Asymmetrical interaction of GroEL and GroES in the ATPase cycle of assisted protein folding

The chaperonins GroEL and GroES of Escherichia coli facilitate protein folding in an adenosine triphosphate (ATP)-dependent reaction cycle. The kinetic parameters for the formation and dissociation of GroEL-GroES complexes were analyzed by surface plasmon resonance. Association of GroES and subsequent ATP hydrolysis in the interacting GroEL toroid resulted in the formation of a stable GroEL:ADP:GroES complex. The complex dissociated as a result of ATP hydrolysis in the opposite GroEL toroid, without formation of a symmetrical GroEL:(GroES)2 intermediate. Dissociation was accelerated by the addition of unfolded polypeptide. Thus, the functional chaperonin unit is an asymmetrical GroEL:GroES complex, and substrate protein plays an active role in modulating the chaperonin reaction cycle.     

Prevalence and risk factors for diabetic retinopathy among Omani diabetics

To determine the minimal contrast dosage required for diagnostic contrast-enhanced three-dimensional (3D) magnetic resonance angiography (MRA) image quality of the pulmonary (PAs) or renal arteries (RAs). In 12 volunteers (10 females, 2 males; mean age 24 years) imaging was performed with 4 different dosages: 0.05, 0.1, 0.2 and 0.3 mmol/kg of body weight (BW) 0.5 M gadolinium (Gd) contrast agent. The PAs and RAs were evaluated separately each in groups of six volunteers. Qualitative and quantitative signal-to-noise ratio (SNR) image analysis was performed. For the PAs, the increases in signal-to-noise ratio were paralleled by increases in image quality ratings. For the PAs, with the use of 0.05 mmol/kg, only 50.3% of all segments were rated diagnostic, whereas with higher dosages the percentage rose to 89.2% for 0.1 mmol/kg, 98.2% for 0.2 mmol/kg. and 99.1% for 0.3 mmol/kg. For the RAs, 0.3 mmol/kg provided no significant increase in singal-to-noise ratio compared to 0.2 mmol/kg (p = 0.4). Only by a dosage of 0.2 and 0.3 mmol/kg, all evaluated segments were diagnostic evaluable. A dose of 0.2 mmol/kg is required for proper assessment of the RAs or PAs.     

Intravascular ultrasound findings in stenting of unprotected left main coronary artery stenosis

We evaluated the role of intravascular ultrasound (IVUS) in 16 patients with unprotected left main coronary artery (LMCA) stenting compared with 80 patients with other (non-LMCA) native coronary artery stenting and found that (1) additional high-pressure or larger size balloon dilations were more frequently performed in LMCA stenting than in non-LMCA stenting (p <0.05) and (2) after IVUS-guided stent implantation, minimum lumen area was > or = 9 mm2 in 88% of patients who underwent LMCA stenting and in 19% of those who underwent non-LMCA stenting (p <0.001). IVUS guidance may be a more important adjunctive imaging modality in the stenting of unprotected LMCA stenoses than in stenting of non-LMCA stenoses.     

Epidemiology of giardiasis and cryptosporidiosis in Jamaica

We report the findings of a cross-sectional epidemiologic study of Giardia lamblia and Cryptosporidum infections in Jamaica. Three hundred twenty eight stool samples from patients less than one to 81 years of age were examined using formalin-ether concentration for G. lamblia, Zeihl-Neelsen staining for Cryptosporidium, and the Prospect rapid enzyme immunoassay (EIA; Alexon, Sunnyvale, CA) for parasite diagnosis. The Prospect Giardia rapid assay detected 17 cases of G. lamblia infection compared with six by formalin-ether concentration. However, the Prospect Cryptosporidum EIA did not increase the rate of detection of Cryptosporidum when compared with Zeihl-Neelsen staining. Cryptosporidum infections were most frequently diagnosed in children less than five years old and prevalence decreased with age. In contrast, the prevalence of giardiasis increased as children became older. There were no associations between the infections and stool consistency, clinical manifestations, or sex of the individuals. The contribution of the parasites to childhood morbidity will depend on accurate laboratory diagnosis.     

LEC18, a dominant Chinese hamster ovary glycosylation mutant synthesizes N-linked carbohydrates with a novel core structure

The dominant Chinese hamster ovary cell glycosylation mutant, LEC18, was selected for resistance to pea lectin (Pisum sativum agglutinin (PSA)). Lectin binding studies show that LEC18 cells express altered cell surface carbohydrates with markedly reduced binding to 125I-PSA and increased binding to 125I-labeled Datura stramonium agglutinin (DSA) compared with parental cells. Desialylated [3H]Glc-labeled LEC18 cellular glycopeptides that did not bind to concanavalin A-Sepharose exhibited an increased proportion of species that were bound to DSA-agarose. Most of these glycopeptides bound to ricin-agarose and were unique to LEC18 cells. This fraction was purified from approximately 10(10) cells and shown by 1H NMR spectroscopy and methylation linkage analysis to contain novel N-linked structures. Digestion of these glycopeptides with mixtures of beta-D-galactosidases and N-acetyl-beta-D-glucosaminidases gave core glycopeptides that, in contrast to cores from parental cells, were mainly not bound to concanavalin A-Sepharose or to PSA-agarose. 1H NMR spectroscopy, matrix-assisted laser desorption ionization/time of flight mass spectrometry, electrospray mass spectrometry, and collision-activated dissociation mass spectrometry showed that the LEC18 core glycopeptides contained a new GlcNAc residue that substitutes the core GlcNAc residues. Methylation linkage analysis of the parent compound provided evidence that the GlcNAc is linked at O-6 to give the following novel, N-linked core structure. [formula: see text]     

Pi (antitrypsin) typing methods. A comparison of acid starch-gel electrophoresis and isoelectric focusing

The Pi typing methods acid starch-gel electrophoresis (ASGE) and isoelectric focusing (IEF) have been compared by three reference laboratories: 564 samples of phenotypes Pi M, MS and MZ were tested in each of the three laboratories with a 96% agreement on initial typing. The discrepancies are recorded and reasons for disagreement discussed. IEF is a reliable method for Pi typing and gives results comparable to those obtained by ASGE.     

Posturography does not test vestibulospinal function

The clinical usefulness of posturography is unknown, despite its costing more than +500 per test in some areas of the United States, including Boston. We cross-sectionally and prospectively studied blinded vestibulo-ocular and vestibulospinal tests from 29 stable patients with chronic vestibular hypofunction; 22 patients were affected bilaterally (BVH), and 7 were affected unilaterally (UVH). Vestibulo-ocular function was assessed by electronystagmographic caloric stimulation and sinusoidal vertical axis rotation gains at 0.05 Hz. Vestibulospinal function was assessed by moving-platform and visualsurround posturography sensory organization tests (SOTs), paced and free gait in a gait laboratory, and clinical tests of timed gait and standing. Posturography SOT moving-platform tests 4 through 6, designed to assess vestibular function, correlated significantly (r < or = 0.72, P > or = 0.01) with vestibulo-ocular tests in 5 of 6 comparisons among BVH patients. Posturography SOT results, however, correlated poorly with other vestibulospinal measures: correlations were statistically significant for only 7 of 18 comparisons with clinical balance and gait function (r < or = 0.69, P > or = 0.01) and with 2 of 12 comparisons for gait laboratory dynamic stability measures (r < or = 0.55, P > or = 0.01) among the BVH patients. When both the platform and visual surround moved (SOT 6), however, correlations were statistically significant with static standing clinical measures (r = 0.51 to 0.69, P < 0.01) and with whole-body maximum moment arm during paced gait (r = 0.55, P < 0.01). Posturography scores for the UVH patients did not significantly correlate with any vestibulo-ocular or other vestibulospinal measures. These data indicate that among patients with BVH posturography SOT scores relate at best modestly with accepted measure of vestibulo-ocular function, less well with clinical measures of balance control, and poorly with dynamic gait-performance measures. We conclude that posturography SOT does not assess vestibulospinal function.