RESUMO
This study describes the chemical characterisation of two extracts (ethyl acetate, EtOAc-E and hexanic, Hx-E) from Libidibia coriaria fruits and their insecticidal properties on Spodoptera frugiperda. The HPLC analysis in EtOAct-E revealed the presence of ellagic acid and phenolic compounds. The CG-MS analysis in Hx-E revealed the presence of Hexadecanoic acid, 11-Methylheptacosane, Dodecanoic acid and Nonacosane as major compounds. The application of both extracts was performed on the dorsal part of each larva via aspersion. The larval mortality, relative growth and emergence percentage of adults were evaluated. The Hx-E caused a 93.33% mortality at 100 mg/mL at 24 h post-application. A minor relative growth with both EtOAc-E (12.64 mg) and Hx-E (7.90 mg) was observed compared with their respective negative controls (methanol = 25.05 mg and tween20 = 24.53 mg). The lowest emergence percentage of adults with the Hx-E (25%) at 50 mg/mL was observed. Libidibia coriaria fruits exhibit insecticidal properties against S. frugiperda.
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Few studies have been conducted in the cooling systems of power plants; they have focused on Naegleria fowleri, leaving a gap in the knowledge of other pathogenic free-living amoebae in this environment. The objective of this study was to determine the occurrence of saline-tolerant pathogenic Acanthamoeba in a geothermal power plant. The identification of isolated amoebae at genus level was carried out, observing their morphological characteristics; the determination of genotype and species of Acanthamoeba was performed via molecular biology (PCR). Water temperature ranged from 18 to 43 °C and conductivity from 4.0 × 104 to 8.7 × 104 µS/cm; this last value was greater than the seawater value. Only five amoeba genera were found. Acanthamoeba was in all the sampling sites, showing high saline tolerance. The high temperature, but mainly high conductivity, were the environmental conditions that determined the presence of pathogenic free-living amoebae in the hot water. All the strains of Acanthamoeba culbertsoni killed the mice, having a mortality of 40 to 100%. Acanthamoeba genotypes T10 and T5 were identified, T10 is rarely isolated from the environment, while T5 is more frequent. This is the first time that genotypes T5 and T10 have been reported in the environment in Mexico.
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Naegleria fowleri is an etiological agent that generates primary amoebic meningoencephalitis; unfortunately, no effective treatment or vaccine is available. The objective of this work was to determine the immunoprotective response of two vaccine antigens, as follows: (i) the polypeptide band of 19 kDa or (ii) a predicted immunogenic peptide from the membrane protein MP2CL5 (Smp145). Both antigens were administered intranasally in mice using cholera toxin (CT) as an adjuvant. The survival rate and immune response of immunized mice with both antigens and challenged with N. fowleri trophozoites were measured in the nose-associated lymphoid tissue (NALT) and nasal passages (NPs) by flow cytometry and enzyme-linked immunosorbent assay (ELISA). We also determined the immunolocalization of both antigens in N. fowleri trophozoites by confocal microscopy. Immunization with the polypeptide band of 19 kDa alone or coadministered with CT was able to confer 80% and 100% of protection, respectively. The immunization with both antigens (alone or coadministered with CT) showed an increase in T and B lymphocytes. In addition, there was an increase in the expression of integrin α4ß1 and IgA in the nasal cavity of protected mice, and the IgA, IgG, and IgM levels were increased in serum and nasal washes. The immunolocalization of both antigens in N. fowleri trophozoites was observed in the plasma membrane, specifically in pseudopod-like structures. The MP2CL5 antigens evaluated in this work were capable of conferring protection which would lead us to consider them as potential candidates for vaccines against meningitis caused by N. fowleri.
Assuntos
Meningite , Naegleria fowleri , Vacinas , Animais , Camundongos , Toxina da Cólera , Imunidade , Imunoglobulina ARESUMO
Improving the lipid profile in milk of cows with the use of soybean grain (Glycine max L.) can be favored in the grazing systems in the dry tropics of Mexico. The objective was to evaluate the milk production, the chemical composition, and the fatty acids profile (FAs) of the milk of cows in continuous grazing and supplemented with and without ground roasted soybean in the dry tropics of Mexico. Ten cows randomly distributed in two equal groups were used. Daily during confinement for milking, the cows individually received the treatments on dry basis T0: supplement with 4.6 kg commercial concentrate® without soybean, T1: supplement with 3.7 kg commercial concentrate® with 380 g of soybean. During the 78 days of the experiment, milk production was measured in all cows, and samples were collected to determine the chemical composition and FAs profile. Milk production, protein, milk total fat, lactose, and non-fat solids did not vary with treatment (p >0.05). Linoleic acid content (C18: 2, cis, cis-∆9, ∆12) increased by 22% in milk fat of cows of the T1 (p Ë0.05). The sum of the mono- and polyunsaturated FAs 29.1%, the ratio of saturated-unsaturated FAs (1.65), and the atherogenicity index (1.71) also improved in the milk of cows supplemented with T1 (p Ë0.05). It was concluded that ground roasted soybean supplement in the diet of grazing dairy cows did not affect production and did improve the lipid profile in milk fat with favorable index to promote human health.
Assuntos
Ácidos Graxos , Leite , Ração Animal/análise , Animais , Bovinos , Dieta/veterinária , Suplementos Nutricionais , Feminino , Lactação , México , Glycine maxRESUMO
Members of the genus Naegleria are free-living amoebae that are widely distributed in water and soil environments. Moreover, Naegleria fowleri is a pathogenic amoeba species that causes a fatal disease in the central nervous system known as primary amoebic meningoencephalitis (PAM) in humans. Since most reported infections due to N. fowleri are reported in recreational waters worldwide, this study was aimed to describe the presence of these amoebic genus in Mexicali Valley irrigation channels of recreational use. A total of nine water samples were collected and processed by triplicate, in nine different sites of the Valley. After filtering and culturing the samples, plates were examined, and the observed amoebae were morphologically identified at the genus level. In addition, the pathogenicity of these amoebic isolates was checked, and molecular characterization was performed by PCR/sequencing. The results revealed the presence of Naegleria spp. in all the channels sampled. Finally, molecular identification confirmed the presence of five different species of Naegleria: N. fowleri, N. australiensis, N. gruberi, N. clarki and N. pagei. The presence of these protists, particularly N. fowleri, should be considered as a potential human health risk in the region.
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The intranasal administration of Naegleria fowleri lysates plus cholera toxin (CT) increases protection against N. fowleri meningoencephalitis in mice, suggesting that humoral immune response mediated by antibodies is crucial to induce protection against the infection. In the present study, we applied a protein analysis to detect and identify immunogenic antigens from N. fowleri, which might be responsible for such protection. A Western blot assay of N. fowleri polypeptides was performed using the serum and nasal washes from mice immunized with N. fowleri lysates, either alone or with CT after one, two, three, or four weekly immunizations and challenged with trophozoites of N. fowleri. Immunized mice with N. fowleri plus CT, after four doses, had the highest survival rate (100%). Nasal or sera IgA and IgG antibody response was progressively stronger as the number of immunizations was increased, and that response was mainly directed to 250, 100, 70, 50, 37, and 19 kDa polypeptide bands, especially in the third and fourth immunization. Peptides present in these immunogenic bands were matched by nano-LC-ESI-MSMS with different proteins, which could serve as candidates for a vaccine against N. fowleri infection.
RESUMO
Many pathogenicity factors are involved in the development of primary amoebic meningoencephalitis (PAM) caused by N fowleri. However, most of them are not exclusive for N fowleri and they have not even been described in other nonpathogenic Naegleria species. Therefore, the objective of this work was to identify differential proteins and protein pattern recognition between Naegleria fowleri and Naegleria lovaniensis using antibodies anti-N fowleri as strategy to find vaccine candidates against meningoencephalitis. Electrophoresis and Western blots conventional and 2-DE were performed for the identification of antigenic proteins, and these were analysed by the mass spectrometry technique. The results obtained in 2-DE gels and Western blot showed very notable differences in spot intensity between these two species, specifically those with relative molecular weight of 100, 75, 50 and 19 kDa. Some spots corresponding to these molecular weights were identified as actin fragment, myosin II, heat shock protein, membrane protein Mp2CL5 among others, with differences in theoretical post-translational modifications. In this work, we found differences in antigenic proteins between both species, proteins that could be used for a further development of vaccines against N fowleri infection.
Assuntos
Antígenos de Protozoários/imunologia , Infecções Protozoárias do Sistema Nervoso Central/imunologia , Meningoencefalite/imunologia , Naegleria fowleri/imunologia , Proteínas de Protozoários/imunologia , Animais , Anticorpos Antiprotozoários/imunologia , Infecções Protozoárias do Sistema Nervoso Central/parasitologia , Proteínas de Membrana/imunologia , Meningoencefalite/parasitologiaRESUMO
Peptide epitopes have been widely used to develop synthetic vaccines and immunotherapies. However, peptide epitopes may exhibit poor absorption or immunogenicity due to their low molecular weights. Conversely, fourth-generation polyamidoamine (G4-PAMAM) dendrimers are nonimmunogenic and relatively nontoxic synthetic nanoparticles that have been used as adjuvants and nanocarriers of small peptides and to improve nasal absorption. Based on this information, we hypothesized that the combination of intranasal immunization and G4-PAMAM dendrimers would be useful for enhancing the antibody responses of HIV-1 gp120 peptide epitopes. Therefore, we first used structural data, peptide epitope predictors and docking and MD simulations on MHC-II to identify two peptide epitopes on the CD4 binding site of HIV-1 gp120. The formation of G4-PAMAM-peptide complexes was evaluated in silico (molecular docking studies using different G4-PAMAM conformations retrieved from MD simulations as well as the MMGBSA approach) and validated experimentally (electrophoresis, 1H NMR and cryo-TEM). Next, the G4-PAMAM dendrimer-peptide complexes were administered intranasally to groups of female BALB/cJ mice. The results showed that both peptides were immunogenic at the systemic and mucosal levels (nasal and vaginal), and G4-PAMAM dendrimer-peptide complexes improved IgG and IgA responses in serum and nasal washes. Thus, G4-PAMAM dendrimers have potential for use as adjuvants and nanocarriers of peptides.
Assuntos
Simulação por Computador , Dendrímeros/química , Proteína gp120 do Envelope de HIV/química , HIV-1/química , HIV-1/imunologia , Modelos Moleculares , Nylons/química , Peptídeos/química , Peptídeos/imunologia , Animais , Feminino , Proteína gp120 do Envelope de HIV/genética , Proteína gp120 do Envelope de HIV/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Peptídeos/genéticaRESUMO
OBJECTIVE: The posterior vagus nerve trunk innervates the entire small intestine, and elucidating its modulatory role in the IgA response was the aim of this study. METHODS: Two groups of six male BALB/c mice underwent sham or posterior subdiaphragmatic vagotomy and were euthanized on the 14th postoperative day; then, the small intestines were dissected. The intestinal fluid was harvested for antibody analysis by ELISA, and cell suspensions from Peyer's patches and lamina propria were prepared for cytofluorometric analysis of plasma cells and T lymphocytes. The CD4+ T cells were labeled for the intracellular IgA-producing interleukins (ILs)-4, -5, -6, and -10; transforming growth factor (TGF)-ß; and the inflammatory cytokines tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and IL-12. In the intestinal tissue samples, myeloperoxidase (MPO) visualization and the enzymatic activity were assessed by immunohistochemistry and ELISA, respectively. The data were analyzed by Student's t test, and the differences were considered significant at p < 0.05. RESULTS: In the vagotomy group, the IgA levels and the CD4+ T cells labeled with mediators that promote IgA secretion, including IL-4 (only at lamina propria), TNF-α, and IFN-γ, were decreased, whereas the lamina propria IgA+ plasma cells and MPO presence/activity were increased; changes in the IgM levels, IgM+ plasma cells, and CD4+ T cells labeled with TGF-ß, which have a role in class switch recombination, were not observed. CONCLUSION: The downmodulating impact of vagotomy on IgA levels may result from defective IgA secretion without affecting class switch recombination, whereas vagotomy evoked a proinflammatory response regarding MPO. These findings may reflect the role of the vagus nerve on the control of the IgA response in the small intestine.
Assuntos
Imunidade nas Mucosas/imunologia , Imunoglobulina A/imunologia , Mucosa Intestinal/imunologia , Intestino Delgado/imunologia , Nervo Vago/fisiologia , Animais , Linfócitos T CD4-Positivos/imunologia , Regulação para Baixo , Mucosa Intestinal/inervação , Intestino Delgado/inervação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Plasmócitos/imunologia , VagotomiaRESUMO
This present study was conducted to evaluate the antimicrobial effects of tannic acid (TA) against isolated ruminal bacteria from adult sheep. Rumen samples were collected from two (2) adults sheep, and a total of nine (9) ruminal bacteria were isolated from the sample. The sensitivity of the ruminal bacteria isolates to 0.63, 1.25, 2.50, 5.00 and 10.00â¯mgâ¯TA/mL of growth medium was determined using clearance zone (CZ) of Kirby-Bauer disc diffusion susceptibility test. There was observable increase in the sensitivity of all bacterial isolates as the level of TA increases. Not all bacterial isolates have the capacity to tolerate more than 1.25â¯mgâ¯TA/mL. The result shows that only 20% of the bacterial isolates had the capacity to tolerate 0.63 and 1.25â¯mg of tannic acid per liter. This concentration of tannic acid would be equivalent to 2% tannin in the diet of ruminant. Our findings shows that increase in concentration of tannic acid completely inhibited the ruminal bacteria from the sheep rumen.
Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Ovinos/microbiologia , Taninos/farmacologia , Animais , Antibacterianos/administração & dosagem , Dieta , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Rúmen/microbiologia , Taninos/administração & dosagemRESUMO
The goal of this study was to identify neuraminidase (NA) residue mutants from human influenza AH1N1 using sequences from 1918 to 2012. Multiple alignment studies of complete NA sequences (5732) were performed. Subsequently, the crystallographic structure of the 1918 influenza (PDB ID: 3BEQ-A) was used as a wild-type structure and three-dimensional (3-D) template for homology modeling of the mutated selected NA sequences. The 3-D mutated NAs were refined using molecular dynamics (MD) simulations (50 ns). The refined 3-D models were used to perform docking studies using oseltamivir. Multiple sequence alignment studies showed seven representative mutations (A232V, K262R, V263I, T264V, S367L, S369N, and S369K). MD simulations applied to 3-D NAs showed that each NA had different active-site shapes according to structural surface visualization and docking results. Moreover, Cartesian principal component analyses (cPCA) show structural differences among these NA structures caused by mutations. These theoretical results suggest that the selected mutations that are located outside of the active site of NA could affect oseltamivir recognition and could be associated with resistance to oseltamivir.
Assuntos
Vírus da Influenza A Subtipo H1N1/enzimologia , Neuraminidase/química , Proteínas Virais/química , Sequência de Aminoácidos , Sítios de Ligação , Ligantes , Modelos Moleculares , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Mutação Puntual , Análise de Componente Principal , Estrutura Terciária de ProteínaRESUMO
A region-wide survey was conducted in the tropical area of Tierra Caliente, State of Guerrero, Mexico to estimate the prevalence of subclinical bovine mastitis (SCM), distribution of mastitis pathogens, and in vitro antimicrobial susceptibility of different mastitis pathogens in dairy farms. In total, 1036 quarter milk samples were obtained from 259 cows at 87 different dairy farms. Collected quarter milk samples were submitted for California Mastitis Test (CMT), bacteriological examination, and testing for antimicrobial susceptibility. Overall prevalence of SCM in the studied area was 20.5 %. Prevalence in the different regions was as follows: 28 % in Arcelia municipality, 21 % in Tlalchapa municipality, 19.4 % in Pungarabato municipality, and 14.3 % in Finch Cutzamala municipality. Of all positive isolates, 97.5 % were Gram-negative bacteria. Moreover, of all positive isolates, 37.5 % were Proteus vulgaris, 25 % Salmonella spp., 12.5 % Enterobacter aerogenes, and 10 % Escherichia coli. Klebsiella pneumonia and E. coli were sensitive for netilmicin antimicrobial. However, E. coli was sensitive for pefloxacin and gentamicin with a sensitivity for pefloxacin for E. aerogenes, while Staphylococci were sensitive for gentamicin and dicloxacillin. It could be concluded that practices such as the implementation of mastitis control programs, improved milking hygiene together with an intramammary treatment with netilmicin, pefloxacin, and gentamicin antimicrobials should be considered for mastitis prevention in the study area of Tierra Caliente, in the tropical area of Guerrero, Mexico.
Assuntos
Resistência Microbiana a Medicamentos , Bactérias Gram-Negativas/isolamento & purificação , Mastite Bovina/epidemiologia , Leite/microbiologia , Animais , Infecções Assintomáticas/epidemiologia , Bovinos , Cidades/estatística & dados numéricos , Indústria de Laticínios , Escherichia coli/isolamento & purificação , Feminino , Mastite Bovina/microbiologia , México/epidemiologia , Testes de Sensibilidade Microbiana , Prevalência , Staphylococcus/isolamento & purificação , Clima TropicalRESUMO
In vitro gas production with and without polyethylene glycol (PEG) of the fruits of Crescentia alata and Guazuma ulmifolia was evaluated, the degradation kinetics of lamb diets with added fruit of the tree was determined, and the ration intake and growth rate of lambs fed these diets were measured. Twenty-five entire male lambs of 23.5 ± 0.44 kg body weight were used and distributed in treatments: T0 (control without fruit); T1 and T2, 15 and 30 % of the fruit of C. alata; and T3 and T4, 15 and 30 % of the fruit of G. ulmifolia. Data variables chemical composition, fermentation kinetic, and digestibility in vitro were analyzed by a completely randomized design and data production response factorials design of five treatments by three evaluation periods. The total phenolic content (TP) (23.0 g/kg DM) was higher (P < 0.01) in the fruits of G. ulmifolia. The addition of PEG increased (P < 0.05) in vitro gas production (156.6 mL/g DM) in fruits of G. ulmifolia. In the fermentation kinetics, the total gas volume was higher (P < 0.01) at T0 (b = 293 mL/g DM), and the rate of degradation (c) but Lag time (t lag) was not different. In animal response, total dry matter intake was higher in lambs that received T4 (1.35 kg), and the daily weight gain and feed conversion did not differ (P > 0.05) among lambs receiving the treatments. Thirty percent G. ulmifolia fruit added in the diet increased dry matter intake and improved feed conversion but did not increase weight gain.
Assuntos
Ração Animal/análise , Dieta/veterinária , Comportamento Alimentar , Frutas , Ovinos/fisiologia , Árvores , Fenômenos Fisiológicos da Nutrição Animal , Animais , Animais Recém-Nascidos/fisiologia , Suplementos Nutricionais/análise , Digestão/fisiologia , Masculino , México , Clima Tropical , Aumento de PesoRESUMO
The toxicity of the curcin on cancer cells allows to consider this protein as the toxic component of an immunotoxin directed to Her2, which is associated with cancer. Reductive amination was proposed to conjugate curcin and an anti-Her2; the binding was tested using Polyacrylamide gel electrophoresis, western blot, and immunocytochemistry. The in vitro cytotoxicity of curcin and the immunotoxin was assessed on breast cancer cell lines SK-BR-3 (Her2(+)) and MDA-MB-231 (Her2(-)). IC50 values for curcin were 15.5 ± 8.3 and 18.6 ± 2.4 µg/mL, respectively, statistically equivalent (p < 0.05). While to the immunotoxin was 2.2 ± 0.08 for SK-BR-3 and 147.6 ± 2.5 µg/mL for MDA-MB-231. These values showed that the immunotoxin was seven times more toxic to the SK-BR-3 than curcin and eight times less toxic to the MDA-MB-231. The immunotoxin composed of curcin and an antibody against Her2 and constructed by reductive amination could be a therapeutic candidate against Her2(+) cancer.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Imunotoxinas/metabolismo , Receptor ErbB-2/imunologia , Proteínas Inativadoras de Ribossomos Tipo 1/metabolismo , Aminação , Linhagem Celular Tumoral , Fenômenos Químicos , Simulação por Computador , Humanos , Imunotoxinas/química , Imunotoxinas/imunologia , Modelos Moleculares , Oxirredução , Conformação Proteica , Proteínas Inativadoras de Ribossomos Tipo 1/químicaRESUMO
C-type lectins are multifunctional sugar-binding molecules expressed on dendritic cells (DCs) and macrophages that internalize antigens for processing and presentation. Macrophage galactose-type lectin 1 (MGL1) recognizes glycoconjugates expressing Lewis X structures which contain galactose residues, and it is selectively expressed on immature DCs and macrophages. Helminth parasites contain large amounts of glycosylated components, which play a role in the immune regulation induced by such infections. Macrophages from MGL1(-/-) mice showed less binding ability toward parasite antigens than their wild-type (WT) counterparts. Exposure of WT macrophages to T. crassiceps antigens triggered tyrosine phosphorylation signaling activity, which was diminished in MGL1(-/-) macrophages. Following T. crassiceps infection, MGL1(-/-) mice failed to produce significant levels of inflammatory cytokines early in the infection compared to WT mice. In contrast, MGL1(-/-) mice developed a Th2-dominant immune response that was associated with significantly higher parasite loads, whereas WT mice were resistant. Flow cytometry and RT-PCR analyses showed overexpression of the mannose receptors, IL-4Rα, PDL2, arginase-1, Ym1, and RELM-α on MGL1(-/-) macrophages. These studies indicate that MGL1 is involved in T. crassiceps recognition and subsequent innate immune activation and resistance.
Assuntos
Antígenos de Helmintos/imunologia , Assialoglicoproteínas/metabolismo , Resistência à Doença/imunologia , Lectinas Tipo C/metabolismo , Macrófagos Peritoneais/metabolismo , Proteínas de Membrana/metabolismo , Transdução de Sinais , Taenia/imunologia , Teníase/imunologia , Acetilgalactosamina/metabolismo , Animais , Assialoglicoproteínas/deficiência , Citocinas/biossíntese , Feminino , Galactose/metabolismo , Glicoconjugados/metabolismo , Imunidade , Espaço Intracelular/metabolismo , Cinética , Lectinas Tipo C/deficiência , Ativação de Macrófagos/imunologia , Proteínas de Membrana/deficiência , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Fosforilação , Fosfotirosina/metabolismo , Ligação Proteica , Solubilidade , Teníase/parasitologiaRESUMO
The nasal mucosa is the first contact with antigens to induce IgA response. The role of this site has rarely been studied. We have shown than intranasal administration with Naegleria fowleri lysates plus Cholera toxin (CT) increased the protection (survival up to 100%) against N. fowleri infection in mice and apparently antibodies IgA and IgG together with polymorphonuclear (PMN) cells avoid the attachment of N. fowleri to apical side of the nasal epithelium. We also observed that nasal immunization resulted in the induction of antigen-specific IgG subclasses (IgG1 and IgG2a) in nasal washes at days 3 and 9 after the challenge and IgA and IgG in the nasal cavity, compared to healthy and infected mice. We found that immunization with both treatments, N. fowleri lysates plus CT or CT alone, increased the expression of the genes for alpha chain, its receptor (pIgR), and it also increased the expression of the corresponding proteins evidenced by the â¼65 and â¼74kDa bands, respectively. Since the production of pIgR, IgA and IgG antibodies, is up-regulated by some factors, we analyzed the expression of genes for IL-10, IL-6, IFN-γ, TNF-α and IL-1ß by using RT-PCR of nasal passages. Immunization resulted in an increased expression of IL-10, IL-6, and IFN-γ cytokines. We also aimed to examine the possible influences of immunization and challenge on the production of inflammatory cytokines (TNF-α and IL-1ß). We observed that the stimulus of immunization inhibits the production of TNF-α compared to the infected group where the infection without immunization causes an increase in it. Thus, it is possible that the coexistence of selected cytokines produced by our immunization model may provide a highly effective immunological environment for the production of IgA, IgG and pIgR as well as a strong activation of the PMN in mucosal effector tissue such as nasal passages.
Assuntos
Toxina da Cólera/administração & dosagem , Citocinas/metabolismo , Isotipos de Imunoglobulinas/metabolismo , Naegleria fowleri/química , Mucosa Nasal/imunologia , Receptores de Imunoglobulina Polimérica/metabolismo , Administração Intranasal , Animais , Western Blotting , Toxina da Cólera/imunologia , Citocinas/genética , Regulação da Expressão Gênica , Cabras , Imunoglobulina A/genética , Imunoglobulina A/metabolismo , Imunoglobulina G/genética , Imunoglobulina G/metabolismo , Isotipos de Imunoglobulinas/genética , Imuno-Histoquímica , Camundongos , Naegleria fowleri/imunologia , Mucosa Nasal/efeitos dos fármacos , Mucosa Nasal/parasitologia , RNA Mensageiro/metabolismo , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Imunoglobulina Polimérica/genéticaRESUMO
The aim of this study was to evaluate the effect of long-chain omega-3 polyunsaturated fatty acid (n-3 PUFA) supplementation on metabolic state and gene expression in subcutaneous adipose tissues of obese adolescents. Obese adolescents (n = 26, 10 girls and 16 boys) aged 12.4 ± 2.1 years were assigned to a 12-week regimen of n-3 PUFA intake. Five times per day, subjects received a food supplement consisting of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (3 g per day, 944 mg EPA, and 2,088 mg DHA). Blood parameters were measured, and subcutaneous adipose tissue biopsies were analyzed to determine gene expression at baseline and after 12 weeks. Student's t test and the Wilcoxon signed-rank test were used to estimate differences in arithmetic means of pre- and post-dietary supplementation for various anthropometric, biochemical, clinical, and gene expression parameters. After 12 weeks, n-3 PUFA consumption was associated with decreased body mass index (29.7 ± 4.6 vs. 27.8 ± 4.4 kg/m(2); P < 0.001), waist circumference (93.2 ± 9.9 vs. 90.5 ± 10.0 cm; P < 0.003), hip circumference (102.9 ± 10.9 vs. 101.1 ± 10.9 cm; P < 0.014), and blood triglyceride levels (220.8 ± 27.4 vs. 99.7 ± 32.7 mg/dL; P < 0.001). Fatty acid supplementation/n3 PUFA supplementation was associated with a downregulated expression of the genes encoding PPARγ and PGC-1α (P < 0.001), and an upregulated expression of the genes encoding PPARα (P < 0.007) and SREBP1 (P < 0.021). The expressions of SOD2 (P < 0.04), CAT (P < 0.001), GPX3 (P < 0.032) and HIF-1α protein also decreased. Our study demonstrated that n-3 PUFA consumption and dietary restriction improved the anthropometric parameters and decreased the triglycerides levels of the adolescents, suggesting a reduction in hypoxia in subcutaneous adipose tissue.
Assuntos
Suplementos Nutricionais , Ácidos Graxos Ômega-3/administração & dosagem , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , PPAR alfa/genética , PPAR gama/genética , Obesidade Infantil , Gordura Subcutânea/efeitos dos fármacos , Adolescente , Criança , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , PPAR alfa/metabolismo , PPAR gama/metabolismo , Obesidade Infantil/dietoterapia , Obesidade Infantil/genética , Obesidade Infantil/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Gordura Subcutânea/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
We analyzed the possible role of glycoconjugates containing α-D-mannose and α-D-glucose residues in adherence of trophozoites to mouse nasal epithelium. Trophozoites incubated with 20 µg of one of three different lectins which preferentially recognized these residues were inoculated intranasally in Balb/c mice. Mouse survival was 40% with Pisum sativum and Canavalia ensiformis and 20% with Galanthus nivalis amebic pretreatment, compared with 0% survival for control animals administered trophozoites without pretreatment. Possibly some of the glycoproteins found in Naegleria fowleri represent an adherence factor. Differences in the saccharide sequences of the Naegleria species, even on the same glycoconjugate structure, could explain the different results corresponding to the distinct pretreatments (C. ensiformis, G. nivalis, and P. sativum). We found a higher expression of glycoconjugates recognized by P. sativum in Naegleria lovaniensis than N. fowleri, probably due to the higher number of oligosaccharides containing an α-1,6-linked fucose moiety expressed on the former species.
Assuntos
Adesão Celular/fisiologia , Glicoconjugados/metabolismo , Manose/química , Naegleria/fisiologia , Mucosa Nasal/metabolismo , Trofozoítos/fisiologia , Animais , Citometria de Fluxo , Glucose , Glicoconjugados/química , Lectinas/química , Lectinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Chemical composition, in vitro gas production with and without polyethyleneglycol (PEG-4000 MW), and in vitro digestibility of dry matter (IVDMD) and organic (IVOMD) foliage from Pithecellobium dulce, Gliricidia sepium and Haematoxylum brasiletto were determined. The preference test was run for 15 days: the first 10 days as adaption period and the 5 days served as evaluation period. It was conducted in ten developing female Creole goats of 6 months old, weighing 14 ± 2.0 kg in order to determine goat preference for any of the three foliages. Productive performance of 35 male creole kids of 6 months old (14 ± 3.0 kg) was also determined by ad libitum feeding of the foliage of the tree: the 30 and 15 % of each of the P. dulce (T1, T2), G. sepium (T3, T4), and H. brasiletto (T5, T6) foliages were added to the experiment diets, while T7 served as control diet that did not contain any foliage. The crude protein (CP), total phenols (TP), condensed tannins (CT), IVDMD, and IVOMD were different among the foliages. The PEG determined the biological activity of the TP and CT of H. brasiletto. Goats preferred to consume the foliage of P. dulce because of its higher content of CP and IVDMD and low content of TP and CT. In the productive response, dry matter intake (DMI) was higher in kids fed T1 diet and was stimulated by higher IVDMD and IVOMD, which resulted in the higher daily weight gain (DWG). The contribution with TP and CT of H. brasiletto to T5 and T6 and the rejection by the animals of G. sepium in T3 and T4 explain the negative effects on the DMI and the DWG. Findings of the study suggested higher kid performance for P. dulce foliage. Possible attributes may include its better CP, low TP and CT, and higher digestibility.
Assuntos
Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Fabaceae/química , Comportamento Alimentar , Cabras/fisiologia , Envelhecimento , Animais , Dieta/veterinária , Digestão , Feminino , Cabras/crescimento & desenvolvimento , Masculino , México , Valor Nutritivo , Especificidade da Espécie , Árvores/química , Clima Tropical , Aumento de PesoRESUMO
Few reports exist on the differences in cell populations or immunological functions between the proximal and distal segments of the small intestine (SI). In the current contribution we analyzed the expression of the polymeric immunoglobulin receptor (pIgR) and alpha chains as well as the density of IgA-producing cells from the proximal and distal intestinal segments from Balb/c mice. Furthermore, by using real-time RT-PCR we quantified the expression of cytokines (TNF-alpha, IFN-gamma, IL-4 and TGF-beta), Toll-like receptor-4 (TLR-4), and the glucocorticoid receptor (GR) involved in pIgR expression in intestinal epithelial cells (IEC). In this study, for the first time it has been demonstrated that the expression of the pIgR as well as alpha chain was greater in the proximal than the distal segment of the small intestine of normal mice. Moreover, we found striking differences in the expression of cytokines at the different intestinal compartments. Whereas the expression of TNF-alpha, IFN-gamma and TGF-beta was higher in lamina propria lymphocytes (LPL) of the distal than proximal segment, it was higher in IEC of the proximal than distal segment. In contrast, the expression of the gene for IL-4 was higher in the LPL of the proximal segment and the IEC of the distal segment. Although the overall expression of TNF-alpha, IL-4, IFN-gamma and TGF-beta was higher in the whole mucosa of the distal than proximal segment, we propose that cytokines produced by epithelial cells (TNF-alpha, IFN-gamma and TGF-beta) autocrinally up-regulate the expression of mRNA for the pIgR. Finally the expression of the GR was higher in the proximal segment, while the expression of the gene for TLR-4 was significantly higher in the IEC of the distal than proximal segment. The higher expression of pIgR found in the proximal segment is probably related to the effect on epithelial cells of the higher production of TNF-alpha, IFN-gamma and TGF-beta, as well as the higher expression of the glucocorticoid receptors. The increased expression of pIgR in the proximal segment appears primarily responsible for the increased secretory IgA levels in the small intestine of mice. These results confirm and extend previous findings supporting the compartmentalization of the intestinal immune system.