Quantitative relationships between microdamage and cancellous bone strength and stiffness.

Microscopic tissue damage (microdamage) is an aspect of bone quality associated with impaired bone mechanical performance. While it is clear that bone tissue submitted to more severe loading has greater amounts of microdamage (as measured through staining), how microdamage influences future mechanical performance of the bone has not been well studied, yet is necessary for understanding the mechanical consequences of the presence of microdamage. Here we determine how stained microdamage generated by a single compressive overload affects subsequent biomechanical performance of cancellous bone. Human vertebral cancellous bone specimens (n=47) from 23 donors (14 males, 9 females, 64-92years of age) were submitted to a compressive overload, stained for microdamage, then reloaded in compression to determine the relationship between the amount of microdamage caused by the initial load and reductions in mechanical performance during the reload. Damage volume fraction (DV/BV) caused by the initial overload was related to reductions in Young's modulus, yield strength, ultimate strength, and yield strain upon reloading (p<0.05, R(2)=0.18-0.34). The regression models suggest that, on average, relatively small amounts of microdamage are associated with large reductions in reload mechanical properties: a 1.50% DV/BV caused by a compressive overload was associated with an average reduction in Young's modulus of 41.0±3.2% (mean±SE), an average reduction in yield strength of 63.1±4.5% and an average reduction in ultimate strength of 52.7±4.0%. Specimens loaded beyond 1.2% (1.2-4.0% apparent strain) demonstrated a single relationship between reload mechanical properties (Young's modulus, yield strength, and ultimate strength) and bone volume fraction despite a large range in amounts of microdamage. Hence, estimates of future mechanical performance of cancellous bone can be achieved using the bone volume fraction and whether or not a specimen was previously loaded beyond ultimate strain. The empirical relationships provided in this study make it possible to estimate the degree of impaired mechanical performance resulting from an observed amount of stained microdamage.

The interaction of calmodulin with alternatively spliced isoforms of the type-I inositol trisphosphate receptor.

A 592-amino acid segment of the regulatory domain of the neuronal type-I inositol 1,4,5-trisphosphate receptor (IP(3)R) isoform (type-I long, amino acids1314-1905) and the corresponding 552-amino acid alternatively spliced form present in peripheral tissues (type-I short, amino acids 1693-1733 deleted) were expressed as glutathione S-transferase fusion proteins. These domains encompass a putative calmodulin (CaM) binding domain and two protein kinase A phosphorylation sites. Both long and short fusion proteins retained the ability to bind CaM in a Ca(2+)-dependent manner as measured by CaM-Sepharose chromatography or a dansyl-CaM fluorescence assay. Both assays indicated that the short fusion protein bound twice the amount of CaM than the long form at saturating concentrations of CaM. In addition, the binding of the short form to CaM-Sepharose was inhibited by phosphorylation with protein kinase A, whereas the binding of the long form was unaffected. Full-length cDNAs encoding type-I long, type-I short, and type-III IP(3)R isoforms were expressed in COS cells, and the Ca(2+) sensitivity of [(3)H]IP(3) binding to permeabilized cells was measured. The type-I long isoform was more sensitive to Ca(2+) inhibition (IC(50) = 0.55 microM) than the type-I short (IC(50) = 5.7 microM) or the type-III isoform (IC(50) = 3 microM). In agreement with studies on the fusion proteins, the full-length type-I short bound more CaM-Sepharose, and this binding was inhibited to a greater extent by protein kinase A phosphorylation than the type-I long IP(3)R. Although type-III IP(3)Rs did not bind directly to CaM-Sepharose, hetero-oligomers of type-I/III IP(3)Rs retained the ability to interact with CaM. We conclude that the deletion of the SII splice site in the type-I IP(3)R results in the differential regulation of the alternatively spliced isoforms by Ca(2+), CaM, and protein kinase A.

Calf sonography for detecting deep venous thrombosis in symptomatic patients: experience and review of the literature.

PURPOSE: We determined the accuracy of sonography in the detection of isolated calf deep venous thrombosis (DVT) and the rate of indeterminate ultrasound examinations in patients with physical signs or symptoms suggestive of DVT. METHODS: We reviewed the medical literature (MEDLINE) to determine the accuracy of sonography and the frequency of indeterminate studies in detecting isolated calf DVT in patients with physical signs or symptoms suggestive of DVT. A meta-analysis was used to derive summary measures of sensitivity, specificity, and accuracy from studies in which 5 or more isolated calf DVT were identified. Frequencies of indeterminate examinations were recorded for studies in which these data were provided, and we pooled these results with our own data for 196 patients. RESULTS: The meta-analysis revealed that sonography correctly identified isolated calf DVT in 49 of 53 extremities (sensitivity, 92.5%; 95% confidence interval, 81.8-97.9%) and correctly identified the absence of calf DVT in 157 of 159 extremities (specificity, 98.7%; 95% confidence interval, 95.5-99. 9%), yielding an accuracy of 97.2% (95% confidence interval, 93.9-99. 0%) for ultrasound examinations considered diagnostic. However, when evaluating our patient population and the literature, we found a substantial number of indeterminate studies (overall rate of 54.6% in 463 extremities), with a wide variation in the reported frequency of indeterminate studies (9.3-82.7%). CONCLUSIONS: Sonography is highly accurate in detecting isolated calf DVT in symptomatic patients, but indeterminate studies occur frequently, with a wide range of reported rates. Each ultrasound laboratory should evaluate its own rate of indeterminate studies.

Biosynthesis of inositol trisphosphate receptors: selective association with the molecular chaperone calnexin.

A prominent labelled polypeptide having the same mobility as type-I inositol trisphosphate receptor (IP(3)R) was immunoprecipitated from WB-cell lysates by antibodies to calnexin, an ER integral membrane chaperone. The identity of this polypeptide was confirmed by re-immunoprecipitation of the radioactive polypeptides released from calnexin-antibody immunoprecipitates with type-I IP(3)R antibody. The interaction of calnexin with newly synthesized type-I IP(3)R was transient and inhibited by treatment of the cells with dithiothreitol or the glucosidase inhibitor N-methyldeoxynojirimicin. In similar experiments, there was no evidence for the binding of type-I IP(3)R to calreticulin, an ER luminal chaperone. Calnexin (but not calreticulin) associated with newly synthesized FLAG (DYKDDDDK epitope)-tagged type-III IP(3)R expressed in COS-7 cells. In order to further define the mechanism of interaction of nascent IP(3)R with chaperones, we have utilized an in vitro rabbit reticulocyte translation assay programmed with RNA templates encoding the six putative transmembrane (TM) domains of IP(3)Rs. In accordance with the known preference of calnexin for monoglucosylated oligosaccharide chains, calnexin antibody preferentially immunoprecipitated a proportion of glycosylated type-I translation product. Addition of glucosidase inhibitors prevented the association of calnexin with in vitro translated type-I TM construct. Using truncated RNA templates we found that calnexin did not associate with the first four TM domains but retained affinity for the construct encoding TM domains 5 and 6, which contains the glycosylation sites. We propose that calnexin is a key chaperone involved in the folding, assembly and oligomerization of newly synthesized IP(3) receptors in the ER.

Clinical outcomes of untreated symptomatic patients with negative findings on sonography of the thigh for deep vein thrombosis: our experience and a review of the literature.

OBJECTIVE: Our objective was to determine whether immediate venography or other additional imaging is necessary in symptomatic patients who have negative findings on sonography of the thigh for deep vein thrombosis (DVT). MATERIALS AND METHODS: We retrospectively evaluated the clinical outcomes of 146 patients with physical signs or symptoms consistent with DVT in whom sonography of the thigh was negative for DVT. We combined our results with those of three prospective studies that evaluated symptomatic patients with suspected DVT in whom sonography of the thigh had negative findings. Propagated DVT (from calf to thigh) and pulmonary emboli were considered adverse outcomes. RESULTS: Only one pulmonary embolus (0.7%) occurred in our patient population. However, one (7.7%) of 13 patients who underwent second examinations because of persistent symptoms developed thigh DVT. Review of the literature found four (0.2%) of 1797 patients subsequently developed pulmonary emboli after undergoing sonography of the thigh that was initially interpreted as having negative findings. Fifteen (12.5%) of 120 patients who underwent second sonographic examinations developed DVT of the thigh. No deaths from pulmonary emboli occurred in patients in our study or patients in the studies published in the medical literature. CONCLUSION: Immediate venography or other additional imaging is not necessary in symptomatic patients in whom sonography of the thigh is negative for DVT, given the exceedingly low risk of a pulmonary embolus. Follow-up sonography is indicated in persistently symptomatic patients to detect propagation of calf DVT into the thigh.

Clinically important pulmonary emboli: does calf vein US alter outcomes?

PURPOSE: To assess whether calf imaging is necessary to identify patients at risk for developing clinically important pulmonary emboli (PE) or propagation of calf deep venous thrombosis (DVT) when the initial thigh ultrasonographic (US) scan is negative for DVT. MATERIALS AND METHODS: The authors retrospectively evaluated the radiology reports from 283 patients (168 female and 115 male patients; mean age, 55.7 years; age range, 1-93 years) in whom US was performed to rule out lower extremity DVT. In all patients, the initial thigh examination was negative for DVT. All patients were classified as to the reason for the examination, risk factors for DVT (including recent surgery), whether they received anticoagulation therapy, and findings on calf US scans. Adverse outcomes were considered a clinically important PE or DVT in the thigh. RESULTS: Only 1.1% of patients (95% CI = 0.2%, 3.1%) had adverse outcomes. Adverse outcomes occurred only in postsurgical patients (P = .028) and were not related to the presence or absence of calf DVT or method of treatment. CONCLUSION: US of the calf is unnecessary at initial evaluation to identify patients at risk of clinically important PE or propagation of DVT into the thigh.

Extravisceral masses in the peritoneal cavity: sonographically guided biopsies in 52 patients.

OBJECTIVE: Our objective was to determine the effectiveness of sonographically guided biopsies of extravisceral masses (masses outside the solid organs) in the peritoneal cavity. MATERIALS AND METHODS: We retrospectively reviewed the results of sonographically guided biopsies of extravisceral masses found in the peritoneal cavity of 52 patients (age range, 25-90 years old; mean age, 52 years) from June 1990 to December 1996. Fifty-one patients underwent biopsy through the abdominal wall, and one patient underwent transvaginal biopsy. Sonographic guidance was obtained using 3.5- to 7.0-MHz vector probes. The size, depth, and sonographic characteristics of the mass and the type of biopsy (aspirate versus core) were determined for all lesions. Pathology reports and clinical courses were reviewed. RESULTS: Placement of the biopsy needle within the lesion was successful in all patients. The mean depth from skin surface to lesion was significantly less (p < .0001) when shown by sonography (2.4 cm) than when shown by CT (3.8 cm). Biopsy results were true-positive for malignancy in 37 patients (no false-positives), true-negative for benign masses in 10 patients, and false-negative for malignancy in three patients (sensitivity, 93%; specificity, 100%; accuracy, 94%). Nondiagnostic samples were obtained in two patients (4%). Treatment was based on diagnostic biopsy results in 43 patients (86%). CONCLUSION: Sonography is an effective alternative to CT in guiding biopsy of extravisceral masses in the peritoneal cavity.

Optical triple-in digital logic using nonlinear optical four-wave mixing.

A new programmable optical processor is proposed for implementing triple-in combinatorial digital logic that uses four-wave mixing. Binary-coded decimal-to-octal decoding is experimentally demonstrated by use of a photorefractive BaTiO(3) crystal. The result confirms the feasibility of the proposed system.