Calcaneofibular ligament may act as a tensioner of peroneal tendons as revealed by a contactless three-dimensional scan system on cadavers.

The ligaments are believed to have a role in stabilizing joints and regulating joint motion. Here, we propose a novel function of the calcaneofibular ligament (CFL), which stabilizes the ankle joint. In human bipedal locomotion, the peroneal muscles maintain mediolateral stability and prevent involuntary ankle inversion. To investigate the functional relationship between the peroneal longus tendon (PLT), brevis tendon (PBT), and CFL, we quantitatively analyzed the positional changes of the tendons by using a contactless three-dimensional optical scan system. Eighteen cadaveric specimens were included in the study. Interestingly, with increased tension of the CFL, the tendons significantly moved toward the lateral direction (P < 0.001), compared with their position when the CFL was detached. The actual lift amount reached 2.0 ± 0.8 mm for the PLT and 1.9 ± 1.0 mm for the PBT. These results strongly suggest that a tensed CFL can lift the peroneal tendons and may act as a "tensioner" for the effective transmission of muscle contraction. This phenomenon contributes to postural control, especially in regaining balance on uneven terrain, and provides a new perspective for the exercise methods or understanding the ankle joint instability due to sprains.

Anatomical variation in the form of inter- and intra-individual laterality of the calcaneofibular ligament.

The lateral ligament complex of the ankle is involved in a large proportion of ankle sprains. The calcaneofibular ligament (CFL) is often involved in severe injuries. The purpose of this study was to evaluate the anatomical variation and laterality of the CFL to improve our understanding of the mechanisms of CFL-related injuries. This study utilized 110 paired ankles from 55 formalin-fixed Japanese cadavers (33 male and 22 female). The length and width of the CFL and the angle created by the CFL and long axis of the fibula (CF angle) were measured after exposing the CFL by careful dissection from the surrounding tissues. The results revealed that each parameter exhibited a wide range of values and showed unique patterns of frequency distribution, among which only the length was normally distributed. Among the parameters, only the CF angle showed no significant correlation with the other parameters. Analysis of laterality revealed that the mean left CF angle was significantly greater than the value on the opposite side (p < 0.05) and that the values of the bilateral CF angle showed no significant correlation at the individual level. The present results revealed not only detailed information regarding the CFL morphology, but also inter- and intra-individual laterality regarding the CFL traveling angle. It is likely that the differences in the quality and quantity of mechanical stress against each leg may have caused this morphologic laterality of the CFL.

Histopathology of tenosynovium in trigger fingers.

Stenosing flexor tenosynovitis, trigger finger, is a common clinical disorder causing painful locking or contracture of the involved digits, and most instances are idiopathic. This problem is generally caused by a size mismatch between the swollen flexor tendon and the thickened first annular pulley. Although hypertrophic pulleys have been histologically and ultrasonographically detected, little is known about the histopathology of the tenosynovium covering the tendons of trigger fingers. We identified chondrocytoid cells that produced hyaluronic acid in 23 (61%) fingers and hypocellular collagen matrix in 32 (84%) fingers around the tenosynovium among 38 specimens of tenosynovium from patients with trigger fingers. These chondrocytoid cells expressed the synovial B cell marker CD44, but not the chondrocyte marker S-100 protein. The incidence of these findings was much higher than that of conventional findings of synovitis, such as inflammatory infiltrate (37%), increased vascularity (37%), hyperplasia of synovial lining cells (21%), or fibrin exudation (5%). We discovered the following distinctive histopathological features of trigger finger: hyaluronic acid-producing chondrocytoid cells originated from fibroblastic synovial B cells, and a hypocellular collagen matrix surrounding the tenosynovium. Thus, an edematous extracellular matrix with active hyaluronic acid synthesis might increase pressure under the pulley and contribute to the progression of stenosis.

Posterior condylar canals and posterior condylar emissary veins-a microsurgical and CT anatomical study.

The posterior condylar canals (PCCs) and posterior condylar emissary veins (PCEVs) are potential anatomical landmarks for surgical approaches through the lateral foramen magnum. We conducted computed tomography (CT) and microsurgical investigation of how PCCs and PCEVs can aid in planning and performing these approaches. We analyzed the microanatomy of PCCs and PCEVs using cadaveric specimens, dry skulls, and CT images. The recognition frequency and geometry of PCCs and PCEVs and their relationships with surrounding structures were evaluated. PCCs were identified in 36 of 50 sides in dry bones and 82 of 100 sides by CT. PCCs had a 3.5-mm mean diameter and a 6.8-mm mean canal length. We classified their courses into four types according to intracranial openings: the sigmoid sinus (SS) type, the jugular bulb (JB) type, the occipital sinus type, and the anterior condylar emissary vein type. In most cases, PCEV originated near the boundary between the SS and JB. PCCs and PCEVs can be useful anatomical landmarks to differentiate the transcondylar fossa approach from the transcondylar approach, thus preventing unnecessary injury of the atlantooccipital joint. They can also be used as landmarks when the jugular foramen (JF) and hypoglossal canal (HGC) are being exposed. The area anterior to the brain stem and the medial part of HGC can be accessed by removal of the lateral foramen magnum medial to PCC. JF and the lateral part of HGC can be accessed by removal of the skull base lateral to PCC without damaging the lateral rim of the foramen magnum.

Generational differences in tooth size in the Japanese population: analysis of cohorts with a generation gap of four to five decades.

PURPOSE: Since early modern times, tooth size has reportedly been increasing in each successive generation. A detailed analysis of these trends can provide meaningful information for elucidating the origin of various problems caused by larger teeth, such as an abnormal dentition and occlusion. By using data from most recent generations, this study aimed to clarify the time course of changes in tooth size in the Japanese. MATERIALS AND METHODS: The dentitions of two Japanese cohorts comprising young individuals born in the 1980s and the 1990s were compared with those of another cohort of Japanese individuals born in the 1940s, approximately half a century earlier. The mesiodistal diameter of the tooth crowns was measured on plaster models and subjected to statistical analyses. RESULTS: A mean difference test revealed that each recent generation showed positive generational differences in the size of more than 50% of the tooth types. In addition, a deviation graph analysis indicated that the degree of change in tooth size varied with the tooth type or sampling site. Principal component analysis clearly showed an increase in tooth size on an individual basis in the more recent generations. CONCLUSIONS: This study revealed positive generational differences in tooth size in the Japanese population. The results may aid in understanding the development of abnormal dentitions and occlusion in recent Japanese populations.

Relationship between humeral geometry and shoulder muscle power among suspensory, knuckle-walking, and digitigrade/palmigrade quadrupedal primates.

Shoulder morphology is functionally related to different patterns of locomotion in primates. To investigate this we performed a quantitative analysis of the relationship between cortical bone thickness (Cbt) of the muscle/tendon attachment site on the humerus and physiological cross-sectional area (PCSA) of the shoulder muscle in primates with different locomotory habits. The deltoid, subscapularis, supraspinatus, and infraspinatus were investigated. A chimpanzee, a gibbon, a baboon, two species of macaque, a lutong, a capuchin, and a squirrel monkey were included in the study. The total length of the humerus was measured and the values were converted into three-dimensional reconstructed data on a computer by computed tomography. The Cbt values were obtained from the volumes divided by the areas of the muscle/tendon attachment sites of the humerus by computer analysis. Muscle mass, muscle fascicle length, and muscle pennation angle were measured and PCSA was calculated using these parameters. A relatively high Cbt and small PCSA were characteristic of the gibbon. The gibbon's high Cbt suggests that passive tension in the muscle/tendon attachment site of suspensory primates (brachiators) may be greater than that of quadrupedal primates, whereas the relatively small PCSA indicates an association with a large amount of internal muscle fascia to endure the passive stress of brachiation. Although chimpanzees undertake some suspensory locomotion, the results for this species resemble those of the digitigrade/palmigrade quadrupedal primates rather than those of the suspensory primate. However, the deltoid and subscapularis in chimpanzee differ from those of the other primates and appear to be affected by the peculiar locomotion of knuckle-walking, i.e. the moment arm of forelimb in chimpanzees is relatively longer than that of digitigrade/palmigrade quadrupedal primates. Hence, a large PCSA in the deltoid and subscapularis may contribute to sustaining the body weight during locomotion. On the other hand, a thin cortical bone relative to a large PCSA was a feature of the cercopithecids, indicating that digitigrade/palmigrade quadrupedal locomotion produces less tension at the muscle/tendon attachment sites compared with that produced by brachiators.

Effects of the femoral offset and the head size on the safe range of motion in total hip arthroplasty.

The purpose of this study was to quantify the effects of femoral offset and head size on range of motion (ROM) after total hip arthroplasty. Modular prostheses were implanted into 11 cadaveric hips using a posterolateral approach and tested for ROM with 3 different offsets and 5 different femoral head sizes. Increasing the femoral offset to 4 and 8 mm resulted in 21.1 degrees and 26.7 degrees of improved flexion, and 13.7 degrees and 21.2 degrees of improved internal rotation, respectively. The ROM improved in a head size-dependent manner primarily because of increasing the jumping distance of the femoral head rather than delaying any impingement. In contrast, the effectiveness of femoral offset was driven by delayed osseous impingement.

Age-related NADPH-diaphorase positive bodies in the lumbosacral spinal cord of aged rats.

In the course of a morphological investigation of age-related changes in the rat spinal cord, using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry, we found abundant NADPH-d positive bodies, which were characteristically expressed in the aged lumbosacral spinal cord. Together with a normally stained fiber network and a few neurons, the dense, spheroidal NADPH-d positive bodies occurred in portions of the sacral dorsal spinal cords, such as the dorsal commissural nucleus, intermediolateral nuclei, and superficial dorsal horn, and were scattered throughout the dorsal white column. These NADPH-d positive bodies were occasionally observed in a fibrous structure. Two morphologically distinctive subsets of NADPH-d positive bodies were noted in the spinal cord of rats aged 8 to 36 months: 1) highly-dense spheroidal shapes with sharp edges; 2) moderately-dense spheroidal or multiangular shapes with a central "core" and a peripheral "halo". The quantitative analysis, particularly the stereological measurement, confirmed a gradual increase in the incidence and size of NADPH-d positive bodies with increasing age. With nNOS immunohistochemistry, no corresponding structures to NADPH-d positive bodies were detected in aged rats; thus NADPH-d activity is not always specific to the NO-containing neural structures. The major distribution of the NADPH-d positive bodies in the aged lumbosacral spinal cord indicates some anomalous changes in the neurite, which might account for a disturbance in the aging pathway of the autonomic and sensory nerve in the pelvic visceral organs.

The spatiotemporal characterization of endplate reoccupation, with special reference to the superposition patterns of the presynaptic elements and the postsynaptic receptor regions during muscle reinnervation.

In this study, an immunohistochemical investigation was carried out to define spatiotemporal characteristics of superposition patterns of the presynaptic elements and the postsynaptic acetylcholine receptor (AChR) sites during the period of endplate regeneration after sciatic nerve crush. The extent of close correspondence of terminal Schwann cell (TSC)-, or axon terminal-, apposing AChR sites was quantitated with three-dimensional images of neuromuscular junctions (NMJs) taken under confocal laser-scanning microscopy. After 3-weeks post-crush (wpc), reoccupation of regenerating TSCs and later arriving axon terminals proceeded within the scope of previously denervated AChR plaques. During this period, the areas of presynaptic elements and the areas of postsynaptic elements were highly correlated. TSCs rapidly reoccupied a greater part of the postsynaptic receptors. In contrast, there was a slower increase of the contact areas of AChR sites overlapped by the axon terminals. Reoccupation by the presynaptic elements at 20 wpc was almost completed in a majority of NMJs, but some anomalous changes still continued to occur in a small proportion of the NMJs (20-30%). Our results suggest that: (a) with gradual increase of the contact areas between presynaptic and postsynaptic elements, imperfect reinnervation and regeneration, due to spatial mismatching or unbalanced growth between presynaptic and postsynaptic elements, result in sporadic remodeling; (b) the difference in superposition patterns between TSCs and axon terminals depends on the ability of making alignment to the endplate gutters in regenerating NMJs; and (c) a complex set of anatomical relationships among the three endplate components affects the process of endplate reoccupation synthetically.

N-myc downstream-regulated gene 1 expression in injured sciatic nerves.

N-myc downstream-regulated gene 1 (NDRG1)/RTP/Drg1/Cap43/rit42/TDD5/Ndr1 is expressed ubiquitously and has been proposed to play a role in growth arrest and cell differentiation. A recent study showed that mutation of this gene is responsible for hereditary motor and sensory neuropathy-Lom. However, the role of this gene in the peripheral nervous system is not fully understood. In our study, rabbit polyclonal antibodies were raised against this gene product and were used to examine changes in its expression over the time course of Wallerian degeneration and ensuing regeneration after crush injury of mouse sciatic nerves. Fluorescent immunohistochemistry showed that NDRG1 was expressed over the intact nerve fibers. Double labeling with a Schwann cell (SC) marker, S-100 protein (S-100), revealed that NDRG1 was localized in the cytoplasm of S-100-positive Schwann cells (SCs). NDRG1 expression was maintained in the early stage of myelin degradation but was then markedly depleted at the end stage of myelin degradation when frequent occurrence of BrdU-labeled SCs was observed (at 7-9 days). The depletion of NDRG1 at this time point was also confirmed by Western blotting analysis. NDRG1 expression finally recovered at the stage of remyelination, with immunoreactivity stronger than that in intact nerves. These findings suggest that NDRG1 may play an important role in the terminal differentiation of SCs during nerve regeneration.

Expression of amyloid precursor protein-like molecule in astroglial cells of the subventricular zone and rostral migratory stream of the adult rat forebrain.

In adult mammals, new neurons in the subventricular zone (SVZ) of the lateral ventricle (LV) migrate tangentially through the rostral migratory stream (RMS) to the olfactory bulb (OB), where they mature into local interneurons. Using a monoclonal antibody for the beta-amyloid precursor protein (APP) (mAb 22C11), which is specific for the amino-terminal region of the secreted form of APP and recognizes all APP isoforms and APP-related proteins, immunoreactivity was detected in specific subpopulations of cells in the SVZ and RMS of the adult rat forebrain. In the SVZ, APP-like immunoreactivity was detected in the ependymal cells lining the LV and some of the subependymal cells. The latter were regarded as astrocytes, because they were positive for the glial markers, S-100 protein (S-100) and glial fibrillary acidic protein (GFAP). APP-like immunoreactive astrocytes exhibited strong labelling of the perinuclear cytoplasm and often possessed a long, fine process similar to that found with radial glia. The process extended to an APP-like immunoreactive meshwork in the RMS that consisted of cytoplasmic processes of astrocytes forming 'glial tubes'. Double-immunofluorescent labelling with a highly polysialylated neural cell adhesion molecule (PSA-NCAM) confirmed that the APP-like immunoreactive astrocytes in the SVZ and meshwork in the RMS made close contact with PSA-NCAM-immunopositive neuroblasts, suggesting an interaction between APP-containing cells and neuroblasts. This region of the adult brain is a useful in vivo model to investigate the role of APP in neurogenesis.

Morphological basis of nitric oxide production and its correlation with the polysialylated precursor cells in the dentate gyrus of the adult guinea pig hippocampus.

Neurogenesis in the hippocampus persist throughout life and precursors of neurons reside in the granule cell layer of the dentate gyrus. Until now, the role of nitric oxide (NO) in the phenomenon has been unclear. By using specific antibodies and a confocal laser scanning microscope, the localization of NO synthase (NOS) was examined in the dentate gyrus of the adult guinea pig in relation with the neuronal precursor marker highly polysialylated neural cell adhesion molecule (PSA-N-CAM). Observation of single immunolabeled sections has revealed that both the PSA-N-CAM- and most NOS-positive cells were localized in the granule cell layer of the dentate gyrus. The former were small in size and showed a punctate, clustered immunoreaction with an irregular cellular margin, whereas the latter showed somewhat diverse cellular profiles. Some NOS-positive neurons had elliptical-like morphology with elongated dendrites, whereas others were small, irregularly shaped and mostly lacking dendritic spines. Double immunolabeling has revealed that NOS-immunoreactivity intermingled, as well as colocalized, with that of PSA-N-CAM, particulary in the granule cell layer. The doubly stained cells were morphologically indistinguishable from PSA-N-CAM single positive cells. These results not only suggest the role of NO production in adult hippocampal neurogenesis, but also indicate that some PSA-N-CAM-expressing neuronal precursors produce NO.

Molecular cloning of two guinea pig liver gap junction proteins, connexin32 and connexin26.

Intercellular coupling of hepatocytes through gap junctions facilitates exchange of small metabolites or ions, and contributes to maintenance of tissue homeostasis. As protein constituents of the liver gap junction channels, connexin32 (Cx32) and Cx26 have been identified. By use of rat cDNA probes, we cloned cDNAs for guinea pig homologs of Cx32 and Cx26, and compared their amino acid sequences with those of other species. The deduced primary structure of guinea pig Cx32 was 283 amino acids long and contained 98% identical amino acids to the rat and human Cx32. Only six amino acid exchanges were detected between the guinea pig and rat Cx32. On the contrary, the deduced amino acid sequence of guinea pig Cx26 (226 amino acids long) was 91 and 89% identical to the rat and human Cx26, respectively. Twenty-one amino acid exchanges were found between the guinea pig and rat, and the divergence was mostly located in cytoplasmic domains of Cx26. These results suggest that Cx26 shows structural diversity between species, while Cx32 is highly conserved.