Microanatomy Around the Facial Nerve Pathway for Microvascular Decompression Surgery Investigated with Correlative Light Microscopy and Block-Face Imaging.

BACKGROUND: Microvascular decompression for hemifacial spasm is performed at the root exit zone. More proximal segments of the facial nerve, defined as the root emerging zone (REmZ), may also be susceptible to neurovascular compression. Consequently, detailed knowledge of the microanatomy around facial nerve fibers at the pontomedullary junction is essential for consistent success of microvascular decompression. METHODS: Five human brainstems obtained from cadavers were investigated using correlative light microscopy and block-face imaging, which obtains arbitrary two-dimensional light microscopic and three-dimensional volume data from a single specimen. The entire facial nerve pathway, including the myelin transition, was evaluated inside and outside the brainstem. RESULTS: Correlative light microscopy and block-face imaging showed the intra-brainstem facial nerve fibers emerging at the brainstem surface deep at the pontomedullary sulcus (REmZ) and coursing along the pontine surface before detaching from the pons (root exit zone). An acute emerging angle significantly increased the surface area with central myelin. The facial nerve bundle formed 1 fasciculus in the portion covered by central myelin but divided into 2 fasciculi in the myelin transitional portion and then into multiple fasciculi more distally. Arteries around the REmZ were often anchored by perforating branches entangled with lower cranial nerves. CONCLUSIONS: Facial nerve fibers are susceptible to vascular compression on emerging onto the deep brainstem surface at the pontomedullary sulcus. The key procedure in microvascular decompression is full dissection of the lower cranial nerves down to the brainstem origin to explore both the root exit zone and the REmZ.

Elucidation of the Anatomical Mechanism of Nodal Skip Metastasis in Superficial Thoracic Esophageal Squamous Cell Carcinoma.

BACKGROUND: Lymph node metastasis (LNM) is a standard mechanism of cancer progression in esophageal squamous cell carcinoma (ESCC). We aimed to clarify the anatomical mechanism of skip nodal metastasis to mediastinal zones by analyzing the relationship between LNM to sentinel zones and lymphatic vessel counts in the muscle layer adjacent to the outer esophagus. METHODS: We examined the surgical records of 287 patients with ESCC who underwent potentially curative surgery (three-field lymphadenectomy) and whole esophagi, including pharynges and stomachs from 10 cadavers, to determine the number of lymphatic vessels in the intra-outer longitudinal muscle layer adjacent to the outer esophagus of the cervical (Ce), upper thoracic, middle thoracic (Mt), lower thoracic (Lt), and abdominal esophagi (Ae). RESULTS: The frequency of LNM to the middle mediastinal and supraclavicular zones, including the Mt and Ce, respectively, was lower than to the upper and lower mediastinal and abdominal zone in patients with superficial and advanced thoracic ESCC. In cadavers, the lymphatic vessel counts of the intra-outer longitudinal muscle layer in the Mt and Ce were significantly lower than those of the Lt and Ae, suggesting that lymphatic flow toward the outside of the Mt and Ce was not more abundant than to other sites. CONCLUSION: Our anatomical data suggested that the absence of intra-muscle lymphatic vessels in the middle mediastinal and supraclavicular zones causes skip LNM in patients with thoracic ESCC. Thus, standard esophagectomy with lymph node dissection, including distant zones, may be appropriate for treating patients with superficial thoracic ESCC.

Loss of VAMP5 in mice results in duplication of the ureter and insufficient expansion of the lung.

BACKGROUND: Vesicle-associated membrane protein 5 (VAMP5) is a member of the SNARE protein family, which regulates the docking and fusion of membrane vesicles within cells. Previously, we reported ubiquitous expression of VAMP5 proteins in various organs except the brain and small intestine. However, the precise roles of VAMP5 in each organ remain unclear. To explore the roles of VAMP5 in vivo, we generated VAMP5 knockout (KO) mice. RESULTS: VAMP5 KO mice showed low birth rate and low body weight. KO embryos grew normally in the uterus, and tended to die around birth. Anatomical analysis revealed that viable KO mice often exhibited duplication of the ureter, and dead KO mice showed insufficient expansion of the lung. VAMP5 was localized in the epithelial cells of the ureter and terminal bronchiole. CONCLUSIONS: VAMP5 KO mice showed a low birth rate and abnormalities of the urinary and respiratory systems. VAMP5 KO mice died around birth, possibly due to defects in vesicoureteral flow and breathing. The results presented could provide a basis for future studies to understand the roles of VAMP5 protein. Developmental Dynamics 247:754-762, 2018. © 2018 Wiley Periodicals, Inc.

A novel imaging method for correlating 2D light microscopic data and 3D volume data based on block-face imaging.

We have developed an imaging method designated as correlative light microscopy and block-face imaging (CoMBI), which contributes to improve the reliability of morphological analyses. This method can collect both the frozen sections and serial block-face images in a single specimen. The frozen section can be used for conventional light microscopic analysis to obtain 2-dimensional (2D) anatomical and molecular information, while serial block-face images can be used as 3-dimensional (3D) volume data for anatomical analysis. Thus, the sections maintain positional information in the specimen, and allows the correlation of 2D microscopic data and 3D volume data in a single specimen. The subjects can vary in size and type, and can cover most specimens encountered in biology. In addition, the required system for our method is characterized by cost-effectiveness. Here, we demonstrated the utility of CoMBI using specimens ranging in size from several millimeters to several centimeters, i.e., mouse embryos, human brainstem samples, and stag beetle larvae, and present successful correlation between the 2D light microscopic images and 3D volume data in a single specimen.

The correlation between femoral sulcus morphology and osteoarthritic changes in the patello-femoral joint.

PURPOSE: The purpose of this study was to reveal the correlation between the type of lesion and the depth of osteoarthritic (OA) changes in the patello-femoral (PF) joint and its bony morphological characteristics using computed tomography (CT) data. METHODS: Eighty-seven cadaveric knees were included in this study with median age of 83 years (62-97). OA depth evaluation was performed following Outerbridge's classification. Patella OA lesions were classified macroscopically using Han's method: type (1) no or minimal lesion, type (2) medial facet lesion without involvement of the ridge, type (3) lateral facet lesion without involvement of the ridge, type (4) lesion involving the ridge only, type (5) medial facet lesion with involvement of the ridge, type (6) lateral facet lesion with involvement of the ridge, and type (7) global lesion. Femoral-side OA lesions in the PF joint were classified using a modified Chang's method. Type (1) no or minimal lesion, type (2) medial facet lesion, type (3) centre of patella groove lesion, type (4) lateral facet lesion, and type (5) global lesion. Whole-body CTs of all cadavers were taken before knee dissection. Using the CT data, patella morphology was evaluated following Wiberg's classification. Femoral sulcus angle (SA), sulcus depth (SD), and sulcus width (SW) were also measured using CT data. RESULTS: The measured SA, SD, and SW were 144.8° ± 7.2°, 7.0 ± 1.6 mm and 3.4 ± 0.3 mm, respectively. When patella OA depth was divided into grades 1-2 (n = 30) and grades 3-4 (n = 57), the SD of grade 1-2 knees was 6.5 ± 1.3 mm, and the SD of grade 3-4 knees was 7.3 ± 1.6 mm, constituting a significant difference (p = 0.01). No significant difference in either SA or SW was observed between the two groups. Patella OA lesion, femoral-side OA lesion, and depth were not affected by SA, SD, or SW. Wiberg's classification also showed no significant correlation with PF-OA. CONCLUSION: Deep SD was significantly correlated with the incidence of severe patella OA. Wiberg's classification, SA, and SW were not correlated with PF-OA. For clinical relevance, there is a risk of PF-OA progression in patients with deep SD, and treatment should be applied accordingly.

Analysis of human cytomegalovirus replication in primary cultured human corneal endothelial cells.

BACKGROUND/AIMS: Since the first case of human cytomegalovirus (HCMV)-induced corneal endotheliitis in which HCMV DNA was detected from the patient's aqueous humour using PCR, the clinical evidence for HCMV endotheliitis has been accumulating. However, it remains to be confirmed whether HCMV can efficiently replicate in corneal endothelial cells. We, therefore, sought to determine whether primary cultured human corneal endothelial cells (HCECs) could support HCMV replication. METHODS: Human foreskin fibroblasts (HFFs) have been shown to be fully permissive for HCMV replication, and are commonly used as an in vitro model for HCMV lytic replication. Therefore, primary cultured HCECs or HFFs were infected with the vascular endotheliotropic HCMV strain TB40/E or laboratory strain Towne. We then compared viral mRNA and protein expression, genome replication and growth between the TB40/E-infected and Towne-infected HCECs and HFFs. RESULTS: When HCECs were infected with TB40/E or Towne, rounded cells resembling owl's eyes as well as viral antigens were detected. Viral mRNA synthesis and protein expression proceeded efficiently in the HCECs and HFFs infected with TB40/E or Towne at a high multiplicity of infection (MOI). Similarly, the viral genome was also effectively replicated, with UL44--a viral DNA polymerase processivity factor--foci observed in the nuclei of HCECs. HCECs produced a substantial number of infectious virions after infection with TB40/E at both a high and low MOI. CONCLUSIONS: Primary cultured HCECs could efficiently support HCMV replication after infection at both a high and low MOI.

A cadaveric study of the cervical nerve roots and spinal segments.

PURPOSE: The purpose of this study was to anatomically measure the width of the cervical nerve root and spinal cord segment in addition to clarifying the anatomical characteristics of the cervical nerve root. METHODS: We assessed 132 cervical nerve roots obtained from 11 cadavers. A total of 11 cervical spines from C3 to C8 were directly evaluated using digital calipers. The patients from whom the cadaveric specimens were obtained ranged from 79 to 90 years of age at the time of death. Four measurements were taken: the width at the entry of the spinal nerve in the vertebral foramen (WE), the maximum width of the spinal nerve (MW) and the length of the spinal segment on the ventral (LV) and dorsal rootlets (LD). RESULTS: The mean values of the WE from C3 to C8 were 5.5, 5.6, 6.0, 5.8, 4.8 and 4.3 mm, respectively. The value of C8 was significantly smaller than that of C3, C4, C5 and C6. The mean values of the MW from C3 to C8 were 5.6, 6.0, 6.4, 6.7, 6.3 and 6.0 mm, respectively. The mean values of the LV from C3 to C8 were 12.1, 12.5, 12.6, 12.7, 11.8 and 10.6 mm, respectively. The value of C8 was significantly narrower than that of C4, C5 and C6. The mean values of the LD from C3 to C8 were 12.1, 13.3, 13.6, 12.2, 11.0 and 10.6 mm, respectively. The value of C8 was significantly narrower than that of C4 and C5. CONCLUSIONS: We anatomically measured the width of cervical nerve roots and spinal segments. The spinal segment of C8 was significantly narrower than some of the roots located in the middle of the cervical spine, and this characteristic continued to the entry of the root in the vertebral foramen, although the difference disappeared at the maximum width point of the root.

Organization of organelles and VAMP-associated vesicular transport systems in differentiating skeletal muscle cells.

Vesicular transport plays an important role in the regulation of cellular function and differentiation of the cell, and intracellular vesicles play a role in the delivery of membrane components and in sorting membrane proteins to appropriate domains in organelles and the plasma membrane. Research on vesicular transport in differentiating cells has mostly focused on neurons and epithelial cells, and few such studies have been carried out on skeletal muscle cells. Skeletal muscle cells have specialized organelles and plasma membrane domains, including T-tubules, sarcoplasmic reticulum, neuromuscular junctions, and myotendinous junctions. The differentiation of skeletal muscle cells is achieved by multiple steps, i.e., proliferation of myoblasts, formation of myotubes by cell-cell fusion, and maturation of myotubes into myofibers. Systematic vesicular transport is expected to play a role in the maintenance and development of skeletal muscle cells. Here, we review a map of the vesicular transport system during the differentiation of skeletal muscle cells. The characteristics of organelle arrangement in myotubes are described according to morphological studies. Vesicular transport in myotubes is explained by the expression profiles of soluble N-ethylmaleimide-sensitive factor attachment protein receptor proteins.

Filamentous structures in skeletal muscle: anchors for the subsarcolemmal space.

In skeletal muscle fibers, intermediate filaments and actin filaments provide structural support to the myofibrils and the sarcolemma. For many years, it was poorly understood from ultrastructural observations that how these filamentous structures were kept anchored. The present study was conducted to determine the architecture of filamentous anchoring structures in the subsarcolemmal space and the intermyofibrils. The diaphragms (Dp) of adult wild type and mdx mice (mdx is a model for Duchenne muscular dystrophy) were subjected to tension applied perpendicular to the long axis of the muscle fibers, with or without treatment with 1% Triton X-100 or 0.03% saponin. These experiments were conducted to confirm the presence and integrity of the filamentous anchoring structures. Transmission electron microscopy revealed that these structures provide firm transverse connections between the sarcolemma and peripheral myofibrils. Most of the filamentous structures appeared to be inserted into subsarcolemmal densities, forming anchoring connections between the sarcolemma and peripheral myofibrils. In some cases, actin filaments were found to run longitudinally in the subsarcolemmal space to connect to the sarcolemma or in some cases to connect to the intermyofibrils as elongated thin filaments. These filamentous anchoring structures were less common in the mdx Dp. Our data suggest that the transverse and longitudinal filamentous structures form an anchoring system in the subsarcolemmal space and the intermyofibrils.

Reconfirmation of the anatomy of the left triangular ligament and the appendix fibrosa hepatis in human livers, and its implication in abdominal surgery.

BACKGROUND: The aim of the present study was to clarify the anatomy between the left triangular ligament (LTL) and the appendix fibrosa hepatis (AFH) in order not to sever the AFH when dissecting the LTL. METHODS: Totals of 43 and 27 cadaveric livers were examined macroscopically and histologically, respectively. RESULTS: The LTL attached itself to the diaphragmatic surface of the AFH through almost all lengths of the AFH. This might be the reason why AFH is so often dissected together with the LTL. There were two types of relation between the LTL and the AFH; in one type, the starting point of the LTL existed on the left liver and in the other type, it was on the AFH. Twenty-five of 27 AFH included remnants of the bile duct and 12 of 25 AFH had comparatively large bile ducts, which was unexceptionally accompanied by the well-developed peribiliary vascular plexus. AFH showed a variety of shapes, such as rectangular (6/43), long triangular (4/43), short triangular (7/43), triangular plus cordlike (11/43), cordlike (12/43) and bifurcated (3/43) types. CONCLUSIONS: As AFH sometimes includes relatively large bile ducts, it is recommended for surgeons to sever the AFH not just simply by electrocautery but by ligating its stump securely.

An integrated teaching method of gross anatomy and computed tomography radiology.

It is essential for medical students to learn and comprehend human anatomy in three dimensions (3D). With this in mind, a new system was designed in order to integrate anatomical dissections with diagnostic computed tomography (CT) radiology. Cadavers were scanned by CT scanners, and students then consulted the postmortem CT images during cadaver dissection to gain a better understanding of 3D human anatomy and diagnostic radiology. Students used handheld digital imaging and communications in medicine viewers at the bench-side (OsiriX on iPod touch or iPad), which enabled "pixel-to-tissue" direct comparisons of CT images and cadavers. Students had lectures and workshops on diagnostic radiology, and they completed study assignments where they discussed findings in the anatomy laboratory compared with CT radiology findings. This teaching method for gross and radiological anatomy was used beginning in 2009, and it yielded strongly positive student perspectives and significant improvements in radiology skills in later clinical courses.

Vesicular transport system in myotubes: ultrastructural study and signposting with vesicle-associated membrane proteins.

Myofibers have characteristic membrane compartments in their cytoplasm and sarcolemma, such as the sarcoplasmic reticulum, T-tubules, neuromuscular junction, and myotendinous junction. Little is known about the vesicular transport that is believed to mediate the development of these membrane compartments. We determined the locations of organelles in differentiating myotubes. Electron microscopic observation of a whole myotube revealed the arrangement of Golgi apparatus, rough endoplasmic reticulum, autolysosomes, mitochondria, and smooth endoplasmic reticulum from the perinuclear region toward the end of myotubes and the existence of a large number of vesicles near the ends of myotubes. Vesicles in myotubes were further characterized using immunofluorescence microscopy to analyze expression and localization of vesicle-associated membrane proteins (VAMPs). VAMPs are a family of seven proteins that regulate post-Golgi vesicular transport via the fusion of vesicles to the target membranes. Myotubes express five VAMPs in total. Vesicles with VAMP2, VAMP3, or VAMP5 were found near the ends of the myotubes. Some of these vesicles are also positive for caveolin-3, suggesting their participation in the development of T-tubules. Our morphological analyses revealed the characteristic arrangement of organelles in myotubes and the existence of transport vesicles near the ends of the myotubes.

Commonly used ACL autograft areas do not correlate with the size of the ACL footprint or the femoral condyle.

PURPOSE: The purpose of this study was to reveal the correlation between the size of the native anterior cruciate ligament (ACL) footprint and the area of commonly used autografts using cadaveric knees. METHODS: Twenty-Four non-paired human cadaver knees were used. The size of the femoral and tibial ACL footprints, length of Blumensaat's line, and the height and area of the lateral wall of the femoral intercondylar notch were photographed and measured with Image J software (National Institution of Health). Simulating an semitendinosus tendon (ST) graft, the ST was cut in half. The bigger half was regarded as the antero-medial (AM) bundle, and the remaining half was regarded as the postero-lateral (PL) bundle. Simulating an semitendinosus and gracilis (ST-G) graft, the bigger half of the ST and G was regarded as the AM bundle, and the smaller half of the ST was regarded as the PL bundle. Each graft diameter was measured, and the graft area was calculated. Simulating a bone-patella tendon-bone (BPTB) graft, a 10-mm wide BPTB graft was harvested and the area calculated. RESULTS: The sizes of the native femoral and tibial ACL footprints were 72.3 ± 24.4 and 134.1 ± 32.4 mm(2), respectively. The length of Blumensaat's line, and the height and area of the lateral wall of the femoral intercondylar notch were 29.5 ± 2.5 mm, 17.7 ± 2.3 mm, and 400.9 ± 62.6 mm(2), respectively. The average areas of the ST, ST-G, and BPTB graft were 52.7 ± 6.3, 64.7 ± 7.6, and 37.1 ± 7.5 mm(2). Both the height and the area of the lateral wall of the femoral intercondylar notch were significantly correlated with the femoral size of the ACL footprint (p = 0.007 and 0.008, respectively). However, no significant correlation was observed between ACL footprint size and autograft size. No significant correlation was observed between autograft size and the size of the lateral wall of the femoral intercondylar notch. CONCLUSION: In ACL reconstruction, if the reconstructed ACL size is determined by the harvested autograft size alone, native ACL size and anatomy are unlikely to be reproduced.

Evaluation of ACL mid-substance cross-sectional area for reconstructed autograft selection.

PURPOSE: The purpose of this study was to compare the size of the native ACL mid-substance cross-sectional area and the size of commonly used autografts. Hypothesis of this study was that the reconstructed graft size with autografts would be smaller than the native ACL size. METHODS: Twelve non-paired human cadaver knees were used. The ACL was carefully dissected, and the mid-substance of the ACL was cross-sectioned parallel to the articular surface of the femoral posterior condyles at 90 degrees of knee flexion. The size of the cross-sectional area of the ACL, and the femoral and tibial footprints were measured using Image J software (National Institute of Health). The semitendinosus tendon (ST) and the gracilis (G) tendon were harvested and prepared for ACL grafts. Simulating an ST graft, the ST was cut in half. The bigger half was regarded as the antero-medial (AM) bundle, and the remaining half was regarded as the postero-lateral (PL) bundle. Simulating an ST-G graft, the bigger half of the ST and G were regarded as the AM bundle, and the smaller half of the ST was regarded as the PL bundle. Each graft diameter was measured, and the graft area was calculated. Simulating a rectangular bone-patella tendon-bone (BPTB) graft, a 10-mm-wide BPTB graft was harvested and the area calculated. RESULTS: The sizes of the ACL mid-substance cross-sectional area, femoral and tibial ACL footprint were 46.9 ± 18.3, 60.1 ± 16.9 and 123.5 ± 12.5 mm(2), respectively. The average areas of the ST, ST-G, and BPTB grafts were 52.0 ± 3.8, 64.4 ± 6.2, and 40.8 ± 6.7 mm(2), respectively. The ST and BPTB grafts showed no significant difference in graft size when compared with the ACL cross-sectional area. CONCLUSION: ST and BPTB autografts were able to reproduce the native size of the ACL mid-substance cross-sectional area. The ST-G graft was significantly larger than the ACL cross-sectional area. For clinical relevance, ST and BPTB grafts are recommended in order to reproduce the native size of the ACL in anatomical ACL reconstruction with autograft.

Anterior cruciate ligament deterioration correlates with patella osteoarthritis.

PURPOSE: The correlation between anterior cruciate ligament (ACL) condition and patella osteoarthritic (OA) changes has not been well reported. The aim of this study was to reveal the correlation between ACL deterioration and the morphology of OA changes in the patella. The hypothesis was that significant correlation between ACL deterioration and patella OA morphology would be revealed in this study. METHODS: Two hundred ninety-one cadaveric knees from 151 cadavers were included in this study with a median age of 83 years (54-98). Knees were opened with a sub-vastus approach and the ACL condition was classified as intact or deteriorated. Patella OA lesions were classified using Han's method: type 1, no or minimal lesion; type 2, medial facet lesion without involvement of the ridge; type 3, lateral facet lesion without involvement of the ridge; type 4, lesion involving the ridge only; type 5, medial facet lesion with involvement of the ridge; type 6, lateral facet lesion with involvement of the ridge; and type 7, global lesion. OA depth evaluation was performed following Outerbridge's classification. Statistical analysis of the collected data was performed using generalized estimating equations (GEE). RESULTS: The ACL was intact in 277 knees and deteriorated in 14 knees. Patella OA lesions were observed as follows: type 1, 29%; type 2, 15%; type 3, 2%; type 4, 12%; type 5, 18%; type 6, 2%; and type 7, 22%. Outerbridge's classification of over grade 2 OA depth was observed in 73.5% of subjects. When patella OA was divided into types 1-4 and types 5-7, ACL deterioration was correlated with the occurrence of type 5-7 patella OA [OR 6.44, 95%CI 2.27-18.25, p = 0.000]. When patella OA was divided into types 1-6 and type 7, ACL deterioration was correlated with the occurrence of type 7 patella OA [OR 6.02, 95%CI 2.57-14.09, p = 0.000]. When patella OA depth was divided into grades 1-3 and grade 4, ACL deterioration was highly correlated with the occurrence of grade 4 patella OA [OR 9.31, 95%CI 2.96-29.33, p = 0.025]. CONCLUSION: ACL deterioration is a strong risk factor of wider and deeper OA changes in the patella. For clinical relevance, subjects with ACL tear should be aware of the progression of patella OA.

ACL footprint size is correlated with the height and area of the lateral wall of femoral intercondylar notch.

PURPOSE: The purpose of this study was to reveal the correlation between the size of the native anterior cruciate ligament (ACL) footprint and the size of the lateral wall of femoral intercondylar notch. METHODS: Eighteen non-paired human cadaver knees were used. All soft tissues around the knee were resected except the ACL. The ACL was cut in the middle, and the femoral bone was cut at the most proximal point of the femoral notch. The ACL was carefully dissected, and the periphery of the ACL insertion site was outlined on both the femoral and tibial sides. An accurate lateral view of the femoral condyle and the tibial plateau was photographed with a digital camera, and the images were downloaded to a personal computer. The size of the femoral and tibial ACL footprints, length of Blumensaat's line, and the height and area of the lateral wall of femoral intercondylar notch were measured with Image J software (National Institution of Health). RESULTS: The sizes of the native femoral and tibial ACL footprints were 84 ± 25.3 and 144.7 ± 35.9 mm(2), respectively. The length of Blumensaat's line and the height and area of the lateral wall of femoral intercondylar notch were 29.4 ± 2.8 mm, 17.1 ± 2.7 mm, and 392.4 ± 86 mm(2), respectively. Both the height and the area of the lateral wall of femoral intercondylar notch were significantly correlated with the size of the ACL footprint on both the femoral and tibial sides. CONCLUSION: For clinical relevance, the height and area of the lateral wall of femoral intercondylar notch can be a predictor of native ACL size prior to surgery. However, the length of Blumensaat's line showed no significant correlation with native ACL size.

Size comparison of ACL footprint and reconstructed auto graft.

PURPOSE: The purpose of this study was to compare the size of native anterior cruciate ligament (ACL) footprints and the size of commonly used auto grafts. The hypothesis was that the reconstructed graft size with auto grafts might be smaller than the native ACL footprint. METHODS: Fourteen non-paired human cadaver knees were used. The semitendinosus tendon (ST) and the gracilis (G) tendon were harvested and prepared for ACL grafts. Simulating an ST graft, the ST was cut in half. The bigger half was regarded as the antero-medial (AM) bundle, and the remaining half was regarded as the postero-lateral (PL) bundle. Simulating an ST-G graft, the bigger half of the ST and G were regarded as the AM bundle, and the smaller half of the ST was regarded as the PL bundle. Each graft diameter was measured, and the graft area was calculated. Simulating a rectangular bone-patella tendon-bone (BPTB) graft, a 10-mm wide BPTB graft was harvested and the area calculated. The ACL was carefully dissected, and the size of the femoral and tibial footprints was measured using Image J software (National Institution of Health). RESULTS: The average areas of the ST, ST-G, and BPTB graft were 52.3 ± 7.3, 64.4 ± 9.2, and 32.7 ± 6.5 mm(2), respectively. The sizes of the native femoral and tibial ACL footprints were 85.4 ± 26.3 and 145.4 ± 39.8 mm(2), respectively. Only the ST-G graft showed no significant difference in graft size when compared with the femoral ACL footprint. CONCLUSION: Only the ST-G auto graft was able to reproduce the native size of the ACL footprint on the femoral side. None of the auto grafts could reproduce the size of the tibial ACL footprint. For clinical relevance, ST-G graft is recommended in order to reproduce the native size of the ACL in anatomical ACL reconstruction with auto graft.

The localization of VAMP5 in skeletal and cardiac muscle.

Vesicle-associated membrane protein 5 (VAMP5) is a member of the SNARE protein family, which is generally thought to regulate the docking and fusion of vesicles with their target membranes. This study investigated the expression and localization of the VAMP5 protein. Immunoblotting analyses detected the VAMP5 protein in skeletal muscle, heart, spleen, lung, liver, and kidney tissue, but not in brain or small intestine tissue. Through the immunofluorescence microscopy of skeletal muscle, we found that the expression level of VAMP5 varies among fibers. Most of the fibers with high expression levels of VAMP5 were categorized as type IIa fibers on the basis of their myosin heavy chain subtypes. In addition, the expression patterns of VAMP5 and glucose transporter 4 (GLUT4) were similar. In cardiac muscle, we determined that VAMP5 was localized to the vicinity of intercalated discs. These results suggest that VAMP5 plays local roles in membrane trafficking in skeletal and cardiac muscle.

Alterations of biochemical marker levels and myonuclear numbers in rat skeletal muscle after ischemia-reperfusion.

Prolonged ischemia-reperfusion results in various damages in skeletal muscle. Following reperfusion, although the damaged muscles undergo regeneration, the precise process and mechanism of regeneration have not yet been fully understood. Here, we show the altered levels of plasma biochemical markers of muscle damage, and the change in myonuclear numbers in adult rat skeletal muscle by ischemia-reperfusion. Male Wistar rats were subjected to unilateral hindlimb ischemia by clamping the anterior tibial artery for 2 h before reperfusion. Both plasma creatine kinase activity and C-reactive protein levels in plasma were increased significantly at 0.5 h of reperfusion and returned to the control level at 24 h. The transverse sectional area of muscle belly of the anterior tibial muscles in ischemic side was significantly decreased by 20 % compared with those in sham-ischemic (control) side at 2 days, and returned to the control level at 5 days of reperfusion. Moreover, the number of interstitial nuclei in the ischemic side were significantly increased at 5-14 days and returned to the control level at 21 days of reperfusion. Central nuclei that are specifically observed in regenerating muscle, appeared at 5 days, reached a peak at 14 days, and disappeared at 28 days of reperfusion. Furthermore, MyoD, a regulatory factor for myogenesis, showed a transient expression at 5 days of reperfusion. These results indicate that, although the size of muscle seems to be recovered by 5 days of reperfusion, the most active muscle regeneration occurs much later, as shown by the increase in central nuclei.

Cadaveric assessment of osteoarthritic changes in the patello-femoral joint: evaluation of 203 knees.

PURPOSE: The purpose of this study was to determine the prevalence, type of lesion, and depth of osteoarthritic (OA) changes in the patello-femoral (PF) joint. METHODS: Two hundred and three cadaveric knees were included in this study with median age of 84 years (54-97). Patella OA lesions were classified using Han's method: (Type 1) no or minimal lesion, (Type 2) medial facet lesion without involvement of the ridge, (Type 3) lateral facet lesion without involvement of the ridge, (Type 4) lesion involvement of the ridge, (Type 5) medial facet lesion with involvement of the ridge, (Type 6) lateral facet lesion with involvement of the ridge, (Type 7) global lesion. Femoral side OA lesions in the PF joint were classified using modified Chang's method: (Type 1) no or minimal lesion, (Type 2) medial facet lesion, (Type 3) centre of patella groove lesion, (Type 4) lateral facet lesion, (Type 5) global lesion. OA depth evaluation was performed following Outerbridge's classification. RESULTS: OA lesions of the patella were observed as follows: (Type 1) 31%, (Type 2) 16%, (Type 3) 3%, (Type 4) 12%, (Type 5) 22%, (Type 6) 2%, (Type 7) 14%. Outerbridge's classification of over Grade 2 OA depth was observed in 75.9% of subjects. Femoral side OA lesions of the PF joint were observed as follows: (Type 1) 42%, (Type 2) 20%, (Type 3) 26%, (Type 4) 2%, (Type 5) 11%. Outerbridge's classification of over Grade 2 OA depth was observed in 58% of subjects. CONCLUSION: Patella OA and femoral side OA in the PF joint occurred mainly on the medial side. Isolated OA in the lateral facet of the PF joint was exceedingly rare. Female subjects had a greater incidence of severe PF-OA than male subjects, and therefore, the physicians should pay attention when they treat the female subjects not to advance the PF-OA.