Characterization of organic matrix macromolecules from the shells of the Antarctic scallop, Adamussium colbecki.

The highly acidic soluble organic matrix (SM) isolated from shells of the Antarctic scallop, Adamussium colbecki, was shown to consist of 1.5% carbohydrate by weight and 12.8% phosphate by weight. Total SM is composed of approximately 31% Asx, 29% Ser, and 18% Gly. Separation of the SM using RP-HPLC yielded a minimum of six protein fractions labeled RP-1 through RP-6 in order of elution off the column. The first fraction, RP-1, was found to be an effective inhibitor of calcium carbonate crystal nucleation in vitro suggesting a role for this protein in the regulation of shell mineralization. A less acidic fraction, RP-3, showed less inhibitory activity and dephosphorylation of RP-1 resulted in almost complete loss of inhibitory activity. Automated Edman degradation was used to sequence peptides generated by chemical cleavage of RP-1. Mild acid hydrolysis yielded peptides with sequences of N-S-G-D-D-D-D-G-G-OH, N-S-G-G-(S,G)-G-OH, and N-S-G-R-G-OH. Cleavage with hydroxylamine yielded peptides of N-D-D-D-D-D-D-D-D-OH, N-L-Y-Y-OH, and N-A-V-G-E-S-D-OH. These data suggest biochemical similarities between this SM and other SM proteins isolated from both calcium carbonate and calcium phosphate biominerals, and presents evidence of a primary domain structure similar to that described for oyster SM and phosphophoryn isolated from rat dentin.

Strength and endurance training of an individual with left upper and lower limb amputations.

PURPOSE: The purpose of this article is to describe the development of a strength and endurance training programme designed to prepare an individual with a left glenohumeral disarticulation and transtibial amputation for a bike trip across the USA. METHOD: The subject was scheduled for training three times per week over a two-month period followed by two times per week for an additional two months. Training consisted of a resistance training circuit using variable resistance machines, cycling using a recumbent stationary bike, and core stability training using stability ball exercises. Changes in strength were assessed using 10 RM tests on the resistance machines and changes in peak VO(2) were monitored utilizing the Cosmed K4b pulmonary function tester. RESULTS: The subject demonstrated a 30.3% gain in peak VO(2). The subject's 10 RM for left single limb leg press increased 36.8% and gains of at least 7.7% were seen for all other muscle groups tested. CONCLUSION: The strength and endurance training programme adapted to compensate for this subject's limb losses was effective in increasing both strength and peak VO(2). Adapting exercise programmes to compensate for limb loss may allow individuals with amputations to participate in physically challenging activities that otherwise may not be available to them.

A facilitative role for carbonic anhydrase activity in matrix vesicle mineralization.

Carbonic anhydrase (CA) which catalyzes the reversible hydrolysis of carbon dioxide is known to be important in osteoclastic bone resorption, however, suggested roles in calcium phosphate mineral formation have not been previously demonstrated. Biochemical evidence is provided for the presence of CA in growth plate matrix vesicles (MV) and the level of activity determined by enzyme assay. Inhibition of CA activity with the specific inhibitor acetazolamide resulted in reduced rates of MV mineralization. Other inhibitor studies showed that MV mineralization was also impaired by 4,4-diisothiocyanatostilbene-2, 2-disulfonic acid (DIDS), a blocker of membrane bicarbonate channels. No evidence was found for the presence of any proton pumps or channels. When acetazolamide and DIDS were combined, their inhibitory effects on MV mineralization were additive. These findings suggest that MV possess a pH regulation system composed of carbonic anhydrase and a putative bicarbonate channel. This system may function in the MV by providing intraluminal buffering capacity. The control of intravesicular pH is important for the stabilization of the acid-labile nucleational core complex and in preventing the build-up of protons during calcium phosphate phase transformations.