Модификация мембран на основе полисульфона с использованием триблоксополимера полиэтиленгликоля и полипропиленгликоля

The phase state, viscosity and optical properties of polysulfone (PSF) solutions in N,N-dimethylacetamide (DMAc) with the addition of block copolymer Pluronic F127 and polyethylene glycol (PEG-4000, Mn = 4000 g-mol-1) were studied. Additives are both pore-forming and hydrophilizing agents. It was found that 18-22 % PSF solutions in DMAc with the Pluronic F127 content t ≥5 wt. % feature a lower critical solution temperature (LCST). Mixed matrix PSF/Pluronic F127 and PSF/PEG membranes were obtained by the phase inversion technique and the comparative studies of their structure and performance were carried out. It was found that the average surface roughness parameters of PSF/Pluronic F127 membranes significantly exceed those of PSF/PEG membranes. The presence of the both additives in the casting solution leads to the effective hydrophilization of the membrane selective layer. It was shown that the increase in the membrane flux and the decrease in the polyvinylpyrrolidone (K-30, Mn = 40000 g-mol-1) rejection coefficient are a result of adding both Pluronic F127 and PEG-4000 to the casting solution. It was shown that PSF/Pluronic F127 membranes are characterized by higher shrinkage resistance and a lower total flux decrease during ultrafiltration of bovine serum albumin solutions. It was found that the antifouling performance of PSF/Pluronic F127 membranes significantly exceeds that of PSF/PEG membranes.Изучено фазовое состояние, вязкостные и оптические свойства растворов полисульфона (ПСФ) в N,N-диметилацетамиде (ДМАА) с добавками блоксополимера Pluronic F127 и полиэтиленгликоля (ПЭГ-4000, Мn = 4000 г-моль-1) в качестве порообразователей и гидрофилизующих агентов. Установлено, что 18-22 %-ные растворы ПСФ в ДМАА с добавкойt ≥5 % Pluronic F127 характеризуются наличием нижней критической температуры смешения (НКТС). Методом инверсии фаз получена серия мембран со смешанной матрицей ПСФ/Pluronic F127 и ПСФ/ПЭГ и проведены сравнительные исследования их структуры и транспортных характеристик. Установлено, что значения параметров средней шероховатости поверхности для мембран ПСФ/Pluronic F127 существенно превышают таковые для мембран ПСФ/ПЭГ. Введение обеих добавок в формовочную композицию приводит к эффективной гидрофилизации селективного слоя мембран. Установлено, что введение в растворы как PluronicF127, так и ПЭГ-4000 приводит к увеличению удельной производительности мембран и снижению коэффициента задерживания по поливинилпирролидону (ПВП К-30, Mn = 40000 г-моль-1). Показано, что мембраны со смешанной матрицей ПСФ/ Pluronic F127 характеризуются более высокой устойчивостью к уплотнению, меньшей степенью уменьшения общего потока при фильтрации растворов бычьего сывороточного альбумина и существенно превосходят по устойчивости к загрязнению мембраны со смешанной матрицей ПСФ/ПЭГ

Hygienic quality of dehydrated aromatic herbs marketed in Southern Portugal

Dehydrated aromatic herbs are highly valued ingredients, widely used at home level and by food processing industry, frequently added to a great number of recipes in the Mediterranean countries. Despite being considered low-moisture products and classified as GRAS, during pre and post-harvesting stages of production they are susceptible of microbial contamination. In Europe an increasing number of food recalls and disease outbreaks associated with dehydrated herbs have been reported in recent years. In this study the microbial quality of 99 samples of aromatic herbs (bay leaves, basil, coriander, oregano, parsley, Provence herbs, rosemary and thyme) collected from retails shops in the region of Algarve (Southern Portugal) was assessed. All the samples were tested by conventional methods and were assayed for the total count of aerobic mesophilic microorganisms, Salmonella spp., Escherichia coli, coagulase-positive staphylococci and filamentous fungi. Almost 50 % of the herbs did not exceed the aerobic mesophilic level of 104 CFU/g. The fungi count regarded as unacceptable (106 CFU/g) was not found in any of the tested herbs, while 84 % of the samples ranged from ≤102 to 104 CFU/g. No sample was positive for the presence of Salmonella spp., Escherichia coli and staphylococci. The results are in compliance with the European Commission criteria although they point out to the permanent need of surveillance on the good standards of handling/cooking practices as well as the importance of avoiding contamination at production, retailing and distribution. The microbiological hazards associated with the pathogenic and toxigenic microbiota of dried herbs remain as a relevant public health issue, due to the fact that they are added to foods not submitted to any following lethal procedure. Control measures should be adopted in order to ensure that all phases of their supply chain respect the food safety standards.FCT: UID/BIA/04325/2019.info:eu-repo/semantics/publishedVersio

The type VII secretion system of <i>Staphylococcus aureus</i> secretes a nuclease toxin that targets competitor bacteria

The type VII protein secretion system (T7SS) plays a critical role in the virulence of human pathogens including Mycobacterium tuberculosis and Staphylococcus aureus. Here we report that the S. aureus T7SS secretes a large nuclease toxin, EsaD. The toxic activity of EsaD is neutralised during its biosynthesis through complex formation with an antitoxin, EsaG, which binds to its C-terminal nuclease domain. The secretion of EsaD is dependent upon a further accessory protein, EsaE, that does not interact with the nuclease domain, but instead binds to the EsaD N-terminal region. EsaE has a dual cytoplasmic/membrane localization and membrane-bound EsaE interacts with the T7SS secretion ATPase, EssC, implicating EsaE in targeting the EsaDG complex to the secretion apparatus. EsaD and EsaE are co-secreted whereas EsaG is found only in the cytoplasm and may be stripped off during the secretion process. Strain variants of S. aureus that lack esaD encode at least two copies of EsaG-like proteins most likely to protect themselves from the toxic activity of EsaD secreted by esaD(+) strains. In support of this, a strain overproducing EsaD elicits significant growth inhibition against a sensitive strain. We conclude that T7SSs may play unexpected and key roles in bacterial competitiveness

Decolonisation of MRSA, S. aureus and E. coli by Cold-Atmospheric Plasma Using a Porcine Skin Model In Vitro

In the last twenty years new antibacterial agents approved by the U.S. FDA decreased whereas in parallel the resistance situation of multi-resistant bacteria increased. Thus, community and nosocomial acquired infections of resistant bacteria led to a decrease in the efficacy of standard therapy, prolonging treatment time and increasing healthcare costs. Therefore, the aim of this work was to demonstrate the applicability of cold atmospheric plasma for decolonisation of Gram-positive (Methicillin-resistant Staphylococcus aureus (MRSA), Methicillin-sensitive Staphylococcus aureus) and Gram-negative bacteria (E. coli) using an ex vivo pig skin model. Freshly excised skin samples were taken from six month old female pigs (breed: Pietrain). After application of pure bacteria on the surface of the explants these were treated with cold atmospheric plasma for up to 15 min. Two different plasma devices were evaluated. A decolonisation efficacy of 3 log10 steps was achieved already after 6 min of plasma treatment. Longer plasma treatment times achieved a killing rate of 5 log10 steps independently from the applied bacteria strains. Histological evaluations of untreated and treated skin areas upon cold atmospheric plasma treatment within 24 h showed no morphological changes as well as no significant degree of necrosis or apoptosis determined by the TUNEL-assay indicating that the porcine skin is still vital. This study demonstrates for the first time that cold atmospheric plasma is able to very efficiently kill bacteria applied to an intact skin surface using an ex vivo porcine skin model. The results emphasize the potential of cold atmospheric plasma as a new possible treatment option for decolonisation of human skin from bacteria in patients in the future without harming the surrounding tissue

Влияние давления при формировании селективного слоя на структуру и свойства динамических композиционных мембран для первапорации

Composite membranes for pervaporation were prepared by forming a selective layer based on cross-linked polyvinyl alcohol (PVA) on the porous membrane-substrate surface in the dynamic mode (via PVA solution ultrafiltration). It was found that the pressure growth results in increasing the thickness of the composite membrane selective layer. Composite membrane contact angle, flux, water content in permeate in ethanol/water (mass ratio 90/10) pervaporation were revealed to have maximum values at 3–4 atm depending on the PVA concentration in the feed solution. It was shown that the revealed dependence of the contact angle, selectivity, and permeability on the pressure of the selective layer formation is due to the compaction of the polymer matrix-substrate under the action of the transmembrane pressure and its relaxation after pressure release. When using elevated pressures (more than 3–4 atm), the relaxation of the polymer matrix causes the microdefect to form as a result of deformation of the selective layer.Композиционные мембраны для первапорации были получены методом формирования селективного слоя на основе сшитого поливинилового спирта (ПВС) на поверхности пористой мембраны-подложки в динамическом режиме (ультрафильтрации растворов ПВС). Установлено, что повышение давления приводит к увеличению толщины селективного слоя композиционных мембран; при этом их контактный угол смачивания, удельная производительность, содержание воды в пермеате в процессе первапорации смеси этанол/вода 90/10 изменяются экстремально с максимумом при 3–4 атм в зависимости от концентрации раствора ПВС. Экстремальная зависимость краевого угла смачивания, коэффициента разделения и проницаемости композиционных мембран объясняется процессами уплотнения полимерной матрицы-подложки под действием трансмембранного давления и ее релаксации после снятия давления. При использовании повышенных давлений (более 3–4 атм) релаксация полимерной матрицы приводит к появлению микродефектов в результате деформации сформированного селективного слоя

Systematic Genetic Nomenclature for Type VII Secretion Systems

CITATION: Bitter, W., et al. 2009. Systematic genetic nomenclature for type VII secretion systems. PLoS Pathogens, 5(10): 1-6, doi: 10.1371/journal.ppat.1000507.The original publication is available at http://journals.plos.org/plospathogensMycobacteria, such as the etiological agent of human tuberculosis, Mycobacterium tuberculosis, are protected by an impermeable cell envelope composed of an inner cytoplasmic membrane, a peptidoglycan layer, an arabinogalactan layer, and an outer membrane. This second membrane consists of covalently linked, tightly packed long-chain mycolic acids [1,2] and noncovalently bound shorter lipids involved in pathogenicity [3–5]. To ensure protein transport across this complex cell envelope, mycobacteria use various secretion pathways, such as the SecA1-mediated general secretory pathway [6,7], an alternative SecA2-operated pathway [8], a twin-arginine translocation system [9,10], and a specialized secretion pathway variously named ESAT-6-, SNM-, ESX-, or type VII secretion [11–16]. The latter pathway, hereafter referred to as type VII secretion (T7S), has recently become a large and competitive research topic that is closely linked to studies of host–pathogen interactions of M. tuberculosis [17] and other pathogenic mycobacteria [16]. Molecular details are just beginning to be revealed [18–22] showing that T7S systems are complex machineries with multiple components and multiple substrates. Despite their biological importance, there has been a lack of a clear naming policy for the components and substrates of these systems. As there are multiple paralogous T7S systems within the Mycobacteria and orthologous systems in related bacteria, we are concerned that, without a unified nomenclature system, a multitude of redundant and obscure gene names will be used that will inevitably lead to confusion and hinder future progress. In this opinion piece we will therefore propose and introduce a systematic nomenclature with guidelines for name selection of new components that will greatly facilitate communication and understanding in this rapidly developing field of research.http://journals.plos.org/plospathogens/article?id=10.1371%2Fjournal.ppat.1000507Publisher's versio

The Staphylococcus aureus Response to Unsaturated Long Chain Free Fatty Acids: Survival Mechanisms and Virulence Implications

Staphylococcus aureus is an important human commensal and opportunistic pathogen responsible for a wide range of infections. Long chain unsaturated free fatty acids represent a barrier to colonisation and infection by S. aureus and act as an antimicrobial component of the innate immune system where they are found on epithelial surfaces and in abscesses. Despite many contradictory reports, the precise anti-staphylococcal mode of action of free fatty acids remains undetermined. In this study, transcriptional (microarrays and qRT-PCR) and translational (proteomics) analyses were applied to ascertain the response of S. aureus to a range of free fatty acids. An increase in expression of the σB and CtsR stress response regulons was observed. This included increased expression of genes associated with staphyloxanthin synthesis, which has been linked to membrane stabilisation. Similarly, up-regulation of genes involved in capsule formation was recorded as were significant changes in the expression of genes associated with peptidoglycan synthesis and regulation. Overall, alterations were recorded predominantly in pathways involved in cellular energetics. In addition, sensitivity to linoleic acid of a range of defined (sigB, arcA, sasF, sarA, agr, crtM) and transposon-derived mutants (vraE, SAR2632) was determined. Taken together, these data indicate a common mode of action for long chain unsaturated fatty acids that involves disruption of the cell membrane, leading to interference with energy production within the bacterial cell. Contrary to data reported for other strains, the clinically important EMRSA-16 strain MRSA252 used in this study showed an increase in expression of the important virulence regulator RNAIII following all of the treatment conditions tested. An adaptive response by S. aureus of reducing cell surface hydrophobicity was also observed. Two fatty acid sensitive mutants created during this study were also shown to diplay altered pathogenesis as assessed by a murine arthritis model. Differences in the prevalence and clinical importance of S. aureus strains might partly be explained by their responses to antimicrobial fatty acids

Genome Sequence of a Lancefield Group C Streptococcus zooepidemicus Strain Causing Epidemic Nephritis: New Information about an Old Disease

Outbreaks of disease attributable to human error or natural causes can provide unique opportunities to gain new information about host-pathogen interactions and new leads for pathogenesis research. Poststreptococcal glomerulonephritis (PSGN), a sequela of infection with pathogenic streptococci, is a common cause of preventable kidney disease worldwide. Although PSGN usually occurs after infection with group A streptococci, organisms of Lancefield group C and G also can be responsible. Despite decades of study, the molecular pathogenesis of PSGN is poorly understood. As a first step toward gaining new information about PSGN pathogenesis, we sequenced the genome of Streptococcus equi subsp. zooepidemicus strain MGCS10565, a group C organism that caused a very large and unusually severe epidemic of nephritis in Brazil. The genome is a circular chromosome of 2,024,171 bp. The genome shares extensive gene content, including many virulence factors, with genetically related group A streptococci, but unexpectedly lacks prophages. The genome contains many apparently foreign genes interspersed around the chromosome, consistent with the presence of a full array of genes required for natural competence. An inordinately large family of genes encodes secreted extracellular collagen-like proteins with multiple integrin-binding motifs. The absence of a gene related to speB rules out the long-held belief that streptococcal pyrogenic exotoxin B or antibodies reacting with it singularly cause PSGN. Many proteins previously implicated in GAS PSGN, such as streptokinase, are either highly divergent in strain MGCS10565 or are not more closely related between these species than to orthologs present in other streptococci that do not commonly cause PSGN. Our analysis provides a comparative genomics framework for renewed appraisal of molecular events underlying APSGN pathogenesis

A single natural nucleotide mutation alters bacterial pathogen host tropism

The capacity of microbial pathogens to alter their host tropism leading to epidemics in distinct host species populations is a global public and veterinary health concern. To investigate the molecular basis of a bacterial host-switching event in a tractable host species, we traced the evolutionary trajectory of the common rabbit clone of Staphylococcus aureus. We report that it evolved through a likely human-to-rabbit host jump over 40 years ago and that only a single naturally occurring nucleotide mutation was required and sufficient to convert a human-specific S. aureus strain into one that could infect rabbits. Related mutations were identified at the same locus in other rabbit strains of distinct clonal origin, consistent with convergent evolution. This first report of a single mutation that was sufficient to alter the host tropism of a microorganism during its evolution highlights the capacity of some pathogens to readily expand into new host species populations

Development of Antifouling Polysulfone Membranes by Synergistic Modification with Two Different Additives in Casting Solution and Coagulation Bath: Synperonic F108 and Polyacrylic Acid

This study deals with the development of antifouling ultrafiltration membranes based on polysulfone (PSF) for wastewater treatment and the concentration and purification of hemicellulose and lignin in the pulp and paper industry. The efficient simple and reproducible technique of PSF membrane modification to increase antifouling performance by simultaneous addition of triblock copolymer polyethylene glycol-polypropylene glycol-polyethylene glycol (Synperonic F108, Mn =14 × 103 g mol−1) to the casting solution and addition of polyacrylic acid (PAA, Mn = 250 × 103 g mol−1) to the coagulation bath is proposed for the first time. The effect of the PAA concentration in the aqueous solution on the PSF/Synperonic F108 membrane structure, surface characteristics, performance, and antifouling stability was investigated. PAA concentrations were varied from 0.35 to 2.0 wt.%. Membrane composition, structure, and topology were investigated by Fourier-transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and scanning electron microscopy (SEM). The addition of PAA into the coagulation bath was revealed to cause the formation of a thicker and denser selective layer with decreasing its pore size and porosity; according to the structural characterization, an interpolymer complex of the two additives was formed on the surface of the PSF membrane. Hydrophilicity of the membrane selective layer surface was shown to increase significantly. The selective layer surface charge was found to become more negative in comparison to the reference membrane. It was shown that PSF/Synperonic F108/PAA membranes are characterized by better antifouling performance in ultrafiltration of humic acid solution and thermomechanical pulp mill (ThMP) process water. Membrane modification with PAA results in higher ThMP process water flux, fouling recovery ratio, and hemicellulose and total lignin rejection compared to the reference PSF/Synperonic F108 membrane. This suggests the possibility of applying the developed membranes for hemicellulose concentration and purification