electrogenic mechanism, endocardial electrogram, and location. RESULTS: Nineteen FATs could be studied in 16 dogs, 12 were automatic, 4 nonautomatic, and 3 incessant. Two dogs had >1 focus. Mean atrial cycle length (CL) was 238.2 +/- 69.2 (SD) milliseconds, mean ventricular CL of 292.7 +/- 72.5 (SD) milliseconds, with atrioventricular block in 6 cases. Mean presystolic atrial activity recorded at the ectopic focus was -39.9 +/- 17.7 (SD) milliseconds. Atrial potentials were fragmented in 11 dogs and were low amplitude in 6 dogs. Sixty-three percent of ectopic foci were distributed within the right atrium (5 crista terminalis, 3 triangle of Koch, 2 tricuspid valve annulus, 1 interatrial septum, and 1 right auricle) and 37% in the pulmonary veins (PVs) (4 right superior PV, 2 left superior PV, and 1 right inferior PV). Persistent atrial fibrillation (AF) and paroxysmal AF were triggered by FATs in 7 dogs (2 with multiple ectopic foci and 4 with at least one PV focus). CONCLUSION AND CLINICAL RELEVANCE: According to our findings, dogs have a predominance of right-sided FAT. The majority of FATs are automatic and can trigger AF, particularly in the case of PV location.

Aerobic training reverses airway inflammation and remodelling in an asthma murine model.

Aerobic training (AT) decreases dyspnoea and exercise-induced bronchospasm, and improves aerobic capacity and quality of life; however, the mechanisms for such benefits remain poorly understood. The aim of the present study was to evaluate the AT effects in a chronic model of allergic lung inflammation in mice after the establishment of airway inflammation and remodelling. Mice were divided into the control group, AT group, ovalbumin (OVA) group or OVA+AT group and exposed to saline or OVA. AT was started on day 28 for 60 min five times per week for 4 weeks. Respiratory mechanics, specific immunoglobulin (Ig)E and IgG(1), collagen and elastic fibres deposition, smooth muscle thickness, epithelial mucus, and peribronchial density of eosinophils, CD3+ and CD4+, IL-4, IL-5, IL-13, interferon-gamma, IL-2, IL-1ra, IL-10, nuclear factor (NF)-kappaB and Foxp3 were evaluated. The OVA group showed an increase in IgE and IgG(1), eosinophils, CD3+, CD4+, IL-4, IL-5, IL-13, NF-kappaB, collagen and elastic, mucus synthesis, smooth muscle thickness and lung tissue resistance and elastance. The OVA+AT group demonstrated an increase of IgE and IgG(1), and reduction of eosinophils, CD3+, CD4+, IL-4, IL-5, IL-13, NF-kappaB, airway remodelling, mucus synthesis, smooth muscle thickness and tissue resistance and elastance compared with the OVA group (p<0.05). The OVA+AT group also showed an increase in IL-10 and IL-1ra (p<0.05), independently of Foxp3. AT reversed airway inflammation and remodelling and T-helper cell 2 response, and improved respiratory mechanics. These results seem to occur due to an increase in the expression of IL-10 and IL-1ra and a decrease of NF-kappaB.

PAS-1, an Ascaris suum Protein, Modulates Allergic Airway Inflammation via CD8+ ãäTCR+ and CD4+ CD25+ FoxP3+ T Cells

We have previously demonstrated that PAS-1, a 200 kDa protein from Ascaris suum, has a potent immunomodulatory effect on humoral and cell-mediated responses induced by APAS-3 (an allergenic protein from A. suum) or unrelated antigens. In this study, we investigated the mechanisms by which PAS-1 is able to induce this effect on an allergic airway inflammation induced by OVA in mice. C57BL/6 mice were adoptively transferred on day 0 with seven different PAS-1-primed cell populations: PAS-1-primed CD19+ or B220+ or CD3+ or CD4+ or CD8+ or CD4+ CD25- or CD4+ CD25+ lymphocytes. These mice were immunized twice with OVA and alum by intraperitoneal route (days 0 and 7) and challenged twice by intranasal route (days 14 and 21). Two days after the last challenge, the airway inflammation was evaluated by antibody levels, cellular migration, eosinophil peroxidase levels, cytokine and eotaxin production, and pulmonary mechanical parameters. Among the adoptively transferred primed lymphocytes, only CD4+ CD25+, CD8+ or the combination of both T cells impaired the production of total IgE and OVA-specific IgE and IgG1 antibodies, eosinophilic airway inflammation, Th2-type cytokines (IL-4, IL-5 and IL-13), eotaxin release and airway hyperreactivity. Moreover, airway recruited cells from CD4+ CD25+ and CD8+ T-cell recipient secreted more IL-10/TGF-â and IFN-ã, respectively. Moreover, we found that PAS-1 expands significantly the number of CD4+ CD25+ FoxP3+ and CD8+ ãäTCR+ cells. In conclusion, these findings demonstrate that the immunomodulatory effect of PAS-1 is mediated by these T-cell subsets.

PAS-1, a protein from Ascaris suum, modulates allergic inflammation via IL-10 and IFN-gamma, but not IL-12.

Helminths and their products have a profound immunomodulatory effect upon the inductive and effector phases of inflammatory responses, including allergy. We have demonstrated that PAS-1, a protein isolated from Ascaris suum worms, has an inhibitory effect on lung allergic inflammation due to its ability to down-regulate eosinophilic inflammation, Th2 cytokine release and IgE antibody production. Here, we investigated the role of IL-12, IFN-gamma and IL-10 in the PAS-1-induced inhibitory mechanism using a murine model of asthma. Wild type C57BL/6, IL-12(-/-), IFN-gamma(-/-) and IL-10(-/-) mice were immunized with PAS-1 and/or OVA and challenged with the same antigens intranasally. The suppressive effect of PAS-1 was demonstrated on the cellular influx into airways, with reduction of eosinophil number and eosinophil peroxidase activity in OVA+PAS-1-immunized wild type mice. This effect well correlated with a significant reduction in the levels of IL-4, IL-5, IL-13 and eotaxin in BAL fluid. Levels of IgE and IgG1 antibodies were also impaired in serum from these mice. The inhibitory activity of PAS-1 was also observed in IL-12(-/-) mice, but not in IFN-gamma(-/-) and IL-10(-/-) animals. These data show that IFN-gamma and IL-10, but not IL-12, play an important role in the PAS-1 modulatory effect.

PAS-1, a protein from Ascaris suum, modulates allergic inflammationvia IL-10 and IFN-c, but not IL-12

Helminths and their products have a profound immunomodulatory effect upon the inductive and effectorphases of inflammatory responses, including allergy. We have demonstrated that PAS-1, a protein isolatedfrom Ascaris suum worms, has an inhibitory effect on lung allergic inflammation due to its abilityto down-regulate eosinophilic inflammation, Th2 cytokine release and IgE antibody production. Here,we investigated the role of IL-12, IFN-c and IL-10 in the PAS-1-induced inhibitory mechanism using amurine model of asthma. Wild type C57BL/6, IL-12 / , IFN-c / and IL-10 / mice were immunized withPAS-1 and/or OVA and challenged with the same antigens intranasally. The suppressive effect of PAS-1was demonstrated on the cellular influx into airways, with reduction of eosinophil number and eosinophilperoxidase activity in OVA + PAS-1-immunized wild type mice. This effect well correlated with a significantreduction in the levels of IL-4, IL-5, IL-13 and eotaxin in BAL fluid. Levels of IgE and IgG1antibodies were also impaired in serum from these mice. The inhibitory activity of PAS-1 was alsoobserved in IL-12 / mice, but not in IFN-c / and IL-10 / animals. These data show that IFN-c andIL-10, but not IL-12, play an important role in the PAS-1 modulatory effect.