RESUMO
Segregation of bacteria based on their metabolic activities in biofilms plays an important role in the development of antibiotic resistance. Mushroom-shaped biofilm structures, which are reported for many bacteria, exhibit topographically varying levels of multiple drug resistance from the cap of the mushroom to its stalk. Understanding the dynamics behind the formation of such structures can aid in design of drug delivery systems, antibiotics, or physical systems for removal of biofilms. We explored the development of metabolically heterogeneous Pseudomonas aeruginosa biofilms using numerical models and laboratory knockout experiments on wild-type and chemotaxis-deficient mutants. We show that chemotactic processes dominate the transformation of slender and hemispherical structures into mushroom structures with a signature cap. Cellular Potts model simulation and experimental data provide evidence that accelerated movement of bacteria along the periphery of the biofilm, due to nutrient cues, results in the formation of mushroom structures and bacterial segregation. Multidrug resistance of bacteria is one of the most threatening dangers to public health. Understanding the mechanisms of the development of mushroom-shaped biofilms helps to identify the multidrug-resistant regions. We decoded the dynamics of the structural evolution of bacterial biofilms and the physics behind the formation of biofilm structures as well as the biological triggers that produce them. Combining in vitro gene knockout experiments with in silico models showed that chemotactic motility is one of the main driving forces for the formation of stalks and caps. Our results provide physicists and biologists with a new perspective on biofilm removal and eradication strategies.
Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Pseudomonas aeruginosa/citologiaRESUMO
Biofilms are surface-attached communities of microorganisms embedded in an extracellular matrix and are essential for the cycling of organic matter in natural and engineered environments. They are also the leading cause of many infections, for example, those associated with chronic wounds and implanted medical devices. The extracellular matrix is a key biofilm component that determines its architecture and defines its physical properties. Herein, we used growth chambers embedded with micropillars to study the net mechanical forces (differential pressure) exerted during biofilm formation in situ. Pressure from the biofilm is transferred to the micropillars via the extracellular matrix, and reduction of major matrix components decreases the magnitude of micropillar deflections. The spatial arrangement of micropillar deflections caused by pressure differences in the different biofilm strains may potentially be used as mechanical signatures for biofilm characterization. Hence, we submit that micropillar-embedded growth chambers provide insights into the mechanical properties and dynamics of the biofilm and its matrix.
Assuntos
Bactérias/crescimento & desenvolvimento , Biofilmes/crescimento & desenvolvimento , PressãoRESUMO
Understanding the formation of electrodynamically interacting assemblies of metal nanoparticles requires accurate computational methods for determining the forces and propagating trajectories. However, since computation of electromagnetic forces occurs on attosecond to femtosecond timescales, simulating the motion of colloidal nanoparticles on milliseconds to seconds timescales is a challenging multi-scale computational problem. Here, we present a computational technique for performing accurate simulations of laser-illuminated metal nanoparticles. In the simulation, we self-consistently combine the finite-difference time-domain method for electrodynamics (ED) with Langevin dynamics (LD) for the particle motions. We demonstrate the ED-LD method by calculating the 3D trajectories of a single 100-nm-diameter Ag nanoparticle and optical trapping and optical binding of two and three 150-nm-diameter Ag nanoparticles in simulated optical tweezers. We show that surface charge on the colloidal metal nanoparticles plays an important role in their optically driven self-organization. In fact, these simulations provide a more complete understanding of the assembly of different structures of two and three Ag nanoparticles that have been observed experimentally, demonstrating that the ED-LD method will be a very useful tool for understanding the self-organization of optical matter.
RESUMO
Vitamin D can be synthesized following exposure to ultraviolet radiation (UVR), ingested in the diet or provided through oral supplementation. The medical literature frequently states that humans obtain most of their vitamin D from sunshine and that UVR exposure is essential to maintain vitamin D levels. A systematic review was conducted to determine the requirement for UVR in maintaining adequate (> 50 nmol L(-1) ) serum 25-hydroxyvitamin D [25(OH)D] levels. Studies reporting serum 25(OH)D during situations of negligible UVR exposure were sought. Forty-one studies (from a search yielding 42 698 articles) with a total of 4211 healthy adults met the inclusion criteria, providing 56 datasets from different population groups. Over 50% of subjects had > 50 nmol L(-1) 25(OH)D in 10 of 19 datasets reporting winter levels in areas with limited UVR. In addition, > 50% of subjects had adequate 25(OH)D levels in four of 12 datasets from polar regions during periods of negligible UVR, one of nine datasets documenting clothing-related minimal UVR and two of eight datasets detailing employment-related minimal UVR. The data demonstrate that many adults maintain adequate serum vitamin D levels despite negligible UVR exposure for several months. However, we acknowledge that preceding UVR exposure leading to vitamin D storage and delayed release may account for this maintenance of adequate serum vitamin D levels. There remains a need for further research on whether UVR exposure is required for longer-term maintenance of adequate vitamin D levels.
Assuntos
Raios Ultravioleta , Deficiência de Vitamina D/prevenção & controle , Vitamina D/análogos & derivados , Vestuário/efeitos adversos , Emprego , Feminino , Humanos , Masculino , Exposição à Radiação , Estações do Ano , Vitamina D/metabolismo , Deficiência de Vitamina D/sangueRESUMO
We developed two dose-response algorithms for P. aeruginosa pool folliculitis using bacterial and lesion density estimates, associated with undetectable, significant, and almost certain folliculitis. Literature data were fitted to Furumoto & Mickey's equations, developed for plant epidermis-invading pathogens: N l = A ln(1 + BC) (log-linear model); P inf = 1-e(-r c C) (exponential model), where A and B are 2.51644 × 107 lesions/m2 and 2.28011 × 10-11 c.f.u./ml P. aeruginosa, respectively; C = pathogen density (c.f.u./ml), N l = folliculitis lesions/m2, P inf = probability of infection, and r C = 4·3 × 10-7 c.f.u./ml P. aeruginosa. Outbreak data indicates these algorithms apply to exposure durations of 41 ± 25 min. Typical water quality benchmarks (≈10-2 c.f.u./ml) appear conservative but still useful as the literature indicated repeated detection likely implies unstable control barriers and bacterial bloom potential. In future, culture-based outbreak testing should be supplemented with quantitative polymerase chain reaction and organic carbon assays, and quantification of folliculitis aetiology to better understand P. aeruginosa risks.
Assuntos
Algoritmos , Foliculite/microbiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/fisiologia , Microbiologia da Água , Humanos , Piscinas , Fatores de TempoRESUMO
AIMS: Membrane distillation bioreactors (MDBR) have potential for industrial applications where wastewater is hot or waste heat is available, but the role of micro-organisms in MDBRs has never been determined, and thus was the purpose of this study. METHODS AND RESULTS: Microbial communities were characterized by bacterial and archaeal 16S and eukaryotic 18S rRNA gene tag-encoded pyrosequencing of DNA obtained from sludge. Taxonomy-independent analysis revealed that bacterial communities had a relatively low richness and diversity, and community composition strongly correlated with conductivity, total nitrogen and bound extracellular polymeric substances (EPS). Taxonomy-dependent analysis revealed that Rubrobacter and Caldalkalibacillus were abundant members of the bacterial community, but no archaea were detected. Eukaryotic communities had a relatively high richness and diversity, and both changes in community composition and abundance of the dominant genus, Candida, correlated with bound EPS. CONCLUSIONS: Thermophilic MDBR communities were comprised of a low diversity bacterial community and a highly diverse eukaryotic community with no archea detected. Communities exhibited low resilience to changes in operational parameters. Specifically, retenatate nutrient composition and concentration was strongly correlated with the dominant species. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides an understanding of microbial community diversity in an MDBR, which is fundamental to the optimization of reactor performance.
Assuntos
Reatores Biológicos/microbiologia , Esgotos/microbiologia , Archaea/isolamento & purificação , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Biodiversidade , Destilação , Eucariotos/classificação , Eucariotos/isolamento & purificação , Filtração , Eliminação de Resíduos LíquidosRESUMO
Pseudomonas aeruginosa is the opportunistic pathogen mostly implicated in folliculitis and acute otitis externa in pools and hot tubs. Nevertheless, infection risks remain poorly quantified. This paper reviews disease aetiologies and bacterial skin colonization science to advance dose-response theory development. Three model forms are identified for predicting disease likelihood from pathogen density. Two are based on Furumoto & Mickey's exponential 'single-hit' model and predict infection likelihood and severity (lesions/m2), respectively. 'Third-generation', mechanistic, dose-response algorithm development is additionally scoped. The proposed formulation integrates dispersion, epidermal interaction, and follicle invasion. The review also details uncertainties needing consideration which pertain to water quality, outbreaks, exposure time, infection sites, biofilms, cerumen, environmental factors (e.g. skin saturation, hydrodynamics), and whether P. aeruginosa is endogenous or exogenous. The review's findings are used to propose a conceptual infection model and identify research priorities including pool dose-response modelling, epidermis ecology and infection likelihood-based hygiene management.
Assuntos
Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/patogenicidade , Dermatopatias Bacterianas/microbiologia , Microbiologia da Água , Surtos de Doenças , Temperatura Alta , Humanos , Higiene , PiscinasRESUMO
Omega 3 polyunsaturated fatty acids (PUFAs) influence metabolism and thermogenesis in non-hibernators. How omega 3 PUFAs influence Arctic Ground Squirrels (AGS) during hibernation is unknown. Prior to hibernation we fed AGS chow composed of an omega 6:3 ratio approximately 1:1 (high in omega 3 PUFA, termed Balanced Diet), or an omega 6:3 ratio of 5:1 (Standard Rodent Chow), and measured the influence of diet on core body temperature (Tb), brown adipose tissue (BAT) mass, fatty acid profiles of BAT, white adipose tissue (WAT) and plasma as well as hypothalamic endocannabinoid and endocannabinoid-like bioactive fatty acid amides during hibernation. Results show feeding a diet high in omega 3 PUFAs, with a more balanced omega 6:3 ratio, increases AGS Tb in torpor. We found the diet-induced increase in Tb during torpor is most easily explained by an increase in the mass of BAT deposits of Balanced Diet AGS. The increase in BAT mass is associated with elevated levels of metabolites DHA and EPA in tissue and plasma suggesting that these omega 3 PUFAs may play a role in thermogenesis during torpor. While we did not observe diet-induced change in endocannabinoids, we do report altered hypothalamic levels of some endocannabinoids, and endocannabinoid-like compounds, during hibernation.
Assuntos
Tecido Adiposo Marrom/metabolismo , Ácidos Graxos Ômega-3/farmacologia , Sciuridae/metabolismo , Termogênese/efeitos dos fármacos , Torpor/efeitos dos fármacos , Animais , Ácidos Graxos Ômega-3/metabolismoRESUMO
A 46-year-old woman with Graves' disease developed infiltrative dermopathy of the thenar eminences. We believe this to be the first reported case of infiltrative dermopathy affecting the thenar eminences, and question whether repetitive occupational injury may have been a contributing factor. There is little published evidence to guide the treatment of infiltrative dermopathy.
Assuntos
Dermatoses do Pé/etiologia , Doença de Graves/complicações , Mixedema/etiologia , Tireotoxicose/tratamento farmacológico , Transtornos Traumáticos Cumulativos/complicações , Progressão da Doença , Feminino , Dermatoses do Pé/patologia , Doença de Graves/tratamento farmacológico , Humanos , Pessoa de Meia-Idade , Mixedema/patologia , Tireotoxicose/patologia , Resultado do TratamentoRESUMO
The structure factor of amorphous solid D(2)O deposited from the vapor at 10 degrees K has been obtained by measuring the neutron diffraction spectrum in the wave vector transfer from 0.8 to 12.3 reciprocal angstroms. The results indicate that the phase investigated is amorphous and has a liquiid-like structure factor. The Fourier-transformed structure e factor yields a real space pair distribution function consistent with local tetrahedral coordination and hydrogen bonding, as in other condensed phases of water. The intramolecular OD separation is 1.00 angstrom; the lack of data for very large wave vector transfer and the expected near equality of the intramolecular DD separation and intermolecular O . . . D separation make it impossible to determine the intramolecular DOD angle with precision. The neutron scattering data presented are complementary to the x-ray diffraction studies of Venkatesh, Rice, antd Narten.
RESUMO
Water vapor that condenses on a metal surface at 10 degrees K forms a noncrystalline phase of estimated density 1.2 grams per cubic centimeter. X-ray diffraction data of high precision and resolution have been analyzed to yield oxygen atom pair correlation functions. The positional correlation in amorphous solid water extends over only a few molecular radii, and the radial distribution of nearneighbor oxygen atoms in amorphous solid water is qualitatively different from that found in the low-pressure ice modifications. Amorphous solid water is a useful material for liquid water models because it can be studied under conditions such that the effects of static disorder and thermal excitation can be separated.
RESUMO
The expression of human immunodeficiency virus (HIV) after T cell activation is regulated by NF-kappa B, an inducible DNA-binding protein that stimulates transcription. Proteins encoded by a variety of DNA viruses are also able to activate expression from the HIV enhancer. To determine how this activation occurs, specific genes from herpes simplex virus type 1 and adenovirus that activate HIV in T lymphoma cells have been identified. The cis-acting regulatory sequences in the HIV enhancer that mediate their effect have also been characterized. The relevant genes are those for ICP0-an immediate-early product of herpes simplex virus type 1-and the form of E1A encoded by the 13S messenger RNA of adenovirus. Activation of HIV by adenovirus E1A was found to depend on the TATA box, whereas herpesvirus ICP0 did not work through a single defined cis-acting element. These findings suggest multiple pathways that can be used to bypass normal cellular activation of HIV, and they raise the possibility that infection by herpes simplex virus or adenovirus may directly contribute to the activation of HIV in acquired immunodeficiency syndrome by mechanisms independent of antigenic stimulation in T cells.
Assuntos
Elementos Facilitadores Genéticos , Genes Virais , HIV/genética , Ativação Linfocitária , Linfócitos T/imunologia , Adenovírus Humanos/genética , Genes Reguladores , HIV/crescimento & desenvolvimento , Humanos , Plasmídeos , Simplexvirus/genética , Transcrição Gênica , Ativação ViralRESUMO
The liquid-vapor interface of a bismuth-gallium mixture (0.2 percent bismuth and 99.8 percent gallium) at 36 degrees C has been studied by grazing incidence x-ray diffraction. The data show, in agreement with thermodynamic arguments, that bismuth is heavily concentrated in the liquid-vapor interface. The x-ray diffraction data are interpreted with the assistance of a simple model that represents the interface as a partial monolayer of bismuth. This analysis leads to the conclusion that the bismuth concentration in the interface is about 80 percent, that there is no significant mixing of gallium and bismuth in the interface, and that the structure function of the interfacial bismuth is like that of supercooled bulk liquid bismuth.
RESUMO
Biofilms are organised aggregates of bacteria that adhere to each other or surfaces. The matrix of extracellular polymeric substances that holds the cells together provides the mechanical stability of the biofilm. In this study, we have applied Brillouin microscopy, a technique that is capable of measuring mechanical properties of specimens on a micrometre scale based on the shift in frequency of light incident upon a sample due to thermal fluctuations, to investigate the micromechanical properties of an active, live Pseudomonas aeruginosa biofilm. Using this non-contact and label-free technique, we have extracted information about the internal stiffness of biofilms under continuous flow. No correlation with colony size was found when comparing the averages of Brillouin shifts of two-dimensional cross-sections of randomly selected colonies. However, when focusing on single colonies, we observed two distinct spatial patterns: in smaller colonies, stiffness increased towards their interior, indicating a more compact structure of the centre of the colony, whereas, larger (over 45 µm) colonies were found to have less stiff interiors.
RESUMO
In Pseudomonas aeruginosa, chemical deconvolution of the pyocyanin voltammetric signal allows its expression to be observed simultaneously with the quorum sensing molecule Pseudomonas quinolone signal (PQS). Such analysis has revealed that PQS might protect pyocyanin from self-oxidation, but also exert a pro-oxidative effect on pyocyanin under oxidative conditions to produce additional redox metabolites.
Assuntos
Pseudomonas aeruginosa/metabolismo , Piocianina/química , Piocianina/metabolismo , Quinolonas/química , Quinolonas/metabolismo , Antioxidantes/química , Antioxidantes/metabolismo , Compostos de Bifenilo/química , Técnicas Eletroquímicas , Oxirredução , Peroxidases/química , Picratos/químicaRESUMO
Vibrio vulnificus is an opportunistic pathogen that exhibits numerous virulence factors, including the secretion of a zinc metalloprotease and the production of a capsule. We have cloned and sequenced a gene from V. vulnificus that is a homologue of the positive transcriptional regulator, luxR, of the lux operon in Vibrio harveyi. This gene encodes a putative, single complete open reading frame designated smcR, which shares greater than 75% nucleotide identity with luxR of V. harveyi. The deduced amino acid sequence of the putative SmcR protein is more than 90% identical and 95% similar to that of LuxR of V. harveyi, suggesting that V. vulnificus possesses a member of the family of signal-response genes recently described in Vibrio cholerae and in Vibrio parahaemolyticus. Our data also demonstrate that, in addition to V. vulnificus, all six Vibrio spp. tested contained genes that hybridized with the luxR probe. We also present evidence that this regulatory protein was inherited from a common ancestor, and that the gene is ancient and widespread in marine Vibrio spp.
Assuntos
Proteínas de Bactérias/genética , Proteínas Repressoras/genética , Transativadores/genética , Vibrio/genética , Sequência de Aminoácidos , Sequência de Bases , DNA Bacteriano/química , DNA Bacteriano/genética , Humanos , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Água do Mar/microbiologia , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
Invertebrate interspecific developmental patterns can be highly variable and, taxonomically, are considered only weakly constrained. Intraspecifically, some invertebrate species possess multiple developmental modes-a condition known as poecilogony. Closer examination of most putative poecilogenous species, however, has not supported poecilogony, but rather has uncovered hidden or cryptic species. The polychaete Streblospio benedicti is a well-known, poecilogenous species found along the coast of North America. We collected mitochondrial cytochrome subunit I DNA sequence data from 88 individuals taken from 11 locations along the Atlantic, Gulf, and Pacific Coasts of the United States to provide a phylogenetic framework from which to interpret intraspecific variation in larval life history and brooding structure morphology in this species. Our results are consistent with a recent revision of the species into two separate species: S. benedicti, a pouched brooding form distributed along the Atlantic and Pacific Coasts, and S. gynobranchiata, a branchiate brooding form in the Gulf of Mexico. Contrary to the redescription, S. benedicti is paraphyletic because the pouched brooding population in Vero Beach, Florida shows strong genetic affinity with Gulf of Mexico populations (S. gynobranchiata). However, S. benedicti is a true poecilogenous species, with both lecithotrophic and planktotrophic individuals possessing identical mitochondrial DNA haplotypes. Crossbreeding experiments further support the molecular phylogeny with reproductive isolation demonstrated between, but not within, the major phylogenetic clades consistent with the previously described species. The genetic break near Vero Beach, Florida, corresponds to a well-known phylogeographic boundary, but the estimated time of separation for the Streblospio spp., approximately 10 million years before present, predates all other known phylogeographic subdivisions in this area. This suggests that biogeographic sundering in this region is a recurrent event. Divergence times within the major Streblospio spp. clades are recent and indicate that changes in larval life history as well as brooding structure morphology are highly plastic and can evolve rapidly.
Assuntos
DNA Mitocondrial/genética , Evolução Molecular , Filogenia , Poliquetos/classificação , Poliquetos/genética , Animais , Complexo IV da Cadeia de Transporte de Elétrons/genética , Meio Ambiente , Geografia , Haplótipos , América do Norte , Poliquetos/fisiologia , ReproduçãoRESUMO
The intraperitoneal administration of thiourea (TU) to mature male rats results in a significant increase in lung vascular permeability to Evans Blue dye (EBD). On the other hand, young, sexually immature rats are resistant to this effect. The increase in lung vascular permeability in response to TU in mature rats is associated with corresponding increases in lung and plasma histamine levels. The correlation of increases in lung and plasma histamine in response to TU is similar to that reported for ammonium salts which produce similar pulmonary edema.