Publications by authors named "Patricia Cunha"

34 Publications

Th17-related mammary immunity, but not a high systemic Th1 immune response is associated with protection against E. coli mastitis.

NPJ Vaccines 2020 Nov 24;5(1):108. Epub 2020 Nov 24.

IHAP, Université de Toulouse, ENVT, INRAE, 31076, Toulouse, France.

Vaccination against bovine mastitis lags behind despite high demand from the dairy industry and margin for efficacy improvement. We previously compared two immunization protocols against E. coli using either only the intramuscular route or a combination of intramuscular and mammary ductal routes, also known as 'prime and pull' strategy. A homologous mammary challenge during the memory phase showed that immunization favorably modified the mastitis course, notably in locally immunized cows in comparison to intramuscular and control adjuvant-only groups. Here, we performed whole-blood profiling through RNA-seq transcriptome and plasma cytokine 15-plex analyses at time points of the E. coli mastitis that showed significant clinical and laboratory differences among the groups. Diminished production of inflammatory cytokines and increased IFNγ were detected in the blood of immunized cows, where a T lymphocyte activation profile was evidenced at 12-h post infection. Acute phase neutropenia was less severe in these cows, and pathways related to neutrophil diapedesis and monocyte activation were also present. Furthermore, three intramammary-immunized cows showing faster healing and shorter mastitis duration had gene profiles that differed from their counterparts, but without any clue for the mastitis susceptibility difference. Inasmuch, when gene expression of CD4 T cells was assessed in mammary tissue, enrichment of IL-17-associated pathways was identified in the quarters of intramammary-immunized cows not only after challenge but also in the control quarters that were not infected. These findings indicate that local immunization mobilizes protective mechanisms that rely on the settlement of type 3 immunity-related CD4 T cells prior to infection.
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http://dx.doi.org/10.1038/s41541-020-00258-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7686320PMC
November 2020

Phytochemical screening and cardiovascular effects of the ethanol extract of mart.

Nat Prod Res 2020 Nov 10:1-5. Epub 2020 Nov 10.

Department of Physiology, Federal University of Sergipe, São Cristóvão, Brazil.

Species of genus are popularly used as anti-inflammatories and in the treatment of renal and respiratory disorders. Although it has been reported that species from the genus induce cardiovascular effects, had not been studied specifically in this respect. However, previous phytochemical studies of demonstrated the presence of compounds which can have potential activity on the cardiovascular system. In this study, phytochemical screening of the ethanol extract of (EEEP) detected polyphenols, but not alkaloids. EEEP caused hypotension, bradycardia and vasorelaxation in rats. The vasorelaxation was attenuated by N-nitro-L-arginine methyl ester (L-NAME) or L-NAME + indomethacin (INDO), but not by INDO alone. Vasorelaxation was also significantly attenuated after endothelium removal or after incubation with high K+, 4-aminopyridine, glibenclamide or tetraethylammonium, but was not affected by pre-contraction with serotonin. Thus, EEEP induces hypotension and endothelium-dependent and independent vasorelaxation, which seems to involve the nitric oxide and K+-channels.
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http://dx.doi.org/10.1080/14786419.2020.1844690DOI Listing
November 2020

Type 3 immunity: a perspective for the defense of the mammary gland against infections.

Vet Res 2020 Oct 15;51(1):129. Epub 2020 Oct 15.

IHAP, Université de Toulouse, INRAE, ENVT, Toulouse, France.

Type 3 immunity encompasses innate and adaptive immune responses mediated by cells that produce the signature cytokines IL-17A and IL-17F. This class of effector immunity is particularly adept at controlling infections by pyogenic extracellular bacteria at epithelial barriers. Since mastitis results from infections by bacteria such as streptococci, staphylococci and coliform bacteria that cause neutrophilic inflammation, type 3 immunity can be expected to be mobilized at the mammary gland. In effect, the main defenses of this organ are provided by epithelial cells and neutrophils, which are the main terminal effectors of type 3 immunity. In addition to theoretical grounds, there is observational and experimental evidence that supports a role for type 3 immunity in the mammary gland, such as the production of IL-17A, IL-17F, and IL-22 in milk and mammary tissue during infection, although their respective sources remain to be fully identified. Moreover, mouse mastitis models have shown a positive effect of IL-17A on the course of mastitis. A lot remains to be uncovered before we can safely harness type 3 immunity to reinforce mammary gland defenses through innate immune training or vaccination. However, this is a promising way to find new means of improving mammary gland defenses against infection.
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http://dx.doi.org/10.1186/s13567-020-00852-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559147PMC
October 2020

Fracture load of CAD/CAM ultrathin occlusal veneers luted to enamel or dentin.

J Esthet Restor Dent 2020 Sep 19. Epub 2020 Sep 19.

Department of Restorative Dentistry, School of Dentistry, Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, Brazil.

Objectives: To evaluate, in vitro, the fracture load of IPS e.max CAD occlusal veneers at thicknesses of either 0.3 or 0.6 mm luted to enamel or dentin.

Materials And Methods: Fifty human molars were randomly distributed into five groups (n = 10): G1 - healthy teeth (control); G2-0.3-mm-thick veneers luted to enamel; G3-0.6-mm-thick veneers luted to enamel; G4-0.3-mm-thick veneers luted to dentin; and G5-0.6-mm-thick veneers luted to dentin. After the luting procedures, the specimens were immersed in distilled water at 37°C for 24 hours and then subjected to mechanical loading (10  cycles at 200 N load). The specimens were subjected to a fracture load test in a universal testing machine. Two-way ANOVA and Tukey's test (α = 0.05) were used to analyze data.

Results: Only the thickness factor was significant (P = .002). Values of fracture load followed by distinct letters represent significant differences (P < .05): G1 (3204 N ± 730) ; G2 (3144 N ± 729) ; G3 (2489 N ± 606) ; G4 (3591 N ± 776) ; and G5 (2770 N ± 598) .

Conclusion: IPS e.max ultrathin occlusal veneers luted to enamel or dentin obtained fracture load comparable to that of the healthy tooth.

Clinical Significance: IPS e.max CAD ultrathin occlusal veneers at 0.3 or 0.6-mm-thick seem to provide good perspectives in relation to the clinical use.
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http://dx.doi.org/10.1111/jerd.12658DOI Listing
September 2020

Effects of high doses of glucocorticoids on insulin-mediated vasodilation in the mesenteric artery of rats.

PLoS One 2020 18;15(3):e0230514. Epub 2020 Mar 18.

Laboratory of Cardiovascular Pharmacology, Department of Physiology, Federal University of Sergipe, Sao Cristovao, Sergipe, Brazil.

Several pathological conditions predict the use of glucocorticoids for the management of the inflammatory response; however, chronic or high dose glucocorticoid treatment is associated with hyperglycemia, hyperlipidemia, and insulin resistance and can be considered a risk factor for cardiovascular disease. Therefore, we investigated the mechanisms involved in the vascular responsiveness and inflammatory profile of mesenteric arteries of rats treated with high doses of glucocorticoids. Wistar rats were divided into a control (CO) group and a dexamethasone (DEX) group, that received dexamethasone for 7 days (2mg/kg/day, i.p.). Blood samples were used to assess the lipid profile and insulin tolerance. Vascular reactivity to Phenylephrine (Phe) and insulin, and O2•-production were evaluated. The intracellular insulin signaling pathway PI3K/AKT/eNOS and MAPK/ET-1 were investigated. Regarding the vascular inflammatory profile, TNF-α, IL-6, IL-1β and IL-18 were assessed. Dexamethasone-treated rats had decreased insulin tolerance test and endothelium-dependent vasodilation induced by insulin. eNOS inhibition caused vasoconstriction in the DEX group, which was abolished by the ET-A antagonist. Insulin-mediated relaxation in the DEX group was restored in the presence of the O2.- scavenger TIRON. Nevertheless, in the DEX group there was an increase in Phe-induced vasoconstriction. In addition, the intracellular insulin signaling pathway PI3K/AKT/eNOS was impaired, decreasing NO bioavailability. Regarding superoxide anion generation, there was an increase in the DEX group, and all measured proinflammatory cytokines were also augmented in the DEX group. In addition, the DEX-group presented an increase in low-density lipoprotein cholesterol (LDL-c) and total cholesterol (TC) and reduced high-density lipoprotein cholesterol (HDL-c) levels. In summary, treatment with high doses of dexamethasone promoted changes in insulin-induced vasodilation, through the reduction of NO bioavailability and an increase in vasoconstriction via ET-1 associated with generation of O2•- and proinflammatory cytokines.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0230514PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7080254PMC
June 2020

IncX4 Plasmid-Mediated in Polymyxin-Resistant from Outpatients in Santa Catarina, Southern Brazil.

Microb Drug Resist 2020 Nov 31;26(11):1326-1333. Epub 2020 Jan 31.

Department of Clinical Analysis, Health Sciences Center, Federal University of Santa Catarina, Florianopolis, Brazil.

Plasmid-mediated polymyxin resistance has become a global health concern, not only because its dissemination has occurred drastically but also because it has begun to be reported in multidrug-resistant (MDR) pathogens. We hereby report microbiological and genomic characteristics of two -positive polymyxin-resistant isolates identified for the first time in community patients, in Santa Catarina, Southern Brazil. strains belonging to ST206 and ST354 and the resistome analysis revealed the presence of clinically important genes responsible for MDR profile. Interestingly, in both polymyxin-resistant strains, genes were carried by IncX4 plasmids, responsible for the worldwide dissemination of type genes. In this regard, plasmid backbones were almost identical to the first IncX4 plasmid reported in Brazil and sharing more than 99.9% identity to IncX4 plasmids from China, also lacking the ISApl1 insertion sequence upstream of . In conclusion, these data confirm the presence of international ST206 and ST354 carrying genes and that the IncX4 plasmids have been key vectors contributing to the endemic status of -positive polymyxin-resistant in Brazil. Also, we described the first known clinical isolate with the gene in Santa Catarina state, Brazil, showing that plasmid-mediated polymyxin resistance has been affecting humans earlier than has been known so far.
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http://dx.doi.org/10.1089/mdr.2019.0203DOI Listing
November 2020

Expansion, isolation and first characterization of bovine Th17 lymphocytes.

Sci Rep 2019 11 6;9(1):16115. Epub 2019 Nov 6.

ISP, INRA, Université de Tours, UMR1282, Nouzilly, France.

Interleukin 17A-producing T helper cells (Th17) are CD4+ T cells that are crucial to immunity to extracellular bacteria. The roles of these cells in the bovine species are poorly defined, because the characterization of bovine Th17 cells lags behind for want of straightforward cultivation and isolation procedures. We have developed procedures to differentiate, expand, and isolate bovine Th17 cells from circulating CD4+ T cells of adult cows. Using polyclonal stimulation with antibodies to CD3 and CD28, we expanded IL-17A-positive CD4+ T cells in a serum-free cell culture medium supplemented with TGF-β1, IL-6 and IL-2. Populations of CD4+ T cells producing IL-17A or IFN-γ or both cytokines were obtained. Isolation of IL-17A-secreting CD4+ T cells was performed by labelling surface IL-17A, followed by flow cytometry cell sorting. The sorted Th17 cells were restimulated and could be expanded for several weeks. These cells were further characterized by cytokine profiling at transcriptomic and protein levels. They produced high amounts of IL-17A and IL-17F, and moderate amounts of IL-22 and IFN-γ. The techniques developed will be useful to characterize the phenotypic and functional properties of bovine Th17 cells.
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http://dx.doi.org/10.1038/s41598-019-52562-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6834651PMC
November 2019

Auditory processing disorder evaluations and cognitive profiles of children with specific learning disorder.

Clin Neurophysiol Pract 2019 31;4:119-127. Epub 2019 May 31.

Faculty of Medicine, Post-Graduation Program in Medical Sciences, University of Brasília, Brasília, DF, Brazil.

Objective: This study investigated the auditory sensory-perceptual level of specific learning disorder (SLD) and explored relationships among neuropsychological assessments for SLD, auditory processing, and short and long latencies of auditory event-related potentials (ERPs).

Methods: Fifteen children (7-14 years old) comprised the control group; 34 children comprised the SLD group. Audiologic assessments included tone audiometry, acoustic immittance measurements, acoustic reflex, central auditory processing, brainstem evoked response audiometry, and long latency potentials (P3 and N2). Children's intelligence levels were assessed with 2 intelligence batteries, 1 verbal and 1 non-verbal, as well as with visuomotor skills.

Results: Multiple regression showed a significant interaction effect of APE tests and P3/N2 over Wechsler Scale performance in freedom of distractibility indexes and multiple subtests. Errors in the Bender Visual Motor Gestalt Test were predicted by lower parental education, lower performance in APE tests: dichotic digits and pediatric/synthetic sentence identification-ipsilateral, and longer P3/N2 latencies, particularly regarding integration and rotation distortions.

Conclusions: Children with altered auditory processing exhibit a specific cognitive profile, including lower verbal and spatial reasoning performance, that is sensitive to parental education level.

Significance: Children with SLD should undergo a complete multimodal examination to identify their specific difficulties and needs.
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http://dx.doi.org/10.1016/j.cnp.2019.05.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6584683PMC
May 2019

Challenging Management of Hepatitis B infection in Ankylosing Spondylitis Patients in an Endemic Area during Immunosuppressive Therapy.

Rev Soc Bras Med Trop 2019 Mar 18;52:e20180386. Epub 2019 Mar 18.

Departamento de Clínica Médica, Residência de Reumatologia, Faculdade de Medicina, Universidade Federal do Amazonas, Amazonas, Brasil.

Hepatitis B infection is a global health issue. When considering patients with rheumatic diseases, this is no different. By using immunosuppressant drugs, such as DMARDs and biologics, viral reactivation is possible, leading to serious consequences on the patient. We report 3 cases of association between ankylosing spondylitis and hepatitis B with the use of immunosuppressant drugs. Case 1 was a patient with previous HBV infection using DMARD. Cases 2 and 3 were patients chronically infected by HBV during immunosuppressant therapy. The management of HBV infection during immunosuppressant therapy is challenging and needs multidisciplinary support.
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http://dx.doi.org/10.1590/0037-8682-0386-2018DOI Listing
March 2019

Devic's syndrome and mixed connective tissue disease: an unusual association.

Acta Reumatol Port 2018 Apr-Jun;43(2):146-150

Hospital Universitário Getúlio Vargas, Universidade Federal do Amazonas, Manaus, Brasil.

Devic's disease or neuromyelitis optica (NMO) is an autoimmune, inflammatory and demyelinating pathology of the central nervous system that affects the optic nerve and the spinal cord. Diagnosis confirmed by imaging, magnetic resonance (MR) and the presence of the anti-aquaporin 4 antibody (anti-AQP4). We describe two cases of patients with mixed connective tissue disease (MCTD) and Devic's disease, who had anti-AQP4 positive and areas with neuroaxis MR abnormalities, showing this rare association.
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February 2019

Microtensile bond strength of CAD/CAM materials to dentin under different adhesive strategies.

Braz Oral Res 2017 Dec 18;31:e109. Epub 2017 Dec 18.

Pontifícia Universidade Católica da Universidade Federal do Rio Grande do Sul - PUC-RS, School of Dentistry, Graduate Program, Porto Alegre, RS, Brazil.

The present study aimed to evaluate the microtensile bond strength (μTBS) of novel computer-aided design/computer-aided manufacturing (CAD/CAM) restorative materials to dentin using different adhesive strategies. Thirty-two crowns were milled using CAD/CAM materials (Vita Mark II as control, Vita Suprinity, Vita Enamic and Lava Ultimate) and luted to dentin using different resin cements (RelyX ARC, RelyX Unicem 2 and RelyX Ultimate). The specimens were stored in 100% relative humidity at 37°C for 24 h and sectioned. The samples (n = 16) with cross-sectional areas of approximately 0.90 mm2, were submitted to a μTBS test in a universal testing machine with a crosshead speed of 0.5 mm/min. The samples were analyzed with SEM to determinate the failure mode. According to 2-way ANOVA and Tukey's test (α=0.05), the interaction effect (material x luting strategy) was significant (p=0.001). Regardless of the luting strategy, a higher μTBS was obtained with Lava Ultimate and Vita Enamic, which were significantly different from Vita Mark II and Vita Suprinity (p<0.05). For Vita Mark II, Vita Suprinity, Vita Enamic, and Lava Ultimate, the μTBS obtained with RelyX Unicem 2 was not significantly different from that obtained with RelyX ARC or RelyX Ultimate. All groups obtained at least three types of failure. The adhesive strategy with self-adhesive resin cement was comparable to conventional resin cement with total-etch or self-etch adhesive techniques in the bond of novel CAD/CAM materials to dentin.
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http://dx.doi.org/10.1590/1807-3107BOR-2017.vol31.0109DOI Listing
December 2017

Escherichia coli mastitis strains: In vitro phenotypes and severity of infection in vivo.

PLoS One 2017 20;12(7):e0178285. Epub 2017 Jul 20.

ISP, INRA, Université François Rabelais de Tours, UMR 1282, Nouzilly, France.

Mastitis remains a major infection of dairy cows and an important issue for dairy farmers and the dairy industry, in particular infections due to Escherichia coli strains. So far, properties specific to E. coli causing mastitis remain ill defined. In an attempt to better understand the properties required for E. coli to trigger mastitis, we used a range of in vitro assays to phenotypically characterize four E. coli strains, including the prototypical E. coli mastitis strain P4, possessing different relative abilities to cause mastitis in a mouse model. Our results indicate that a certain level of serum resistance might be required for colonization of the mammary gland. Resistance to neutrophil killing is also likely to contribute to a slower clearance of bacteria and higher chances to colonize the udder. In addition, we show that the four different strains do induce a pro-inflammatory response by mammary epithelial cells but with different intensities. Interestingly, the prototypical mastitis strain P4 actually induces the less intense response while it is responsible for the most severe infections in vivo. Altogether, our results suggest that different strategies can be used by E. coli strains to colonize the mammary gland and cause mastitis.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0178285PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5519002PMC
September 2017

Prevalence of dyspeptic symptoms and heartburn of adults in Belo Horizonte, Brazil.

Arq Gastroenterol 2017 Jan-Mar;54(1):46-50

Programa de Pós-Graduação, Faculdade de Ciências Médicas de Minas Gerais, MG, Brasil.

Background: - Medical literature has shown dyspepsia and heartburn-related symptoms occur among 15% to 40% of the population. These symptoms can occur at any age and are more prevalent in women.

Objective: - Investigate the prevalence of dyspeptic symptoms and heartburn among individuals over 18.

Methods: - Individuals over 18 were randomly selected in public venues in Belo Horizonte/MG to participate. A standardized questionnaire that included questions related to social-demographic characteristics, eating habits, digestive symptoms, medical appointments, medications, exams, previous surgeries and comorbidities was applied. A questionnaire about functional dyspepsia diagnosis (Rome III) was also applied.

Results: - A total of 548 individuals were interviewed. Among these, 58.4% were women, 59.3% were white, 55.9% were single and the average age was 36 years. Within this group, 376 individuals (68.6%) declared to have some symptom and/or use medication to relieve dyspepsia symptoms, and for these patients were applied the Rome III questionnaire. Based on the diagnostic criteria for the questionnaire proposed by the Rome III consensus, the symptom of postprandial fullness was reported by 6.7% of the individuals, early satiety (3.5%) and epigastric pain (10.6%). The overlap of these symptoms was very frequent. The prevalence of functional dyspepsia was 10.6% (postprandial discomfort syndrome (8.2%) and epigastric pain syndrome (2.4%). Among all participants, 52.5% reported heartburn, and 11.1% presented this symptom at least once a week. The most used drug was omeprazole.

Conclusion: - The prevalence of dyspeptic symptoms and heartburn among a Brazilian adult urban population is similar to those described in other countries.
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http://dx.doi.org/10.1590/S0004-2803.2017v54n1-09DOI Listing
April 2017

Metabolomic profiling of breast tumors using ductal fluid.

Int J Oncol 2016 Dec 13;49(6):2245-2254. Epub 2016 Oct 13.

Lombardi Comprehensive Cancer Center and Department of Oncology, Georgetown University Medical Center, Georgetown University, Washington DC, USA.

Identification of new biomarkers for breast cancer remains critical in order to enhance early detection of the disease and improve its prognosis. Towards this end, we performed an untargeted metabolomic analysis of breast ductal fluid using an ultra-performance liquid chromatography coupled with a quadrupole time-of-light (UPLC-QTOF) mass spectrometer. We investigated the metabolomic profiles of breast tumors using ductal fluid samples collected by ductal lavage (DL). We studied fluid from both the affected breasts and the unaffected contralateral breasts (as controls) from 43 women with confirmed unilateral breast cancer. Using this approach, we identified 1560 ions in the positive mode and 538 ions in the negative mode after preprocessing of the UPLC‑QTOF data. Paired t-tests applied on these data matrices identified 209 ions (positive and negative modes combined) with significant change in intensity level between affected and unaffected control breasts (adjusted p-values <0.05). Among these, 83 ions (39.7%) showed a fold change (FC) >1.2 and 66 ions (31.6%) were identified with putative compound names. The metabolites that we identified included endogenous metabolites such as amino acid derivatives (N-Acetyl-DL-tryptophan) or products of lipid metabolism such as N-linoleoyl taurine, trans-2-dodecenoylcarnitine, lysophosphatidylcholine LysoPC(18:2(9Z,12Z)), glycerophospholipids PG(18:0/0:0), and phosphatidylserine PS(20:4(5Z,8Z,11Z,14Z). Generalized LASSO regression further selected 21 metabolites when race, menopausal status, smoking, grade and TNM stage were adjusted for. A predictive conditional logistic regression model, using the LASSO selected 21 ions, provided diagnostic accuracy with the area under the curve of 0.956 (sensitivity/specificity of 0.907/0.884). This is the first study that shows the feasibility of conducting a comprehensive metabolomic profiling of breast tumors using breast ductal fluid to detect changes in the cellular microenvironment of the tumors and shows the potential for this approach to be used to improve detection of breast cancer.
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http://dx.doi.org/10.3892/ijo.2016.3732DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5117995PMC
December 2016

Innate and Adaptive Immunity Synergize to Trigger Inflammation in the Mammary Gland.

PLoS One 2016 21;11(4):e0154172. Epub 2016 Apr 21.

ISP, INRA, Université Tours, Nouzilly, France.

The mammary gland is able to detect and react to bacterial intrusion through innate immunity mechanisms, but mammary inflammation can also result from antigen-specific adaptive immunity. We postulated that innate and adaptive immune responses could synergize to trigger inflammation in the mammary gland. To test this hypothesis, we immunized cows with the model antigen ovalbumin and challenged the sensitized animals with either Escherichia coli lipopolysaccharide (LPS) as innate immunity agonist, ovalbumin as adaptive immunity agonist, or both agonists in three different udder quarters of lactating cows. There was a significant amplification of the initial milk leukocytosis in the quarters challenged with the two agonists compared to leukocytosis in quarters challenged with LPS or ovalbumin alone. This synergistic response occurred only with the cows that developed the ovalbumin-specific inflammatory response, and there were significant correlations between milk leukocytosis and production of IL-17A and IFN-γ in a whole-blood ovalbumin stimulation assay. The antigen-specific response induced substantial concentrations of IL-17A and IFN-γ in milk contrary to the response to LPS. Such a synergy at the onset of the reaction of the mammary gland suggests that induction of antigen-specific immune response with bacterial antigens could improve the initial immune response to infection, hence reducing the bacterial load and contributing to protection.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0154172PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4839698PMC
April 2017

MicroRNA analysis of breast ductal fluid in breast cancer patients.

Int J Oncol 2016 May 10;48(5):2071-8. Epub 2016 Mar 10.

Lombardi Comprehensive Cancer Center and Department of Oncology, Georgetown University Medical Center, Georgetown University, Washington, DC, USA.

Recent studies suggest that microRNAs show promise as excellent biomarkers for breast cancer; however there is still a high degree of variability between studies making the findings difficult to interpret. In addition to blood, ductal lavage (DL) and nipple aspirate fluids represent an excellent opportunity for biomarker detection because they can be obtained in a less invasive manner than biopsies and circumvent the limitations of evaluating blood biomarkers with regards to tissue of origin specificity. In this study, we have investigated for the first time, through a real-time PCR array, the expression of 742 miRNAs in the ductal lavage fluid collected from 22 women with unilateral breast tumors. We identified 17 differentially expressed miRNAs between tumor and paired normal samples from patients with ductal breast carcinoma. Most of these miRNAs have various roles in breast cancer tumorigenesis, invasion and metastasis, therapeutic response, or are associated with several clinical and pathological characteristics of breast tumors. Moreover, some miRNAs were also detected in other biological fluids of breast cancer patients such as serum (miR-23b, -133b, -181a, 338-3p, -625), plasma (miR-200a), and breast milk (miR-181a). A systems biology analysis of these differentially expressed miRNAs points out possible pathways and cellular processes previously described as having an important role in breast cancer such as Wnt, ErbB, MAPK, TGF-β, mTOR, PI3K-Akt, p53 signaling pathways. We also observed a difference in the miRNA expression with respect to the histological type of the tumors. In conclusion, our findings suggest that miRNA analysis of breast ductal fluid is feasible and potentially very useful for the detection of breast cancer.
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http://dx.doi.org/10.3892/ijo.2016.3435DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4809650PMC
May 2016

IL-17A Is an Important Effector of the Immune Response of the Mammary Gland to Escherichia coli Infection.

J Immunol 2016 Jan 18;196(2):803-12. Epub 2015 Dec 18.

UMR1282 Infectiologie et Santé Publique, Institut National de la Recherche Agronomique, F-37380 Nouzilly, France; and UMR1282 Infectiologie et Santé Publique, Université François-Rabelais de Tours, F-37000 Tours, France

The cytokine IL-17A has been shown to play critical roles in host defense against bacterial and fungal infections at different epithelial sites, but its role in the defense of the mammary gland (MG) has seldom been investigated, although infections of the MG constitute the main pathology afflicting dairy cows. In this study, we showed that IL-17A contributes to the defense of the MG against Escherichia coli infection by using a mouse mastitis model. After inoculation of the MG with a mastitis-causing E. coli strain, the bacterial load increased rapidly, triggering an intense influx of leukocytes into mammary tissue and increased concentrations of IL-6, IL-22, TNF-α, and IL-10. Neutrophils were the first cells that migrated intensely to the mammary tissue, in line with an early production of CXCL2. Depletion of neutrophils induced an increased mammary bacterial load. There was a significant increase of IL-17-containing CD4(+) αβ T lymphocyte numbers in infected glands. Depletion of IL-17A correlated with an increased bacterial colonization and IL-10 production. Intramammary infusion of IL-17A at the onset of infection was associated with markedly decreased bacterial numbers, decreased IL-10 production, and increased neutrophil recruitment. Depletion of CD25(+) regulatory T cells correlated with a decreased production of IL-10 and a reduced bacterial load. These results indicate that IL-17A is an important effector of MG immunity to E. coli and suggest that an early increased local production of IL-17A would improve the outcome of infection. These findings point to a new lead to the development of vaccines against mastitis.
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http://dx.doi.org/10.4049/jimmunol.1500705DOI Listing
January 2016

Antigen-Specific Mammary Inflammation Depends on the Production of IL-17A and IFN-γ by Bovine CD4+ T Lymphocytes.

PLoS One 2015 16;10(9):e0137755. Epub 2015 Sep 16.

INP, ENVT, Université de Toulouse, Toulouse, France; UMR1225, Interactions Hôte Agents Pathogènes, INRA, Toulouse, France.

Intramammary infusion of the antigen used to sensitize cows by the systemic route induces a local inflammation associated with neutrophil recruitment. We hypothesize that this form of delayed type hypersensitivity, which may occur naturally during infections or could be induced intentionally by vaccination, can impact the outcome of mammary gland infections. We immunized cows with ovalbumin to identify immunological correlates of antigen-specific mammary inflammation. Intraluminal injection of ovalbumin induced a mastitis characterized by a prompt tissue reaction (increase in teat wall thickness) and an intense influx of leukocytes into milk of 10 responder cows out of 14 immunized animals. The magnitude of the local inflammatory reaction, assessed through milk leukocytosis, correlated with antibody titers, skin thickness test, and production of IL-17A and IFN-γ in a whole-blood antigen stimulation assay (WBA). The production of these two cytokines significantly correlated with the magnitude of the milk leukocytosis following the ovalbumin intramammary challenge. The IL-17A and IFN-γ production in the WBA was dependent on the presence of CD4+ cells in blood samples. In vitro stimulation of peripheral blood lymphocytes with ovalbumin followed by stimulation with PMA/ionomycin allowed the identification by flow cytometry of CD4+ T cells producing either IL-17A, IFN-γ, or both cytokines. The results indicate that the antigen-specific WBA, and specifically IL-17A and IFN-γ production by circulating CD4+ cells, can be used as a predictor of mammary hypersensitivity to protein antigens. This prompts further studies aiming at determining how Th17 and/or Th1 lymphocytes modulate the immune response of the mammary gland to infection.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0137755PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4573518PMC
May 2016

Investigating the contribution of IL-17A and IL-17F to the host response during Escherichia coli mastitis.

Vet Res 2015 Jun 11;46:56. Epub 2015 Jun 11.

INRA, UMR1282, Infectiologie et Santé Publique, F-37380, Nouzilly, France.

Mastitis remains a major disease of cattle with a strong impact on the dairy industry. There is a growing interest in understanding how cell mediated immunity contributes to the defence of the mammary gland against invading mastitis causing bacteria. Cytokines belonging to the IL-17 family, and the cells that produce them, have been described as important modulators of the innate immunity, in particular that of epithelial cells. We report here that expression of IL-17A and IL-17F genes, encoding two members of the IL-17 family, are induced in udder tissues of cows experimentally infected with Escherichia coli. The impact of IL-17A on the innate response of bovine mammary epithelial cells was investigated using a newly isolated cell line, the PS cell line. We first showed that PS cells, similar to primary bovine mammary epithelial cells, were able to respond to agonists of TLR2 and to LPS, provided CD14 was added to the culture medium. We then showed that secretion of CXCL8 and transcription of innate immunity related-genes by PS cells were increased by IL-17A, in particular when these cells were stimulated with live E. coli bacteria. Together with data from the literature, these results support the hypothesis that IL-17A and IL-17 F could play an important role in mediating of host-pathogen interactions during mastitis.
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http://dx.doi.org/10.1186/s13567-015-0201-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4462179PMC
June 2015

Ocular Perfusion Pressure and Pulsatile Ocular Blood Flow in Normal and Systemic Hypertensive Patients.

J Curr Glaucoma Pract 2015 Jan-Apr;9(1):16-9. Epub 2015 Jan 15.

Department of Ophthalmology, Mayo Clinic, Jacksonville Florida, USA.

Purpose: Glaucomatous neuropathy can be a consequence of insufficient blood supply, increase in intraocular pressure (IOP), or other risk factors that diminish the ocular blood flow. To determine the ocular perfusion pressure (OPP) in normal and systemic hypertensive patients.

Materials And Methods: One hundred and twenty-one patients were enrolled in this prospective and comparative study and underwent a complete ophthalmologic examination including slit lamp examination, Goldmann applanation tonometry, stereoscopic fundus examination, and pulsatile ocular blood flow (POBF) measurements. The OPP was calculated as being the medium systemic arterial pressure (MAP) less the IOP. Only right eye values were considered for calculations using Student's t-test.

Results: The mean age of the patients was 57.5 years (36-78), and 68.5% were women. There was a statistically significant difference in the OPP of the normal and systemic hypertensive patients (p < 0.05). The difference in the OPP between these groups varied between 8.84 and 17.9 mm Hg.

Conclusion: The results of this study suggest that although the systemic hypertensive patients have a higher OPP in comparison to normal patients, this increase does not mean that they also have a higher OBF (as measured by POBF tonograph). This may be caused by chronic changes in the vascular network and in the blood hemodynamics in patients with systemic hypertension. How to cite this article: Kanadani FN, Figueiredo CR, Miranda RM, Cunha PLT, Kanadani TCM, Dorairaj S. Ocular Perfusion Pressure and Pulsatile Ocular Blood Flow in Normal and Systemic Hypertensive Patients. J Curr Glaucoma Pract 2015;9(1):16-19.
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http://dx.doi.org/10.5005/jp-journals-10008-1177DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4741145PMC
March 2016

Interaction between obesity-related genes, FTO and MC4R, associated to an increase of breast cancer risk.

Mol Biol Rep 2013 Dec 4;40(12):6657-64. Epub 2013 Oct 4.

Cell Biology, Embriology and Genetics Department (UFSC, BEG), Federal University of Santa Catarina, Florianópolis, Brazil.

Breast cancer (BC) is a complex disease and obesity is a well-known risk factor for its development, especially after menopause. Several studies have shown Single Nucleotide Polymorphisms (SNPs) linked to overweight and obesity, such as: rs1121980 (T/C) and rs9939609 (A/T) in Fat Mass and Obesity Associated gene (FTO) and rs17782313 (T/C) in Melanocortin 4 Receptor gene (MC4R). Thus, we aimed to investigate the association between these obesity-related SNPs and BC risk. One hundred BC patients and 148 healthy women from Santa Catarina, Brazil entered the study. SNPs were genotyped using Taqman assays. For statistical analyses SNPStats and SPSS softwares were used. Association analyses were performed by logistic regression and were adjusted for age and Body mass index (BMI). Multiple SNPs inheritance models (log-additive, dominant, recessive, codominant) were performed to determine odds ratios (ORs), assuming 95 % confidence interval (CI) and P value = 0.05 as the significance limit. When analyzed alone, FTO rs1121980 and rs9939609 did not show significant associations with BC development, however MC4R rs17782313 showed increased risk for BC even after adjustments (P-value = 0.032). Interestingly, the interaction of FTO and MC4R polymorphisms showed a powerful association with BC. We observed a 4.59-fold increased risk for woman who have the allele combination C/T/C (FTO rs1121980/FTO rs9939609/MC4R rs17782313) (P-value = 0.0011, adjusted for age and BMI). We found important and unpublished associations between these obesity-related genes and BC risk. These associations seem to be independent of their effect on BMI, indicating a direct role of the interaction between FTO and MC4R polymorphisms in BC development.
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http://dx.doi.org/10.1007/s11033-013-2780-3DOI Listing
December 2013

Elevated C-reactive protein and spontaneous bacterial peritonitis in children with chronic liver disease and ascites.

J Pediatr Gastroenterol Nutr 2014 Jan;58(1):96-8

*Department of Pediatric Emergency †Department of Pediatric Hepatology ‡Department of Pediatric Emergency Department, Children's Institute, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.

Objectives: The aims of this study were to compare laboratory indices of spontaneous bacterial peritonitis (SBP) and noninfected ascites in children with chronic liver disease and to determine the infectious agents involved in SBP.

Methods: The medical records of 90 children with chronic liver disease and ascites studied between January 2005 and August 2011 were reviewed for laboratory data of diagnostic significance in SBP. Standard laboratory tests included blood cell count, coagulation indices, liver and renal function tests, C-reactive protein (CRP), serum sodium concentration, serum albumin, and serum cultures. Ascitic fluid obtained from 152 paracentesis procedures was assayed for cytology, Gram stains, neutrophil counts, and bacteriological cultures.

Results: The SBP group manifested significantly lower albumin levels and elevated CRP levels, prothrombin times, international normalized ratios, and leukocyte number (P<0.05 in each case). CRP was shown to be an independent variable in the prediction of SBP. Values of serum creatinine, sodium concentration, urea, total bilirubin and differential leukocyte shift were comparable in SBP and noninfected ascites. Streptococcus pneumoniae was the most prevalent infectious agent in the ascitic fluid (44%).

Conclusions: CRP may be useful in early detection and monitoring of SBP in children with liver disease.
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http://dx.doi.org/10.1097/MPG.0000000000000177DOI Listing
January 2014

Cardiovascular effects induced by linalool in normotensive and hypertensive rats.

Z Naturforsch C J Biosci 2013 May-Jun;68(5-6):181-90

Department of Physiology, Federal University of Sergipe, São Cristóvio - SE, Brazil.

Linalool is a monoterpene alcohol and constituent of several Brazilian aromatic medicinal plants, popularly used against hypertension. Cardiovascular effects induced by linalool were evaluated. In normotensive rats, (+/-)-linalool [1, 5, 10, and 20 mg/kg body weight (BW); intravenous (i.v.)]-induced hypotension was associated with tachycardia, which was attenuated by atropine (2 mg/kg BW) and N(G)-nitro-L-arginine methyl ester (20 mg/kg BW), but was not modified after indomethacin (5 mg/kg BW) administration. In hypertensive rats, linalool [200 mg/kg BW; oral (v.o.)] reduced blood pressure without changing the heart rate. In intact rings of rat mesenteric artery precontracted with 10 microM phenylephrine, linalool (from 6.4 x 10(-6) to 6.4 x 10(-3) M) induced relaxations in a concentration-dependent manner [E(max) = (115 +/- 13)%] that were not changed after atropine administration [E(max) = (105 +/- 2)%], and were not different from those obtained in endothelium-denuded rings precontracted with phenylephrine [E(max) = (108 +/- 7)%] or 80 mM KCl [E(max) = (113 +/- 7)%] or tetraethylammonium incubation [E(max) = (105 +/- 12)%]. Linalool (1.9 x 10(-3) M) antagonized the contractions induced by CaCl2 (3 x 10(-6)-10(-2) M) (maximal inhibition, 81%). Furthermore, linalool inhibited the contractions induced by 10 microM phenylephrine or 20 mM caffeine. In conclusion, these results demonstrate that linalool reduces blood pressure probably due to a direct effect on the vascular smooth muscle leading to vasodilation.
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August 2013

Efficacy and tolerability of a gatifloxacin/prednisolone acetate fixed combination for topical prophylaxis and control of inflammation in phacoemulsification: a 20-day-double-blind comparison to its individual components.

Clinics (Sao Paulo) 2013 Jun;68(6):834-9

Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, Ophthalmology, São Paulo/SP, Brazil.

Objective: To compare the efficacy and tolerability of a fixed combination of 0.3% gatifloxacin and 1% prednisolone(ZypredH) versus the individual components used separately (Zymar® and Predfort® ) for infection prophylaxis and inflammation control after cataract surgery with intraocular lens implantation [corrected].

Methods: A prospective, randomized, double-blind, parallel-group study of 108 patients who underwent phacoemulsification and intraocular lens implantation was conducted. After random assignment, 47 eyes received the fixed combination of topical 0.3% gatifloxacin/1% prednisolone drops, and 61 eyes received the same doses of the individual components as separate solutions four times a day for 15 days. Baseline and postoperative assessments were made on postoperative days 1, 7, 15, and 20.

Results: All objective (best corrected visual acuity, sign of active ocular inflammation, central and incisional corneal edema, the number of cells per high-power field in the anterior chamber, and intraocular pressure) and subjective (eye pain, photophobia, burning sensation, itching, and foreign body sensation) criteria of efficacy were similar in both groups, with no significant differences. Group I included 47 eyes that received the fixed combination of gatifloxacin/prednisolone acetate eye drops and a placebo eye drop solution. Group II included 61 eyes that were treated with 0.3% gatifloxacin and 1% prednisolone acetate eye drops separately. The intraocular pressure was slightly higher in Group II (p<0.05).

Conclusion: Treatment with the fixed-dose combination of gatifloxacin/prednisolone eye drops was as effective as the non-fixed combination in preventing infection and controlling inflammation after phacoemulsification and intraocular lens implantation.
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http://dx.doi.org/10.6061/clinics/2013(06)18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3674263PMC
June 2013

Differential response of bovine mammary epithelial cells to Staphylococcus aureus or Escherichia coli agonists of the innate immune system.

Vet Res 2013 Jun 11;44:40. Epub 2013 Jun 11.

INRA, UMR1282, ISP, F 37380, Nouzilly, France.

Mastitis caused by Escherichia coli and Staphylococcus aureus is a major pathology of dairy cows. To better understand the differential response of the mammary gland to these two pathogens, we stimulated bovine mammary epithelial cells (bMEC) with either E. coli crude lipopolysaccharide (LPS) or with S. aureus culture supernatant (SaS) to compare the transcriptomic profiles of the initial bMEC response. By using HEK 293 reporter cells for pattern recognition receptors, the LPS preparation was found to stimulate TLR2 and TLR4 but not TLR5, Nod1 or Nod2, whereas SaS stimulated TLR2. Biochemical analysis revealed that lipoteichoic acid, protein A and α-hemolysin were all present in SaS, and bMEC were found to be responsive to each of these molecules. Transcriptome profiling revealed a core innate immune response partly shared by LPS and SaS. However, LPS induced expression of a significant higher number of genes and the fold changes were of greater magnitude than those induced by SaS. Microarray data analysis suggests that the activation pathways and the early chemokine and cytokine production preceded the defense and stress responses. A major differential response was the activation of the type I IFN pathway by LPS but not by SaS. The higher upregulation of chemokines (Cxcl10, Ccl2, Ccl5 and Ccl20) that target mononuclear leucocytes by LPS than by SaS is likely to be related to the differential activation of the type I IFN pathway, and could induce a different profile of the initial recruitment of leucocytes. The MEC responses to the two stimuli were different, as LPS was associated with NF-κB and Fas signaling pathways, whereas SaS was associated with AP-1 and IL-17A signaling pathways. It is noteworthy that at the protein level secretion of TNF-α and IL-1β was not induced by either stimulus. These results suggest that the response of MEC to diffusible stimuli from E. coli and S. aureus contributes to the onset of the response with differential leucocyte recruitment and distinct inflammatory and innate immune reactions of the mammary gland to infection.
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http://dx.doi.org/10.1186/1297-9716-44-40DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3686618PMC
June 2013

T helper 17-associated cytokines are produced during antigen-specific inflammation in the mammary gland.

PLoS One 2013 16;8(5):e63471. Epub 2013 May 16.

Infectiology and Public Health Research Unit, Institut National de la Recherche Agronomique, Nouzilly, France.

Infectious mastitis cuts down milk production profitability and is a major animal welfare problem. Bacteria-induced inflammation in the mammary gland (MG) is driven by innate immunity, but adaptive immunity can modulate the innate response. Several studies have shown that it is possible to elicit inflammation in the MG by sensitization to an antigen subsequently infused into the lumen of the gland. The objective of our study was to characterize the inflammation triggered in the MG of cows sensitized to ovalbumin, by identifying the cytokines and chemokines likely to play a part in the reaction. Among immunized cows, responders mobilized locally high numbers of leukocytes. An overexpression of the genes encoding IL-17a, IL-17F, IL-21, IL-22 and INF-γ was found in milk cell RNA extracts in the early phase of the inflammatory response. At the protein level, IL-17A was detected in milk as soon as the first sampling time (8 h post-challenge), and both IL-17A and IFN-γ concentrations peaked at 12 to 24 h post-challenge. In mammary tissue from challenged quarters, overexpression of the genes encoding IL-17A, IL-17F, IL-21, IL-22, IL-26 and IFN-γ was observed. Neutrophil-attracting chemokines (CXCL3 and CXCL8) were found in milk, and overexpressed transcripts of chemokines attracting lymphocytes and other mononuclear leukocytes (CXCL10, CCL2, CCL5, CCL20) were detected in mammary tissue. Expression of IL-17A, as revealed by immunohistochemistry, was located in epithelial cells, in leukocytes in the connective tissue and in association with the epithelium, and in migrated alveolar leukocytes of challenged quarters. Altogether, these results show that antigen-specific inflammation in the MG was characterized by the production of IL-17 and IFN-γ. The orientation of the inflammatory response induced by the antigen-specific response has the potential to strongly impact the outcome of bacterial infections of the MG.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0063471PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3656053PMC
December 2013

Genetic susceptibility to S. aureus mastitis in sheep: differential expression of mammary epithelial cells in response to live bacteria or supernatant.

Physiol Genomics 2012 Apr 14;44(7):403-16. Epub 2012 Feb 14.

Université de Toulouse, Institut National Polytechnique (INP), École Nationale Vétérinaire de Toulouse (ENVT), Unité Mixte de Recherche (UMR)1225, Interactions Hôtes - Agents Pathogènes (IHAP), Toulouse, France.

Staphylococcus aureus is a prevalent pathogen for mastitis in dairy ruminants and is responsible for both clinical and subclinical mastitis. Mammary epithelial cells (MEC) represent not only a physical barrier against bacterial invasion but are also active players of the innate immune response permitting infection clearance. To decipher their functions in general and in animals showing different levels of genetic predisposition to Staphylococcus in particular, MEC from ewes undergoing a divergent selection on milk somatic cell count were stimulated by S. aureus. MEC response was also studied according to the stimulation condition with live bacteria or culture supernatant. The early MEC response was studied during a 5 h time course by microarray to identify differentially expressed genes with regard to the host genetic background and as a function of the conditions of stimulation. In both conditions of stimulation, metabolic processes were altered, the apoptosis-associated pathways were considerably modified, and inflammatory and immune responses were enhanced with the upregulation of il1a, il1b, and tnfa and several chemokines known to enhance neutrophil (cxcl8) or mononuclear leukocyte (ccl20) recruitment. Genes associated with oxidative stress were increased after live bacteria stimulation, whereas immune response-related genes were higher after supernatant stimulation in the early phase. Only 20 genes were differentially expressed between Staphylococcus spp-mastitis resistant and susceptible animals without any clearly defined role on the control of infection. To conclude, this suggests that MEC may not represent the cell type at the origin of the difference of mastitis susceptibility, at least as demonstrated in our genetic model. Supernatant or heat-killed S. aureus produce biological effects that are essentially different from those induced by live bacteria.
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http://dx.doi.org/10.1152/physiolgenomics.00155.2011DOI Listing
April 2012

Repertoire of Escherichia coli agonists sensed by innate immunity receptors of the bovine udder and mammary epithelial cells.

Vet Res 2012 Feb 13;43:14. Epub 2012 Feb 13.

INRA, UMR 1282 Infectiologie et Santé Publique, F-37380 Nouzilly, France.

Escherichia coli is a frequent cause of clinical mastitis in dairy cows. It has been shown that a prompt response of the mammary gland after E. coli entry into the lumen of the gland is required to control the infection, which means that the early detection of bacteria is of prime importance. Yet, apart from lipopolysaccharide (LPS), little is known of the bacterial components which are detected by the mammary innate immune system. We investigated the repertoire of potential bacterial agonists sensed by the udder and bovine mammary epithelial cells (bMEC) during E. coli mastitis by using purified or synthetic molecular surrogates of bacterial agonists of identified pattern-recognition receptors (PRRs). The production of CXCL8 and the influx of leucocytes in milk were the readouts of reactivity of stimulated cultured bMEC and challenged udders, respectively. Quantitative PCR revealed that bMEC in culture expressed the nucleotide oligomerization domain receptors NOD1 and NOD2, along with the Toll-like receptors TLR1, TLR2, TLR4, and TLR6, but hardly TLR5. In line with expression data, bMEC proved to react to the cognate agonists C12-iE-DAP (NOD1), Pam3CSK4 (TLR1/2), Pam2CSK4 (TLR2/6), pure LPS (TLR4), but not to flagellin (TLR5). As the udder reactivity to NOD1 and TLR5 agonists has never been reported, we tested whether the mammary gland reacted to intramammary infusion of C12-iE-DAP or flagellin. The udder reacted to C12-iE-DAP, but not to flagellin, in line with the reactivity of bMEC. These results extend our knowledge of the reactivity of the bovine mammary gland to bacterial agonists of the innate immune system, and suggest that E. coli can be recognized by several PRRs including NOD1, but unexpectedly not by TLR5. The way the mammary gland senses E. coli is likely to shape the innate immune response and finally the outcome of E. coli mastitis.
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http://dx.doi.org/10.1186/1297-9716-43-14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3305352PMC
February 2012

Chemical composition and vasorelaxant effect induced by the essential oil of Lippia alba (Mill.) N.E. Brown. (Verbenaceae) in rat mesenteric artery.

Indian J Pharmacol 2011 Nov;43(6):694-8

Departamento de Fisiologia, Universidade Federal de Sergipe, São Cristóvão - SE, Brazil.

Objectives: To investigate the chemical composition and vasorelaxant effect of the essential oil of Lippia alba (EOLA) in rat mesenteric artery.

Material And Methods: Chemical composition of EOLA was investigated by gas chromatography-mass spectrometry (GC/MS). Vasorelaxant effect was evaluated in vitro in rat superior mesenteric artery rings.

Results: GC/MS analysis revealed the presence of 19 compounds, with geranial (48.58%) and neral (35.42%) being the major constituents. In intact rings precontracted with phenylephrine (Phe: 1 μM), EOLA (100-1000 μg/mL) induced relaxation, where the maximal effect (Emax) was 110.8 ± 10.8%. This effect was not modified after endothelium removal (Emax = 134.8 ± 16.5%), after tetraethylammonium (TEA) (Emax = 117.2 ± 4.96%), or in rings precontracted with KCl (80 mM) (Emax = 112.6 ± 6.70%). In addition, EOLA was able to inhibit the contraction caused by CaCl(2) and produced a small but significant (P<0.05) additional effect (from 70.5 ± 3.4 to 105.3 ± 13.5%, n = 5) on the maximal relaxation of nifedipine (NIF: 10 μM).

Conclusions: The results demonstrated that EOLA induces endothelium-independent vasorelaxation, which appears to be caused, at least in part, by blocking Ca(2+) influx through voltage-operated Ca(2+) channels.
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http://dx.doi.org/10.4103/0253-7613.89828DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3229787PMC
November 2011

Staphylococcal-associated molecular patterns enhance expression of immune defense genes induced by IL-17 in mammary epithelial cells.

Cytokine 2011 Dec 17;56(3):749-59. Epub 2011 Oct 17.

INRA, UR1282 Infectiologie Animale et Santé Publique (IASP), F-37380 Nouzilly, France.

Interleukin-17A (IL-17A) and IL-17F have been shown to mediate a crucial crosstalk between the immune system and various epithelial tissues, stimulating various defensive mechanisms to bacterial infections. A number of studies have characterized the response to IL-17A and IL-17F of epithelial cells from airways, intestine, and skin, but not from the mammary gland. To evaluate the potential contribution of IL-17 to the immune defense of the mammary gland, we analyzed the effects of recombinant bovine IL-17A and IL-17F on primary bovine mammary epithelial cells (MEC) by quantitative PCR and ELISA. We found expression (mRNA) of the two components of the IL-17 receptor complex, IL-17RA and IL-17RC, in mammary tissue and MEC in vitro. The expression of a number of genes encoding cytokines, chemokines and proteins endowed with antibacterial activities was increased by IL-17A, and to a lesser extent by IL-17F, but the magnitude of responses was modest. As expected, responses were augmented by the combination of IL-17A or IL-17F with TNF-α. Interestingly, responses of a few of the tested genes, such as IL8, CCL20, iNOS, and CfB, were augmented by the combination of IL-17A with staphylococcal lipoteichoic acid or muramyl dipeptide, bacterial agonists of the innate immune system. This can be interpreted as indicating that IL-17A and IL-17F are tailored to exert their full potential in a septic environment. MEC responses were characterized by the expression of chemokines targeting not only neutrophils (CXCL3 and CXCL8) but also mononuclear leucocytes (CCL2, CCL20). Production of IL-6 was low and the inflammatory cytokines TNF-α and IL-1β were expressed (mRNA) but proteins were not secreted. Altogether, our results suggest that IL-17A and IL-17F have a potential to modulate the mammary gland immune response to mastitis-causing pathogens.
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http://dx.doi.org/10.1016/j.cyto.2011.09.020DOI Listing
December 2011