The crystal structure of human procathepsin K

Cathepsin K is a cysteine protease present in human osteoclasts that plays an important role in bone resorption. Cathepsin K is synthesized as an inactive proenzyme and activated under conditions of low pH. Autoproteolytic processing of the N-terminal 99 amino acid propeptide produces the active, mature form of cathepsin K. It is presumed that the activation of procathepsin K in vivo occurs in the bone resorption pit, which has a low-pH environment. We have determined the structure of human procathepsin K at 2.8 A resolution. The structure of the mature enzyme domain within procathepsin K is virtually identical to that of mature cathepsin K. The fold of the propeptide of procathepsin K is similar to that observed in procathepsins B and L despite differences in length and sequence. A portion of the propeptide occupies the active site cleft of cathepsin K. Hydrophobic interactions, salt bridges, and hydrogen-bonding interactions are observed in the structure of the propeptide and between the propeptide and the mature enzyme of procathepsin K. These interactions suggest an explanation for the stability of the proenzyme. The structure of procathepsin K contributes to an understanding of the molecular basis of inhibition by the propeptide portion of the molecule and activation of this important member of the cysteine protease family.     

Generation of tumor immunity by bone marrow-derived dendritic cells correlates with dendritic cell maturation stage

The mutational pattern of IgVH and IgVL genes from synovial tissue B cell hybridomas (n = 8) of patients (n = 4) with rheumatoid arthritis (RA) was analysed, which had been produced by the electrofusion technique without prior in vitro stimulation. The molecular data were correlated with immunohistopathological data and parameters of local disease activity. The IgVH genes of the B cell hybridomas belonged to the VH3 family (DP42; DP47, n = 2; DP53), the VH1 family (DP75), the VH4 family (DP71) and the VH5 family (DP73); 7/7 IgVH genes showed somatic mutations, the R/S ratio (CDR) was > 3 in 4/7 IgVH genes and the mean R/S ratio of all IgVH genes was 9.3 (CDR) and 1.0 (FR), suggesting an antigen-dependent selection. The IgVL/lambda genes belonged to the Vlambda1 family (DPL2, DPL5, DPL8nf), the Vlambda2 family (DPL11, n = 2) and to the Vlambda6 family (IGLV6S1); 6/6 IgVL genes showed somatic mutations, the R/S ratio (CDR) was > 3 in 3/6 IgVL genes and the mean R/S ratio of all IgVL was 3.0 (CDR) and 2.3 (FR), suggesting an antigen-dependent selection. The synovial tissue exhibited germinal centres in the follicles (3/4), with the unique distribution of Ki-M4+ follicular dendritic cells and Ki-67+ proliferating cells and a dominance of IgA+ plasma cells (3/3). All patients were positive for RF in serum and exhibited severe local symptoms (swelling 4/4; warmth 4/4; effusion 2/4), whereas the hybridomas were negative for RF. Since B cell hybridomas showed hypermutation and affinity selection for IgVH and IgVL/lambda genes and the patients exhibited severe local symptoms with germinal centres in synovial tissue, this study indicates that an antigen-driven process is behind the B cell expansion in the synovial tissue of clinically affected joints. These mutated B hybridomas were negative for RF, thus suggesting that antigens different from RF are also involved in the local B cell expansion and in the chronic synovitis of RA.     

Clinical profile of commotio cordis: an under appreciated cause of sudden death in the young during sports and other activities

We describe a glucose sensor based on a mutant glucose/galactose binding protein (GGBP) and phase-modulation fluorometry. The GGBP from Escherichia coli was mutated to contain a single cysteine residue at position 26. When labeled with a sulfhydryl-reactive probe 2-(4'-iodoacetamidoanilino)naphthalene-6-sulfonic acid, the labeled protein displayed a twofold decrease in intensity in response to glucose, with a dissociation constant near 1 microM glucose. The ANS-labeled protein displayed only a modest change in lifetime, precluding lifetime-based sensing of glucose. A modulation sensor was created by combining ANS26-GGBP with a long-lifetime ruthenium (Ru) metal-ligand complex on the surface of the cuvette. Binding of glucose changed the relative intensity of ANS26-GGBP and the Ru complex, resulting in a dramatic change in modulation at a low frequency of 2.1 MHz. Modulation measurements at 2.1 MHz were shown to accurately determine the glucose concentration. These results suggest an approach to glucose sensing with simple devices.     

Recovery of health related quality of life in the year after radical prostatectomy: early experience

PURPOSE: We determined the temporal course of patient return to baseline quality of life after treatment with radical prostatectomy for early stage prostate cancer. MATERIALS AND METHODS: After establishing a longitudinal observational database of men undergoing radical retropubic prostatectomy we used established, validated quality of life questionnaires (RAND 36-Item Health Survey and University of California, Los Angeles Prostate Cancer Index) to document changes in general and disease specific health related quality of life (HRQOL). We assessed 90 patients at baseline before surgery and then at 3-month intervals for 1 year postoperatively. Logistic regression was used to explore predictors of the return to baseline. RESULTS: After prostatectomy patients had a significant decrease in all domains of HRQOL. Return to baseline was rapid in the general and bowel domains with at least two-thirds to three-fourths of patients reaching pretreatment levels within 6 months of surgery. Return to baseline was slower in the urinary and sexual function domains with 61 and 31% of the men, respectively, reaching pretreatment levels by 1 year after surgery. Of those who reached baseline the average intervals for the bowel, sexual and urinary domains were 5, 6 and 7 months, respectively. Married and white patients were more likely to achieve a return to baseline HRQOL during year 1 postoperatively. However, education level was inversely associated with the likelihood of returning to baseline. CONCLUSIONS: During the year after radical prostatectomy for early stage prostate cancer patient quality of life steadily improved. By 3 months postoperatively 30 to 40% of the patients had already recovered baseline levels of physical, mental and social functioning, and by 6 months more than 70% had reached baseline in the general HRQOL domains. By 12 months after surgery 86 to 97% of the patients had returned to baseline levels in each domain. Each domain continued to improve throughout the year. For the patients who reached baseline general HRQOL during followup average recovery time was 5 to 6 months.     

A strategy for sampling on a sphere applied to 3D selective RF pulse design

Conventional constant angular velocity sampling of the surface of a sphere results in a higher sampling density near the two poles relative to the equatorial region. More samples, and hence longer sampling time, are required to achieve a given sampling density in the equatorial region when compared with uniform sampling. This paper presents a simple expression for a continuous sample path through a nearly uniform distribution of points on the surface of a sphere. Sampling of concentric spherical shells in k-space with the new strategy is used to design 3D selective inversion and spin-echo pulses. These new 3D selective pulses have been implemented and verified experimentally.