Neurotransmission, Vasculogenesis, and Osteogenesis Activities are Altered in the Aging Temporomandibular Joint of the Senescence-Accelerated Prone 8 Mouse Model.

BACKGROUND: Alterations in neurotransmission, vasculogenesis, and osteogenesis pathways that may play pivotal roles in age-related changes in the temporomandibular joint (TMJ) are poorly understood. PURPOSE: This study aimed to measure the associations between gene and protein profiles in senescence-accelerated prone 8 (SAMP8) mice. STUDY DESIGN: The investigators designed and used 3 groups of 2 mouse models: 1) early aging SAMP8 at 24 weeks of age and control SAMR1 at 12 and 24 weeks (each stage n = 12). PREDICTOR/EXPOSURE/INDEPENDENT VARIABLE: The independent variable was investigated using 3 mouse models: an early aging mouse model and a control mouse model (12 and 24 weeks). MAIN OUTCOME VARIABLE(S): The primary outcome variables were CGRP, VEGF-A, CD31, LYVE-1, osteocalcin, osteopontin, type I and II collagen, and MMP-2. The secondary outcome variables were histological characteristics. COVARIATES: Not applicable. ANALYSES: The gene and protein expression profiles of neurotransmitters, vasculogenesis, and osteogenesis were identified by quantitative real-time polymerase chain reaction and dot blot analysis, respectively. The cellular localization of these events was verified by in situ hybridization and immunohistochemistry. Bivariate statistics were computed for each of the outcome variables. Statistical significance was set to a P value < .05. RESULTS: The expression of CGRP mRNA in the bony mandibular condyle (BMC) of SAMP8 mice (SAMP8, 3.3 ± 0.39 vs SAMR1, 0.001 ± 0.0001) was high at 24 weeks of age (24 weeks) (P < .001). Higher numbers of cells positive percentage for CGRP (MF, SAMP8, 28.67 ± 1.60 vs SAMR 1, 6.36 ± 1.10; CMC, 27.5 ± 2.12 vs 9.00 ± 1.21; BMC, 31.31 ± 2.81 vs 7.85 ± 1.14) and VEGF-A (MF, 34.43 ± 2.45 vs 14.01 ± 1.28; MD, 32.69 ± 1.86 vs 8.00 ± 0.91; CMC, 36.60 ± 2.05 vs 14.19 ± 1.25 BMC 36.49 vs 12.59 ± 1.41) antibodies were found in the 24 weeks TMJ (P < .01). CONCLUSIONS AND RELEVANCE: The neurotransmitter, vasculogenesis, and osteogenesis pathways are associated with TMJ aging in the SAMP8 mouse model. In the future, the SAMP8 mouse model may prove to be a robust model for identifying molecular and biochemical events underlying the effects of feeding, occlusal changes, and tooth loss in the aging TMJ.

Expression of neurotransmitters, vasculogenesis markers and myosin heavy chain isoforms in the masseter muscle of senescence-accelerated mouse prone 8 mice.

BACKGROUND: Learning and memory deficits and pathologic changes in the hippocampus caused by toothlessness and soft diet feeding are related to reduced masseter muscle (MM) function. OBJECTIVE: Myosin heavy chain (MyHC) isoform expression in the MM also changes under different chewing conditions. The neurotransmitter calcitonin gene-related peptide (CGRP) and vascular endothelial growth factor A (VEGF-A) are involved in MM formation. However, the relationship between CGRP, VEGF-A and MyHC isoforms in the MM in the senescence-accelerated mouse prone 8 (SAMP8) strain, a model of learning and memory deficits, remains unclear. METHODS: Changes in CGRP, VEGF-A, vasculogenesis marker and MyHC isoform mRNA expression in the MMs of ageing SAMP8 and senescence-accelerated mouse resistant 1 (SAMR1) mice was investigated through quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization. RESULTS: qRT-PCR revealed obviously high CGRP levels in the SAMP8 mouse MM (p < .001). MyHC-IId/x mRNA expression in the MM was higher in 24-week-old SAMP8 mice than 24-week-old SAMR1 mice (p < .001) but lower in slow-MyHC SAMP8 mice than SAMR1 mice (p < .001). CGRP mRNA was observed on the muscle fibres of the SAMP8 mouse MM but not the SAMR1 mouse MM through in situ hybridization. Principal component analysis (PCA) revealed strong positive contributions of SAMP8-MyHC-IId/x, SAMP8-CGRP, SAMR1-MyHC-emb, SAMR1-CGRP, SAMR1-VEGF-A, SAMR1-CD31, SAMP8-VEGF-A, and SAMP8-CD31 in the MM at 12 and 24 weeks. CONCLUSION: Calcitonin gene-related peptide is also key for the MyHC-IId/x and slow-MyHC patterns in the MMs of SAMP8 mice.

Structural and CBCT analysis of mandibular canal microvessels expressing neurotransmitters in human cadavers.

PURPOSE: This study focused on the detailed structure of microvessels of the neurotransmitter-positive vasa nervorum of the inferior alveolar nerve, vein, and artery in the mandibular canal (MC) to obtain information for improved safety in dental treatments. We also observed the detailed structure of the MC from the mental foramen to the mandibular foramen using cone-beam computed tomography (CBCT). METHODS: In this study, mandibles from 45 sides of 23 human cadavers aged 76-104 years were examined by microscopy, immunohistochemistry, and CBCT analysis. These data were further evaluated by principal component analysis (PCA). RESULTS: The microvessels of the vasa nervorum with calcitonin gene-related peptide- and neuropeptide Y-positive reactions were classified into 5 types: large (4.19%, 28/667); irregular large (7.35%, 49/667), numerous intermediate (29.23%, 195/667), irregular intermediate (29.23%, 195/667), and scattered fine (30.0%, 200/667) microvessels. The MC showed various structures from the 3rd molar to the premolars and was also classified into three types, including complete (57.0%, 228/400), partial (33.8%, 135/400), and unclear (9.2%, 37/400), from the mandibular foramen to the mental foramen. PCA results revealed that developed capillaries were mainly localized in the molar region. CONCLUSIONS: Fine microvessels of the vasa nervorum expressing neurotransmitters are present from the molar to premolar region, which is key information for mandibular dental treatments. The different microvessel structures also indicate differences in specific characteristics between dentulous and edentulous cadavers regarding oral surgical and implant treatments.

Three-dimensional structure of the facial canal and related blood vessels and nerves in the temporal bone.

PURPOSE: There are only limited anatomical data on nerves, veins, and arteries in the temporal bone. More detailed anatomical data are required to improve planning of treatments targeting the temporal bone region. Herein, we performed a detailed analysis of the facial canal (FC) and the related carotid artery and vein. METHODS: We examined the bony structure of the middle ear and FC, jugular foramen, and carotid canal in 30 Japanese elderly donor cadavers. Three-dimensional reconstruction of the canal structure was achieved using cone beam computed tomography, while macroscopic and histological analyses were also performed. RESULTS: The FC form was classified as either straight (28%) or bent (72%). There were significant differences in the diameter of the FC and the distance between the internal jugular vein, other FC branches, and the FC. Principal component analysis (PCA) was performed for the FC using 29 factors. Two principal components significantly explained 30.9% (component 1, 18.6%; component 2, 12.3%) of the FC. Histological observation showed numerous ganglion cells and shrunken neurons in the geniculate ganglion of the facial nerve of elderly samples. CONCLUSION: FC diameter is an important contributor to the relationship between the FC and the jugular foramen. The FC and the internal jugular vein are located close to each other, which is useful information for the trans-canal surgery of the otology. Furthermore, the geniculate ganglion contains numerous ganglion cells and shrunken neurons, which may affect the FC structure during bone matrix remodeling with aging.

Relationship between the thoracic asymmetry in standing position and the asymmetry of ankle moment in the frontal plane during gait.

[Purpose] We aimed to investigate the relationship of thoracic asymmetry in standing position with asymmetry of the internal ankle moment in the frontal plane during gait. [Participants and Methods] The following measurements were recorded in 22 healthy adult males using a 3D motion analyzer and force plates: thoracic lateral deviation, asymmetrical ratios of the upper and lower thoracic shape, internal ankle moment in the frontal plane, mediolateral deviations of the center of mass and center of pressure. [Results] In the standing position, the thorax was deviated to the left relative to the pelvis, and the upper and lower thoracic shapes were asymmetrical. During gait, significant lateralities were observed in the internal ankle moment in the frontal plane, mediolateral deviations of the center of mass and the center of pressure. Significant positive correlations were observed between the asymmetrical ratio of the lower thoracic shape and both the asymmetry of the internal ankle moment in the frontal plane and the mediolateral deviation of the center of pressure. [Conclusion] These results suggest that thoracic asymmetry is associated with mediolateral control of the ankle during gait.

Relationship between the bilateral ratios of the thoracic shape and electromyographic activity of the thoracic and lumbar iliocostalis muscles during thoracic lateral translation.

[Purpose] This study aimed to determine the relationship between thoracic lateral deviation, the bilateral ratio of the thoracic shape, and the bilateral ratio of the thoracic and lumbar iliocostalis muscles during resting sitting and thoracic lateral translation. [Participants and Methods] We included 23 healthy adult males in the study. The measurement tasks were resting sitting and thoracic lateral translation relative to the pelvis. The thoracic lateral deviation and bilateral ratio of the upper and lower thoracic shapes were measured using three-dimensional motion capture. The bilateral ratio of the thoracic and lumbar iliocostalis muscles were measured using the surface electromyographic recording. [Results] The bilateral ratio of the lower thoracic shape was significantly positively correlated with the thoracic translation distance and the bilateral ratio of the thoracic and iliocostalis muscles. In addition, the bilateral ratio of the thoracic iliocostalis muscles was significantly negatively correlated with the bilateral ratios of the lower thoracic shape and lumbar iliocostalis muscles. [Conclusion] Our findings showed that the asymmetry of the lower thoracic shape is associated with left lateral deviation of the thorax at rest and thoracic translation distance. In addition, the thoracic and lumbar iliocostalis muscle activity differed between the left and right translations.

Relationship between scapular elevation exercises with different alignments and activity of the trapezius and levator scapulae muscles.

[Purpose] This study aimed to examine whether scapular elevation exercises in sitting positions with different alignments lead to contractions of the trapezius and levator scapulae muscles. [Participants and Methods] The participants were 25 males, measured in four sitting positions with different alignments. Spine alignment was assessed by measuring the head protrusion, upper thoracic spine tilt, and pelvic tilt angles. Upper limb alignment was evaluated using the scapula tilt angle, scapula rotation angle, and distance between scapular spinous processes. Scapular elevation exercises were measured, and the thickness of the trapezius and levator scapulae muscles were measured in resting and elevated positions, with changes in muscle thickness. [Results] The trapezius muscle thickness was greater in the sitting position with less thoracic spine tilt and scapula tilt angles. Conversely, the levator scapulae muscle thickness was greater in the sitting position with more thoracic spine tilt and scapula tilt angles. [Conclusion] Scapular elevation exercises induce separate contractions of the trapezius and levator scapulae muscles by modifying the alignment of the spine and upper limbs.

Anorectal Transplantation: The First Long-term Success in a Canine Model.

OBJECTIVE: Anorectal transplantation is a challenging procedure but a promising option for patients with weakened or completely absent anorectal function. SUMMARY BACKGROUND DATA: We constructed a canine model of anorectal transplantation, evaluated the long-term outcomes, and controlled rejection and infection in allotransplantation. METHODS: In the pudendal nerve function study, 6 dogs were randomly divided into 2 groups, transection and anastomosis, and were compared with a control using anorectal manometry, electromyography, and histological examination. In the anorectal transplantation model, 4 dogs were assigned to 4 groups: autotransplant, allotransplant with immunosuppression, allotransplant without immunosuppression, and normal control. Long-term function was evaluated by defecography, videography, and histological examination. RESULTS: In the pudendal nerve function study, anorectal manometry indicated that the anastomosis group recovered partial function 6 months postoperatively. Microscopically, the pudendal nerve and the sphincter muscle regenerated in the anastomosis group. Anorectal transplantation was technically successful with a 3-stage operation: colostomy preparation, anorectal transplantation, and stoma closure. The dog who underwent allotransplantation and immunosuppression had 2 episodes of mild rejection, which were reversed with methylprednisolone and tacrolimus. The dog who underwent allotransplantation without immunosuppression had a severe acute rejection that resulted in graft necrosis. Successful dogs had full defecation control at the end of the study. CONCLUSIONS: We describe the critical role of the pudendal nerve in anorectal function and the first long-term success with anorectal transplantation in a canine model. This report is a proof-of-concept study for anorectal transplantation as a treatment for patients with an ostomy because of anorectal dysfunction.

GIT1 is an untolerized autoantigen involved in immunologic disturbance of spermatogenesis.

Male haploid cells, spermatids and spermatozoa, that appear after the establishment of immune tolerance express novel cell surface and intracellular proteins that can be recognized as foreign antigens by the self-immune system. However, these germ cells do not normally evoke a pathological immune response. The immune-privileged micro-circumstance in testis involving the blood-testis-barrier formed by Sertoli cells protects these germ cells from autoimmune attack. We recently found that immunization with heat shock protein family A member 4-like (HSPA4L), one of the new differentiation antigens of haploid cells, induced experimental autoimmune orchitis (EAO) in A/J male mice. In this study, we focused on G protein-coupled receptor kinase interacting protein-1 (GIT1), another haploid cell-specific differentiation antigen, to investigate whether GIT1 is a target autoantigen for EAO induction. GIT1 emulsified with complete Freund's adjuvant was injected subcutaneously into the mice inguinal region once on day 0 and again on day 14, and the optimum condition of EAO induction was determined. Mice immunized with 200 µg GIT1 showed significantly higher incidence of EAO than that of immunization with other concentrations. In particular, significant lymphocytic inflammation and extensive aspermatogenesis were observed in these mice at 120 days after the first immunization. These findings indicate that GIT1 is also a target antigen that induces EAO, like HSPA4L.

High-Pressure Carbon Monoxide and Oxygen Mixture is Effective for Lung Preservation.

(1) Background: Heme oxygenase-1 (HO-1) degrades heme and generates carbon monoxide (CO), producing various anti-inflammatory, anti-oxidative, and anti-apoptotic effects. This study aimed to confirm the effects of CO on the ischemia-reperfusion injury (IRI) of donor lungs using a high-pressure gas (HPG) preservation method. (2) Methods: Donor rat and canine lungs were preserved in a chamber filled with CO (1.5 atm) and oxygen (O2; 2 atm) and were ventilated with either CO and O2 mixture (CO/O2 group) or air (air group) immediately before storage. Rat lungs were subjected to heterotopic cervical transplantation and evaluated after reperfusion, whereas canine lungs were subjected to allogeneic transplantation and evaluated. (3) Results: Alveolar hemorrhage in the CO/O2 group was significantly milder than that in the air group. mRNA expression levels of HO-1 remained unchanged in both the groups; however, inflammatory mediator levels were significantly lower in the CO/O2 group than in the air group. The oxygenation of graft lungs was comparable between the two groups, but lactic acid level tended to be higher in the air group. (4) Conclusions: The HO-1/CO system in the HPG preservation method is effective in suppressing IRI and preserving donor lungs.

Short-term ethanol exposure causes imbalanced neurotrophic factor allocation in the basal forebrain cholinergic system: a novel insight into understanding the initial processes of alcohol addiction.

Alcohol ingestion affects both motor and cognitive functions. One brain system that is influenced by ethanol is the basal forebrain (BF) cholinergic projection system, which projects to diverse neocortical and limbic areas. The BF is associated with memory and cognitive function. Our primary interest is the examination of how regions that receive BF cholinergic projections are influenced by short-term ethanol exposure through alterations in the mRNA levels of neurotrophic factors [nerve growth factor/TrkA, brain-derived neurotrophic factor/TrkB, and glial-derived neurotrophic factor (GDNF)/GDNF family receptor α1]. Male BALB/C mice were fed a liquid diet containing 5 % (v/v) ethanol. Pair-fed control mice were maintained on an identical liquid diet, except that the ethanol was isocalorically substituted with sucrose. Mice exhibiting signs of ethanol intoxication (stages 1-2) were used for real-time reverse transcription-polymerase chain reaction analyses. Among the BF cholinergic projection regions, decreased levels of GDNF mRNA and increased levels of TrkB mRNA were observed in the basal nucleus, and increased levels of TrkB mRNA were observed in the cerebral cortex. There were no significant alterations in the levels of expression of relevant neurotrophic factors in the septal nucleus and hippocampus. Given that neurotrophic factors function in retrograde/anterograde or autocrine/paracrine mechanisms and that BF cholinergic projection regions are neuroanatomically connected, these findings suggested that an imbalanced allocation of neurotrophic factor ligands and receptors is an initial phenomenon in alcohol addiction. The exact mechanisms underlying this phenomenon in the BF cholinergic system are unknown. However, our results provide a novel notion for the understanding of the initial processes in alcohol addiction.

A rat model for acute limb ischemia using microsized gelatin beads.

OBJECTIVE: Acute limb ischemia (ALI) is a rapid decrease in lower limb blood flow due to acute occlusion of peripheral arteries or bypass grafts. This study aimed to establish an ALI model using microsized gelatin beads and to investigate the pathophysiological conditions. METHODS: Male Sprague-Dawley rats were anesthetized, and a low or high dose of microsized gelatin beads was administered into the left femoral artery on days 0 and 7. A control, that is, normal saline (NS) group in which NS was administered in the left femoral artery, a femoral artery cut (FAC) group in which the left femoral artery was cut, and a sciatic nerve cut (SNC) group in which the left sciatic nerve was cut were prepared. After 21 days, the temperature changes and the muscle weights in the lower limbs were measured. To assess nerve damage, the L1-6 sympathetic ganglia were immunostained with activating transcription factor 3 (ATF3) antibody. RESULTS: In the Low-dose, High-dose, and FAC groups, a decrease in temperature was predominantly observed in the left limb. In the High-dose and SNC groups, the weight of the soleus muscle and extensor digitorum longus in the left limb decreased; however, no weight changes were observed in the Low-dose and FAC groups. Conversely, the weight of the gastrocnemius muscle significantly decreased in the Low-dose, High-dose, FAC, and SNC groups. In the High-dose and SNC groups, the number of ATF3-positive cells in the sympathetic ganglia significantly increased, and in the Low-dose, a small number of ATF3-positive cells were observed. However, ATF3-positive cells were rarely observed in the FAC and NS groups. CONCLUSION: We established an ALI rat model using microsized gelatin beads. The results of this study suggest that autonomic neuropathy in ALI is related to both muscle damage and peripheral neuropathy.

Effects of female bone marrow transplantation on male reproductive organs.

Graft-versus-host disease (GVHD), an adverse effect after bone marrow transplantation (BMT), may affect male reproductive function. It is hypothesized that a sex-mismatched BMT induces GVHD in male reproductive organs because female immune cells are not immunologically tolerant to specific antigens of the male organs. However, this hypothesis has not been experimentally verified using male (M) recipient animals following BMT from the female (F) donors. Therefore, the aim of the present study is to examine whether the female BMT to males (F→M group) induces some GVHD reactions in the testis and the other male reproductive organs. The results showed that no inflammation was found in recipients of the male BMT to males (M→M group), whereas significant inflammatory cell responses lasting for at least 4 months were induced in testis, epididymis, prostate and preputial gland in some mice of F→M group. The most severe lesion was found in the preputial gland, in which lymphocytic inflammation was accompanied by loss of glandular acini, thickening of the interstitum and increased cytokines such as TNF-α and IFN-γ. Western blot analyses revealed that sera from the F→M group reacted with various antigens of the male reproductive organs. These results indicate that transplanted female immune cells may recognize the male reproductive organs as immunologically foreign ones and induce chronic GVHD, which may affect male reproductive function.

Early postnatal maternal separation causes alterations in the expression of ß3-adrenergic receptor in rat adipose tissue suggesting long-term influence on obesity.

The effects of early postnatal maternal deprivation on the biological characteristics of the adipose tissue later in life were investigated in the present study. Sprague-Dawley rats were classified as either maternal deprivation (MD) or mother-reared control (MRC) groups. MD was achieved by separating the rat pups from their mothers for 3h each day during the 10-15 postnatal days. mRNA levels of mitochondrial uncoupling protein 1 (UCP-1), ß3-adrenergic receptor (ß3-AR), and prohibitin (PHB) in the brown and white adipose tissue were determined using real-time RT-PCR analysis. UCP-1, which is mediated through ß3-AR, is closely involved in the energy metabolism and expenditure. PHB is highly expressed in the proliferating tissues/cells. At 10 weeks of age, the body weight of the MRC and MD rats was similar. However, the levels of the key molecules in the adipose tissue were substantially altered. There was a significant increase in the expression of PHB mRNA in the white adipose tissue, while the ß3-AR mRNA expression decreased significantly, and the UCP-1 mRNA expression remained unchanged in the brown adipose tissue. Given that these molecules influence the mitochondrial metabolism, our study indicates that early postnatal maternal deprivation can influence the fate of adipose tissue proliferation, presumably leading to obesity later in life.

Morphological evidence of an altered process of synaptic transcytosis in adult rats exposed to ethanol.

AIMS: The effects of ethanol exposure on synaptic structure were investigated in the nucleus of solitary tract (NST) in rats, using the horse-radish peroxidase (HRP) method. METHODS: Eight-week-old experimental rats were allowed free access to a liquid diet containing ethanol for 3 weeks, while controls were given an isocaloric diet. Some of the control and experimental animals were given an injection of wheat germ agglutinin conjugated with HRP (WGA-HRP) into the vagus nerve toward the end of the treatment period. After the treatment, the neuropil region of the NST was examined under an electron microscope. RESULTS: We observed that a few terminals were characterized by deep indentation of axodendritic membranes into the post-synaptic neurons. This appeared to be similar to that commonly seen in exocrine glands. Interestingly, the indented portion often contained various sizes of vacuoles and flattened cisternae. HRP-reaction product (RP) transported to terminals was recognized easily as an electron-dense lysosomal substance when lead citrate staining was omitted. Terminals containing HRP-RP also revealed quite a similar structure with indentation of axodendritic membranes as described earlier. The results are considered to confirm that terminals forming 'apocrine-like structures' observed in the ethanol-fed animals with no injection of WGA-HRP originate from afferent fibers of the vagus nerve. CONCLUSION: The present study suggests the possibility that the alteration of the synaptic structure induced by ethanol exposure can lead to the neuronal transcytosis of materials including proteins which is different from the normal vesicular exocytosis involved in chemical synaptic transmission.

Glucose infusion suppresses acute restraint stress-induced peripheral and central sympathetic responses in rats.

BACKGROUND: Acute restraint stress (RS) induces sympathetic activation such as elevating plasma catecholamines, resulting in increase in blood glucose. We aimed to investigate whether glucose infusion affects the RS-induced sympathetic responses. METHODS: Plasma catecholamines were measured by high-performance liquid chromatography with electrochemical detection. Blood glucose levels were measured with a glucometer and a glucose assay kit. Cardiac parameters were measured by echocardiographic and hemodynamic analysis. Prostanoid levels in the paraventricular nucleus of hypothalamus (PVN) microdialysates were measured by liquid chromatography-ion trap tandem mass spectrometry analysis. RESULTS: RS significantly increased plasma noradrenaline and adrenaline. Intravenous infusion of a 5% glucose solution significantly attenuated the RS-induced elevation of plasma adrenaline but did not alter the plasma noradrenaline. Glucose administration during RS suppressed the progression of cardiac impairment by attenuating the decline rates in left ventricular diastolic, end-diastolic volume, stroke volume, fractional shortening, and ejection fraction. Both Intravenous and intracerebroventricular infusion of glucose solution significantly attenuated the RS-induced elevation of thromboxane B2 (TxB2) (a metabolite of TxA2) levels in the PVN but did not alter prostaglandin E2 levels in the PVN. CONCLUSION: Our results demonstrate that glucose infusion suppresses RS-induced elevation of plasma adrenaline and left ventricular dysfunction. In the brain, glucose infusion suppresses RS-induced production of TxA2 in the PVN.

The effect of simulation-based education before a cadaver dissection course.

Although the methods for medical education continue to evolve due to the development of medicines, the cadaver dissection course still plays a fundamental role. The cadaver dissection course allows students to learn to handle instruments correctly while actively exploring three-dimensional anatomy. However, dissection comes with the risk of accidental injury. In recent years, the number of classes offered for the cadaver dissection course has decreased while the amount of knowledge required in clinical medicine has increased. Simulation-based education (SBE) has been proven to be an effective educational method that enhances the development of practical skills by integrating learners' knowledge and skills. This study aimed to investigate the effect of SBE as a preparatory education course when taken prior to a medical student's enrollment in the cadaver dissection course. In the present study, an SBE assuming practical cadaver dissection course was performed in the Clinical Simulation Center. The frequency of injury rates per 1000 h of cadaver dissection course was significantly less in 2017 and 2018 compared to that in 2016. Two years after the implementation of the SBE, average student self-efficacy scores and written examination scores significantly increased, whereas self-contentment scores were relatively unchanged. The results showed that the implementation of SBE decreased the incidence of injuries and improved students' overall self-efficacy scores and increased acquisition of knowledge evident on written examination score. Therefore, SBE as a preparatory education course may effectively promote the combined development of dissection skills and anatomical knowledge in the subsequent fundamental cadaver dissection course.

Characteristic expression of CGRP and osteogenic and vasculogenic markers in the proximal and distal regions of the rib during male mouse development.

BACKGROUND: Neuropeptide calcitonin gene-related peptide (CGRP) is a neurotransmitter related to vasculogenesis and osteogenesis during bone formation and organ development. From the foetal period to the postnatal period, the thorax, which is necessary for lung respiration, forms. The thorax exhibits the same cartilage ossification as the bones of the extremities, but a specific system within the thorax exists as costal cartilage after birth. The relationship among CGRP, osteogenesis and vasculogenic markers in the two rib locations during thorax formation is not fully understood. MATERIALS AND METHODS: In our study, male mice were used to provide ribs under different development conditions on various embryonic days (E12. 5, E14.5, and E17.5) and postnatal days (P1 and P5). The mRNA expression levels of CGRP, vascular endothelial growth factor (VEGF-A), type 1 collagen (Col1a-1), type 2 collagen (Col2a1), neuropeptide Y (NPY), osteocalcin (OCN) and osteopontin (OPN) were analysed by qRT-PCR. We also analysed the mRNA expression of CGRP, VEGF-A and OPN by in situ hybridization. Multivariate modelling with principal component analysis (PCA) was performed to estimate the interactions among the quantitative real-time RT-PCR data. RESULTS: The mRNA expression levels of CGRP, VEGF-A, Col2a, Col1a-1, OCN, and NPY in the male mouse rib gradually increased during development. An antisense probe for CGRP mRNA was strongly detected in the central region of the mouse rib at E12.5 and the hypertrophic and ossification zones at E17.5 by in situ hybridization. VEGF-A was also located in the same region as CGRP at E12.5 and E17.5. OPN was strongly detected at the rib formation stage from E14.5 to E17.5. The expression of CGRP also differed between the proximal and distal regions of the rib at E17.5. As demonstrated by in situ hybridization, CGRP continuously participates in cartilage formation in the distal regions of the rib after birth. The PCA revealed that the mRNA expression of CGRP was related to that of Col1a-1 and VEGF-A during rib formation. CONCLUSION: This study shows that CGRP is involved in vascular and bone formation during rib development and may also be involved in cartilage formation after birth. The findings suggest that CGRP may temporarily participate in bone formation and continuously participate in cartilage formation in the rib, which may also be related to the formation of the anterior thoracic wall after birth.

Site dependent elastic property of human iliotibial band and the effect of hip and knee joint angle configuration.

The iliotibial band (ITB) is the lateral thickening of the fascia lata. The ITB has been extensively studied for its relevance to injury, but not much is known about its elastic properties. We aimed to investigate the site- and joint angle-dependence of ITB elasticity. We tested twelve healthy males (22-30 years; in vivo) and twelve male cadavers (69-93 years; cadaver). The Young's modulus of the ITB was measured in the longitudinal direction at five sites (over the proximal, middle, and distal bellies of the vastus lateralis (VL), superior border of the patella, and between femur and tibia) of the right limb, by ultrasound shear wave elastography (in vivo) and the tensile test (cadaver). Joint angle-dependence was also studied for nine different positions (knee angles at 0, 25, 90˚ x hip angles at 0, 40, 90˚) (in vivo). Over VL, the ITB was more compliant at the distal (17.6-190.1 kPa; in vivo, 219.4 ± 68.8 MPa; cadaver, mean ± SD) than other sites (24.2-221.4 kPa, 337.9-362.7 MPa). The ITB at the superior border of the patella and between femur and tibia was stiffer in vivo (31.8-271.8 and 50.9-208.8 kPa), while it was more compliant in cadavers (113.4 ± 63.7 and 130.4 ± 73.7 MPa), compared to other sites. The ITB became stiffer associated with increasing hip extension angle and knee flexion angle, and the hip remarkably affecting the values regardless of site (in vivo). Our findings have clinical significance with respect to the site- and joint angle-dependence of ITB-related overuse injury.

Morphological and mechanical properties of the human triceps surae aponeuroses taken from elderly cadavers: Implications for muscle-tendon interactions.

The human triceps surae (two gastrocnemii and soleus) has aponeuroses in the proximal and distal aspects, the latter of which insert into the calcaneus by sharing the common Achilles tendon. These tendinous tissues are known to have elasticity and upon muscle contraction the aponeurosis is stretched both longitudinally (along the muscle's line of action) and transversely. Higher aponeurosis transverse deformability has been documented, but there is a paucity of information on the morphology and mechanical properties of human aponeurosis. This study aimed to identify morphological and mechanical characteristics of the human triceps surae aponeuroses. Twenty-five triceps surae muscle-tendon units were procured from 13 human donors (formalin fixed, 6 males, 7 females) aged 67-91 years. Specimens of aponeuroses were excised from the eight regions (posterior and anterior regions of the gastrocnemius medialis and lateralis, medial and lateral parts of soleus; proximal, middle, and distal sites each, 2-4 cm × 2-4 cm). Aponeurosis thickness was measured using a digital caliper. Uniaxial tensile tests were implemented to determine the mechanical properties of specimens loaded longitudinally (along the muscle's line of action) and transversely. The aponeurosis thickness showed significant differences between muscles and sites, while Young's modulus showed direction-dependent (longitudinal vs. transverse) differences within sites. Results show different morphology and mechanical properties of aponeuroses between synergist muscles. The reason for site-dependent differences in stiffness is due to a reduced aponeurosis thickness rather than a reduction in the material property. The anisotropic elastic feature (differences between longitudinal and transverse directions) of the aponeuroses was more pronounced than previous in vivo findings, suggesting inherent material design of the aponeurosis that matches three-dimensional contractile behavior of muscle fibers.