Biosynthesis and assembly of alcohol oxidase, a peroxisomal matrix protein in methylotrophic yeasts: a review

Alcohol oxidase (AO) catalyses the first step of methanol metabolism in yeasts. In vivo the enzyme is compartmentalized in special cell compartments, called peroxisomes. The enzyme along with the organelles are induced during growth of methylotrophic yeasts on methanol as the sole carbon source. Like all other peroxisomal matrix proteins, AO is encoded by a nuclear gene. Expression of the protein is regulated by a repression/derepression mechanism, but also by induction. Inactive monomeric precursor protein is synthesized in the cytosol and subsequently imported post-translationally into peroxisomes without further processing. Assembly into the active homo-octameric enzyme and binding of the prosthetic group flavin adenine dinucleotide occurs inside the organelle. When enhanced concentration of octameric alcohol oxidase are present in the organelles, the enzyme may form a crystalloid. Oligomerization is not dependent on translocation of AO precursors into their target organelle since octameric, active AO is detected in the cytosol and nucleus of peroxisome-deficient mutants of Hansenula polymorpha: at high expression rates large cytosolic AO crystalloids are formed, which occasionally are also encountered inside the nucleus of such mutants. This paper summarizes recent findings and views on the mechanisms involved in synthesis, import, assembly and crystallization of this important peroxisomal enzyme.     

Dynamic behaviour of viologen-activated nanostructured TiO2: correlation between kinetics of charging and coloration

The dynamic response of viologen-activated nanostructured titanium dioxide has been studied by means of electrical and electro-optical measurements. We show that the state of charge of the semiconductor network is the key factor mediating between the electrode potential and colouration of viologen. Theoretically, we relate the electrode potential to the statistics of occupancy of both TiO₂ nanoparticles and oxidized viologen molecules attached to the surface, on the assumption of quasi-equilibrium of Fermi levels in these contacting phases. Experimentally, we determine the statistical function from steady-state measurements (electrochemical impedance spectroscopy) of the capacitance of the semiconductor film. From this understanding we explain the main features that correlate the simultaneous voltammetry and transmittance responses. Finally, the redox process of viologen is resolved separately from the TiO₂ response by means of transmittance data.     

A method for predicting the probability of business network profitability

In the design phase of business collaboration, it is desirable to be able to predict the profitability of the business-to-be. Therefore, techniques to assess qualities such as costs, revenues, risks, and profitability have been previously proposed. However, they do not allow the modeler to properly manage uncertainty with respect to the design of the considered business collaboration. In many real collaboration projects today, uncertainty regarding the business’ present or future characteristics is so significant that ignoring it becomes problematic. In this paper, we propose an approach based on the predictive, probabilistic architecture modeling framework (P²AMF), capable of advanced and probabilistically sound reasoning about profitability risks. The P²AMF-based approach for profitability risk prediction is also based on the e³-value modeling language and on the object constraint language. The paper introduces the prediction and modeling approach, and a supporting software tool. The use of the approach is illustrated by means of a case study originated from the Stockholm Royal Seaport smart city project.     

Measures of larval lake whitefish length and abundance as early predictors of year-class strength in Lake Michigan

Many lake whitefish stocks in Lake Michigan have experienced substantial declines in growth and condition since the 1990s. Reduced growth and condition could result in reduced quality or quantity of eggs produced by spawning females, which in turn could negatively impact recruitment. We evaluated the potential for reduced recruitment by measuring early life stage density and length, and we discuss the utility of these measures as early indicators of lake whitefish year-class strength. Overall, mean larval density (number per 1000 m³ ± SE) in Lake Michigan was greater in 2006 (373.7 ± 28.3) than in 2005 (16.6 ± 24.8); whereas, mean length (mm ± SE) of larval lake whitefish was smaller in 2006 (12.87 ± 0.07) than in 2005 (14.38 ± 0.13). The ratio of zooplankton to fish density did not show an expected relationship with larval fish density or length. Rather, variation in larval density was best explained by a multiple-regression model that included larval length, spring wind intensity, and adult stock density as predictor variables. Our results suggest that the density of larval lake whitefish is not directly regulated by temperature or zooplankton density at the time of emergence, but that a potential for density-dependent regulation exists when larval emergence rates are high. We conclude that the observed declines in growth and condition of adult lake whitefish are not resulting in substantial reductions in recruitment.     

Peroxisome biogenesis and degradation in yeast: a structure/function analysis

In yeast, peroxisomes are the site of specific catabolic pathways that characteristically include hydrogen peroxide producing oxidases and catalase. During the last 10 years, much progress has been made in unravelling the molecular mechanisms involved in the biogenesis of this organelle. At present, 23 different genes (PEX genes) have been identified that are involved in different aspects of peroxisome biogenesis (e.g., proliferation, formation of the peroxisomal membrane, import of matrix proteins). The principles of peroxisome degradation are still much less understood. Recently, the first yeast mutants affected in this process have become available and used to clone corresponding genes by functional complementation. In this paper, an overview is presented of the research on yeast peroxisomes, focusing on recent achievements in the molecular aspects of peroxisome development, function, and turnover.     

Hairline lowering during foreheadplasty

Many patients seeking rejuvenation of their foreheads have high hairlines and are troubled by the prospect that surgery will worsen the deformity. Hairline elevation occurs in both coronal and endoscopic foreheadplasty techniques and is at least part of the reason that many surgeons do not always recommend these procedures when otherwise indicated. Although a pretrichial "hairline" incision prevents hairline retro-displacement, it results in forehead shortening only and not true hairline lowering. When a pretrichial incision is used in combination with a posterior scalp advancement flap, however, true hairline lowering is possible. Experience with this technique encompasses 27 procedures performed over a 5-year period. Patients ranged in age from 35 to 71 years. A significant improvement was demonstrable in all cases and corresponded with a high degree of patient satisfaction. No serious complications were seen. A high hairline must be recognized as the source of both a disproportionate and aged appearance. The ability to lower the hairline and place it in a more proportionate, youthful relationship with the rest of the face adds a new dimension to foreheadplasty equal to or greater in importance to an overall improved appearance as other maneuvers typically performed during the procedure. For many patients, the benefits of hairline lowering far outweigh the trade-off of a more anteriorly situated, and possibly more visible, scar.     

Characterization of the Hansenula polymorpha CPY gene encoding carboxypeptidase Y

We have isolated the Hansenula polymorpha CPY gene encoding carboxypeptidase Y (Hp-CPY). The deduced amino acid sequence revealed that Hp-CPY consists of 541 amino acids and has a calculated Mr of 60,793. The protein is highly similar to Saccharomyces cerevisiae CPY (61.8% identity). At the N-terminus of Hp-CPY signals for the entry into the secretory pathway and subsequent sorting to the vacuole were identified. Immunocytochemically, using monospecific antibodies raised against Hp-CPY, the protein was localized to the vacuole. On Western blots, a diffuse protein band was observed in extracts of H. polymorpha cells, suggesting that the protein is glycosylated. This was confirmed by endoglycosidase H treatment, which resulted in a strong reduction of the apparent Mr of the protein. We have investigated the effect of CPY deletion on the degradation of peroxisomes, an autophagous process that occurs when the organelles become redundant for growth. In deltacpy cells peroxisomal proteins were degraded in the vacuole as efficiently as in wild-type H. polymorpha cells, indicating that CPY is not a major proteinase in this pathway.     

Model-driven approach to enterprise interoperability at the technical service level

Enterprise Interoperability is the ability of enterprises to interoperate in order to achieve their business goals. Although the purpose of enterprise interoperability is determined at the business level, the use of technical (IT) services to support business services implies that interoperability solutions at both the business and technical level should be aligned. This paper introduces and demonstrates the suitability of an approach based on model transformations to automate enterprise interoperability. We start by considering that a set of enterprises are willing to interoperate in the context of their individual goals. The interactions necessary for their cooperation are then properly captured in terms of a so-called choreography. Our approach allows a choreography to be mapped and transformed to an orchestration, which defines the operation of the actual technical services of the interoperating enterprises. The paper discusses the technical challenges of implementing the transformation, and illustrates our approach with two application scenarios.     

Cuticular Hydrocarbon Phenotypes Do not Indicate Cryptic Species in Fungus-Growing Termites (Isoptera: Macrotermitinae)

In several termite species, distinct differences in the composition of cuticular hydrocarbons among colonies correspond to high genetic divergence of mitochondrial DNA sequences. These observations suggest that hydrocarbon phenotypes represent cryptic species. Different cuticular hydrocarbon phenotypes also are found among colonies of fungus-growing termites of the genus Macrotermes. To determine if these hydrocarbon differences in Macrotermes also indicate cryptic species, we sequenced the mitochondrial CO I gene from species in West and East Africa. Among individuals of a supposed species but belonging to different cuticular hydrocarbon phenotypes, the genetic distances are much smaller than distances between species. Unlike what has been observed in other termites, Macrotermes hydrocarbon phenotypes do not represent cryptic species. Our findings suggest fundamental differences in the evolution and/or function of cuticular hydrocarbons among different termite lineages. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.