Ultrasound imaging of the distal radioulnar joint: a new method to assess ulnar radial translation in forearm rotation.

A cross-sectional reliability study was conducted with 23 normal participants to establish normal values, and the repeatability and validity of distal radioulnar joint translation measurements using ultrasound imaging. Static transverse images of maximal supination, neutral and maximal pronation were examined to assess translation, using a method consistent with the rheumatoid arthritis subluxation ratio. Translation while gripping a 1 kg weight in supinated and pronated positions was then compared with non-gripping translation. There was significantly more ulnar radial translation found with pronation than supination, when compared with neutral. Gripping in pronation did not produce statistically significant changes in translation, whereas the changes produced by gripping in supination were significant. Internal consistency was deemed very high and the rheumatoid arthritis subluxation ratio values measured using ultrasound imaging were consistent with previously documented values measured by computerized tomography. This study demonstrated that translational movement of the distal radioulnar joint can be reliably detected in healthy participants using ultrasound imaging. This may reduce dependency on other imaging modalities to diagnose distal radioulnar joint instability. LEVEL OF EVIDENCE: 2.

Intrinsically disordered protein.

Proteins can exist in a trinity of structures: the ordered state, the molten globule, and the random coil. The five following examples suggest that native protein structure can correspond to any of the three states (not just the ordered state) and that protein function can arise from any of the three states and their transitions. (1) In a process that likely mimics infection, fd phage converts from the ordered into the disordered molten globular state. (2) Nucleosome hyperacetylation is crucial to DNA replication and transcription; this chemical modification greatly increases the net negative charge of the nucleosome core particle. We propose that the increased charge imbalance promotes its conversion to a much less rigid form. (3) Clusterin contains an ordered domain and also a native molten globular region. The molten globular domain likely functions as a proteinaceous detergent for cell remodeling and removal of apoptotic debris. (4) In a critical signaling event, a helix in calcineurin becomes bound and surrounded by calmodulin, thereby turning on calcineurin's serine/threonine phosphatase activity. Locating the calcineurin helix within a region of disorder is essential for enabling calmodulin to surround its target upon binding. (5) Calsequestrin regulates calcium levels in the sarcoplasmic reticulum by binding approximately 50 ions/molecule. Disordered polyanion tails at the carboxy terminus bind many of these calcium ions, perhaps without adopting a unique structure. In addition to these examples, we will discuss 16 more proteins with native disorder. These disordered regions include molecular recognition domains, protein folding inhibitors, flexible linkers, entropic springs, entropic clocks, and entropic bristles. Motivated by such examples of intrinsic disorder, we are studying the relationships between amino acid sequence and order/disorder, and from this information we are predicting intrinsic order/disorder from amino acid sequence. The sequence-structure relationships indicate that disorder is an encoded property, and the predictions strongly suggest that proteins in nature are much richer in intrinsic disorder than are those in the Protein Data Bank. Recent predictions on 29 genomes indicate that proteins from eucaryotes apparently have more intrinsic disorder than those from either bacteria or archaea, with typically > 30% of eucaryotic proteins having disordered regions of length > or = 50 consecutive residues.

Sexual orientation and eating psychopathology: the role of masculinity and femininity.

OBJECTIVE: Previous research suggests that eating disorders are related to homosexuality in men, although links with female sexual orientation are less clear. Appearance factors have generally been implicated in this relationship. However, previous studies have failed to consider the role of femininity, even though evidence suggests that this is a more critical factor than sexual preference. The aim of this study was to consider the relationship between gender-role orientation and eating psychopathology in nonclinical men and women of different sexual orientations. METHOD: One hundred university students (40 homosexual; 60 heterosexual) completed the Bem Sex Role Inventory and the Eating Attitudes Test. RESULTS: For the group as a whole, there were links between femininity and high levels of eating psychopathology, whereas masculinity was associated with relatively healthy eating-related attitudes and behaviors. When considering the role of sexual orientation, these links were specific to homosexual men and women. CONCLUSIONS: In relation to homosexual men and women, the results support a model where femininity might be seen as a specific risk factor for eating disorders, whereas masculinity is likely to be a protective factor. Methodological and conceptual implications are discussed.

Gordonia amicalis sp. nov., a novel dibenzothiophene-desulphurizing actinomycete.

The taxonomic position of a dibenzothiophene-desulphurizing soil actinomycete was established using a polyphasic taxonomic approach. The organism, strain IEGMT, was shown to have chemical and morphological properties typical of members of the genus Gordonia. The tested strain formed a distinct phyletic line within the evolutionary radiation occupied by the genus Gordonia, with Gordonia alkanivorans DSM 44369T, Gordonia desulfuricans NCIMB 40816T and Gordonia rubropertincta DSM 43197T as the most closely related organisms. Strain IEGMT has a range of phenotypic properties that distinguish it from representatives of all of the validly described species of Gordonia. It was also sharply distinguished from the type strains of Gordonia desulfuricans and Gordonia rubropertincta on the basis of DNA-DNA relatedness data. The combined genotypic and phenotypic data show that strain IEGMT merits recognition as a new species of Gordonia. The name proposed for the new species is Gordonia amicalis; the type strain is IEGMT (= DSM 44461T = KCTC 9899T).

Gordonia desulfuricans sp. nov., a benzothiophene-desulphurizing actinomycete.

The taxonomic position of two actinomycetes isolated from soil was established using a polyphasic approach. The organisms, designated 213ET and 213F, were found to have chemical and morphological properties consistent with their assignment to the genus Gordonia. Nearly complete sequences of the 16S rDNA genes of the two strains were determined following the isolation and direct sequencing of the amplified genes. The tested strains were found to have identical 16S rDNA sequences and formed a phylogenetic line within the evolutionary radiation occupied by the genus Gordonia that was most closely related to Gordonia rubropertincta DSM 43197T. However, DNA-DNA relatedness data showed that strain 213ET and Gordonia rubropertincta DSM 43197T belonged to distinct genomic species. Strains 213ET and 213F also shared an identical phenotypic profile which distinguished them from representatives of validly described Gordonia species. The combined genotypic and phenotypic data show that strains 213ET and 213F merit recognition as a new species of Gordonia. The name proposed for the new species is Gordonia desulfuricans, for which the type strain is 213ET (= NCIMB 40816T).

Desulphurisation of benzothiophene and dibenzothiophene by actinomycete organisms belonging to the genus Rhodococcus, and related taxa.

Desulphurising enzymes remove the sulphur moiety from an organosulphur molecule leaving the carbon skeleton intact. Two kinds of desulphurisation reaction are recognised. The dibenzothiophene (DBT)-specific pathway desulphurises DBT to inorganic sulphite and 2-hydroxybiphenyl (HBP), and the benzothiophene (BTH)-specific pathway desulphurises BTH to 2-(2'-hydroxyphenyl)ethan 1-al (HPEal) and probably inorganic sulphite. The DBT-desulphurisation pathway was originally identified in Rhodococcus erythropolis strain IGTS8 (ATCC 53968), and the BTH-desulphurisation pathway in Gordonia sp. strain 213E (NCIMB 40816). These organisms do not further metabolise the organic product of desulphurisation. In this article current knowledge of the biochemistry and genetics of the desulphurisation enzymes is reviewed. The need for separate, DBT- and BTH-specific desulphurisation routes is rationalised in terms of the chemical differences between the two compounds. The desulphurisation pathway is compared with other microbial DBT-degrading enzyme systems. Finally some comments are made concerning the application of desulphurisation enzymes for fuel desulphurisation and on the relevance of these enzymes to the ecology of the mycolata (sensu Chun et al, 1996).

Characterization of the desulfurization genes from Rhodococcus sp. strain IGTS8.

Rhodococcus sp. strain IGTS8 possesses an enzymatic pathway that can remove covalently bound sulfur from dibenzothiophene (DBT) without breaking carbon-carbon bonds. The DNA sequence of a 4.0-kb BstBI-BsiWI fragment that carries the genes for this pathway was determined. Frameshift and deletion mutations established that three open reading frames were required for DBT desulfurization, and the genes were designated soxABC (for sulfur oxidation). Each sox gene was subcloned independently and expressed in Escherichia coli MZ1 under control of the inducible lambda pL promoter with a lambda cII ribosomal binding site. SoxC is an approximately 45-kDa protein that oxidizes DBT to DBT-5,5'-dioxide. SoxA is an approximately 50-kDa protein responsible for metabolizing DBT-5,5'-dioxide to an unidentified intermediate. SoxB is an approximately 40-kDa protein that, together with the SoxA protein, completes the desulfurization of DBT-5,5'-dioxide to 2-hydroxybiphenyl. Protein sequence comparisons revealed that the predicted SoxC protein is similar to members of the acyl coenzyme A dehydrogenase family but that the SoxA and SoxB proteins have no significant identities to other known proteins. The sox genes are plasmidborne and appear to be expressed as an operon in Rhodococcus sp. strain IGTS8 and in E. coli.

Evaluation of steady-state kinetic parameters for enzymes solubilized in water-in-oil microemulsion systems.

1. Equations are derived for the steady-state kinetics of substrate conversion by enzymes confined within the water-droplets of water-in-oil microemulsion systems. 2. Water-soluble substrates initially confined within droplets that do not contain enzyme are assumed to be converted into product only after they enter enzyme-containing droplets via the inter-droplet exchange process. 3. Hyperbolic (Michaelis-Menten) kinetics are predicted when the substrate concentration is varied in microemulsions of fixed composition. Both kcat. and Km are predicted to be dependent on the size and concentration of the water-droplets in the microemulsion. 4. The predicted behaviour is shown to be supported by published experimental data. A physical interpretation of the form of the rate equation is presented. 5. The rate equation for an oil-soluble substrate was derived assuming a pseudo-two-phase (oil & water) model for the microemulsion. Both kcat. and Km are shown to be independent of phi aq. Km is larger than the aqueous solution value by a factor approximately equal to the oil/water partition coefficient of the substrate. The validity of the rate equation is confirmed by published data.

Analysis of the inactivation of liver alcohol dehydrogenase during storage in Aerosol-OT/isooctane microemulsions.

Changes in the enzymatic properties of horse liver alcohol dehydrogenase (HLADH; EC 1.1.1.1) were studied as a function of incubation time in Aerosol-OT/isooctane microemulsions. The enzyme was characterized by fluorimetric binding studies of the inhibitor isobutyramide to the binary complex, HLADH-NADH and by determination of Km,app and Vmax,app values for cyclohexanone. The Km,app values for cyclohexanone and the Kd,app for isobutyramide stay constant throughout a 48-h incubation, whereas the Vmax,app and the total number of inhibitor binding sites decrease. Thus the inactivation process previously described corresponds to progressive loss of functional sites, while the properties of the remaining functional sites are unchanged. If no co-enzyme is added to the system, the enzyme loses catalytic activity within less than an hour, but if co-enzyme is added, a fraction of the HLADH enzyme population retains enzyme activity over a long period of time. Hence the presence of bound co-enzyme significantly inhibits the process(es) leading to inactivation of the enzyme in the microemulsions.

Kinetics of bilirubin oxidation catalysed by bilirubin oxidase in a water-in-oil microemulsion system.

1. Bilirubin oxidase can catalyse the oxidation of its primary substrate, bilirubin, in a water-in-oil microemulsion, which consists of discrete nanometer-diameter water droplets dispersed in a continuous water-immiscible oil medium. The droplets are stabilized by a monolayer of the surfactant, cetyltrimethylammonium bromide present at the oil/water interface. 2. Spectroscopic evidence is presented to show that bilirubin solubilized in this system is located mainly in the surfactant layer, in a form accessible to the enzyme molecule. 3. Studies are presented on the enzyme-catalysed rate of bilirubin oxidation in this system, as a function of temperature, pH, water content, and substrate and enzyme concentrations. 4. The main conclusions are that the enzyme can efficiently oxidise bilirubin in microemulsions of low water content. The reaction obeys Michaelis-Menten kinetics. The optimal pH for the catalysis is 8.0. The efficiency of catalysis decreases sharply as the water content increases.