RESUMO
Generation 4 of polyamidoamine dendrimer (G4-PAMAM) has several biological effects due to its tridimensional globular structure, repetitive branched amides, tertiary amines, and amino-terminal subunit groups liked to a common core. G4-PAMAM is cytotoxic due to its positive charges. However, its cytotoxicity could increase in cancer cells due to the excessive intracellular negative charges in these cells. Furthermore, this work reports G4-PAMAM chemical structural characterization using UHPLC-QTOF-MS/MS (LC-MS) by electrospray ionization to measure its population according to its positive charges. Additionally, the antiproliferative effects and intracellular localization were explored in the HMC-1 and K-562 cell lines by confocal microscopy. The LC-MS results show that G4-PAMAM generated multivalent mass spectrum values, and its protonated terminal amino groups produced numerous positive charges, which allowed us to determine its exact mass despite having a high molecular weight. Additionally, G4-PAMAM showed antiproliferative activity in the HMC-1 tumor cell line after 24 h (IC50 = 16.97 µM), 48 h (IC50 = 7.02 µM) and 72 h (IC50 = 5.98 µM) and in the K-562 cell line after 24 h (IC50 = 15.14 µM), 48 h (IC50 = 14.18 µM) and 72 h (IC50 = 9.91 µM). Finally, our results showed that the G4-PAMAM dendrimers were located in the cytoplasm and nucleus in both tumor cell lines studied.
Assuntos
Dendrímeros/farmacologia , Leucemia/tratamento farmacológico , Leucemia/metabolismo , Nylons/farmacologia , Antineoplásicos/análise , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Cromatografia Líquida , Dendrímeros/análise , Dendrímeros/farmacocinética , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Humanos , Concentração Inibidora 50 , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Células K562 , Leucemia/patologia , Nylons/análise , Nylons/farmacocinética , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Distribuição TecidualRESUMO
Background: Adenoviruses are highly contagious pathogens which cause respiratory disease particularly in children; they may induce severe disease in infants. Human neutrophil peptides (HNPs) have been found to exhibit antiadenoviral activity. Thus, we have investigated HNPs in nasal aspirates (NAs) of children suffering from adenoviral common cold. Objective: To investigate the release of HNP-1-4 in adenovirus infection and the relationship with self-limiting upper respiratory tract infections. Methods: Nasal aspirate samples (n=14) were obtained from children (aged 6-12 years) infected with adenovirus between June 2012 and December 2015. Control samples were taken 4 weeks after infection when the children were asymptomatic. Levels of HNPs were measured using an enzyme-linked immunosorbent assay (ELISA). Results: There were increased levels of HNP-1, -3, and -4, but not HNP-2, in nasal aspirates (NAs) during adenovirus infections compared to healthy specimens (p ≤ 0.01). Moreover, there was also increase in the neutrophil count, which is a known cell source of HNPs. Conclusion: Our finding supports the involvement of HNP-1, -3, and -4 in naturally occurring cold in children infected with adenovirus. Because of their known antiviral properties, it is tempting to hypothesize that HNPs might play a protective role in adenovirus-induced respiratory disease; however, this remains to be shown.
Assuntos
Infecções por Adenovirus Humanos/metabolismo , Defensinas/metabolismo , Neutrófilos/metabolismo , Criança , Feminino , Humanos , MasculinoRESUMO
The present study of Mexican high school football players on the offensive squad had the aim of determining the relation between their anthropometric profile, position on the team, and performance in competition. The study included 41 players from 15 to 18 years of age, studying and playing football at one of two high schools. Anthropometric measurements were taken (weight, height, diameter and perimeter of muscle areas) to determine the body mass index, body composition and somatotype. The results show that the anthropometric profile is related to performance (good/very good or poor/deficient), and that this relation depends on the position of the player. For linemen, good performance was associated with being tall, heavy and with a relatively high body fat percentage, and poor/deficient performance with a significantly lower height, lower weight and lower body fat percentage. For receivers and running backs, height did not appear to influence performance. For running backs, good performance was associated with heavy weight and a high body fat percentage. However for receivers, good performance was associated with a lower body fat percentage. The average somatotype for linemen was mesomorph-endomorph, for running backs meso-ectomorph, and for receivers mesomorph-ectomorph.
El objetivo de este trabajo fue determinar la relación entre los perfiles antropométricos de jugadores juveniles mexicanos de futbol americano y su desempeño en la competición, tomando en cuenta su posición ofensiva en el juego. Se estudiaron 41 jugadores de 15 a 18 años de edad, de nivel bachillerato de dos instituciones educativas. Se registraron medidas antropométricas (peso, estatura, pliegues, diámetros y perímetros) para determinar el índice de masa corporal, la composición corporal y el somatotipo. Nuestros resultados muestran que el perfil antropométrico de los jugadores estuvo relacionado con su desempeño (bueno/muy bueno o malo/deficiente) en su posición ofensiva en el juego. La estatura fue un factor importante en el caso de los linieros, ya que los clasificados como malos/deficientes para esa posición, tuvieron una talla significativamente más baja que los buenos/muy buenos; sin embargo, ésta no parece influir como criterio para clasificar a receptores y corredores. Se encontró que los linieros ofensivos fueron los jugadores más altos y pesados y con mayor porcentaje de grasa, seguidos de los receptores y por último de los corredores. En esta posición, los malos/deficientes fueron los de menor peso, y los buenos/muy buenos fueron los de mayor porcentaje graso, al igual que en la posición de corredores. Sin embargo en los receptores la relación se invierte, ya que los buenos/muy buenos fueron los de menor porcentaje graso. El somatotipo medio por posición fue mesomorfo-endomorfo para los linieros, mesomorfo-ectomorfo para los receptores y meso-ectomorfo para los corredores.
Assuntos
Humanos , Masculino , Adolescente , Composição Corporal , Desempenho Atlético , Futebol Americano , Somatotipos , Índice de Massa Corporal , Estudos Transversais , MéxicoRESUMO
The impact of intermittent fasting versus ad libitum feeding during Salmonella typhimurium infection was evaluated in terms of duodenum IgA levels, bacterial clearance and intestinal and extra-intestinal infection susceptibility. Mice that were intermittently fasted for 12 weeks or fed ad libitum were infected with S. typhimurium and assessed at 7 and 14 days post-infection. Next, we evaluated bacterial load in the faeces, Peyer's patches, spleen and liver by plate counting, as well as total and specific intestinal IgA and plasmatic corticosterone levels (by immunoenzymatic assay) and lamina propria IgA levels in plasma cells (by cytofluorometry). Polymeric immunoglobulin receptor, α- and J-chains, Pax-5 factor, pro-inflammatory cytokine (tumour necrosis factor-α and interferon-γ) and anti-inflammatory cytokine (transforming growth factor-ß) mRNA levels were assessed in mucosal and liver samples (by real-time PCR). Compared with the infected ad libitum mice, the intermittently fasted infected animals had (1) lower intestinal and systemic bacterial loads; (2) higher SIgA and IgA plasma cell levels; (3) higher mRNA expression of most intestinal parameters; and (4) increased or decreased corticosterone levels on day 7 and 14 post-infection, respectively. No contribution of liver IgA was observed at the intestinal level. Apparently, the changes following metabolic stress induced by intermittent fasting during food deprivation days increased the resistance to S. typhimurium infection by triggering intestinal IgA production and presumably, pathogen elimination by phagocytic inflammatory cells.
Assuntos
Duodeno/imunologia , Jejum , Imunoglobulina A/imunologia , Plasmócitos/imunologia , Infecções por Salmonella/imunologia , Salmonella typhimurium/imunologia , Estresse Fisiológico/imunologia , Animais , Carga Bacteriana , Corticosterona/sangue , Citocinas/genética , Citocinas/metabolismo , Duodeno/microbiologia , Fezes/microbiologia , Regulação da Expressão Gênica , Imunidade nas Mucosas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fator de Transcrição PAX5/genética , Fator de Transcrição PAX5/metabolismoRESUMO
El objetivo de este estudio es analizar la composición corporal y el somatotipo de judokas participantes en el campeonato escolar de España. Fueron estudiados 57 niños de categoría infantil (13,33±0,66 años) y 40 niños de categoría cadete (15,18±0,74 años) (n=97), mediante un fraccionamiento corporal tetracompartimental y el análisis del somatotipo según el método de Heath-Carter. En categoría infantil obtuvimos una media de 48,11% de masa muscular, 25,49% de masa residual, 14,80% de masa grasa y 11,56% de masa ósea. En categoría cadete, 47.67% de masa muscular, 25,25% de masa residual, 16,46% de masa grasa y 10,57% de masa ósea. Se encontraron diferencias significativas únicamente en el compartimiento de masa ósea entre ambos grupos. El somatotipo medio para ambas categorías es Ectomorfo Balanceado.
The aim of the study was to analyze body composition and somatotype of the judokas participants at the Spanish Junior Championship. We analyzed 97 male judokas (n= 97): 57 junior (13.33±0.66 years) and 40 cadets (15.18±0.74 years), through a Tetracompartimental Fractionation and the somatotype through a Heath-Carter method. In junior category, 48.11% of muscle mass, 25.49% of residual mass, 14.80% of fat mass and 11.56% of bone mass. In cadet category, 47.67% of muscle mass, 25.25% of residual mass, 16.46% of fat mass and 10.57% of bone mass. Significant differences were found only in the bone compartment between both groups. The average somatotype for both categories was Ectomorph Balanced.
Assuntos
Humanos , Masculino , Somatotipos , Composição Corporal , Artes Marciais , AntropometriaRESUMO
New molecular markers of cancer had emerged with novel applications in cancer prevention and therapeutics, including for breast cancer of unknown causes, which has a high impact on the health of women worldwide. The purpose of this research was to determine protein and mRNA expression of synaptic vesicle 2 (SV2) isoforms A, B and C in breast cancer cell lines. Cultured cell lines MDA-MB-231, SKBR3, T47D were lysed and their protein and mRNA expression analyzed by real-time PCR and western blot technique, respectively. SV2A, B proteins were identified in non-tumor (MCF-10A) and tumor cell lines (MDA-MB-231 and T47D) while SV2C only was found in the T47D cell line. Furthermore, the genomic expression was consistent with protein expression for a such cell line, but in MDA-MB-231 there was no SV2B genomic expression, and the SV2C mRNA and protein were not found in the non tumoral cell line. These findings suggest a possible cellular transdifferentiation to neural character in breast cancer, of possible relevance to cancer development, and point to possible use of SV2 as molecular marker and a vehicle for cancer treatment with botulinum toxin.
Assuntos
Neoplasias da Mama/metabolismo , Mama/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Western Blotting , Neoplasias da Mama/genética , Células Cultivadas , Feminino , Humanos , Glicoproteínas de Membrana/genética , Proteínas do Tecido Nervoso/genética , Isoformas de Proteínas , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Involvement of cytochrome P450 genes (CYPs) in breast cancer (BCa) may differ between populations, with expression patterns affected by tumorigenesis. This may have an important role in the metabolism of anticancer drugs and in the progression of cancer. The aim of this study was to determine the mRNA expression patterns of four cytochrome P450 genes (CYP2W1, 3A5, 4F11 and 8A1) in Mexican women with breast cancer. Real- time PCR analyses were conducted on 32 sets of human breast tumors and adjacent non-tumor tissues, as well as 20 normal breast tissues. Expression levels were tested for association with clinical and pathological data of patients. We found higher gene expression of CYP2W1, CYP3A5, CYP4F11 in BCa than in adjacent tissues and only low in normal mammary glands in our Mexican population while CYP8A1 was only expressed in BCa and adjacent tissues. We found that Ki67 protein expression was associated with clinicopathological features as well as with CYP2W1, CYP4F11 and CYP8A1 but not with CYP3A5. The results indicated that breast cancer tissues may be better able to metabolize carcinogens and other xenobiotics to active species than normal or adjacent non-tumor tissues.
Assuntos
Neoplasias da Mama/enzimologia , Citocromo P-450 CYP3A/biossíntese , Sistema Enzimático do Citocromo P-450/biossíntese , Neoplasias da Mama/genética , Citocromo P-450 CYP3A/genética , Sistema Enzimático do Citocromo P-450/genética , Família 2 do Citocromo P450 , Família 4 do Citocromo P450 , Feminino , Humanos , Antígeno Ki-67/biossíntese , México , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The immune-suppression caused by acute stress can be reduced by a regular practice of moderate exercise which is known to modulate the expression of secretory-IgA. This antibody is essential for protection against infections and maintenance of homeostasis at the mucosal level. In order to explore the effects of moderate exercise on secretory-IgA production in ileum of the small intestine, 2 groups of mice were submitted to this protocol for 6 months, an exercise group and a sedentary group. After sacrifice, levels of secretory-IgA in intestinal fluid and levels of adrenal hormones in serum were determined by enzyme immunoenzymatic assay. IgA-plasma cells in lamina propria were evaluated by flow cytometry. Transcriptional mRNA expression in mucosa of alpha-chain, J-chain, pIgR and cytokines (Interleukin-2, -4, -6, -10, transforming growth factor-beta, interferon-gamma and tumor necrosis factor) were determined by RT-PCR. In comparison with sedentary mice, moderate exercised mice displayed an up-regulating effect on the production of secretory-IgA and IgA-plasma cells, on the expression of all mRNA transcripts from secretory-IgA associated proteins, and on all cytokines tested. However, serum levels of adrenal hormones were not altered. Future studies on secretory-IgA production are necessary to support the substantive effect of moderate exercise on protection and homeostasis at the intestinal level.
Assuntos
Íleo/imunologia , Imunoglobulina A Secretora/imunologia , Condicionamento Físico Animal/fisiologia , Esforço Físico/imunologia , Animais , Íleo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Esforço Físico/fisiologiaRESUMO
Breast cancer (BCa) is the leading type of cancer in Mexican women. Genetic factors, such as single nucleotide polymorphisms (SNP) of P450 system, have been reported in BCa. In this report, and for the first time in the literature, we analyzed the rs3735684 (7021 G>A), rs11553651 (15016 G>T) and rs56195291 (60020 C>G) polymorphisms in the CYP2W1, 4F11 and 8A1 genes in patients with BCa and in healthy Mexican women to identify a potential association between these polymorphisms and BCa risk. Patients and controls were used for polymorphism analysis using an allelic discrimination assay with TaqMan probes and confirmed by DNA sequencing. Links with clinic-pathological characteristics were also analyzed. Statistical analysis was performed using the standard χ2 or Fisher exact test statistic. No significant differences were observed in the distributions of CYP2W1 (OR 8.6, 95%CI 0.43-172.5 P>0.05; OR 2.0, 95%CI 0.76-5.4, P>0.05) and CYP4F11 (OR 0.3, 95%CI 0.01-8.4 P>0.05) genotypes between the patients and controls. Only the CYP8A1 CC genotype was detected in patients with BCa and the controls. All polymorphism frequencies were in Hardy-Weinberg Equilibrium (HWE) in the controls (P>0.05). We found a significant association between BCa risk and smoking, use of oral contraceptives or hormonal replacement therapy (HRT), obesity, hyperglycemia, chronic diseases, family history of cancer and menopausal status in the population studied (P<0.05). Tobacco, oral contraceptive or HRT, chronic diseases and obesity or overweight were strongly associated with almost eight, thirty-five, nine and five-fold increased risk for BCa. Tobaco, obesity and hyperglycemia significantly increased the risk of BCa in the patients carrying variant genotypes of CYP2W1 (P<0.05). These results indicate that the CYP2W1 rs3735684, CYP4F11 rs11553651 and CYP8A1 rs56195291 SNPs are not a key risk factor for BCa in Mexican women. This study did not detect an association between the CYP2W1, 4F11 and 8A1 genes polymorphisms and BCa risk in a Mexican population. However, some clinico-pathological risk factors interact with CYP2W1 genotypes and modifies susceptibility to BCa.
Assuntos
Neoplasias da Mama/genética , Sistema Enzimático do Citocromo P-450/genética , Idoso , Sequência de Bases , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/radioterapia , Anticoncepcionais Orais , Família 2 do Citocromo P450 , Família 4 do Citocromo P450 , Terapia de Reposição de Estrogênios , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Hiperglicemia , Menopausa , México , Pessoa de Meia-Idade , Obesidade , Polimorfismo de Nucleotídeo Único , Fatores de Risco , Análise de Sequência de DNA , FumarRESUMO
According to previous reports, intranasal administration of the Cry1Ac protein alone or with amoebic lysates increases protection against Naegleria fowleri meningoencephalitis in mice, apparently by eliciting IgA responses in the nasal mucosa. In the current study, we performed an immunohistochemical analysis of IgA in the nasal mucosa of mice immunized intranasally with Cry1Ac, and amoebic lysates or a combination of both. The animals were sacrificed 24 h after the last immunization or after an intranasal lethal challenge with N. fowleri. Our results indicate that all of the intranasal immunizations provoked an increase in areas with metaplasia in the olfactory epithelium, allowing for secretion of IgA. As a result, IgA antibodies were found interacting with trophozoites in the nasal lumen, and there was a marked increase of IgA in the metaplasic epithelium. On the other hand in nonimmunized mice trophozoites were observed invading the nasal mucosa, which was not the case for immunized mice. Our results suggest that intranasal immunization provokes cellular changes in the olfactory epithelium, leading to greater protection against N. fowleri that is probably caused by an increased secretion of IgA. The increased IgA response induced in the nasal mucosa by immunization probably impedes both amoebic adhesion and subsequent invasion of the parasite to the nasal epithelium.
Assuntos
Amebíase/imunologia , Proteínas de Bactérias/imunologia , Toxinas Bacterianas/imunologia , Endotoxinas/imunologia , Proteínas Hemolisinas/imunologia , Imunização , Imunoglobulina A Secretora/análise , Meningoencefalite/imunologia , Naegleria fowleri/imunologia , Mucosa Olfatória/imunologia , Adjuvantes Imunológicos , Administração Intranasal , Amebíase/parasitologia , Animais , Anticorpos Antiprotozoários/análise , Antígenos de Protozoários/imunologia , Toxinas de Bacillus thuringiensis , Masculino , Meningoencefalite/parasitologia , Metaplasia , Camundongos , Camundongos Endogâmicos BALB C , Mucosa Nasal/imunologia , Mucosa Nasal/parasitologia , Proteínas Recombinantes/imunologiaRESUMO
It is well known that Entamoeba histolytica can cause damage to its host by means of enzymes, cytotoxins and perforins that affect different types of cells. In this work we investigated the effect of different Entamoeba histolytica trophozoites products over the secretion of antibodies by mice Peyer's patches using a reverse hemolytic assay. Results showed that amoebic glycoproteins inhibited the secretion of antibodies by lymphocytes from mice Peyer's patches. In all cases the decrease in antibody secretion was not due to cell death. The molecules responsible for this effect were shown to be high molecular weight immunogenic glycoproteins without enzymatic activity. The fact that amoebic glycoproteins interfere with plasmatic cells function could be another mechanism of evasion of the immune response by Entamoeba histolytica, although we still do not know how these amoebic products cause the inhibition.
Assuntos
Células Produtoras de Anticorpos/efeitos dos fármacos , Entamoeba histolytica/química , Glicoproteínas/farmacologia , Nódulos Linfáticos Agregados/citologia , Proteínas de Protozoários/farmacologia , Animais , Formação de Anticorpos/efeitos dos fármacos , Células Produtoras de Anticorpos/imunologia , Glicoproteínas/química , Glicoproteínas/isolamento & purificação , Técnica de Placa Hemolítica , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Proteínas de Protozoários/química , Proteínas de Protozoários/isolamento & purificaçãoRESUMO
Because the correct diagnosis of indeterminate leprosy (IL) requires the finding of acid-fast bacilli in skin lesions from clinically and histopathologically suggestive cases, it is necessary to develop a reliable method for this purpose. This paper presents a simple procedure, available to every general laboratory, which consists in obtaining 2 suspensions: SI, by mincing and grinding the tissue in phosphate-buffered saline; and SII, after treating SI with NaOH solution and digesting with trypsin. In 22 IL skin biopsies, bacilli were directly observed in only 3 with the Ziehl-Neelsen (ZN) stain; and with the peroxidase-antiperoxidase method it was impossible to differentiate between nonspecific precipitate and true positive reactions. In contrast, 18 positive results from the same 22 samples were obtained when both SI and SII were evaluated with ZN stain. The logarithmic bacterial index was also increased in at least 7 cases.