Diagnostic performance of knee physical exam and participant-reported symptoms for MRI-detected effusion-synovitis among participants with early or late stage knee osteoarthritis: data from the Osteoarthritis Initiative.

OBJECTIVE: Evaluate the diagnostic performance of knee physical exam findings and participant-reported symptoms for MRI-detected effusion-synovitis (ES) among knees with early and late-stage osteoarthritis (OA). DESIGN: The Osteoarthritis Initiative (OAI) is a longitudinal study of participants with or at risk for knee OA. Two samples with MRI readings were available: 344 knees with early OA (312 participants) and 216 with late-stage OA (186 participants). Trained examiners performed bulge sign (BS) and patellar tap (PT) exams, and participants reported on knee swelling and pain with leg straightening. Effusion-synovitis on 3T non-contrast MRI was scored using the MRI Osteoarthritis Knee Score (MOAKS). Diagnostic performance of physical exam findings and symptoms was estimated with bootstrapped confidence intervals. RESULTS: For the early OA sample, the highest sensitivity for medium/large effusion-synovitis was achieved with a positive finding for any of the physical exam maneuvers and/or participant-reported symptoms (81.0 [95% CI: 70.0, 91.3]). Both knee symptoms in combination had a prevalence of 11.7% and yielded the highest estimated positive predictive value (PPV) (50.0 [95% CI: 34.2, 66.7]) and likelihood ratio positive (LR+) (5.2 [95% CI: 2.9, 9.7]). In late-stage OA knees, exam findings and symptoms provided minimal information beyond the prevalence. CONCLUSION: Patient report of both symptoms, or at least one positive exam finding and at least one symptom, could be used to identify knees at increased risk of effusion-synovitis in knees with early stage OA, either for screening purposes in clinical evaluation, or for study sample enrichment with an inflammatory phenotype; diagnostic performance was not sufficiently high for clinical diagnostic purposes.

Fine mapping a major obesity locus (jObes1) using a Berlin Fat Mouse × B6N advanced intercross population.

BACKGROUND/OBJECTIVES: The Berlin Fat Mouse Inbred line 860 is a model for juvenile obesity. Previously, a recessive major effect locus (jObes1) on chromosome 3 between 34 and 44 Mb has been found to be responsible for 39% of the variance of total fat mass at 10 weeks in a (BFMI860 x C57BL/6NCrl) F2 population. The aim of this study was fine mapping of the jObes1 locus. SUBJECTS/METHODS: An advanced intercross line (AIL) was generated from the initial F2 mapping population. Three hundred and forty-four male mice of generation 28 were excessively phenotyped and genotyped using the MegaMuga mouse chip containing 22 164 informative single-nucleotide polymorphisms. Expression of candidate genes was investigated in gonadal adipose tissue, liver and whole brain from mice of different genotype classes. Classical genetic complementation tests were performed to test candidate genes. RESULTS: The high mapping resolution of the AIL reduced the confidence interval for jObes1 from 10 to 0.37 Mb between 36.48 and 36.85 Mb. This region was highly significantly (logarithm (base 10) of odds (LOD) score after Benjamini and Hochberg correction (LOD(BH))>50) associated with total fat mass starting at puberty (6 weeks). Male homozygous carriers of the jObese1 BFMI allele had 3 g more fat than the other genotypes. Surprisingly, this genotype class showed lower body mass until weaning at 3 weeks (LOD(BH)=3.2). The mapped interval contains four genes. Bbs7, the most likely candidate gene that also caused obesity in the complementation test was differentially expressed in all tissues examined, whereas the neighboring cyclin A2 (Ccna2) gene showed differential expression in gonadal adipose tissue. CONCLUSIONS: Using an AIL, the confidence interval for jObes1 could be 27-fold reduced by finding chromosomal recombinations. Although Bbs7 is the most likely obesity gene in the jObes1 region, neighboring genes cannot be entirely excluded. Further examinations are needed to enlighten the mechanism leading to physiological consequences on body mass and fat mass in juvenile animals.

High Variability of Insulin Sensitivity in Closely Related Obese Mouse Inbred Strains.

Obesity is one of several risk factors for insulin resistance and type 2 diabetes. Here we examined males of 6 obese mouse inbred lines derived from the Berlin Fat Mouse (BFM) outbred population with respect to insulin sensitivity and factors of the metabolic syndrome with focus on the skeletal muscle as a major target of insulin dependent glucose uptake.Males were kept on a rodent standard diet and several approaches were carried out to address insulin sensitivity, adiposity and lipids in the serum. Transcript and protein levels of several genes in the insulin signalling pathway were measured. 2 of the lines, BFMI860-12 and in particular BFMI861-S1, showed a markedly reduced insulin sensitivity already at the age of 20 weeks. BFMI861-S1 mice also displayed elevated liver triglyceride levels as a sign of lipid overload and ectopic fat storage. The analysis of the insulin signalling pathway in skeletal muscle provided evidence for low insulin receptor (INSR) and normal glucose 4 transporter (GLUT4) protein amounts in BFMI861-S1 mice, while BFMI860-12 mice showed increased INSR and very low GLUT4 protein amounts. Interestingly, the sublines BFMI860-S2 and BFMI861-S2, which are highly related to the former 2 lines, respectively, were inconspicuously insulin sensitive. The expected few genetic differences among the BFMI lines facilitate the identification of causal genetic variation. This study identified 2 mouse lines with different impairments of insulin signalling. These lines resemble useful models for studying mechanisms leading to the pathophysiology of the metabolic syndrome, in particular insulin resistance.

Quantification of lumen stenoses with known dimensions by conventional angiography and computed tomography: implications of using conventional angiography as gold standard.

BACKGROUND: Quantitative coronary angiography (QCA) has inherent limitations for displaying complex vascular anatomy, yet it remains the gold standard for stenosis quantification. OBJECTIVE: To investigate the accuracy of stenosis assessment by multi-detector computed tomography (MDCT) and QCA compared to known dimensions. METHODS: Nineteen acrylic coronary vessel phantoms with precisely drilled stenoses of mild (25%), moderate (50%) and severe (75%) grade were studied with 64-slice MDCT and digital flat panel angiography. Fifty-seven stenoses of circular and non-circular shape were imaged with simulated cardiac motion (60 bpm). Image acquisition was optimised for both imaging modalities, and stenoses were quantified by blinded expert readers using electronic callipers (for MDCT) or lumen contour detection software (for QCA). RESULTS: Average difference between true and measured per cent diameter stenosis for QCA was similar compared to MDCT: 7 (+/-6)% vs 7 (+/-5)% (p=0.78). While QCA performed better than MDCT in stenoses with circular lumen (mean error 4 (+/-3)% vs 7 (+/-6)%, p<0.01), MDCT was superior to QCA for evaluating stenoses with non-circular geometry (mean error 10 (+/-7)% vs 7 (+/-5)%, p<0.05). In such lesions, QCA underestimated the true diameter stenosis by >20% in 9 of 27 (33%) vs 1 of 29 (3%) in lumen with circular geometry. CONCLUSIONS: QCA often underestimates diameter stenoses in lumen with non-circular geometry. Compared to QCA, MDCT yields mildly greater measurement errors in perfectly circular lumen but performs better in non-circular lesions. These findings have implications for using QCA as the gold standard for stenosis quantification by MDCT.

Two-dimensional roughening of adsorbate islands in thermodynamic equilibrium.

Equilibrium fluctuations of islands of adsorbed O atoms on Ru(0001) were investigated by scanning tunneling microscopy (STM), density functional theory calculations (DFT) and Monte Carlo (MC) simulations. Very ramified (2 x 2)-O islands were observed by high-speed STM that point to complex interactions between the O atoms. The DFT calculations show that, in addition to pairwise attractive interactions between third-nearest neighbors, a repulsive three-body interaction exists between these. MC simulations that include three-body interactions reproduce the observed ordering behavior.

Perioperative beta-blocker withdrawal and mortality in vascular surgical patients.

OBJECTIVE: Our purpose was to determine the effect of postoperative beta-blocker withdrawal on mortality and cardiovascular events after vascular surgery. METHODS: Detailed data were collected on perioperative cardiovascular medication use and discontinuation and cardiovascular risk factors among consecutive major vascular surgical procedures at two university hospitals. RESULTS: A total of 140 patients received beta-blockers preoperatively. Mortality in the 8 patients who had beta-blockers discontinued postoperatively (50%) was significantly greater than in 132 patients who had beta-blockers continued (1.5%, odds ratio 65.0, P<.001). The effect of beta-blocker discontinuation was unaffected by adjustment by stratification for risk factors (all P< or =.01), for contraindications to restarting beta-blockers (P = .006), and by multivariable analyses adjusting for potential confounders (adjusted odds ratio 17.0, P =.01). beta-Blocker discontinuation also was associated with increased cardiovascular mortality (0% vs 29%, P =.005) and postoperative myocardial infarction (odds ratio 17.7, P =.003). CONCLUSION: Discontinuing beta-blockers immediately after vascular surgery may increase the risk of postoperative cardiovascular morbidity and mortality.

A tool for quantifying hip flexor function during gait.

An index to measure the change in hip flexor function after hip surgery was developed in the Motion Analysis Laboratory at Gillette Children's Specialty Healthcare (GCSH). The hip flexor index (HFI) utilizes principal component analysis applied to five kinematic and kinetic variables collected during normal acquisition of gait data. A single index number is derived that describes accurately the overall hip function. Initial clinical validation was carried out using a data set of 23 normal controls and six patients with a diagnosis of cerebral palsy. The patients were all independent ambulators who had undergone orthopedic hip surgery. The controls were used to establish patterns of interdependency between selected gait parameters and form a basis for the index number. The HFI based surgical outcome was compared to the subjective rating of six clinicians. The clinicians were versed in gait analysis and were blinded to the HFI based outcome. The HFI measure of post-operative change in hip function was found to correspond well with the subjective clinical evaluation. A valid tool such as the HFI can be used to objectify clinical impressions of change in hip function, and can thereby assist researchers with statistical and outcome analysis of interdependent and redundant gait variables.

The D1 C-terminal processing protease of photosystem II from Scenedesmus obliquus. Protein purification and gene characterization in wild type and processing mutants.

Polypeptide D1 of the photosystem II reaction center of oxygenic photosynthesis is expressed in precursor form (pre-D1), and it must be proteolytically processed at its C terminus to enable assembly of the manganese cluster responsible for photosynthetic water oxidation. A rapid and highly sensitive enzyme-linked immunosorbent assay-based microtiter plate method is described for assaying this D1 C-terminal processing protease. A protocol is described for the isolation and purification to homogeneity of the enzyme from the green alga, Scenedesmus obliquus. Amino acid sequence information on the purified protease was used to clone the corresponding gene, the translated sequence of which is presented. A comparison of the gene product with homologous proteases points to a region of conserved residues that likely corresponds to the active site of a new class of serine protease. The LF-1 mutant strain of Scenedesmus (isolated by Dr. Norman Bishop) is incapable of processing pre-D1. We show here that the C-terminal processing protease gene in this strain contains a single base deletion that causes a frame shift and a premature stop of translation within the likely active site of the enzyme. A suppressor strain, LF-1-RVT-1, which is photoautotrophic and capable of processing pre-D1 has a nearby single base insertion that restores the expression of active enzyme. These observations provide the first definitive proof that the enzyme isolated is responsible for in vivo proteolytic processing of pre-D1 and that no other protease can compensate for its loss.

Single core polypeptide in the reaction center of the photosynthetic bacterium Heliobacillus mobilis: structural implications and relations to other photosystems.

The gene for a reaction center core polypeptide from the anoxygenic photosynthetic bacterium Heliobacillus mobilis was cloned and sequenced. The deduced amino acid sequence consists of 609 residues with a molecular mass of 68 kDa. An adjacent open reading frame is not transcribed under our experimental conditions. No evidence for a second related reaction center core gene was found. The primary sequence of the reaction center protein (P800 protein) shows a high percentage of sequence identity to photosystem I in a cysteine-containing loop, which is the putative binding site of the iron-sulfur center FX and in the preceding hydrophobic region. Our data imply a homodimeric organization of the reaction center. This is fundamentally different from photosystem I and most other photosynthetic reaction centers, where the reaction center core is composed of two similar but nonidentical subunits.

Role of the carboxy terminus of polypeptide D1 in the assembly of a functional water-oxidizing manganese cluster in photosystem II of the cyanobacterium Synechocystis sp. PCC 6803: assembly requires a free carboxyl group at C-terminal position 344.

The D1 polypeptide of the photosystem II (PSII) reaction center is synthesized as a precursor polypeptide which is posttranslationally processed at the carboxy terminus. It has been shown in spinach that such processing removes nine amino acids, leaving Ala344 as the C-terminal residue [Takahashi, M., Shiraishi, T., & Asada, K. (1988) FEBS Lett. 240, 6-8; Takahashi, Y., Nakane, H., Kojima, H., & Satoh, K. (1990) Plant Cell Physiol. 31, 273-280]. We show here that processing on the carboxy side of Ala344 also occurs in the cyanobacterium Synechocystis 6803, resulting in the removal of 16 amino acids. By constructing a deletion strain of Synechocystis 6803 that lacks the three copies of the psbA gene encoding D1, we have developed a system for generating psbA mutants. Using this system, we have constructed mutants of Synechocystis 6803 that are modified in the region of the C-terminus of the D1 polypeptide. Characterization of these mutants has revealed that (1) processing of the D1 polypeptide is blocked when the residue after the cleavage site is changed from serine to proline (mutant Ser345Pro) with the result that the manganese cluster is unable to assemble correctly; (2) the C-terminal extension of 16 amino acid residues can be deleted with little consequence either for insertion of D1 into the thylakoid membrane or for assembly of D1 into a fully active PSII complex; (3) removal of only one more residue (mutant Ala344stop) results in a loss of assembly of the manganese cluster; and (4) the ability of detergent-solubilized PSII core complexes (lacking the manganese cluster) to bind and oxidize exogenous Mn2+ by the secondary donor, Z+, is largely unaffected in the processing mutants (the Ser345Pro mutant of Synechocystis 6803 and the LF-1 mutant of Scenedesmus obliquus) and the truncation mutant Ala344stop. Our results are consistent with a role for processing in regulating the assembly of the photosynthetic manganese cluster and a role for the free carboxy terminus of the mature D1 polypeptide in the ligation of one or more manganese ions of the cluster.

The last decades and matrimonial changes.

PIP: This paper focuses on changes in fertility, marriage patterns, cohabitation, divorce, and separation in Western European countries over the past few decades. Major trends discussed include the decline in fertility and marriage rates and the increase in nonmarital cohabitation, divorce, and separation.^ieng

Protein sequences and redox titrations indicate that the electron acceptors in reaction centers from heliobacteria are similar to Photosystem I.

Photosynthetic reaction centers isolated from Heliobacillus mobilis exhibit a single major protein on SDS-PAGE of 47 000 Mr. Attempts to sequence the reaction center polypeptide indicated that the N-terminus is blocked. After enzymatic and chemical cleavage, four peptide fragments were sequenced from the Heliobacillus mobilis apoprotein. Only one of these sequences showed significant specific similarity to any of the protein and deduced protein sequences in the GenBank data base. This fragment is identical with 56% of the residues, including both cysteines, found in highly conserved region that is proposed to bind iron-sulfur center Fx in the Photosystem I reaction center peptide that is the psaB gene product. The similarity to the psaA gene product in this region is 48%. Redox titrations of laser-flash-induced photobleaching with millisecond decay kinetics on isolated reaction centers from Heliobacterium gestii indicate a midpoint potential of -414 mV with n = 2 titration behavior. In membranes, the behavior is intermediate between n = 1 and n = 2, and the apparent midpoint potential is -444 mV. This is compared to the behavior in Photosystem I, where the intermediate electron acceptor A1, thought to be a phylloquinone molecule, has been proposed to undergo a double reduction at low redox potentials in the presence of viologen redox mediators. These results strongly suggest that the acceptor side electron transfer system in reaction centers from heliobacteria is indeed analogous to that found in Photosystem I. The sequence similarities indicate that the divergence of the heliobacteria from the Photosystem I line occurred before the gene duplication and subsequent divergence that lead to the heterodimeric protein core of the Photosystem I reaction center.

Protein sequences and redox titrations indicate that the electron acceptors in reaction centers from heliobacteria are similar to Photosystem I.

Photosynthetic reaction centers isolated from Heliobacillus mobilis exhibit a single major protein on SDS-PAGE of 47 000 Mr. Attempts to sequence the reaction center polypeptide indicated that the N-terminus is blocked. After enzymatic and chemical cleavage, four peptide fragments were sequenced from the Heliobacillus mobilis apoprotein. Only one of these sequences showed significant specific similarity to any of the protein and deduced protein sequences in the GenBank data base. This fragment is identical with 56% of the residues, including both cysteines, found in the highly conserved region that is proposed to bind iron-sulfur center FX in the Photosystem I reaction center peptide that is the psaB gene product. The similarity to the psaA gene product in this region is 48%.Redox titrations of laser-flash-induced photobleaching with millisecond decay kinetics on isolated reaction centers from Heliobacterium gestii indicate a midpoint potential of -414 mV with n=2 titration behavior. In membranes, the behavior is intermediate between n=1 and n=2, and the apparent midpoint potential is -444 mV. This is compared to the behavior in Photosystem I, where the intermediate electron acceptor A1, thought to be a phylloquinone molecule, has been proposed to undergo a double reduction at low redox potentials in the presence of viologen redox mediators.These results strongly suggest that the acceptor side electron transfer system in reaction centers from heliobacteria is indeed analogous to that found in Photosystem I. The sequence similarities indicate that the divergence of the heliobacteria from the Photosystem I line occurred before the gene duplication and subsequent divergence that lead to the heterodimeric protein core of the Photosystem I reaction center.

Isolation of a photoactive photosynthetic reaction center-core antenna complex from Heliobacillus mobilis.

A photoactive reaction center-core antenna complex was isolated from the photosynthetic bacterium Heliobacillus mobilis by extraction of membranes with Deriphat 160c followed by differential centrifugation and sucrose density gradient ultracentrifugation. The purified complex contained a Mr 47,000 polypeptide(s) that bound both the primary donor (P800) and approximately 24 antenna bacteriochlorophylls g. Time-resolved fluorescence emission spectroscopy indicated that the antenna bacteriochlorophylls g are active in energy transfer to P800, exhibiting a decay time of 25 ps. The complex contained 1.4 menaquinones, 9 Fe, and 3 labile S2- per P800. The complex was photoactive with an exponential decay time of 14 ms for P800+ yet showed no EPR-detectable Fe-S center signal in the g less than or equal to 2.0 region, either by chemical reduction to -600 mV or by illumination of reduced samples. The complex is similar to photosystem I of oxygen-evolving photosynthetic systems in that both the primary donor and a core antenna are bound to the same pigment-protein complex.