Hirohiko Aoyama

Potential Risks of Femoral Tunnel Drilling Through the Far Anteromedial Portal: A Cadaveric Study

PURPOSE The purpose of this study was to estimate the potential risks when drilling femoral tunnels through the far anteromedial portal in double-bundle anterior cruciate ligament reconstruction in cadaveric knees. METHODS Ten cadaveric knees were used. We drilled the anteromedial bundle (AMB) and posterolateral bundle (PLB) through the far anteromedial portal at 3 different knee flexion angles: 70 degrees, 90 degrees, and 110 degrees. We measured the shortest distance to the common peroneal nerve and the posterior articular cartilage of the lateral femoral condyle and the femoral tunnel length. RESULTS At 70 degrees, the distance to the nerve was less than 10 mm in 7 AMB cases and in 9 PLB cases, and the distance to the cartilage was less than 10 mm in all the AMB and PLB cases. At 90 degrees, the distance to the nerve was less than 10 mm in 1 AMB and 5 PLBs, and the distance to the cartilage was less than 10 mm in 2 AMBs and all the PLBs. On the other hand, at 110 degrees , the distance to the nerve was greater than 10 mm in all the AMBs and PLBs, and the distance to the cartilage did not exceed 10 mm in just 2 of the PLBs. CONCLUSIONS In our cadaveric study we found that the low knee flexion angles when drilling femoral tunnels through the far anteromedial portal might have the potential risks of damage to the common peroneal nerve and the posterior articular cartilage, and the risks would be decreased at higher degrees of knee flexion. However, we found there was a 20% risk of damage to the cartilage while drilling the PLB at 110 degrees. CLINICAL RELEVANCE High knee flexion angles are recommended to avoid damage to the nerve and the cartilage when drilling femoral tunnels through the far anteromedial portal in double-bundle anterior cruciate ligament reconstruction.

The developmental fate of the rostral/caudal half of a somite for vertebra and rib formation: experimental confirmation of the resegmentation theory using chick-quail chimeras.

To determine whether resegmentation of somites forms the axial skeleton, we traced the development of the rostral and the caudal half of a somite during skeletogenesis in chick-quail chimeras by replacing the rostral or caudal half of a newly formed chick somite with that of a quail somite. The rostral half-somite transplant formed the caudal half of the vertebral body, the entire spinous process and the distal rib, while the caudal half-somite transplant formed the rostral half of vertebral body, the rostral half of spinous process, the vertebral arch, the transverse process and the entire rib. These findings confirm the resegmentation theory except the spinous process and the distal rib.

Developmental Plasticity of the Prospective Dermatome and the Prospective Sclerotome Region of an Avian Somite

The ventro‐medial wall of a somite gives rise to the sclerotome and then to cartilaginous axial skeleton, while the dorso‐lateral wall differentiates into the dermomyotome to form dermal mesenchyme and muscle. Although previous studies suggested pluri‐potency of somite cell differentiation, apparent pluri‐potency may be the result of migration of predetermined cells. To investigate whether the developmental fate of any region is determined, I isolated fragments of a region of a quail somite and transplanted them into chick embryos. When a fragment of the ventral wall of a quail somite, the prospective sclerotome, was transplanted into a chick embryo between the ectoderm and a newly formed somite, the transplanted quail cells were shown to form myotome and mesenchyme in 4‐day chimera embryos and to form muscle and dermal tissue in 9‐day chimeras. On the other hand, when a fragment of the dorsal wall of a quail somite, the prospective dermomyotome, was transplanted into a chick embryo between the neural tube and a newly formed somite, the graft gave rise to mesenchyme around the neural tube and notochord and then to vertebral cartilage. Thus the developmental fate of a region of a somite was shown not to be determined at the time of somite segmentation, confirming previous observations.

Do peanut agglutinin receptors on somites control the behavior of neural cells

Peanut agglutinin (PNA) receptors are expressed in the caudal halves of sclerotomes in chick embryos after 3 days of incubation (stages 19–20 of Hamburger & Hamilton). The neural crest cells forming dorsal root ganglia (DRG) and motor nerves appear to avoid PNA positive regions and concentrate into rostral halves of sclerotomes. To investigate the role of PNA receptors in gangliogenesis and nerve growth, we examined PNA binding ability in quail sclerotomes and in chick‐quail chimeric embryos made by transplanting quail somites to chick embryos, comparing the development of DRG, motor nerves and sclerotomes. PNA did not bind to any part of the somites of 4.5‐day quail embryos, although dorsal root ganglia and motor nerves appeared only in the rostral halves of sclerotomes as in chick embryos. Moreover, in spite of no PNA binding ability of the transplanted quail somite in 4.5‐day chick‐quail chimeric embryos, DRG and motor nerves derived from chick tissues appeared only in the rostral halves of the sclerotomes derived from these somites. Thus, both quail and chick neural crest cells and motor nerves recognized the difference between the rostral and caudal halves of sclerotomes of quail embryos in the absence of PNA binding ability, indicating that PNA binding site on somite cells does not support the selective neural crest migration and nerve growth.

Heterogeneity of neurogenic responses in intra- and extrameningeal arteries of dogs

1 . Neurogenic responses to transmural electrical stimulation were examined in endothelium‐denuded extrameningeal (vertebral and carotid) and intrameningeal (spinal, basilar and middle cerebral) arteries isolated from dogs. 2 . In the extrameningeal arteries, transmural electrical stimulation produced a phasic contraction. This contraction was abolished by tetrodotoxin, prazosin and guanethidine. However, α,β‐methylene ATP and NG‐nitro‐L‐arginine (1‐NOARG) had no significant effect on the contractile responses. 3 . In the intrameningeal arteries, the neurogenic responses to electrical stimulation were composed of a transient contraction and relaxation. The transient contraction was selectively inhibited by guanethidine or after desensitization of P2x‐purinoceptors with α,β‐methylene ATP. L‐NOARG abolished the relaxation but not the contraction induced by electrical stimulation. Prazosin had no effect on either neurogenic response. 4 . Noradrenaline produced a large contraction in the extrameningeal arteries which was selectively inhibited by prazosin. α,β‐Methylene ATP produced neither contraction nor inhibition of the response to noradrenaline in the extrameningeal arteries. 5 . In the intrameningeal arteries, α,β‐methylene ATP produced a greater contraction than noradrenaline. The response to α,β‐methylene ATP was selectively abolished by desensitization of P2X‐purinoceptors with α,β‐methylene ATP itself. The contractile response to noradrenaline was inhibited by rauwolscine but not by prazosin. 6 . ATP produced endothelium‐dependent relaxations in the extrameningeal and intrameningeal arteries, which were attenuated by endothelium removal. 7 . NADPH diaphorase‐positive fibres were dense in the middle cerebral and basilar arteries but rare or absent in the spinal artery. In the extrameningeal arteries diaphorase‐positive traces were observed in the vasa vasorum. 8 . The present findings indicate that the neurogenic responses of intrameningeal arteries of dogs are composed of NO‐ergic and sympathetic purinergic components, while the extrameningeal arteries tested produced only sympathetic adrenergic responses, suggesting that regional heterogeneity may be associated with a sudden transition in innervation and receptor expression at the meninx.

Body wall morphogenesis: Limb-genesis interferes with body wall-genesis via its influence on the abaxial somite derivatives

The vertebrate body wall is regionalized into thoracic and lumbosacral/abdominal regions that differ in their morphology and developmental origin. The thoracic body wall has ribs and intercostal muscles, which develops from thoracic somites, whereas the abdominal wall has abdominal muscles, which develops from lumbosacral somites without ribs cage. To examine whether limb-genesis interferes with body wall-genesis, and to test the possibility that limb generation leads to the regional differentiation, an ectopic limb was induced in the thoracic region by transplanting prospective limb somatopleural mesoderm of Japanese quail between the ectoderm and somatopleural mesoderm of the chick prospective thoracic region. This ectopic limb generation induced the somitic cells to migrate into the ectopic limb mesenchyme to become its muscles and caused the loss of distal thoracic body wall (sterno-distal rib and distal intercostal muscle), without causing any significant effect on the more proximal region (proximal rib, vertebro-distal rib and proximal intercostal muscle). According to a new primaxial-abaxial classification, the proximal region is classified as primaxial and the distal region, as well as limb, is classified as abaxial. We demonstrated that ectopic limb development interfered with body wall development via its influence on the abaxial somite derivatives. The present study supports the idea that the somitic cells give rise to the primaxial derivatives keeping their own identity and fate, whereas they produce the abaxial derivatives responding to the lateral plate mesoderm.

Somite development without influence of the surface ectoderm in the chick embryo: The compartments of a somite responsible for distal rib development

In the development of the somite, signals from neighboring tissues have been suggested to play critical roles. We have found that when interaction between the ectoderm and the somite is blocked by inserting a piece of polyethylene terephatalate film between them in 2‐day‐chicken embryo, one of the derivatives of somite, the distal rib, did not form. We examined somite development after the operation, to know the correlation between somite development and distal rib formation. In the operated embryo, the dermomyotome was medio‐laterally shorter than in the normal embryo, and Pax3 and Sim1 expressions that are seen in the lateral part of normal dermomyotomes were not found, suggesting that the lateral part of the dermomyotome was missing. Although the sclerotome appeared to be normal in its histology and Pax1 expression pattern in the operated embryo, we could not detect the expression of either Scleraxis nor γ‐FBP that are expressed in the cells around the boundaries between the adjacent dermomyotomes in normal embryos. Thus, under the influence of surface ectoderm, the lateral part of dermomyotome and/or the mesenchyme around rostral and caudal edges of dermomyotomes are suggested to play an important role in the distal rib development.

Elevated Intraocular Pressure, Optic Nerve Atrophy, and Impaired Retinal Development in ODAG Transgenic Mice

PURPOSE In an earlier study, a cDNA was cloned that showed abundant expression in the eye at postnatal day (P)2 but was downregulated at P10; it was named ODAG (ocular development-associated gene). Its biological function was examined by generating and analyzing transgenic mice overexpressing ODAG (ODAG Tg) in the eye and by identifying ODAG-binding proteins. METHODS Transgenic mice were generated by using the mouse Crx promoter. EGFP was designed to be coexpressed with transgenic ODAG, to identify transgene-expressing cells. Overexpression of ODAG was confirmed by Northern and Western blot analysis. IOP was measured with a microneedle technique. The eyes were macroscopically examined and histologically analyzed. EGFP expression was detected by confocal microscope. Proteins associated with ODAG were isolated by pull-down assay in conjugation with mass spectrometry. RESULTS Macroscopically, ODAG Tg exhibited gradual protrusion of the eyeballs. The mean IOP of ODAG Tg was significantly higher than that of wild-type (WT) littermates. Histologic analysis exhibited optic nerve atrophy and impaired retinal development in the ODAG Tg eye. EGFP was expressed highly in the presumptive outer nuclear layer and weakly in the presumptive inner nuclear layer in the ODAG Tg retina. Rab6-GTPase-activating protein (Rab6-GAP) and its substrate, Rab6, were identified as ODAG-binding proteins. CONCLUSIONS Deregulated expression of ODAG in the eye induces elevated intraocular pressure and optic nerve atrophy and impairs retinal development, possibly by interfering with the Rab6/Rab6-GAP-mediated signaling pathway. These results provide new insights into the mechanisms regulating ocular development, and ODAG Tg would be a novel animal model for human diseases caused by ocular hypertension.

Radiation dose reduction at MDCT with iterative reconstruction for prenatal diagnosis of skeletal dysplasia: preliminary study using normal fetal specimens.

OBJECTIVE The purpose of this study was to investigate to what degree the radiation dose can be reduced without affecting the ability to evaluate normal fetal bones at MDCT with iterative reconstruction. MATERIALS AND METHODS Fifteen normal fetal specimens immersed in containers (30- and 35-cm diameter) were scanned with a 64-MDCT scanner, with tube voltage of 100 kVp and tube current of 600, 300, 150, 100, and 50 mA. Images were subjected to adaptive statistical iterative reconstruction (ASIR). The fetal dose was measured using glass dosimeters. We calculated the relative ratio of the dose at 600 mA. Image quality was evaluated on maximum-intensity-projection and volume-rendering images. Two radiologists recorded the visualization scores of five regions. Images at 600 mA were considered to be standard. RESULTS With the 30-cm-diameter container, the fetal dose was 10.15 mGy (relative ratio, 100%) at a tube current of 600, 51% at 300, 25% at 150, 17% at 100, and 9% at 50 mA. With the 35-cm-diameter container the fetal dose was 10.01 mGy (relative ratio, 100%) at 600, 47% at 300, 24% at 150, 17% at 100, and 8% at 50 mA. Visual evaluation showed that in both containers, with ASIR 90%, there was a statistically significant difference between 50-and 600-mA images (p<0.01) but not between 600-mA images and those acquired at 100, 150, and 300 mA (p=0.08-1.00). CONCLUSION The fetal radiation dose for the evaluation of normal fetal bones can be reduced by 83% with ASIR 90%.

Regulation of Cell Number in Formation of the Dorsal Root Ganglion Revealed by Transplantation of Quail Neural Crest Cells into Chick Embryos

Neural crest cells appear transiently in early embryogenesis on the dorsal surface of the neural tube and subsequently migrate along specific pathways. Some migrate to between the neural tube and somites, aggregating to form the rudiments of dorsal root ganglia (DRG). The size of DRG at a given somite level is almost constant in all chick embryos. To determine the mechanisms controlling the size of DRG, we transplanted neural crest cells of 2.5‐day‐old quail embryos into 2.5‐day‐old chick embryos between the neural tube and the somites, and examined the size of DRG in these chimeric embryos with extra neural crest cells 2 days after the operation, when natural cell death in DRG had not yet occurred. The DRG on the operated side were composed of both chick and quail cells in various proportions. The cell numbers of these chimeric DRG were almost the same as those of the normal DRG on the opposite side. That is, there were significantly fewer chick cells in chimeric DRG than in DRG composed of only chick cells on the opposite unoperated side. This finding indicates that the size of DRG is not determined in migrating neural crest cells but is regulated by the circumstances.