An extracytoplasmic function sigma factor operon regulates Myxococcus xanthus developmental gene expression

Myxococcus xanthus is a Gram-negative soil bacterium that undergoes multicellular development when high-density cells are starved on a solid surface. Expression of the 4445 gene, predicted to encode a periplasmic protein, commences 1.5 h after the initiation of development and requires starvation and high density conditions. Addition of crude or boiled supernatant from starving high-density cells restored 4445 expression to starving low-density cells. Addition of L-threonine or L-isoleucine to starving low-density cells also restored 4445 expression, indicating that the high-density signaling activity present in the supernatant might be composed of extracellular amino acids or small peptides. To investigate the circuitry integrating these starvation and high-density signals, the cis- and trans-acting elements controlling 4445 expression were identified. The 4445 transcription start site was determined by primer extension analysis to be 58 by upstream of the predicted translation start site. The promoter region contained a consensus sequence characteristic of e&barbelow;xtrac&barbelow;ytoplasmic f&barbelow;unction (ECF) sigma factor-dependent promoters, suggesting that 4445 expression might be regulated by an ECF sigma factor-dependent pathway, which are known to respond to envelope stresses. The small size of the minimum regulatory region, identified by 5′-end deletion analysis as being only 66 by upstream of the transcription start site, suggests that RNA polymerase could be the sole direct regulator of 4445 expression. To identify trans-acting negative regulators of 4445 expression, a strain containing a 4445-lacZ was mutagenized using the Himar1-tet transposon. The four transposon insertions characterized mapped to an operon encoding a putative ECF sigma factor, ecfA; an anti-sigma factor, reaA; and a negative regulator, reaB. The reaA and the reaB mutants expressed 4445 during growth and development at levels almost 100-fold higher than wild type, indicating that these genes encode negative regulators. The ecfA mutant expressed 4445-lacZ at basal levels, indicating that ecfA is a positive regulator. High Mg2+ concentrations over-stimulated this ecfA pathway possibly due to the depletion of exopolysaccharides and assembled type IV pili. These data indicate that the ecfA operon encodes a new regulatory stress pathway that integrates and transduces starvation and cell density cues during early development and is also responsive to cell-surface alterations.^

ANÁLISE MICROSCÓPICA E DA RUGOSIDADE DE SUPERFÍCIE DE FIOS ORTODÔNTICOS ESTÉTICOS, ANTES E APÓS DEFLEXÃO

O objetivo deste estudo foi avaliar a topografia de superfície dos fios estéticos, antes e após teste de deflexão. A amostra foi composta por 70 corpos de prova de fios 0,014 redondos, sendo 10 de cada uma das marcas comerciais avaliadas: Orthocosmetic Elastinol (Masel), Flexy Super Elastic Esthetic (Orthometric), InVu (TP Orthodontics) e ProForm Nitanium (Ortho Organizers) fios de NiTi revestidos por Teflon®; Optis (TP Orthodontics) fio de resina reforçado por fibra de vidro ou FRP; Niticosmetic (Tecnident) fio de NiTi revestido por resina epoxídica; e Nitinol SE (3M Unitek) fio de NiTi superelástico, usado para controle. A topografia de superfície de cada fio foi avaliada por rugosímetro e por microscópio óptico, antes e após ser submetido a ensaio de deflexão, no lado em que a força foi aplicada e no lado oposto a este. Cada fio foi defletido em 3,1mm, a uma velocidade de 1mm/min, com célula de carga de 5N a 36⁰C + 1⁰C. A análise de variância a três critérios (p<0,05) mostrou diferença significante entre os fios e o teste de Tukey mostrou que o fio Optis (TP Orthodontics) apresentou aumento nos parâmetros de rugosidade Ra, Rt e Rz, após a deflexão. O fio Niticosmetic (Tecnident) apresentou aumento na rugosidade média (Ra). O fio InVu (TP Orthodontics) foi o único que mostrou aumento na rugosidade no lado em que a força foi aplicada. A análise visual por meio de microscopia óptica revelou alterações na superfície em todos os fios estéticos após o teste de deflexão, desde delaminações do revestimento, observadas nos fios Orthocosmetic Elastinol e InVu, riscos permanentes na superfície, como visto nos fios Flexy Super Elastic Esthetic, Niticosmetic e ProForm Nitanium, e até mesmo fratura incompleta, no fio Optis. Concluiu-se que o fio Niticosmetic apresentou topografia de superfície similar ao fio metálico, e os demais fios estéticos apresentaram maior rugosidade e alterações visuaisna superfície.

The beneficiation of steel making slags

The thesis examines the possibilities for the beneficiation of steel making slags by using mineral processing methods. Chemical and Mineralogical investigations have been carried out by SEM and EP}ffi to determine the most suitable separation methods in terms of crystal size, chemical composition and surface properties. Magnetic separation was applied in connection with size reductions for the extraction of the metallic iron prills and other iron containing phases and the results were related to the feed size and operating conditions. The behaviour of the slags in flotation tests was studied with respect to the recovery and grade. It was found that the free lime presence in the slags caused a high acid consumption of both weak and strong acids. It also reacted with acids and consequently produced a white precipitate, (CaS04 for H2 S04 ). The poor response of the phases to the flotation by different types of collector was found to be due to surface alteration caused by the free lime. The flocculation tests were carried out at the natural pH of the slags to prevent surface alterations. Settling tests were done to determine the suitable flocculants for the separation tests. The effect of the settling period, flocculant concentration, conditioning period and number of cleaning cycles were determined to optimize the separation tests. The discussion brings together this study with previous theoretically based work cited in the literature to elucidate the factor5governing the utilisation of steel making slags.

Pathogen-Specific Adaptations to Conserved Signaling Pathways in Cryptococcus neoformans

Cryptococcus neoformans is an opportunistic fungal pathogen that causes significant disease worldwide. Even though this fungus has not evolved specifically to cause human disease, it has a remarkable ability to adapt to many different environments within its infected host. C. neoformans adapts by utilizing conserved eukaryotic and fungal-specific signaling pathways to sense and respond to stresses within the host. Upon infection, two of the most significant environmental changes this organism experiences are elevated temperature and high pH.

Conserved Rho and Ras family GTPases are central regulators of thermotolerance in C. neoformans. Many GTPases require prenylation to associate with cellular membranes and function properly. Using molecular genetic techniques, microscopy, and infection models, I demonstrated that the prenyltransferase, geranylgeranyl transferase I (GGTase I) is required for thermotolerance and pathogenesis. Using fluorescence microscopy, I found that only a subset of conserved GGTase I substrates requires this enzyme for membrane localization. Therefore, the C. neoformans GGTase I may recognize its substrate in a slightly different manner than other eukaryotic organisms.

The alkaline response transcription factor, Rim101, is a central regulator of stress-response genes important for adapting to the host environment. In particular, Rim101 regulates cell surface alterations involved in immune avoidance. In other fungi, Rim101 is activated by alkaline pH through a conserved signaling pathway, but this pathway had yet been characterized in C. neoformans. Using molecular genetic techniques, I identified and analyzed the conserved members of the Rim pathway. I found that it was only partially conserved in C. neoformans, missing the components that sense pH and initiate pathway activation. Using a genetic screen, I identified a novel Rim pathway component named Rra1. Structural prediction and genetic epistasis experiments suggest that Rra1 may serve as the Rim pathway pH sensor in C. neoformans and other related basidiomycete fungi.

To explore the relevance of Rim pathway signaling in the interaction of C neoformans with its host, I characterized the Rim101-regulated cell wall changes that prevent immune detection. Using HPLC, enzymatic degradation, and cell wall stains, I found that the rim101Δ mutation resulted in increased cell wall chitin exposure. In vitro co-culture assays demonstrated that increased chitin exposure is associated with enhanced activation of macrophages and dendritic cells. To further test this association, I demonstrated that other mutant strains with increased chitin exposure induce macrophage and dendritic cell responses similar to rim101Δ. We used primary macrophages from mutant mouse lines to demonstrate that members of both the Toll-like receptor and C-type lectin receptor families are involved in detecting strains with increased chitin exposure. Finally, in vivo immunological experiments demonstrated that the rim101Δ strain induced a global inflammatory immune response in infected mouse lungs, expanding upon our previous in vivo rim101Δ studies. These results demonstrate that cell wall organization largely determines how fungal cells are detected by the immune system.

Minimal alterations on the enamel surface by micro-abrasion: in vitro roughness and wear assessments

Objective: To evaluate the in vitro changes on the enamel surface after a micro-abrasion treatment promoted by different products. Material and Methods: Fifty (50) fragments of bovine enamel (15 mm × 5 mm) were randomly assigned to five groups (n=10) according to the product utilized: G1 (control)= silicone polisher (TDV), G2= 37% phosphoric acid (3M/ESPE) + pumice stone (SS White), G3= Micropol (DMC Equipment), G4= Opalustre (Ultradent) and G5= Whiteness RM (FGM Dental Products). Roughness and wear were the responsible variables used to analyze these surfaces in four stages: baseline, 60 s and 120 s after the micro-abrasion and after polishing, using a Hommel Tester T1000 device. After the tests, a normal distribution of data was verified, with repeated ANOVA analyses (p?0.05) which were used to compare each product in different stages. One-way ANOVA and Tukey tests were applied for individual comparisons between the products in each stage (p?0.05). Results: Means and standard deviations of roughness and wear (µm) after all the promoted stages were: G1=7.26(1.81)/13.16(2.67), G2=2.02(0.62)/37.44(3.33), G3=1.81(0.91)/34.93(6.92), G4=1.92(0.29)/38.42(0.65) and G5=1.98(0.53)/33.45(2.66). At 60 seconds, all products tended to produce less surface roughness with a variable gradual decrease over time. After polishing, there were no statistically significant differences between the groups, except for G1. Independent of the product utilized, the enamel wear occurred after the micro-abrasion. Conclusions: In this in vitro study, enamel micro-abrasion presented itself as a conservative approach, regardless of the type of the paste compound utilized. These products promoted minor roughness alterations and minimal wear. The use of phosphoric acid and pumice stone showed similar results to commercial products for the micro-abrasion with regard to the surface roughness and wear.

Changes in neutrophil surface receptor expression, degranulation, and respiratory burst activity after moderate- and high-intensity exercise

Intense exercise stimulates the systemic release of a variety of factors that alter neutrophil surface receptor expression and functional activity. These alterations may influence resistance to infection after intense exercise. The aim of this study was to examine the influence of exercise intensity on neutrophil receptor expression, degranulation (measured by plasma and intracellular myeloperoxidase concentrations), and respiratory burst activity. Ten well-trained male runners ran on a treadmill for 60 min at 60% [moderate-intensity exercise (MI)] and 85% maximal oxygen consumption [high-intensity exercise (HI)]. Blood was drawn immediately before and after exercise and at 1 h postexercise. Immediately after HI, the expression of the neutrophil receptor CD16 was significantly below preexercise values (P < 0.01), whereas MI significantly reduced CD35 expression below preexercise values (P < 0.05). One hour after exercise at both intensities, there was a significant decline in CD11b expression (P < 0.05) and a further decrease in CD16 expression compared with preexercise values (P < 0.01). CD16 expression was lower 1 h after HI than 1 h after MI (P < 0.01). Immediately after HI, intracellular myeloperoxidase concentration was less than preexercise values (P < 0.01), whereas plasma myeloperoxidase concentration was greater (P < 0.01), indicating that HI stimulated neutrophil degranulation. Plasma myeloperoxidase concentration was higher immediately after HI than after MI (P < 0.01). Neutrophil respiratory burst activity increased after HI (P < 0.01). In summary, both MI and HI reduced neutrophil surface receptor expression. Although CD16 expression was reduced to a greater extent after HI, this reduction did not impair neutrophil degranulation and respiratory burst activity.

In vivo confocal microscopy of the ocular surface : from bench to bedside

In vivo confocal microscopy (IVCM) is an emerging technology that provides minimally invasive, high resolution, steady-state assessment of the ocular surface at the cellular level. Several challenges still remain but, at present, IVCM may be considered a promising technique for clinical diagnosis and management. This mini-review summarizes some key findings in IVCM of the ocular surface, focusing on recent and promising attempts to move “from bench to bedside”. IVCM allows prompt diagnosis, disease course follow-up, and management of potentially blinding atypical forms of infectious processes, such as acanthamoeba and fungal keratitis. This technology has improved our knowledge of corneal alterations and some of the processes that affect the visual outcome after lamellar keratoplasty and excimer keratorefractive surgery. In dry eye disease, IVCM has provided new information on the whole-ocular surface morphofunctional unit. It has also improved understanding of pathophysiologic mechanisms and helped in the assessment of prognosis and treatment. IVCM is particularly useful in the study of corneal nerves, enabling description of the morphology, density, and disease- or surgically induced alterations of nerves, particularly the subbasal nerve plexus. In glaucoma, IVCM constitutes an important aid to evaluate filtering blebs, to better understand the conjunctival wound healing process, and to assess corneal changes induced by topical antiglaucoma medications and their preservatives. IVCM has significantly enhanced our understanding of the ocular response to contact lens wear. It has provided new perspectives at a cellular level on a wide range of contact lens complications, revealing findings that were not previously possible to image in the living human eye. The final section of this mini-review provides a focus on advances in confocal microscopy imaging. These include 2D wide-field mapping, 3D reconstruction of the cornea and automated image analysis.

Automated 3D mapping & shape analysis of the lateral ventricles via fluid registration of multiple surface-based atlases

We developed and validated a new method to create automated 3D parametric surface models of the lateral ventricles, designed for monitoring degenerative disease effects in clinical neuroscience studies and drug trials. First we used a set of parameterized surfaces to represent the ventricles in a manually labeled set of 9 subjects' MRIs (atlases). We fluidly registered each of these atlases and mesh models to a set of MRIs from 12 Alzheimer's disease (AD) patients and 14 matched healthy elderly subjects, and we averaged the resulting meshes for each of these images. Validation experiments on expert segmentations showed that (1) the Hausdorff labeling error rapidly decreased, and (2) the power to detect disease-related alterations monotonically improved as the number of atlases, N, was increased from 1 to 9. We then combined the segmentations with a radial mapping approach to localize ventricular shape differences in patients. In surface-based statistical maps, we detected more widespread and intense anatomical deficits as we increased the number of atlases, and we formulated a statistical stopping criterion to determine the optimal value of N. Anterior horn anomalies in Alzheimer's patients were only detected with the multi-atlas segmentation, which clearly outperformed the standard single-atlas approach.

TUNPOP: A simulation of the dynamics and structure of the yellowfin tuna stock and surface fishery of the Eastern Pacific Ocean

ENGLISH: Three distinct versions of TUNP0P, an age-structured computer simulation model of the eastern Pacific yellowfin tuna, Thunnus albacores, stock and surface tuna fishery, are used to reveal mechanisms which appear to have a significant effect on the fishery dynamics. Real data on this fishery are used to make deductions on the distribution of the fish and to show how that distribution might influence events in the fishery. The most important result of the paper is that the concept of the eastern Pacific yellowfin tuna stock as a homogeneous unit is inadequate to represent the recent history of the fishery. Inferences are made on the size and distribution of the underlying stock as well as its potential yield to the surface fishery as a result of alterations in the level and distribution of the effort. SPANISH: Se han empleado tres versiones diferentes de TUNP0P, un modelo de simulación de la computadora (basado en la estructura de la edad) de la población y la pesca epipelágica del atún aleta amarilla, Tbunnus albacares, del Pacífico oriental, para revelar los mecanismos que parecen tener un efecto importante en la dinámica pesquera. Se emplean los datos verdaderos de esta pesca para hacer deducciones sobre la distribución de los peces y para mostrar cómo puede influir esta distribución en los eventos de pesca. La conclusión más importante de este estudio es que el concepto de que la población del aleta amarilla del Pacífico oriental es una unidad homogénea, es inadecuado para representar la historia reciente de pesca. Se teoriza sobre la talla y distribución de la población subyacente como también sobre su producción potencial en la pesca epipelágica al cambiar el nivel y distribución del esfuerzo.

Surface-enhanced Raman scattering studies on aggregated silver nanoplates in aqueous solution

Four different sizes of citrate-protected silver nanoplates with the corresponding in-plane dipole resonance band at 530, 619, 778, and 858 nm, respectively, are synthesized for surface-enhanced Raman scattering (SERS) study. Their aggregation behaviors are monitored by use of UV-vis spectroscopy. During the aggregation process, a marked red shift of the in-plane dipole resonance of silver nanoplates is observed, whereas other resonance modes of them only have small alterations in the site or intensity. Aggregated silver nanoplates can serve as active SERS substrates with an enhancement factor of about 4.5 x 10(5) using 2-aminothiophenol as a probing molecule. The SERS performance of silver nanoplates is even superior to the commonly used Lee-Meisel silver colloid, making them very attractive for SERS applications.

Effects Of Oil On Digestive Cells In Mussels - Quantitative Alterations In Cellular And Lysosomal Structure

Structural changes were observed in the digestive tubule epithelial cells of Mytilus edulis following long-term exposure to the water accommodated fraction (WAF) of North Sea crude oil (30 μg · l−1 total oil derived aromatic hydrocarbons). The changes observed involved a reduction in the height of the digestive cells beyond that demonstrated in a normal feeding cycle. In addition there was a loss of the normal synchrony of the digestive cells to a point where nearly all the tubules exhibited an appearance similar to that which is usually termed ‘reconstituting’. These alterations were quantified using an image analysis technique and the mean height of the digestive cells used as an index of digestive function or state. Long-term exposure also induced a radical alteration of the structure of secondary lysosomes within the digestive cells, resulting in the formation of large lysosomes, believed to be autolysosomes. Stereological analyses showed that these lysosomes are reduced in numbers and greatly increased in volume in comparison with controls. There is a concomitant increase in surface area of lysosomes per unit volume of digestive cell compared with control conditions. These alterations are indicative of fundamental changes in secondary lysosomal function involving an autophagic response to oil derived hydrocarbons. which would contribute to the reduction of digestive cell cytoplasm. These cellular alterations are discussed in terms of their use as indices of cell injury, in response to oil.

Poly(epsilon-caprolactone) and poly(epsilon-caprolactone)-polyvinylpyrrolidone-iodine blends as ureteral biomaterials: characterisation of mechanical and surface properties, degradation and resistance to encrustation in vitro.

This study describes the physicochemical properties and in vitro resistance to encrustation of solvent cast films composed of either poly(epsilon-caprolactone) (PCL), prepared using different ratios of high (50,000) to low (4000) (molecular weight) m.wt., or blends of PCL and the polymeric antimicrobial complex, poly(vinylpyrrolidone)-iodine (PVP-I). The incorporation of PVP-I offered antimicrobial activity to the biomaterials. Films were characterised in terms of mechanical (tensile analysis, dynamic mechanical thermal analysis) and surface properties (dynamic contact angle analysis, scanning electron microscopy), whereas degradation (at 37degreesC in PBS at pH 7.4) was determined gravimetrically. The resistance of the films to encrustation was evaluated using an in vitro encrustation model. Reductions in the ratio of high:low-m.wt. PCL significantly reduced the ultimate tensile strength, % elongation at break and the advancing contact angle of the films. These effects were attributed to alterations in the amorphous content and the more hydrophilic nature of the films. Conversely, there were no alterations in Young's modulus, the viscoelastic properties and glass-transition temperature. Incorporation of PVP-I did not affect the mechanical or rheological properties of the films, indicative of a limited interaction between the two polymers in the solid state. Manipulation of the high:low m.wt. ratio of PCL significantly altered the degradation of the films, most notably following longer immersion periods, and resistance to encrustation. Accordingly, maximum degradation and resistance to encrustation was observed with the biomaterial composed of 40:60 high:low m.wt. ratios of PCL; however, the mechanical properties of this system were considered inappropriate for clinical application. Films composed of either 50:50 or 60:40 ratio of high:low m.wt. PCL offered an appropriate compromise between physicochemical properties and resistance to encrustation. This study has highlighted the important usefulness of degradable polymer systems as ureteral biomaterials

Surface modification of poly(ε-caprolactone) using a dielectric barrier discharge in atmospheric pressure glow discharge mode

The role of roughening and functionalization processes involved in modifying the wettability of poly(e-caprolactone) (PCL) after treatment by an atmospheric pressure glow discharge plasma is discussed. The change in the ratio of Cdouble bond; length as m-dashO/C–O bonds is a significant factor influencing the wettability of PCL. As the contact angle decreases, the level of Cdouble bond; length as m-dashO bonds tends to rise. Surface roughness alterations are the driving force for lasting increases in wettability, while the surface functional species are shorter lived. We can approximate from ageing that the increase in wettability for PCL after plasma treatment is 55–60% due to roughening and 40–45% due to surface functionalization for the plasma device investigated.

FASCIOLA-HEPATICA - TEGUMENTAL SURFACE CHANGES IN ADULT AND JUVENILE FLUKES FOLLOWING TREATMENT IN-VITRO WITH THE SULFOXIDE METABOLITE OF TRICLABENDAZOLE (FASINEX)

The effects of the novel benzimidazole, triclabendazole (Fasinex, Ciba-Geigy), in its active sulphoxide metabolite form (TCBZ-SX), on the tegumental surface of Fasciola hepatica has been examined in vitro. The tegument of adult flukes incubated in TCBZ-SX (50 mug/ml) appeared swollen and blebbed after only 6 h. In addition, progressive spine loss at the oral cone was evident following 12 h treatment. After 24 h, the tegumental syncytium and spines had completely sloughed away, leaving an exposed basal lamina and empty spine sockets. Juvenile flukes (3 weeks old) also demonstrated tegumental alterations after treatment with TCBZ-SX (20 mug/ml). The syncytium became extremely roughened and corrugated on both dorsal and ventral surfaces after only 3 h. Following 6- and 9-h incubations, there were many deep furrows, which were especially pronounced on the ventral surface, and by 18 h, the juvenile tegument was severely disrupted, especially on the ventral surface. In all cases, the effects were more marked than in the previous incubation periods. The results confirm the potent activity of triclabendazole against F. hepatica and suggest that the tegument of adult and juvenile flukes may be a target organ for this important fasciolicide.

Sequence stratigraphy related distribution of diagenetic alterations In Miocene deltaic and shoreface arenites, the Suez Rift, EGYPT.

The distribution of eogenetic alterations in shoreface-offshore and coarse-grained deltaic, calcarenite to hybrid arenites of the Mheiherrat Formation (lower Rudeis), Early Miocene, the Gulf of Suez, Egypt) can be constrained within a sequence stratigraphic framework. The bioclast-rich, shoreface (trangressive systems tract; TST) and shoreface (highstand systems tract; HST) arenites, particularly those below the parasequence boundaries and maximum flooding surface, are cemented by grain-coating microcrystalline, circumgranular isopacheous acicular and columnar, and coarse-crystalline calcite (δ18OVPDB = -3.6 to -0.3 ‰; δ13CVPDB = -2.3 to -0.7 ‰), non-Ferro an dolomite (δ18OVPDB = -3.9 to +0.9‰; δ13CVPDB = -2.5 ‰ to -0.7 ‰), and pyrite. Zeolite, palygorskite and gypsum occur in the HST shoreface arenites, being enhanced by aird climatic condations. The coarse-grained deltaic LST deposits are pervasively cemented by coarse-crystalline, pore-filling calcite and small amounts of microcrystalline calcite (δ18OVPDB = -4.4 to -2.3 ‰; δ13CVPDB = -2.8 to -1.3 ‰) and non-ferroan dolomite (δ18OVPDB = -4.8 to -2.5 ‰; δ13CVPDB = -3.3 to -1.5 ‰). Thus, this study demonstrates that changes in pore-water chemistry, which induced changes in the texture, composition and extent of cementation in the Miocene arenites was controlled by changes in the relative sea level and by the paleo-climatic conditions during deposition of the HST arenites.

Sequence stratigraphy related distribution of diagenetic alterations In Miocene deltaic and shoreface arenites, the Suez Rift, EGYPT.. Available from: https://www.researchgate.net/publication/264545153_Sequence_stratigraphy_related_distribution_of_diagenetic_alterations_In_Miocene_deltaic_and_shoreface_arenites_the_Suez_Rift_EGYPT [accessed Apr 15, 2015].