An algorithm for extracting the network geometry of three-dimensional collagen gels

The geometric structure of a biopolymer network impacts its mechanical and biological properties. In this paper, we develop an algorithm for extracting the network architecture of three-dimensional (3d) fluorescently labeled collagen gels, building on the initial work of Wu et al., (2003) . Using artificially generated images, the network extraction algorithm is then validated for its ability to reconstruct the correct bulk properties of the network, including fiber length, persistence length, cross-link density, and shear modulus.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75476/1/j.1365-2818.2008.02141.x.pd

Three-dimensional reconstruction of fibrin clot networks from stereoscopic intermediate voltage electron microscope images and analysis of branching

Fibrin polymerizes to produce branching fibers forming a three-dimensional network, which has been difficult to visualize by conventional microscopy. Three-dimensional images of whole clots at high resolution were obtained from stereo-pair intermediate-voltage electron micrographs. Computer software was developed to produce three-dimensional reconstructions of the networks in the form of a pattern of links that connect branching junctions. Network parameters were measured and analyzed to characterize the clots quantitatively. Models in which all links were moved to the origin, while preserving their orientation, allowed visualization of some network parameters and facilitated comparison of networks. Fibrin clots formed in three different conditions were analyzed and compared by these methods. Clots formed in 0.20 M saline buffer consist of fibers of uniform size, and most of the branching junctions consist of three links. Fibrin clots formed in 0.05 M saline buffer are made up of very large diameter fiber bundles with far fewer branching junctions and correspondingly longer links. Clots formed in 0.40 M saline buffer consist of very fine fibers with numerous branching junctions and very short links. In summary, the extent of lateral aggregation is directly related to the distance between branching junctions and inversely related to the total number of branching junctions. These observations must be considered in defining possible mechanisms of fibrin branching

A study of fibrin clot structure under various ionic conditions using stereoscopic IVEM images

Fibrin clots are formed by the conversion of fibrinogen into fibrin monomers which assemble to produce two-stranded protofibrils that aggregate to form fibers. These fibers may also aggregate laterally with other fibers to form larger fiber bundles. Investigations of fibrin clot structures under various ionic conditions using SEM showed dramatic differences in fiber morphology and clot structure. Fibrin clots formed under various ionic conditions were investigated by the examination of stereoscopic IVEM images. This technique provides greater depth discrimination and higher resolution images of clot ultrastructure. Details of fiber association and branching are of particular interest.Purified human fibrinogen was prepared at a concentration of 0.5 mg/ml in 0.4M, 0.2M, and 0.05M buffers (0.05M Tris-HCI, pH 7.4, with 2mM CaCl2). Fibrinogen solutions were mixed with thrombin to a final concentration of 0.3U/ml and aiiquots placed onto Formvar and carbon coated grids. After clotting for 1 hr. at room temperature, clots were fixed with 3% glutaraldehyde in 0.1 M Na-cacodylate buffer, pH 7.2, for 5 min. Grids were kept constantly moist to avoid collapse or syneresis.</jats:p

Methicillin-resistant Staphylococcus aureus strains in New York City hospitals: inter-hospital spread of resistant strains of type 88

A survey of methicillin-resistant strains of Staphylococcus aureus received for phage typing indicated a marked increase of resistant strains received in 1982 and 1983. Of 62 hospitals in New York City which sent strains for phage typing, 35 had methicillin-resistant isolates. A significant development was the presence of strains of the same phage type at several hospitals, indicating a possible inter-hospital spread of these strains. Among strains present at several hospitals, the largest group was of experimental phage type 88. Strains of type 88 were received from 23 hospitals, representing 56% of all methicillin-resistant strains received from New York City hospitals. Strains of type 88 were resistant to all antistaphylococcal antibiotics, with the exception of vancomycin, and represented a major source of nosocomial infections at 13 hospitals. As experimental phage 88 is not routinely used for typing in U.S. laboratories, the nationwide distribution of strains of type 88 is difficult to assess.</jats:p

Expression of parafibromin in major renal cell tumors

Parafibromin, encoded by HRPT2 gene, is a recently identified tumor suppressor. Complete and partial loss of its expression have been observed in hyperparathyroidism-jaw tumor (HPT-JT), parathyroid carcinoma, breast carcinoma, lung carcinoma, gastric and colorectal carcinoma. However, little has been known about its expression in renal tumors. In order to study the expression of parafibromin in a series of the 4 major renal cell tumors - clear cell renal cell carcinoma (ccRCC), papillary renal cell carcinoma (pRCC), chromophobe renal cell carcinoma (chRCC) and oncocytoma, one hundred thirty nine renal tumors including 61 ccRCCs, 37 pRCCs, 22 chRCCs and 19 oncocytomas were retrieved and used for the construction of renal tissue microarrays (TMAs). The expression of parafibromin was detected by immunohistochemical method on the constructed TMAs. Positive parafibromin stains are seen in 4 out of 61 ccRCCs (7%), 7 out of 37 pRCCs (19%), 12 out of 23 chRCCs (52%) and all 19 oncocytomas (100%). Parafibromin expression varies significantly (P<8.8×10(−16)) among the four major renal cell tumors and were correlated closely with tumor types. No correlation of parafibromin expression with tumor staging in ccRCCs, pRCCs and chRCCs, and Fuhrman nuclear grading in ccRCCs and pRCCs was seen. In summary, parafibromin expression was strongly correlated with tumor types, which may suggest that it plays a role in the tumorigenesis in renal cell tumors

Expression of parafibromin in major renal cell tumors.

Parafibromin, encoded by HRPT2 gene, is a recently identified tumor suppressor. Complete and partial loss of its expression have been observed in hyperparathyroidism-jaw tumor (HPT-JT), parathyroid carcinoma, breast carcinoma, lung carcinoma, gastric and colorectal carcinoma. However, little has been known about its expression in renal tumors. In order to study the expression of parafibromin in a series of the 4 major renal cell tumors - clear cell renal cell carcinoma (ccRCC), papillary renal cell carcinoma (pRCC), chromophobe renal cell carcinoma (chRCC) and oncocytoma. One hundred thirty nine renal tumors including 61 ccRCCs, 37 pRCCs, 22 chRCCs and 19 oncocytomas were retrieved and used for the construction of renal tissue microarrays (TMAs). The expression of parafibromin was detected by immunohistochemical method on the constructed TMAs. Positive parafibromin stains are seen in 4 out of 61 ccRCCs (7%), 7 out of 37 pRCCs (19%), 12 out of 23 chRCCs (52%) and all 19 oncocytomas (100%). Parafibromin expression varies significantly (P\u3c 8.8 x10-16) among the four major renal cell tumors and were correlated closely with tumor types. No correlation of parafibromin expression with tumor staging in ccRCCs, pRCCs and chRCCs, and Fuhrman nuclear grading in ccRCCs and pRCCs. In summary, parafibromin expression was strongly correlated with tumor types, which may suggest that it plays a role in the tumorigenesis in renal cell tumors

Temporal expression of 8 growth factors in tendon-to-bone healing in a rat supraspinatus model

Growth factors play an important role in supraspinatus tendon to bone healing. The objective of this study was to evaluate the temporal expression of eight different growth factors in tendon to bone healing in an animal model. We hypothesize that growth factors exhibit unique temporal profiles that correlate to specific stages in the acute process of the supraspinatus tendon. To test this hypothesis, rats underwent bilateral supraspinatus tendon detachment and repair. Animals were sacrificed at 1, 2, 4, 8 and 16 week time points. Immunohistochemical staining was done using antibodies for bFGF, BMP-12, BMP-13, BMP-14, COMP, CTGF, PDGF-B, and TGF-β1. Immunoassays showed an increase in the expression of all growth factors at 1 week followed by a return to control or undetectable levels by 16 weeks in both the insertion and midsubstance. Future studies will investigate the different impacts of growth factor expression in tendon to bone healing