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High density sickle cell erythrocyte core membrane skeletons demonstrate slow temperature dependent dissociation.

We have previously demonstrated that slow dissociation of HDSS membrane skeletons in high ionic strength Triton X-100 buffer was related to a posttranslational modification in beta-actin, in which a disulfide bridge was formed between cysteine 284 and cysteine 373[Shartava et al: J Cell Bio 128:805, 1995]. These previous dissociation assays were limited to two homozygous (SS) sickle cell patients and a single temperature (37 degrees C). In the current work, we have expanded the SS subjects to 9 and have carried out dissociation assays at 0, 24, 30, 34, and 37 degrees C. At 0 degrees C there was limited dissociation of spectrin and actin from normal(AA), low density sickle cell(LDSS), and high density sickle cell (HDSS) core skeleton up to 24 hr. The first order rate constants for dissociation of spectrin, at 0 degrees C, was 0.030-0.035 x 10-4 sec-1 for AA,LDSS, and HDSS core skeletons. However at 24, 30, 34, and 37 degrees C the rate of dissociation of spectrin from HDSS core skeletons was significantly slower than the rate of dissociation from AA core skeletons. Having determined the first order rate constants for spectrin dissociation at these specified temperatures, we then asked whether dithiothreitol (DTT) would hasten the dissociation of core skeletons. The presence of DTT caused the rate of dissociation of the HDSS membrane skeleton to become statistically indistinguishable from the rate of dissociation of AA membrane skeletons. This is consistent with the suggestion that reversible thiol oxidation is responsible for the slow dissociation of the HDSS membrane skeleton.

Actins↗

Effects of ultrasonic skeletonization on internal thoracic and gastroepiploic arteries for coronary artery bypass grafting.

OBJECTIVE: There are few data available on the effect of ultrasonic skeletonization with the harmonic scalpel on internal thoracic artery (ITA) and gastroepiploic artery (GEA) vessel function. METHODS: Rings of segments of the skeletonized ITA, pedicled ITA, skeletonized GEA, and pedicled GEA were studied. Arterial segments were treated with high KCl and norepinephrine (NE) to obtain smooth muscle contractions. Endothelium-dependent and independent vasorelaxant potencies in 10(-6)mol/l NE-pre-constricted arteries were assessed by acetylcholine (ACh), and isosorbide dinitrate (ISDN) and diltiazem, respectively. RESULTS: There were no differences in contractile potencies induced by high KCl and NE between the rings cut from skeletonized and pedicled grafts. The rings from skeletonized and pedicled vessels also showed equal sensitivity to ISDN and diltiazem. However, the rings from pedicled grafts showed greater relaxation responses to ACh than rings from skeletonized grafts. CONCLUSION: Ultrasonic complete skeletonization with the harmonic scalpel may retain smooth muscle function of skeletonized grafts, whereas endothelial function of ultrasonic skeletonized grafts may be significantly compromised.

Acetylcholine↗

Conformation and elasticity of the isolated red blood cell membrane skeleton.

We studied the structure and elasticity of membrane skeletons from human red blood cells (RBCs) during and after extraction of RBC ghosts with nonionic detergent. Optical tweezers were used to suspend individual cells inside a flow chamber, away from all surfaces; this procedure allowed complete exchange of medium while the low-contrast protein network of the skeleton was observed by high resolution, video-enhanced differential interference-contrast (DIC) microscopy. Immediately following extraction in a 5 mM salt buffer, skeletons assumed expanded, nearly spherical shapes that were uncorrelated with the shapes of their parent RBCs. Judging by the extent of thermal undulations and by their deformability in small flow fields, the bending rigidity of skeletons was markedly lower than that of either RBCs or ghosts. No further changes were apparent in skeletons maintained in this buffer for up to 40 min at low temperatures (T less than 10 degrees C), but skeletons shrank when the ionic strength of the buffer was increased. When the salt concentration was raised to 1.5 M, shrinkage remained reversible for approximately 1 min but thereafter became irreversible. When maintained in 1.5 M salt buffer for longer periods, skeletons continued to shrink, lost flexibility, and assumed irregular shapes: this rigidification was irreversible. At this stage, skeletons closely resembled those isolated in standard bulk preparations. We propose that the transformation to the rigid, irreversibly shrunken state is a consequence of spectrin dimer-dimer reconnections and that these structural rearrangements are thermally activated. We also measured the salt-dependent size of fresh and bulk extracted skeletons. Our measurements suggest that, in situ, the spectrin tethers are flexible, with a persistence length of approximately 10 nm at 150 mM salt.

Elasticity↗

Skeletonization of bilateral internal thoracic artery grafts lowers the risk of sternal infection in patients with diabetes.

OBJECTIVE: Deep sternal wound infection is a dreaded complication of coronary artery bypass surgery, particularly in patients with diabetes. This study determines whether skeletonization of internal thoracic artery conduits compared with pedicled harvesting reduces the risk of deep sternal wound infection in patients with diabetes undergoing bilateral internal thoracic artery grafting. METHODS: We reviewed prospectively gathered data on all patients who have undergone coronary artery bypass grafting and received bilateral internal thoracic artery grafts at our institution since 1990. We compared patients with diabetes who received skeletonized (n = 79) versus conventional pedicled (n = 36) internal thoracic artery conduits. RESULTS: The proportion of patients taking insulin (19.0% vs 14.0% for skeletonized vs conventional grafts, respectively, P =.6) or oral hypoglycemic agents (68.4% vs 69.4%, P =.9), as well as the prevalence of type I diabetes (2.5% vs 8.3%, P =.18), were similar in both groups. Patients who received skeletonized grafts were more likely to receive a free rather than an in situ right internal thoracic artery graft (93.7% vs 30.6%, P <.001). The prevalence of deep sternal wound infection was significantly lower in patients who received skeletonized grafts compared with patients who received conventional grafts (1.3% vs 11.1%, P =.03). Patients in the skeletonized group were also less likely to develop any (superficial or deep) sternal wound infection postoperatively (5.1% vs 22.2%, P =.03). There was no significant difference in the prevalence of deep sternal wound infection between patients with diabetes who received skeletonized internal thoracic arteries and patients without diabetes who underwent conventional internal thoracic artery grafting (n = 578) (1.2% vs 1.6%, respectively, P =.8). CONCLUSIONS: Skeletonization of internal thoracic artery conduits lowers the risk of deep sternal wound infection in patients with diabetes undergoing bilateral internal thoracic artery grafting. We no longer consider diabetes a contraindication to bilateral internal thoracic artery grafting, provided the internal thoracic arteries are skeletonized.

Adult↗

Ultrastructure and immunocytochemistry of the isolated human erythrocyte membrane skeleton.

Isolated skeletons from human erythrocyte ghosts were studied using immunogold labeling; negative staining; and quick-freeze, deep-etch, rotary replication with Pt/C (QFDERR). Isolated skeletons visualized by QFDERR were similar to the negatively stained skeletons in that the proteins spectrin, actin, and ankyrin could be easily distinguished. However, the quick-frozen skeletons had two fewer filaments (4.2 +/- 0.7) at an actin junction. Immunogold labeling of skeletons with site-specific spectrin antibodies not only confirmed the designation of these filaments as spectrin molecules, but indicated that about 30% of spectrin filaments form non-actin junctions consistent with the hexameric organization of these filaments. Many of the filaments displayed a striking banding pattern indicative of underlying substructure. Isolated skeletons prepared by QFDERR also showed evidence of laterally associated spectrin filaments. These associations, as well as many hexamer junctions, are lost during negative staining. Negative staining also apparently caused approximately 21% of the spectrin filaments to separate into their monomeric subunits. These results indicate that the surface tension imposed during negative staining of isolated skeletons can cause a loss of interactions normally present in the intact membrane skeleton.

Erythrocyte Membrane↗

Study of human erythrocyte membrane protein interactions by selective solubilization of Triton-skeletons.

The membrane skeleton, responsible for shape and mechanical properties of the red cell, was purified by the Triton extraction procedure in presence of 5 mM, 150 mM or 600 mM NaCl. The proportion of spectrin, protein 4.1 and actin present in erythrocyte skeletons does not depend on the molarity of NaCl used. In contrast ankyrin, protein band 3 and protein 4.2 are removed from skeletons as the ionic strength increased. Solubilization assays of membrane skeletons were used to study protein interactions inside the skeleton. Solubilization was performed by Tris, a non-selective disruptive reagent, or by p-mercuribenzene sulfonic acid (PMBS), which principally release spectrin and actin. Tris action was assessed by calculation of the percentage of solubilized proteins, which increased proportionally with Tris molarity. PMBS action was kinetically determined as the decrease in skeleton turbidity. With these two reagents, we observed a lower dissociation of skeletons prepared with high ionic strength buffer. Erythrocyte pretreatment with okadaic acid, an inhibitor of serine-threonine phosphatases, revealed a phosphorylation-induced skeleton gelation and a better resistance to Tris-solubilization.

Actins↗

Skeletonization of gastroepiploic artery graft in off-pump coronary artery bypass grafting: early clinical and angiographic assessment.

BACKGROUND: Recently skeletonization has been recognized as an alternative to pedicle harvesting of the internal thoracic artery as a technique that increases the length and caliber size of the graft compared with pedicled internal thoracic artery grafts; however, this is not yet popular for harvesting the gastroepiploic artery (GEA). We report here our experience of skeletonized GEA grafting in off-pump coronary artery bypass grafting with early clinical and angiographic results. The purpose of this study was to evaluate skeletonization of GEA grafting in off-pump coronary artery bypass grafting with a large patient volume. METHODS: One hundred sixty-eight patients including 131 men and 37 women (mean age, 65 years; range, 45 to 87 years) underwent the skeletonized GEA grafting in off-pump coronary artery bypass grafting. These patients represent 41% (168 of 407 patients) of those who underwent off-pump coronary artery bypass grafting operations during the same period. We used the GEA graft of choice in patients with right coronary artery lesion. Skeletonization was performed in a unique manner we developed. RESULTS: There were no in-hospital deaths among the study patients. One patient had a perioperative myocardial infarction, which was considered a result of vasospasm of the GEA graft. None of the other patients had severe morbidity. The patency rate of the skeletonized GEA graft was 98.1% (151 of 154 distal anastomoses). CONCLUSIONS: This study suggests that skeletonization of the GEA graft can enlarge its caliber size and improve its flow capacity. In addition, the acceptable early clinical and angiographic outcome suggests that use of the skeletonized GEA graft in off-pump coronary artery bypass grafting surgery is safe and effective.

Aged↗

Endothelial integrity of ultrasonically skeletonized internal thoracic artery: morphological analysis with scanning electron microscopy.

OBJECTIVE: The skeletonized internal thoracic artery (ITA) has several advantages over a pedicled one in coronary artery bypass grafting (CABG). An ultrasonic scalpel makes ITA skeletonization easy and speedy, however, the ultrasonic energy that is transmitted to the artery itself can occasionally injure the endothelium. Therefore, the endothelial integrity of the ultrasonically skeletonized ITA is a major concern related to this technique. The purpose of this study is to assess the endothelial integrity of the ultrasonically skeletonized ITA. METHODS: We skeletonized the left ITA with an ultrasonic scalpel in nine patients who underwent CABG, and thereafter the terminal portion of this artery was subjected to a morphological study. The endothelial integrity of this artery was morphologically assessed using scanning electron microscopy, and the results were compared to that of the left ITA skeletonized with fine scissors. RESULTS: All ITA specimens showed a completely confluent endothelium, and no endothelial injury was observed by the scanning electron microscopic study. CONCLUSIONS: The skeletonization of the ITA with an ultrasonic scalpel had no deleterious effect on the endothelium. This morphological study confirmed the safety and the reliability of this technique, and we therefore recommend its clinical use in the skeletonization of the ITA for CABG.

Endothelium, Vascular↗

Skeletonization versus pedicle preparation of the radial artery with and without the ultrasonic scalpel.

BACKGROUND: The radial artery (RA) is increasingly used for myocardial revascularization because of its presumed advantageous long-term patency rates. The vessel can be harvested as a pedicle or skeletonized. The aim of this study was to compare the skeletonization technique with pedicle preparation using either an ultrasonic scalpel or scissors. METHODS: Forty consecutive patients with coronary artery disease undergoing complete arterial revascularization were included in the study. In 20 patients the RAs were prepared using scissors and clips (group 1: skeletonization; group 2: pedicle). In another 20 patients the arteries harvested were prepared using an ultrasonic scalpel (group 3: skeletonization; group 4: pedicle). The RA was treated with papaverine to prevent spasm of the vessel during and after harvesting. Tissue specimens of each RA were taken to analyze endothelial morphology by scanning electron microscopy. After implantation of the RA, graft perfusion was measured with a flow probe. RESULTS: Harvesting the RA as a skeletonized vessel took more time as compared with pedicle preparation (group 1 vs group 2: 37.1 +/- 3.5 minutes vs 24.4 +/- 3.9 minutes; p < 0.001 and group 3 vs group 4: 31.1 +/- 3.5 minutes vs 25.6 +/- 3.7 minutes; p < 0.01). The number of hemostatic titanium clips was similarly higher in group 1 as opposed to group 2 (58.7 +/- 7.1 vs 38.7 +/- 7.1; p < 0.01). However, there was no difference between groups 3 and 4 (p = 0.086). The length of the RA after skeletonization with scissors and clips was 20.8 +/- 1.5 cm in contrast with 19.1 +/- 0.9 cm (p < 0.01) after dissection as a pedicle. In the groups using the ultrasonic scalpel, there was no difference in graft length (p = 0.062). Mean blood flow through the graft after establishing the proximal anastomosis was similar among all groups (groups 1, 2, 3, and 4: 50 +/- 20.1 mL/min, 53.8 +/- 24.3 mL/min, 56.3 +/- 25.1 mL/min, and 51.8 +/- 23 mL/min, respectively). Scanning electron microscopy demonstrated endothelial damage in all patients in groups 1, 2, and 3 and in 7 patients of group 4. Most endothelial lesions were minor except in group 3 in which 1 of 5 endothelial lesions were severe. Statistically significant differences was found between groups 1 and 2, and 3 and 4 with respect to the degree of endothelial damage (p < 0.01). CONCLUSIONS: Skeletonization using scissors and clips is more time consuming and technically more difficult, but yield significantly longer grafts. Skeletonization with an ultrasonic scalpel did not result in additional length and was more frequently associated with severe endothelial damage. Pedicle preparation using scissors or an ultrasonic scalpel is much simpler and faster, and does not jeopardize endothelial integrity.

Humans↗

Ultrastructure of the intact skeleton of the human erythrocyte membrane.

Filamentous skeletons were liberated from isolated human erythrocyte membranes in Triton X-100, spread on fenestrated carbon films, negatively stained, and viewed intact and unfixed in the transmission electron microscope. Two forms of the skeleton were examined: (a) basic skeletons, stripped of accessory proteins with 1.5 M NaCl so that they contain predominantly polypeptide bands 1, 2, 4.1, and 5; and (b) unstripped skeletons, which also bore accessory proteins such as ankyrin and band 3 and small plaques of residual lipid. Freshly prepared skeletons were highly condensed. Incubation at low ionic strength and in the presence of dithiothreitol for an hour or more caused an expansion of the skeletons, which greatly increased the visibility of their elements. The expansion may reflect the opening of spectrin from a compact to an elongated disposition. Expanded skeletons appeared to be organized as networks of short actin filaments joined by multiple (5-8) spectrin tetramers. In unstripped preparations, globular masses were observed near the centers of the spectrin filaments, probably corresponding to complexes of ankyrin with band 3 oligomers. Some of these globules linked pairs of spectrin filaments. Skeletons prepared with a minimum of perturbation had thickened actin protofilaments, presumably reflecting the presence of accessory proteins. The length of these actin filaments was highly uniform, averaging 33 +/- 5 nm. This is the length of nonmuscle tropomyosin. Since there is almost enough tropomyosin present to saturate the F-actin, our data support the hypothesis that tropomyosin may determine the length of actin protofilaments in the red cell membrane.

Actin Cytoskeleton↗

Skeleton pruning by contour partitioning with discrete curve evolution.

In this paper, we introduce a new skeleton pruning method based on contour partitioning. Any contour partition can be used, but the partitions obtained by Discrete Curve Evolution (DCE) yield excellent results. The theoretical properties and the experiments presented demonstrate that obtained skeletons are in accord with human visual perception and stable, even in the presence of significant noise and shape variations, and have the same topology as the original skeletons. In particular, we have proven that the proposed approach never produces spurious branches, which are common when using the known skeleton pruning methods. Moreover, the proposed pruning method does not displace the skeleton points. Consequently, all skeleton points are centers of maximal disks. Again, many existing methods displace skeleton points in order to produces pruned skeletons.

Algorithms↗

In vitro reassembly of vesicular stomatitis virus skeletons.

Vesicular stomatitis virus (VSV) has been disrupted with nonionic detergent plus 0.5 M NaCl under conditions which result in solubilization of the viral glycoprotein (G), matrix protein (M), and lipids, leaving the nucleocapsid in a highly extended state. Dialysis of these suspensions to remove NaCl was found to result in reassociation of nucleocapsids with M protein. Reassociated structures were highly condensed and similar in appearance to "native" VSV skeletons produced by extraction of virions with detergent at low ionic strength. For instance, electron microscopic analysis revealed that, like "native" skeletons, "reassembled" skeletons were cylindrical in shape, with diameters in the range of 51.0 to 55.0 nm and cross-striations spaced approximately 6.0 nm apart along the length of the structure. Like native skeletons, reassembled skeletons were found by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to contain the viral N and M proteins, but they lacked the glycoprotein entirely. Both native and reassembled skeletons were found to be capable of in vitro RNA-dependent RNA synthesis (transcription). In vivo skeleton assembly required the presence of M protein and nucleocapsids. No skeleton-like structures were formed by dialysis of nucleocapsids in the absence of M protein or of M protein in the absence of nucleocapsids. These results provide strong support for the view that the VSV M protein plays a functional role in condensing the viral nucleocapsid in vitro and raise the possibility that it may play a similar role in vivo.

Capsid↗

Ontogenetic scaling of hydrostatic skeletons: geometric, static stress and dynamic stress scaling of the earthworm lumbricus terrestris

Soft-bodied organisms with hydrostatic skeletons range enormously in body size, both during the growth of individuals and in the comparison of species. Therefore, body size is an important consideration in an examination of the mechanical function of hydrostatic skeletons. The scaling of hydrostatic skeletons cannot be inferred from existing studies of the lever-like skeletons of vertebrates and arthropods because the two skeleton types function by different mechanisms. Hydrostats are constructed of an extensible body wall in tension surrounding a fluid or deformable tissue under compression. It is the pressurized internal fluid (rather than the rigid levers of vertebrates and arthropods) that enables the maintenance of posture, antagonism of muscles and transfer of muscle forces to the environment. The objectives of the present study were (1) to define the geometric, static stress and dynamic stress similarity scaling hypotheses for hydrostatic skeletons on the basis of their generalized form and function, and (2) to apply these similarity hypotheses in a study of the ontogenetic scaling of earthworms, Lumbricus terrestris, to determine which parameters of skeletal function are conserved or changed as a function of body mass during growth (from 0.01 to 8 g). Morphometric measurements on anesthetized earthworms revealed that the earthworms grew isometrically; the external proportions and number of segments were constant as a function of body size. Calculations of static stresses (forces per cross-sectional area in the body wall) during rest and dynamic stresses during peristaltic crawling (calculated from measurements of internal pressure and body wall geometry) revealed that the earthworms also maintained static and dynamic stress similarity, despite a slight increase in body wall thickness in segment 50 (but not in segment 15). In summary, the hydrostatic skeletons of earthworms differ fundamentally from the rigid, lever-like skeletons of their terrestrial counterparts in their ability to grow isometrically while maintaining similarity in both static and dynamic stresses.

Journal Article↗

Skeletonized gastroepiploic artery for off-pump coronary artery bypass grafting.

BACKGROUND: Skeletonized arterial grafting may reduce the risk of graft spasm and may improve graft patency. Previously we reported a pilot study of skeletonized gastroepiploic artery (GEA) grafting with favorable results. Skeletonized GEA harvesting with an ultrasonic scalpel has now become our routine procedure. In this report, we compare the early clinical outcomes of skeletonized versus pedicled GEA grafting to assess the safety and benefit of use of skeletonized GEA in coronary artery bypass grafting. METHODS: Between July 2002 and October 2003, the GEA was used as a conduit for isolated off-pump coronary artery bypass grafting in 105 patients. Of these, 21 patients (group P) received pedicled GEA and 59 patients (group S) received skeletonized GEA grafts (excluding 25 patients whose results were reported in the pilot study). The perioperative and early follow-up data were prospectively collected and compared. RESULTS: No graft injury was found in either group. The preoperative characteristics were similar in the two groups except that group S had a smaller body surface area (1.64 +/- 0.16 m 2 in group S versus 1.73 +/- 0.16 m 2 in group P, P <.05) and a significant number of patients with diabetes (36/59, 61.0% versus 7/21, 33.3%, P <.05). The number of distal anastomoses was 4.3 < 1.0 versus 3.9 +/- 0.9 ( P = not significant [NS]). An in situ GEA composite graft was constructed in 8 (13.6%) of the patients in group S and none of the patients in group P ( P = NS). There was one hospital death due to infection in group S. Otherwise, there were no cases of low output syndrome or postoperative myocardial infarction in either group. During early postoperative follow-up, no angina recurrence or myocardial infarction was found. CONCLUSION: The GEA can be skeletonized safely with an ultrasonic scalpel. Skeletonization enables a wider variety of choices in the use of GEA grafting.

Adult↗

Binding of mutagens by fractions of the cell wall skeleton of lactic acid bacteria on mutagens.

The binding effect of cells and cell fractions, cell wall skeleton, cytoplasm, whole cells, and cell wall skeleton treated by lysozyme and alpha-amylase at 37 degrees C for 5 h, on Trp-P-1 (3-amino-1,4-dimethyl-[5H]pyrido [4,3-b]indole) and Trp-P-2 (3-amino-1-methyl-[5H]-pyrido[4,3-b]indole) were investigated. The cell and cell wall skeleton of Streptococcus cremoris Z-25 had greater binding activity, but cytoplasm and extract of cell wall skeleton did not bind Trp-P-1 and Trp-P-2. When the cells or cell wall skeleton were treated with lysozyme and alpha-amylase, unbound Trp-P-1 and Trp-P-2 concentrations were greater than that of the untreated control. It is possible that cell walls may be involved in the binding of mutagenic pyrolyzates to lactic acid bacteria. The cell wall skeleton of S. cremoris Z-25, Lactobacillus acidophilus IFO 13951, and Bifidobacterium bifidum IFO 14252 showed binding of Trp-P-1, 2-amino-6-methyldipyrido(1,2-a:3',2'- d)imidazole, 2-amino-5-phenylpyridine, 2-amino-3-methylimidazo(4,5-f)quinoline, 2-amino-3,4-dimethylimidazo(4,5-f) quinoline, and 2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline. The cell wall skeleton of S. cremoris group and Streptococcus lactis also showed the binding activity with A N-nitrosodimethylamine. The binding of Trp-P-1 to cell walls was very high, and the binding of mutagenic pyrolyzates was variable with different bacterial species. The peptidoglycan complex and polysaccharides liberated from cell wall skeleton of S. cremoris Z-25 showed strong binding of Trp-P-2. Peptidoglycans has a binding effect of about 19.86 micrograms/mg; polysaccharides had a binding effect of 14 micrograms/mg.

Bifidobacterium↗

Supernumerary lumbar vertebrae in human skeletons at the Galloway Osteological Collection of Makerere University, Kampala.

OBJECTIVE: To find out the number of other skeletons in the Galloway collection that have a variation of six lumbar vertabrae. DESIGN: A descriptive cross-sectional study. SETTING: Department of Anatomy, Faculty of Medicine, Makerere University, Kampala. MATERIALS: Five hundred and ninety one skeletons in the Galloway Osteological Collection housed in the basement of the Department of Anatomy, Faculty of Medicine, Makerere University. INTERVENTIONS: The skeletons were examined. Those found to have six lumbar vertebrae were scrutinised further. A recount was done of the cervical and thoracic vertebrae in order to ascertain the identity and number of the lumbar vertebrae. Some of the skeletons were x-rayed and photographed to illustrate the finding and the manner in which the transverse processes of the sixth lumbar vertebrum articulated with the alar of the sacrum. RESULTS: Out of the 591 skeletons examined, twenty (4%) were found to have a sixth lumbar vertebrum. In seven (35%) of these, the transverse processes of the sixth lumbar vertebrum were found to be bilaterally articulated with the alar of th sacrum and in another five (25%) only the left was. In the remaining eight (40%), the transverse processes did not articulate with the alar of the sacrum at all. Right unilateral articulation between the transverse process and the alar of the sacrum was not observed in any of the twenty skeletons that had six lumbar vertebrae. CONCLUSION: In the Galloway osteological collection there is a small number of skeletons in which the number of lumbar vertabrae is six instead of 5 as is normally the case. In most of them the transverse processes of th sixth lumbar vertebrum articulate with the alar of the sacrum through a synovial joint (bilateral or unilateral). The above observations have raised several questions in the mind of the author regarding the mechanism of causation of the variation and its clinical and functional implications.

Anatomy↗

Stabilization and remodeling of the membrane skeleton during lens fiber cell differentiation and maturation.

Actin filaments are integral components of the plasma membrane-associated cytoskeleton (membrane skeleton) and are believed to play important roles in the determination of cell polarity, shape, and membrane mechanical properties, however the roles of actin regulatory proteins in controlling the assembly, stability, and organization of actin filaments in the membrane skeleton are not well understood. Tropomodulin is a tropomyosin and actin-binding protein that stabilizes tropomyosin-actin filaments by capping their pointed ends and is associated with the spectrin-actin membrane skeleton in erythrocytes, skeletal muscle cells, and lens fiber cells, a specialized epithelial cell type. In this study, we have investigated the role of tropomodulin and other membrane skeleton components in lens fiber cell differentiation and maturation. Our results demonstrate that tropomodulin is expressed concomitantly with lens fiber cell differentiation and assembles onto the plasma membrane only after fiber cells have begun to elongate and form apical-apical contacts with the undifferentiated epithelium. In contrast, other membrane skeleton components, spectrin, actin, and tropomyosin, are constitutively expressed and assembled on the plasma membranes of both undifferentiated and differentiated fiber cells. Tropomodulin, but not other membrane skeleton components, is also enriched at a novel structure at the apical and basal ends of newly elongated fiber cells at the fiber cell-epithelium and fiber cell-capsule interface, respectively. Once assembled, tropomodulin and its binding partners, tropomyosin and actin, remain membrane-associated and are not proteolyzed during fiber cell maturation and aging, despite proteolysis of alpha-spectrin and other cytoskeletal filament systems such as microtubules and intermediate filaments. We propose that actin filament stabilization by tropomodulin, coupled with partial proteolysis of other cytoskeletal components, represents a programmed remodeling of the lens membrane skeleton that may be essential to maintain plasma membrane integrity and transparency of the extremely elongated, long-lived cells of the lens. The unique localization of tropomodulin at fiber cell tips further suggests a new role for tropomodulin at cell-cell and cell-substratum contacts; this may be important for cell migration and/or adhesion during differentiation and morphogenesis.

Actins↗

The density of long limb bones and the percentage ash weight of the skeleton of Macaca mulatta.

The gravimetric density of humeri, radii, femora and tibiae from a series of 274 male and female skeletons of rhesus monkeys, Macaca mulatta, was determined for fetal, young and adult periods. The ages of 171 of the animals were known: they ranged from 57 days of gestation to 13.6 years; the ages of an additional 103 skeletons were estimated. The mean density of the fetal bones was found to increase linearly with age and was higher for males than females, and higher for the superior than for the inferior limb bones. During the young period the pattern of increase in density can be represented by a power-type curve, and the density is significantly higher in females than in males and in superior than in inferior limb bones. The densities of the long limb bones of the adult skeletons show a slight, but not significant, negative trend with increasing age. In this age group the mean densities are higher for males than females and higher for the superior than for the inferior limb bones. The percentage ash weight was determined for the total skeleton and for 21 subdivisions of 23 postnatal skeletons with estimated ages. The skull and long limb bones were found to have higher mean percentage ash weights than the vertebral segments and the sternum. Both the density and the percentage ash weight of the Macaca mulatta skeletons examined exceed those found in our earlier studies of the human skeleton.

Age Factors↗