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1.
Sci Rep ; 13(1): 8041, 2023 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-37198168

RESUMO

Microorganisms do not live as dispersed single cells but rather they form aggregates with extracellular polymeric substances at interfaces. Biofilms are considered efficient life forms because they shield bacteria from biocides and collect dilute nutrients. This is a big concern in industry since the microorganisms can colonize a wide range of surfaces, accelerating material deterioration, colonizing medical devices, contaminating ultrapure drinking water, increasing energy costs and creating focus of infection. Conventional biocides that target a specific component of the bacteria are not effective in the presence of biofilms. Efficient biofilm inhibitors are based on a multitarget approach interacting with the bacteria and the biofilm matrix. Their rationale design requires a thorough understanding of inhibitory mechanisms that are still largely lacking today. Herein we uncover via molecular modelling the inhibition mechanism of cetrimonium 4-OH cinnamate (CTA-4OHcinn). Simulations show that CTA-4OH micelles can disrupt symmetric and asymmetric bilayers, representative of inner and outer bacterial membranes, following three stages: adsorption, assimilation, and defect formation. The main driving force for micellar attack is electrostatic interactions. In addition to disrupting the bilayers, the micelles work as carriers facilitating the trapping of 4OH cinnamate anions within the bilayer upper leaflet and overcoming electrostatic repulsion. The micelles also interact with extracellular DNA (e-DNA), which is one of the main components of biofilms. It is observed that CTA-4OHcinn forms spherical micelles on the DNA backbone; which hinders their ability to pack. This is demonstrated by modelling the DNA along the hbb histone-like protein, showing that in the presence of CTA-4OHcinn, DNA does not pack properly around hbb. The abilities of CTA-4OHcinn to cause cell death through membrane disruption and to disperse a mature, multi-species biofilm are also confirmed experimentally.


Assuntos
Desinfetantes , Micelas , Cetrimônio/farmacologia , Biofilmes , Bactérias , DNA/farmacologia , Desinfetantes/farmacologia , DNA Bacteriano/farmacologia
2.
Int J Mol Sci ; 23(21)2022 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-36362433

RESUMO

Neuroendocrine neoplasms (NENs) are relatively rare neoplasms occurring predominantly in the gastrointestinal tract and pancreas. Their heterogeneity poses challenges for diagnosis and treatment. There is a paucity of markers for characterisation of NEN tumours. For routine diagnosis, immunohistochemistry of the NEN-specific markers CgA and synaptophysin and the proliferation marker Ki-67 are used. These parameters, however, are qualitative and lack the capacity to fully define the tumour phenotype. Molecules of epithelial-mesenchymal transition (EMT) are potential candidates for improved tumour characterisation. Using qRT-PCR, we measured mRNA levels of 27 tumour markers, including 25 EMT-associated markers, in tumour tissue and matched non-tumour tissues for 13 patients with pancreatic NENs. Tissue from patients with three different grades of tumour had distinctly different mRNA profiles. Of the 25 EMT-associated markers analysed, 17 were higher in G3 tissue relative to matched non-tumour tissue, including CD14, CD24, CD31, CD44, CD45, CD56, CK6, CK7, CK13, CK20, NSE, CDX2, CgA, DAXX, PCNA, laminin and Ki-67. The differences in levels of seven EMT-associated markers, Ki-67, DAXX, CD24, CD44, vimentin, laminin and PDX1 plus CgA and NSE (neuroendocrine markers) enabled a distinct molecular signature for each tumour grade to be generated. EMT molecules differentially expressed in three tumour grades have potential for use in tumour stratification and prognostication and as therapeutic targets for treatment of neuroendocrine cancers, following validation with additional samples.


Assuntos
Neoplasias Pancreáticas , Humanos , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/análise , Transição Epitelial-Mesenquimal/genética , Antígeno Ki-67 , Laminina , Neoplasias Pancreáticas/patologia , RNA Mensageiro
3.
Biotechnol Bioeng ; 119(8): 2031-2045, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35441370

RESUMO

Microalgae are important renewable feedstock to produce biodiesel and high-value chemicals. Different wavelengths of light influence the growth and metabolic activities of algae. Recent research has identified the light-sensing proteins called photoreceptors that respond to blue or red light. Structural elucidations of algal photoreceptors have gained momentum over recent years. These include channelrhodopsins, PHOT proteins, animal-like cryptochromes, and blue-light sensors utilizing flavin-adenine dinucleotide proteins. Pulsing light has also been investigated as a means to optimize energy inputs into bioreactors. This study summarizes the current structural and functional basis of photoreceptor modulation to optimize the growth, production of carotenoids and other high-value metabolites from microalgae. The review also encompasses novel photobioreactor designs that implement different light regimes including light wavelengths and time to optimize algal growth and desired metabolite profiles for high-value products.


Assuntos
Microalgas , Fotobiorreatores , Animais , Biocombustíveis , Criptocromos , Plantas
4.
Microbiol Resour Announc ; 11(5): e0101821, 2022 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-35420464

RESUMO

Here, we describe the genome of Lelliottia sp. strain WAP21, which was isolated from the soil of canola fields in Australia. The genome has a size of 4.9 Mbp and 4,583 predicted genes, with some potential pathways for metabolism of various carbon sources and metal acquisition.

5.
J Colloid Interface Sci ; 610: 785-795, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-34876264

RESUMO

HYPOTHESIS: Molecular interactions between 4-OH-cinnamate and cetrimonium in solution result in improved adsorption of the cinnamate on mild steel, developing a protective mechanism against the diffusion of corrosive chloride to the oxide surface. Fundamental understanding of this mechanism should allow new design routes for the development of eco-friendly corrosion inhibitors. EXPERIMENTS: Via classic molecular dynamics, simulations were carried out for cetrimonium and 4-OH-cinnamate in aqueous solutions at different ionic strengths and the results were validated with experimental SAXS data. Self-aggregation of cetrimonium 4-OH-cinnamate on a hydrated hematite surface was then simulated and results were compared with cryo-TEM imaging for the same compound. Finally, the effect of the adsorbed aggregates on chloride diffusion to the oxide surface was modelled. FINDINGS: Simulations showed the encapsulation of 4-OH-cinnamate into cetrimonium micelles, consistent with experiments. The newly formed micelles adsorb onto a hydrated iron oxide surface by forming hydrogen bonds between their carboxylate outer-shell groups and the surface hydroxyls. As the adsorbate concentrations increase, there is a morphological transition from spherical to wormlike adsorbed aggregates. The wormlike structure can block chloride ions, demonstrating a synergistic inhibitory mechanism between both cetrimonium and 4-OH-cinnamate. Encapsulation and delivery of active compounds to certain targets, such as carcinogenic tumors, have been well studied in biochemistry research, we demonstrate that the same mechanism can be applied to the design of efficient corrosion inhibitors, optimizing their delivery to the metal surface.


Assuntos
Cinamatos , Micelas , Cetrimônio , Compostos de Cetrimônio , Compostos Férricos , Espalhamento a Baixo Ângulo , Tensoativos , Difração de Raios X
6.
BMC Gastroenterol ; 21(1): 194, 2021 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-33906633

RESUMO

BACKGROUND: Neuroendocrine neoplasms (NENs) are a complex group of tumours that occur in many organs. Routinely used IHC markers for NEN diagnosis include CgA, synaptophysin, Ki67 and CD56. These have limitations including lack of correlation to clinical outcomes and their presence in non-tumour tissue. Identification of additional markers and more quantitative analyses of tumour tissue has the potential to contribute to improved clinical outcomes. We used qRT-PCR to profile the expression levels of a panel of markers in tumour and matched non-tumour tissue from a patient with a G1 pancreatic neuroendocrine tumour. Differences in mRNA levels between tumour and non-tumour tissue were compared with IHC analyses of the same sample. CASE PRESENTATION: An elderly man presented with lower abdominal pain for 6 months. Histological analysis identified a low grade, well differentiated pancreatic endocrine neoplasm. Twenty-seven tumour markers for neuroendocrine status, proliferation, stem cell phenotype, angiogenesis, epithelial to mesenchymal transition, cell adhesion, differentiation and tumour suppression were selected from previous studies and mRNA levels of these markers were measured in tumour and adjacent non-tumour tissue sample using qRT-PCR. IHC was carried out on the same tissue to detect the corresponding marker proteins. Of the markers analysed, seven showed higher mRNA levels in tumour relative to non-tumour tissue while thirteen had lower expression in tumour relative to non-tumour tissue. Substantial differences in mRNA levels were a gain of CgA, CD56, ß-catenin, CK20, PDX1 and p53 and loss of Ki67, PCAD, CK7, CD31, MENA, ECAD, EPCAM, CDX2 and CK6. Comparison of qRT-PCR data with IHC showed correlation between fifteen markers. CONCLUSION: Our study is unique as it included matched controls that provided a comparative assessment for tumour tissue analysis, whereas many previous studies report tumour data only. Additionally, we utilised qRT-PCR, a relatively quantitative diagnostic tool for differential marker profiling, having the advantage of being reproducible, fast, cheap and accurate. qRT-PCR has the potential to improve the defining of tumour phenotypes and, in combination with IHC may have clinical utility towards improving tumour stratification or distinguishing tumour grades. The results need to be validated with different grades of NENs and related to clinical outcomes.


Assuntos
Transição Epitelial-Mesenquimal , Tumores Neuroendócrinos , Idoso , Biomarcadores Tumorais/genética , Humanos , Imuno-Histoquímica , Masculino , Tumores Neuroendócrinos/genética , RNA Mensageiro
7.
Epigenomics ; 10(4): 419-431, 2018 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-29561170

RESUMO

AIM: To investigate epigenomic changes in pregnancy and early postpartum in women with and without type 2 diabetes. METHODS: Dimethylation of histones H3K4, H3K9, H3K27, H3K36 and H3K79 was measured in white blood cells of women at 30 weeks pregnancy, at 8-10 and 20 weeks postpartum and in never-pregnant women. RESULTS: Dimethylation levels of all five histones were different between women in pregnancy and early postpartum compared with never-pregnant women and were different between women with and without type 2 diabetes. CONCLUSION: Histone methylation changes are transient in pregnancy and early postpartum and may represent normal physiological responses to hormones. Different epigenomic profiles in women with type 2 diabetes mellitus may correlate with hormonal responses, leading to high risk pregnancy outcomes.


Assuntos
Diabetes Mellitus Tipo 2/genética , Epigênese Genética , Gravidez em Diabéticas/genética , Adulto , Feminino , Código das Histonas , Histonas/metabolismo , Humanos , Metilação , Pessoa de Meia-Idade , Projetos Piloto , Período Pós-Parto/genética , Gravidez
8.
Int Arch Allergy Immunol ; 172(2): 106-115, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28268214

RESUMO

BACKGROUND: Peanuts are still one of the highest contributors to anaphylactic deaths after ingestion of a food allergen. At the molecular level, interactions between peanut allergens and the intestinal epithelium are largely unexplored. Previous findings by our research group demonstrated that the major peanut allergens, i.e., Ara h 1, Ara h 2, Ara h 3, and Ara h 6, were able to cross the Caco-2 human cell culture model of the intestinal epithelium. This research broadened our investigation to identify the mechanisms by which the Caco-2 monolayers uptake peanut allergens, specifically by endocytosis. Here, we aim to increase our understanding of allergen-epithelial interactions and, more broadly, the pathway from allergen to allergy. METHODS: The human Caco-2 cell culture model was exposed to peanut extract and a combination of confocal microscopy and inhibition studies were used to identify the endocytotic mechanisms of peanut allergens in intestinal epithelia. RESULTS: Our findings demonstrate that the peanut allergens Ara h 1 and Ara h 2 are transported through intestinal epithelia initially via early endosomes using multiple endocytotic mechanisms. From there, they are then transported to late endosomes and ultimately to lysosomes. CONCLUSIONS: These novel findings provide insight into the allergen-epithelial interactions of peanut allergens with the intestinal epithelium. Consequently, this opens the possibility of the use of these endocytotic pathways as targets for inhibitors in therapeutic development and preventative measures for peanut allergy in the future.


Assuntos
Albuminas 2S de Plantas/metabolismo , Antígenos de Plantas/metabolismo , Endossomos/metabolismo , Glicoproteínas/metabolismo , Mucosa Intestinal/metabolismo , Lisossomos/metabolismo , Hipersensibilidade a Amendoim/imunologia , Proteínas de Plantas/metabolismo , Antígenos de Plantas/imunologia , Células CACO-2 , Endocitose , Glicoproteínas/imunologia , Humanos , Mucosa Intestinal/ultraestrutura , Proteínas de Membrana , Microscopia Confocal , Proteínas de Plantas/imunologia
9.
Microbiology (Reading) ; 162(12): 2017-2028, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27902405

RESUMO

Electromicrobiology has gained momentum in the last 10 years with advances in microbial fuel cells and the discovery of microbial nanowires (MNWs). The list of MNW-producing micro-organisms is growing and providing intriguing insights into the presence of such micro-organisms in diverse environments and the potential roles MNWs can perform. This review discusses the MNWs produced by different micro-organisms, including their structure, composition and mechanism of electron transfer through MNWs. Two hypotheses, metallic-like conductivity and an electron hopping model, have been proposed for electron transfer and we present a current understanding of both these hypotheses. MNWs not only are poised to change the way we see micro-organisms but also may impact the fields of bioenergy, biogeochemistry and bioremediation; hence, their potential applications in these fields are highlighted here.

10.
Langmuir ; 32(45): 11717-11727, 2016 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-27728769

RESUMO

Docosahexaenoic acid (DHA) is the most abundant polyunsaturated omega-3 fatty acid found in mammalian neuronal cell membranes. Although DHA is known to be important for neuronal cell survival, little is know about how DHA interacts with phospholipid bilayers. This study presents a detailed quartz crystal microbalance with dissipation monitoring (QCM-D) analysis of free DHA interactions with individual and mixed phospholipid supported lipid bilayers (SLB). DHA incorporation and subsequent changes to the SLBs viscoelastic properties were observed to be concentration-dependent, influenced by the phospholipid species, the headgroup charge, and the presence or absence of calcium ions. It was observed that 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) SLBs incorporated the greatest amount of DHA concentration, whereas the presence of phospholipids, phosphatidylserine (PS), and phosphatidylinositol (PI) in a POPC SLB significantly reduced DHA incorporation and changed the SLBs physicochemical properties. These observations are hypothesized to be due to a substitution event occurring between DHA and phospholipid species. PS domain formation in POPC/PS 8:2 SLBs was observed in the presence of calcium ions, which favored DHA incorporation to a similar level as for a POPC only SLB. The changes in SLB thickness observed with different DHA concentrations are also presented. This work contributes to an understanding of the physical changes induced in a lipid bilayer as a consequence of its exposure to different DHA concentrations (from 50 to 200 µM). The capacity of DHA to influence the physical properties of SLBs indicates the potential for dietary DHA supplementation to cause changes in cellular membranes in vivo, with subsequent physiological consequences for cell function.


Assuntos
Ácidos Docosa-Hexaenoicos/análise , Bicamadas Lipídicas/análise , Bicamadas Lipídicas/química , Técnicas de Microbalança de Cristal de Quartzo/métodos , Cálcio/química , Ácidos Docosa-Hexaenoicos/química , Fosfatidilcolinas/química
11.
Arch Biochem Biophys ; 611: 51-57, 2016 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-27317042

RESUMO

Zinc is essential for a wide variety of cellular processes in all cells. It is a critical dietary nutrient, particularly in the early stages of life. In the early neonatal period, adequate sources of zinc can be obtained from breast milk. In rare circumstances, the mammary gland produces zinc deficient milk that is potentially lethal for exclusively breast-fed infants. This can be overcome by zinc supplementation to the infant. Alterations to key zinc transporters provide insights into the mechanisms of cellular zinc homeostasis. The bioavailability of zinc in food depends on the presence of constituents that may complex zinc. In many countries, zinc deficiency is a major health issue due to poor nourishment. Young children are particularly affected. Zinc deficiency can impair immune function and contributes to the global burden of infectious diseases including diarrhoea, pneumonia and malaria. Furthermore, zinc deficiency may extend its influence across generations by inducing epigenetic effects that alter the expression of genes. This review discusses the significance of adequate zinc nutrition in infants, factors that influence zinc nutrition, the consequences of zinc deficiency, including its contribution to the global burden of disease, and addresses some of the knowledge gaps in zinc biology.


Assuntos
Fenômenos Fisiológicos da Nutrição do Lactente , Zinco/sangue , Zinco/química , Disponibilidade Biológica , Aleitamento Materno , Ciências da Nutrição Infantil , Pré-Escolar , Dieta , Suplementos Nutricionais , Exantema/fisiopatologia , Homeostase , Humanos , Lactente , Recém-Nascido , Leite Humano , Estado Nutricional , Zinco/deficiência
12.
J Clin Endocrinol Metab ; 101(6): 2396-404, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27045797

RESUMO

CONTEXT: Lifestyle factors mediate epigenetic changes that can cause chronic diseases. Although animal and laboratory studies link epigenetic changes to diabetes, epigenetic information in women with gestational diabetes (GDM) and type 2 diabetes is lacking. OBJECTIVE: This study sought to measure epigenetic markers across pregnancy and early postpartum and identify markers that could be used as predictors for conversion from GDM to type 2 diabetes. DESIGN: Global histone H3 dimethylation was measured in white blood cells at three time points: 30 wk gestation, 8-10 wk postpartum, and 20 wk postpartum, from four groups of women with and without diabetes. SETTING AND PARTICIPANTS: A total of 39 participants (six to nine in each group) were recruited including: nondiabetic women; women with GDM who developed postpartum type 2 diabetes; women with GDM without postpartum type 2 diabetes; and women with type 2 diabetes. MAIN OUTCOME MEASURE: Percentages of dimethylation of H3 histones relative to total H3 histone methylation were compared between diabetic/nondiabetic groups using appropriate comparative statistics. RESULTS: H3K27 dimethylation was 50-60% lower at 8-10 and 20 wk postpartum in women with GDM who developed type 2 diabetes, compared with nondiabetic women. H3K4 dimethylation was 75% lower at 8-10 wk postpartum in women with GDM who subsequently developed type 2 diabetes compared with women who had GDM who did not. CONCLUSIONS: The percentage of dimethylation of histones H3K27 and H3K4 varied with diabetic state and has the potential as a predictive tool to identify women who will convert from GDM to type 2 diabetes.


Assuntos
Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Gestacional/genética , Epigênese Genética , Histonas/genética , Adulto , Metilação de DNA , Diabetes Mellitus Tipo 2/genética , Progressão da Doença , Feminino , Marcadores Genéticos , Humanos , Pessoa de Meia-Idade , Gravidez
13.
Antonie Van Leeuwenhoek ; 109(3): 475-80, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26754102

RESUMO

Extracellular pili-like structures (PLS) produced by cyanobacteria have been poorly explored. We have done detailed topographical and electrical characterisation of PLS in Nostoc punctiforme PCC 73120 using transmission electron microscopy (TEM) and conductive atomic force microscopy (CAFM). TEM analysis showed that N. punctiforme produces two separate types of PLS differing in their length and diameter. The first type of PLS are 6-7.5 nm in diameter and 0.5-2 µm in length (short/thin PLS) while the second type of PLS are ~20-40 nm in diameter and more than 10 µm long (long/thick PLS). This is the first study to report long/thick PLS in N. punctiforme. Electrical characterisation of these two different PLS by CAFM showed that both are electrically conductive and can act as microbial nanowires. This is the first report to show two distinct PLS and also identifies microbial nanowires in N. punctiforme. This study paves the way for more detailed investigation of N. punctiforme nanowires and their potential role in cell physiology and symbiosis with plants.


Assuntos
Nanofios/ultraestrutura , Nostoc/ultraestrutura , Nostoc/fisiologia
14.
Genes Nutr ; 10(5): 487, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26319140

RESUMO

Two cases of zinc deficiency in breastfed neonates were investigated where zinc levels in the mothers' milk were reduced by more than 75 % compared to normal. The objective of this study was to find the molecular basis of the maternal zinc deficiency condition. Significant reductions in mRNA expression and protein levels of the zinc transporters SLC30A5 and SLC30A6 were found in maternal tissue, suggesting a causal link to the zinc-deficient milk. Novel splice variants of the SLC30A6 transcript were detected. No modifications were found in coding regions, or in transcription binding sites of promoter regions or in 5' and 3' untranslated regions of both transporters in lymphoblasts and fibroblasts isolated from both mothers. Altered DNA methylation in SLC30A5 at two CpG sites was detected and may account for the reduced levels of SLC30A5 mRNA and protein in lymphoblasts. Reduced SLC30A6 mRNA and protein levels in lymphoblasts may be secondary to reduced SLC30A5 expression, as they function as a heterodimer in zinc transport. In conclusion, two cases of zinc deficiency are linked to low levels of the SLC30A5 and SLC30A6 zinc transporters. These two zinc transporters have not been previously associated with zinc deficiency in milk.

15.
Antonie Van Leeuwenhoek ; 108(5): 1213-25, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26319534

RESUMO

Identification of extracellular conductive pilus-like structures (PLS) i.e. microbial nanowires has spurred great interest among scientists due to their potential applications in the fields of biogeochemistry, bioelectronics, bioremediation etc. Using conductive atomic force microscopy, we identified microbial nanowires in Microcystis aeruginosa PCC 7806 which is an aerobic, photosynthetic microorganism. We also confirmed the earlier finding that Synechocystis sp. PCC 6803 produces microbial nanowires. In contrast to the use of highly instrumented continuous flow reactors for Synechocystis reported earlier, we identified simple and optimum culture conditions which allow increased production of nanowires in both test cyanobacteria. Production of these nanowires in Synechocystis and Microcystis were found to be sensitive to the availability of carbon source and light intensity. These structures seem to be proteinaceous in nature and their diameter was found to be 4.5-7 and 8.5-11 nm in Synechocystis and M. aeruginosa, respectively. Characterization of Synechocystis nanowires by transmission electron microscopy and biochemical techniques confirmed that they are type IV pili (TFP) while nanowires in M. aeruginosa were found to be similar to an unnamed protein (GenBank : CAO90693.1). Modelling studies of the Synechocystis TFP subunit i.e. PilA1 indicated that strategically placed aromatic amino acids may be involved in electron transfer through these nanowires. This study identifies PLS from Microcystis which can act as nanowires and supports the earlier hypothesis that microbial nanowires are widespread in nature and play diverse roles.


Assuntos
Microcystis , Nanofios , Synechocystis , Biologia Computacional/métodos , Proteínas de Fímbrias/química , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/metabolismo , Fímbrias Bacterianas/fisiologia , Fímbrias Bacterianas/ultraestrutura , Microcystis/fisiologia , Microcystis/ultraestrutura , Microscopia de Força Atômica , Modelos Moleculares , Conformação Proteica , Synechocystis/fisiologia , Synechocystis/ultraestrutura
16.
Cell Physiol Biochem ; 33(6): 1758-77, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24923432

RESUMO

BACKGROUND/AIMS: Allergen absorption by epithelia may play an important role in downstream immune responses. Transport mechanisms that can bypass Peyer's patches include transcellular and paracellular transport. The capacity of an allergen to cross via these means can modulate downstream processing of the allergen by the immune system. The aim of this study was to investigate allergen-epithelial interactions of peanut allergens with the human intestinal epithelium. METHODS: We achieved this using the human Caco-2 cell culture model, exposed to crude peanut extract. Western and immunofluorescence analysis were used to identify the cellular and molecular changes of peanut extract on the intestinal epithelium. RESULTS: Following exposure of Caco-2 cells to peanut extract, binding of the peanut allergens Ara h 1 and Ara h 2 to the apical cellular membrane and transcytosis across the monolayers were observed. Additionally, the co-localisation of the transmembrane tight junction proteins occludin, JAM-A and claudin-1, with the intracellular adhesion protein ZO-1 was modified. CONCLUSION: Disruption of Caco-2 barrier integrity through tight junction disruption may enable movement of peanut proteins across the intestinal epithelium. This accounts for peanut's increased allergenicity, compared to other food allergens, and provides an explanation for the potency of peanut allergens in immune response elicitation.


Assuntos
Arachis/química , Mucosa Intestinal/efeitos dos fármacos , Extratos Vegetais/farmacologia , Junções Íntimas/efeitos dos fármacos , Albuminas 2S de Plantas/metabolismo , Alérgenos/metabolismo , Antígenos de Plantas/metabolismo , Transporte Biológico , Western Blotting , Células CACO-2 , Moléculas de Adesão Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Claudina-1/metabolismo , Glicoproteínas/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Proteínas de Membrana , Microscopia Confocal , Modelos Biológicos , Ocludina/metabolismo , Permeabilidade/efeitos dos fármacos , Extratos Vegetais/metabolismo , Proteínas de Plantas/metabolismo , Ligação Proteica , Receptores de Superfície Celular/metabolismo , Junções Íntimas/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
17.
J Nutr Biochem ; 25(4): 377-87, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24485600

RESUMO

Adequate amounts of copper in milk are critical for normal neonatal development, however the mechanisms regulating copper supply to milk have not been clearly defined. PMC42-LA cell cultures representative of resting, lactating and suckled mammary epithelia were used to investigate the regulation of the copper uptake protein, CTR1. Both the degree of mammary epithelial differentiation (functionality) and extracellular copper concentration greatly impacted upon CTR1 expression and its plasma membrane association. In all three models (resting, lactating and suckling) there was an inverse correlation between extracellular copper concentration and the level of CTR1. Cell surface biotinylation studies demonstrated that as extracellular copper concentration increased membrane associated CTR1 was reduced. There was a significant increase in CTR1 expression (total and membrane associated) in the suckled gland model in comparison to the resting gland model, across all copper concentrations investigated (0-50 µM). Regulation of CTR1 expression was entirely post-translational, as quantitative real-time PCR analyses showed no change to CTR1 mRNA between all models and culture conditions. X-ray fluorescence microscopy on the differentiated PMC42-LA models revealed that organoid structures distinctively accumulated copper. Furthermore, as PMC42-LA cell cultures became progressively more specialised, successively more copper accumulated in organoids (resting

Assuntos
Proteínas de Transporte de Cátions/metabolismo , Cobre/metabolismo , Células Epiteliais/metabolismo , Lactação/metabolismo , Glândulas Mamárias Humanas/metabolismo , Proteínas de Transporte de Cátions/genética , Diferenciação Celular , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Células Cultivadas , Cobre/farmacocinética , Transportador de Cobre 1 , Feminino , Humanos , Glândulas Mamárias Humanas/citologia , Prolactina/metabolismo
18.
FEMS Microbiol Ecol ; 86(2): 149-71, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23710564

RESUMO

Metal homoeostasis in cyanobacteria is based on uptake and export systems that are controlled by their own regulators. This study characterises the zinc uptake (Znu) system in Nostoc punctiforme. The system was found to comprise of three subunits in an ACB operon: a Zn(2+)-binding protein (ZnuA18), a transmembrane domain (ZnuB) and an ATPase (ZnuC). These proteins are encoded within the znu operon regulated by a zinc uptake transcription repressor (Zur). Interestingly, a second Zn(2+)-binding protein (ZnuA08) was also identified at a distal genomic location. Interactions between components of the ZnuACB system were investigated using knockouts of the individual genes. The znuA08(-), znuA18(-), znuB(-) and znuC(-) mutants displayed overall reduced znuACB transcript levels, suggesting that all system components are required for normal expression of znu genes. Zinc uptake assays in the Zn(2+)-binding protein mutant strains showed that the disruption of znuA18 had a greater negative effect on zinc uptake than disruption of znuA08. Complementation studies in Escherichia coli indicated that both znuA08 and znuA18 were able to restore zinc uptake in a znuA(-) mutant, with znuA18 permitting the highest zinc uptake rate. The N. punctiforme zur was also able to complement the E. coli zur(-) mutant.


Assuntos
Nostoc/genética , Nostoc/metabolismo , Zinco/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Adenosina Trifosfatases/genética , Proteínas de Transporte de Cátions/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Transporte de Íons , Mutação , Óperon
19.
Genes Nutr ; 8(5): 475-86, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23378263

RESUMO

Zinc is an essential trace element required for enzyme catalysis, gene regulation and signal transduction. Zinc absorption takes place in the small intestine; however, the mechanisms by which cells accumulate zinc are not entirely clear. Zip1 (SLC39A1) is a predicted transmembrane protein that is postulated, but not conclusively proven to mediate zinc influx in gut cells. The aim of this study was to investigate a role for hZip1 in mediating zinc uptake in human enterocytes. Both hZip1 mRNA and protein were detected in human intestinal tissue. In non-differentiated Caco-2 human gut cells, hZip1 was partially localised to the endoplasmic reticulum. In contrast, in differentiated Caco-2 cells cultured in extracellular matrix, the hZip1 protein was located in proximity to the apical microvilli. Lack of surface antibody binding and internalisation indicated that hZip1 was not present on the plasma membrane. Functional studies to establish a role for hZip1 in cellular zinc accumulation were carried out using (65)Zn. In Caco-2 cells harbouring an hZip1 overexpression construct, cellular zinc accumulation was enhanced relative to the control. Conversely, Caco-2 cells with an hZip1 siRNA construct showed reduced zinc accumulation. In summary, we show that the Caco-2 cell differentiation endorses targeting of hZip1 to a region near the apical domain. Given the absence of hZip1 at the apical plasma membrane, we propose that hZip1 may act as an intracellular sensor to regulate zinc homoeostasis in human gut cells.

20.
Environ Monit Assess ; 185(3): 2231-41, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22707148

RESUMO

Surface soil (0-20 cm) samples (n = 143) were collected from vegetable, maize, and paddy farmland used for commercial crops in Liaoning, China. Sixteen priority polycyclic aromatic hydrocarbons (PAHs) listed in US Environmental Protection Agency were analyzed by high-performance liquid chromatography using a fluorescence detector. The soil concentrations of the 16 PAH ranged from 50 to 3,309 ng/g with a mean of 388 ng/g. The highest concentration of total PAHs found in soil of the vegetable farmland was 448 ng/g in average, followed by maize and paddy with total PAHs of 391 and 331 ng/g, respectively. Generally, the low molecular weight PAHs were more predominant than the high molecular weight PAHs in most of the soils. The evaluation of soil PAH contamination based on the Canadian criterion indicated that only naphthalene, phenanthrene, and pyrene were over the target values in several sampling sites. Isomer pair ratios and principal component analysis indicated that biomass and coal combustion were the main sources of PAHs in this area. And the average value of total B[a]Peq concentration in vegetable soils was higher than paddy and maize soils. We suggest that biomass burning should be abolished and commercial farming should be carried out far from the highways to ensure the safety of food products derived from commercial farming.


Assuntos
Agricultura , Monitoramento Ambiental , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes do Solo/análise , Solo/química , China , Poluição Ambiental/estatística & dados numéricos , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Medição de Risco , Poluentes do Solo/toxicidade
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