Vermiform Appendix During the Repackaging Process from Umbilical Herniation to Fixation onto the Right Posterior Abdomen: A Study of Human Fetal Horizontal Sections.

The anatomical position of the vermiform appendix varies among adults, and these variations are responsible for differences in the symptoms of appendicitis. However, to date no study has examined how and when these variations occur during fetal development. The present study examined horizontal sections of 27 midterm fetuses (crown rump length [CRL] 38-97 mm, gestational age approximately 8-15 weeks). There were 10 fetuses (CRL 56 mm or more) in which the cecum and appendix were in a posterosuperior site near the right kidney (postmigration phase), and 12 fetuses (CRL 39-72 mm) in which the ileocecal junction and appendix remained on the visceral surface of the liver in the anterior or anterolateral abdominal cavity (migration phase, after physiological umbilical herniation). Analysis of the 12 fetuses in the migration phase indicated that the appendix extended inferiorly in eight fetuses and superiorly in four fetuses. Likewise, a "preileal" appendix (a morphology in which the distal part of the appendix was in front of the terminal ileum) was present in eight of these fetuses. Extension of the appendix superiorly or inferiorly during the migration phase seems unrelated to the topographical relationship of the appendix with the terminal ileum at the postmigration phase in fetuses and in adults. Conversely, it seems likely that a retroileal appendix leads to a coiled appendix behind the ileocecal junction. "Guidance" by the liver surface seemed to be important for posterior migration, which ended with the ascent of the liver. Clin. Anat., 33:667-677, 2020. © 2019 Wiley Periodicals, Inc.

Fetal Development of Human Oral Epithelial Pearls With Special Reference to Their Stage-Dependent Changes in Distribution.

OBJECTIVE: To access detailed distribution and age-dependent changes of oral epithelial pearls. DESIGN: Investigation and analysis with human fetal serial sections. SETTING: Institute of Embryology. METHODS: This study examined serial frontal sections of the upper and lower jaws of 19 human fetuses at 12 to 18 weeks and of the lower jaws of four late-stage fetuses. RESULTS: The upper jaw contained more than 20 midline and more than 60 lateral pearls greater than 20 µm in diameter, whereas the lower jaw contained fewer than 30 pearls of the same size. Midline pearls in the upper jaw were often cylindrical or rugby-ball shaped, whereas all pearls in the lower jaw were small and spherical. Epithelial pearls in the upper jaw started developing along the upper midline until 12 weeks; lateral pearls and additional midline pearls (or strictly, paramedian pearls) developed until 15 weeks. In the lower jaw, however, pearl development started at 18 weeks and was almost always from the dental lamina. Some of the fetuses assessed had an open nasopalatine canal without a duct, but there was no fibrous connection between this canal and pearls. Similarly, the lip frenulum or incisive suture was not connected with these pearls. CONCLUSION: The timing and sequence of development suggest that postfusion rupture of the palate by midline pearls was unlikely.

High-frequency pulsed low-level diode laser therapy accelerates wound healing of tooth extraction socket: An in vivo study.

BACKGROUND AND OBJECTIVE: This study aimed to evaluate the effects of high-frequency pulsed (HiFP) low-level laser therapy (LLLT) on early wound healing of tooth extraction sockets in rats. STUDY DESIGN/MATERIALS AND METHODS: Bilateral maxillary first molars were extracted from 6-week-old Sprague-Dawley rats. Sockets on the right were treated by HiFP low-level diode laser irradiation (904-910 nm); the left sides served as unirradiated controls. LLLT (0.28 W, 30 kHz, 200-ns pulse, 0.6% duty cycle, 61.2 J/cm2 total power density) was employed immediately after extraction and every 24 hours thereafter. The maxillae including the sockets were resected 3 or 7 days after extraction. Soft-tissue healing was evaluated on days 0, 3, and 7. The bone mineral content (BMC), bone volume (BV), and bone mineral density (BMD) of the extraction sockets were evaluated by microcomputed tomography, and histomorphometric analysis was carried out on day 7. Real-time PCR analysis of osteogenic marker expression and immunohistochemical detection of proliferating cell nuclear antigen (PCNA)-positive cells were performed on day 3. RESULTS: Compared with control sites, the un-epithelialized areas of the extracted sites were significantly reduced by irradiation (P = 0.04), and the BMC, BV, and BMD of laser-treated sites were significantly increased (P = 0.004, 0.006, and 0.009, respectively). On day 7, the mean height of newly formed immature woven bone was higher in laser-treated sites (P = 0.24). On day 3, laser-treated sites showed significantly higher osteocalcin mRNA expression (P = 0.04) and PCNA-positive cell numbers (P = 0.01). CONCLUSION: HiFP low-level diode laser irradiation enhanced soft- and hard-tissue healing of tooth extraction sockets. Lasers Surg. Med. 48:955-964, 2016. © 2016 Wiley Periodicals, Inc.

Cricoarytenoid Articulation in Elderly Japanese With Special Reference to Morphology of the Synovial Tissue.

OBJECTIVE: To clarify composite fibers and cells in the synovial tissues of the cricoarytenoid joint (CA joint). METHODS: Routine histology and immunohistrochemistry using sagittal or nearly sagittal sections obtained from 18 elderly cadaveric specimens. RESULTS: The CA joint capsule was thin and contained few elastic fibers. A limited supportive ligament, namely, a thickened fascia of the posterior cricoarytenoid muscles, was sometimes evident on the lateral aspect of the CA joint. However, even in the weaker medial aspect of the joint, no marked destruction of the synovial tissues was found. The CA joint always contained synovial folds--a short medial fold and long lateral folds--but these contained no or few macrophages, lymphocytes, and blood capillaries. In 2 exceptional specimens showing inflammatory cell infiltration in the submucosal tissue of the larynx, the macrophage-rich area extended toward the capsule and medial synovial fold. CONCLUSIONS: The lateral aspect of the CA joint was likely to be supported mechanically by the muscle-associated tissues. Strong support of the arytenoid by muscles might reduce the degree of CA joint injury with age. However, some patients with hoarseness due to mucosal inflammation of the larynx might have accompanying synovitis and subsequent cartilage injury in the CA joint.

The facilitatory effects of hyperbaric oxygen treatment on membrane bone wound healing in a rat calvarial defect model.

We examined the effect of hyperbaric oxygen (HBO2) treatment on bone wound healing in a rat calvarial defect. Critical-sized defects were created in the calvaria of adult Wistar rats. The animals were divided into four groups--HBO2, normobaric oxygen, hyperbaric air, and no treatment. Treatments were performed five days a week, for two weeks. Micro-computerized tomography and histological analysis were used to evaluate the bone defects. Regenerated bone areas were calculated as the percentage of new bone in the cross-sectional area of defect. The new bone cross-sectional area was significantly greater in the HBO2 group than in the other groups. There were no significant differences in the numbers of nucleated cells in the new bone areas. Although new bone volume per defect volume was significantly greater in the HBO2 group than in the other groups, no significant differences in bone mineral density in the new bone area were observed. These findings indicate the facilitatory role of HBO2 treatment on bone wound healing in the rat calvarial bone defect, and it does not appear to have any negative effects on bone maturity. We propose that HBO2 treatment would be useful in promoting bone regeneration following injury in the orofacial region.

CXCR4/CXCL12 signaling impacts enamel progenitor cell proliferation and motility in the dental stem cell niche.

Dental stem cells are located at the proximal ends of rodent incisors. These stem cells reside in the dental epithelial stem cell niche, termed the apical bud. We focused on identifying critical features of a chemotactic signal in the niche. Here, we report that CXCR4/CXCL12 signaling impacts enamel progenitor cell proliferation and motility in dental stem cell niche cells. We report cells in the apical bud express CXCR4 mRNA at high levels while expression is restricted in the basal epithelium (BE) and transit-amplifying (TA) cell regions. Furthermore, the CXCL12 ligand is present in mesenchymal cells adjacent to the apical bud. We then performed gain- and loss-of-function analyses to better elucidate the role of CXCR4 and CXCL12. CXCR4-deficient mice contain epithelial cell aggregates, while cell proliferation in mutant incisors was also significantly reduced. We demonstrate in vitro that dental epithelial cells migrate toward sources of CXCL12, whereas knocking down CXCR4 impaired motility and resulted in formation of dense cell colonies. These results suggest that CXCR4 expression may be critical for activation of enamel progenitor cell division and that CXCR4/CXCL12 signaling may control movement of epithelial progenitors from the dental stem cell niche.

Nucleotide-sugar transporter SLC35D1 is critical to chondroitin sulfate synthesis in cartilage and skeletal development in mouse and human.

Proteoglycans are a family of extracellular macromolecules comprised of glycosaminoglycan chains of a repeated disaccharide linked to a central core protein. Proteoglycans have critical roles in chondrogenesis and skeletal development. The glycosaminoglycan chains found in cartilage proteoglycans are primarily composed of chondroitin sulfate. The integrity of chondroitin sulfate chains is important to cartilage proteoglycan function; however, chondroitin sulfate metabolism in mammals remains poorly understood. The solute carrier-35 D1 (SLC35D1) gene (SLC35D1) encodes an endoplasmic reticulum nucleotide-sugar transporter (NST) that might transport substrates needed for chondroitin sulfate biosynthesis. Here we created Slc35d1-deficient mice that develop a lethal form of skeletal dysplasia with severe shortening of limbs and facial structures. Epiphyseal cartilage in homozygous mutant mice showed a decreased proliferating zone with round chondrocytes, scarce matrices and reduced proteoglycan aggregates. These mice had short, sparse chondroitin sulfate chains caused by a defect in chondroitin sulfate biosynthesis. We also identified that loss-of-function mutations in human SLC35D1 cause Schneckenbecken dysplasia, a severe skeletal dysplasia. Our findings highlight the crucial role of NSTs in proteoglycan function and cartilage metabolism, thus revealing a new paradigm for skeletal disease and glycobiology.

Coexistence of elastic fibers with hyaluronic acid in the human urethral sphincter complex: a histological study.

PURPOSE: To promote the prevention and treatment of urethral sphincteric dysfunction, we examined the distribution of elastic fibers around the urethral sphincter complex and the histological localization of hyaluronic acid in relation to elastic fiber architecture. MATERIALS AND METHODS: Using elastica-Masson staining as well as biotinated hyaluronic acid binding protein, we examined specimens of the urethral sphincter complex obtained from 14 elderly Japanese cadavers, including 10 men and 4 women. As a control, we also observed other striated muscles in male cadavers. RESULTS: Elastic fibers were densely distributed throughout the submucosal and smooth muscle layers along the entire length of the male urethra, including the prostatic urethra. The levator ani fascia and rhabdosphincter also contained abundant elastic fibers. An intramuscular elastic net was seen in the rhabdosphincter but not in other striated muscles. Strong staining for hyaluronic acid was evident in the submucosa and smooth muscle sphincter of the urethra but not in the levator ani fascia or rhabdosphincter, suggesting that elastic fibers and hyaluronic acid might interact at the former sites. Gender related differences in the distribution of elastic fibers and hyaluronic acid were noted with a much lower density of elastic fibers and hyaluronic acid staining in women than in men. CONCLUSIONS: Urethral sites where elastic fibers and hyaluronic acid coexist could be targeted for the prevention and treatment of urethral sphincteric insufficiency. These findings should improve our understanding of the human urethral sphincter complex.

An immunohistochemical study of matrix components in primary and secondary cartilages of embryonic chick skull.

OBJECTIVES: This study aimed to compare the extracellular matrix of primary cartilage with the secondary cartilage of chicks using immunohistochemical analyses in order to understand the features of chick secondary chondrogenesis. METHODS: Immunohistochemical analysis was performed on the extracellular matrix of quadrate (primary), squamosal, surangular, and anterior pterygoid secondary cartilages using various antibodies targeting the extracellular matrix of cartilage and bone. RESULTS: The localization of collagen types I, II, and X, versican, aggrecan, hyaluronan, link protein, and tenascin-C was identified in the quadrate cartilage, with variations within and between the regions. Newly formed squamosal and surangular secondary cartilages showed simultaneous immunoreactivity for all molecules investigated. However, collagen type X immunoreactivity was not observed, and there was weak immunoreactivity for versican and aggrecan in the anterior pterygoid secondary cartilage. CONCLUSIONS: The immunohistochemical localization of extracellular matrix in the quadrate (primary) cartilage was comparable to that of long bone (primary) cartilage in mammals. The fibrocartilaginous nature and rapid differentiation into hypertrophic chondrocytes, which are known structural features of secondary cartilage, were confirmed in the extracellular matrix of squamosal and surangular secondary cartilages. Furthermore, these tissues appear to undergo developmental processes similar to those in mammals. However, the anterior pterygoid secondary cartilage exhibited unique features that differed from primary and other secondary cartilages, suggesting it is formed through a distinct developmental process.

Elastic fiber-mediated enthesis in the human middle ear.

Adaptation to constant vibration (acoustic oscillation) is likely to confer a specific morphology at the bone-tendon and bone-ligament interfaces at the ear ossicles, which therefore represent an exciting target of enthesis research. We histologically examined (i) the bone attachments of the tensor tympani and stapedius muscles and (ii) the annular ligament of the incudostapedial joint obtained from seven elderly donated cadavers. Notably, both aldehyde-fuchsin and elastic-Masson staining demonstrated that the major fibrous component of the entheses was not collagen fibers but mature elastic fibers. The positive controls for elastic fiber staining were the arterial wall elastic laminae included in the temporal bone materials. The elastic fibers were inserted deeply into the type II collagen-poor fibrocartilage covering the ear ossicles. The muscle tendons were composed of an outer thin layer of collagen fibers and an inner thick core of elastic fibers near the malleus or stapes. In the unique elastic fiber-mediated entheses, hyaluronan, versican and fibronectin were expressed strongly along the elastic fibers. The hyaluronan seemed to act as a friction-reducing lubricant for the elastic fibers. Aggrecan was labeled strongly in a disk- or plica-like fibrous mass on the inner side of the elastic fiber-rich ligament, possibly due to compression stress from the ligament. Tenascin-c was not evident in the entheses. The elastic fiber-mediated entheses appeared resistant to tissue destruction in an environment exposed to constant vibration. The morphology was unlikely to be the result of age-related degeneration.

Comparison between continuous and discontinuous multiple vertebral compression fractures.

BACKGROUND: The majority of multiple vertebral fractures (MVFs) occur under high-energy conditions; however, some cases occurring under minor-energy conditions exist. Fractures in successive vertebrae (continuous type) and in skipped vertebrae (discontinuous type) can exist. PURPOSE: The objective of this study was to compare and evaluate the cause, level of injury, and relationship to osteoporosis between continuous and discontinuous MVFs. METHODS: We studied 77 subjects (173 vertebrae) who had presented with acute back pain between September 2007 and April 2010 and who received diagnoses of fresh MVFs through magnetic resonance imaging. Subjects with continuous and discontinuous fractures were evaluated based on age, sex, bone mineral density (BMD), level of affected vertebrae, and cause of injury. RESULTS: Subjects with discontinuous MVFs were significantly older and comprised more female patients. Mean BMD, measured by dual-emission X-ray absorptiometry, was 0.70 and 0.58 g/cm(3) for the continuous and discontinuous MVFs, respectively, demonstrating a significant difference. Of 34 patients with discontinuous MVFs, 32 (94%) exhibited vertebral fractures in the thoracolumbar junction. In subjects with continuous MVFs, the MVFs of 19 (44%) subjects were caused by high-energy trauma, whereas mild trauma and unknown cause were identified in 14 (41%) and 13 (38%) subjects with discontinuous MVFs, respectively. CONCLUSIONS: Discontinuous MVFs generally caused by mild outer force, and often occurred at the thoracolumbar junction. Continuous MVFs, frequently, were caused by high-energy trauma.

Transient appearance of tyrosine hydroxylase immunoreactive cells in the midline epithelial seam of the human fetal secondary palate.

OBJECTIVE: Transient immunoreactivity for tyrosine hydroxylase, which mediates the conversion of the amino acid L-tyrosine to dihydroxyphenylalanine, in the midline epithelial seam between the bilateral palatal shelves was investigated in human fetuses. MATERIALS AND METHODS: Horizontal or frontal paraffin sections of two human fetuses at 9 and 15 weeks of gestation were used to examine the distribution of tyrosine hydroxylase-immunoreactive cells in regions of the entire head other than the brain. Immunohistochemical staining for S100 protein, calretinin, cytokeratin 14, and vimentin was examined using adjacent or near sections. RESULTS: Tyrosine hydroxylase-immunoreactive cells were large and densely distributed in the midline epithelial seam at the site of palatal fusion in fetuses at 9 weeks but not in fetuses at 15 weeks, in which the midline epithelial seam had already disappeared. No expression of S100 protein, calretinin, or vimentin was detected, but the midline epithelial seam was positive for cytokeratin 14. Tyrosine hydroxylase immunoreactivity was not detected in epithelia during the process of palatal fusion in mice from E 14.0 to 15.0. CONCLUSIONS: These findings indicate that tyrosine hydroxylase-immunoreactive cells in the midline epithelial seams are nonneural epithelial cells and suggest that the tyrosine hydroxylase is a novel factor involved in normal palatal formation, especially the fate of the midline epithelial seam in humans.

An in situ hybridization study of syndecan family during the late stages of developing mouse molar tooth germ.

Expression of syndecan-1, 2, 3, and 4 mRNAs during the late stages of tooth germ formation was investigated by in situ hybridization, using [35S]-UTP-labeled cRNA probes. Syndecan-1 mRNA was mainly expressed in the stellate reticulum and stratum intermedium as well as at the cervical region of dental papilla/dental follicle during E18.5-P3.0. Expression in the dental epithelium was enhanced during the postnatal periods, which was supported by real-time RT-PCR analysis. These spatiotemporal expression patterns may suggest specific roles of syndecan-1 in tooth formation such as tooth eruption or root formation. Syndecan-3 mRNA expression became evident in odontoblasts at E18.5, but compared to collagen type I mRNA, which was strongly expressed at this stage, syndecan-3 expression in odontoblast was restricted in mature odontoblasts beneath the cusps during the postnatal periods. This result was also supported by real-time RT-PCR analysis, and indicated that syndecan-3 may be involved in the progress of dentinogenesis rather than in the initiation of it. Syndecan-4 mRNA roughly showed comparable expression patterns to those of syndecan-3. Syndecan-2 mRNA did not show significant expression during the experimental period, but real-time RT-PCR analysis suggested that syndecan-2 expression might be enhanced with hard tissue formation.

Functional analysis of CTRP3/cartducin in Meckel's cartilage and developing condylar cartilage in the fetal mouse mandible.

CTRP3/cartducin, a novel C1q family protein, is expressed in proliferating chondrocytes in the growth plate and has an important role in regulating the growth of both chondrogenic precursors and chondrocytes in vitro. We examined the expression of CTRP3/cartducin mRNA in Meckel's cartilage and in condylar cartilage of the fetal mouse mandible. Based on in situ hybridization studies, CTRP3/cartducin mRNA was not expressed in the anlagen of Meckel's cartilage at embryonic day (E)11.5, but it was strongly expressed in Meckel's cartilage at E14.0, and then reduced in the hypertrophic chondrocytes at E16.0. CTRP3/cartducin mRNA was not expressed in the condylar anlagen at E14.0, but was expressed in the upper part of newly formed condylar cartilage at E15.0. At E16.0, CTRP3/cartducin mRNA was expressed from the polymorphic cell zone to the upper part of the hypertrophic cell zone, but was reduced in the lower part of the hypertrophic cell zone. CTRP3/cartducin-antisense oligodeoxynucleotide (AS-ODN) treatment of Meckel's cartilage and condylar anlagen from E14.0 using an organ culture system indicated that, after 4-day culture, CTRP3/cartducin abrogation induced curvature deformation of Meckel's cartilage with loss of the perichondrium and new cartilage formation. Aggrecan, type I collagen, and tenascin-C were simultaneously immunostained in this newly formed cartilage, indicating possible transformation from the perichondrium into cartilage. Further, addition of recombinant mouse CTRP3/cartducin protein to the organ culture medium with AS-ODN tended to reverse the deformation. These results suggest a novel function for CTRP3/cartducin in maintaining the perichondrium. Moreover, AS-ODN induced a deformation of the shape, loss of the perichondrium/fibrous cell zone, and disorder of the distinct architecture of zones in the mandibular condylar cartilage. Additionally, AS-ODN-treated condylar cartilage showed reduced levels of mRNA expression of aggrecan, collagen types I and X, and reduced BrdU-incorporation. These results suggest that CTRP3/cartducin is not only involved in the proliferation and differentiation of chondrocytes, but also contributes to the regulation of mandibular condylar cartilage.

Fetal head anomaly restricted to the eye, the mandible, and the pterygoid process of the sphenoid: a histological study.

A report on an unusual combination of anomalies in the head of a female fetus. The authors examined whole body semiserial paraffin sections of a female fetus (155 mm CRL; ∼18 weeks of gestation), with a particular focus on the head region. Cranial autonomic ganglia, nasal olfactory cells, and the orbital muscle were investigated using immunohistochemistry for tyrosine hydroxylase, vasoactive intestinal peptide, calretinin, and smooth muscle actin expression. The surface gross anatomy of the fetus appeared normal. The left eyeball lacked a lens (the eyeballs were otherwise normal). The orbital muscle was very thick and located in the anterolateral side of the extraocular muscles. Conversely, the extraocular muscles made a cluster in the superoposterior side of the orbit. The infratemporal fossa was small due to the bulky, transversely extended lateral pterygoid process in contrast to the small coronoid process of the mandible. The bilateral mandibular bases overlapped at the midline symphysis. The thin orbitosphenoid and thick alisphenoid provided an almost flat, anterior cranial base. Nasal olfactory cells and cranial autonomic ganglia appeared to be normal. No major anomaly was observed in the brain. Because of the changes in topographical anatomy, the orbital muscle probably lost its normal bony attachment and appeared to push the extraocular muscles superoposteriorly. A gene function redundancy rather than mutation may explain the present restricted anomalies in the mandible and pterygoid process.

An in situ hybridization study of decorin and biglycan mRNA in mouse osteoblasts in vivo.

In situ hybridization of decorin and biglycan mRNA, principal members of small leucine-rich proteoglycan, was performed using [35S]-labeled RNA probes, in the context of the hypothesis that they show different expression patterns associated with osteoblast differentiation in mice. We adopted two ossifying sites that can clearly follow the developmental process of bone formation: ossifying tympanic ring and developing bone collar of mandibular condylar cartilage. Decorin mRNA was expressed in osteoblasts of developing tympanic ring at E14.0, as well as of developing bone collar at E15.0, but biglycan mRNA was not, indicating decorin mRNA was expressed earlier in newly differentiating osteoblasts than biglycan. With maturation of osteoblasts, biglycan mRNA became expressed and maintained its expression both in the outer region (periosteum) and in the interior region (endosteum) of bone. By contrast, decorin mRNA expression was maintained in the outer region but diminished in the interior region. These results indicate that decorin and biglycan show differential expression patterns in differentiating osteoblasts and play specific roles in bone formation.

An in situ hybridization study of the Syndecan family in the developing condylar cartilage of fetal mouse mandible.

Mandibular condylar cartilage is a representative secondary cartilage, differing from primary cartilage in various ways. Syndecan is a cell-surface heparan sulfate proteoglycan and speculated to be involved in chondrogenesis and osteogenesis. This study aimed to investigate the expression patterns of the syndecan family in the developing mouse mandibular condylar cartilage. At embryonic day (E)13.0 and E14.0, syndecan-1 and -2 mRNAs were expressed in the mesenchymal cell condensation of the condylar anlage. When condylar cartilage was formed at E15.0, syndecan-1 mRNA was expressed in the embryonic zone, wherein the mesenchymal cell condensation is located. Syndecan-2 mRNA was mainly expressed in the perichondrium. At E16.0, syndecan-1 was expressed from fibrous to flattened cell zones and syndecans-2 was expressed in the lower hypertrophic cell zone. Syndecan-3 mRNA was expressed in the condylar anlage at E13.0 and E13.5 but was not expressed in the condylar cartilage at E15.0. It was later expressed in the lower hypertrophic cell zone at E16.0. Syndecan-4 mRNA was expressed in the condylar anlage at E14.0 and the condylar cartilage at E15.0 and E16.0. These findings indicated that syndecans-1 and -2 could be involved in the formation from mesenchymal cell condensation to condylar cartilage. The different expression patterns of the syndecan family in the condylar and limb bud cartilage suggest the functional heterogeneity of chondrocytes in the primary and secondary cartilage.

The third vascular route of the inner ear or the canal of Cotugno: Its topographical anatomy, fetal development, and contribution to ossification of the otic capsule cartilage.

Three vascular routes to the inner ear are known: (a) through the internal acoustic meatus with the vestibulocochlear nerve; (b) from the endolymphatic duct aperture; and (c) along the canal of Cotugno (CC) inserted into the vestibular part of the ear from the superior or brain side. The third is believed to contain only veins. Examinations of 33 human embryos and fetuses at 6-40 weeks demonstrated that (a) the CC appeared as a recess of epidural mesenchymal tissues at the superior aspect of the otic capsule cartilage in embryos and it was inserted deeply to issue multiple peripheral divisions inferolaterally and posteriorly at midterm; (b) the CC consistently passed through a ring of the superior or anterior semicircular canal and contained both, the arteries from the vestibulocochlear nerve origin at the midbrain and the vein draining into the sigmoid sinus or petrosal sinuses; and (c) the CC appeared not to contribute to ossification of the otic capsule cartilage but, after endochondral ossification of the internal ear, woven bone development occurred along a smooth interface of the CC with the ossified ear. In contrast, another interface between the developing bone and the residual cartilage of the otic capsule was rough and wavy with many short bony columns, called osseous globules. In addition, the endolymphatic duct accompanied veins but no arteries. Our results show that the CC is a major vascular route to the vestibular part of the otic capsule cartilage, but its role appears to be limited after ossification.

Association between underlying disease and serious events on a world cruise ship: A prospective cohort study.

BACKGROUND: This study aimed to clarify the effects of underlying diseases on clinical outcomes of patients aboard a world cruise ship. METHODS: This prospective cohort study included patients who sought physician consultations at an onboard clinic on a 105-day world cruise (September-December 201X) on a ship chartered by a Japanese travel agency. Multivariable logistic regression analysis was performed to ascertain whether any concurrent disease, such as hypertension, was associated with additional onboard treatment by the primary physician or serious events, including unexpected final disembarkation, temporary disembarkation for hospitalization ashore, shore-side referral, and onboard clinic admission. RESULTS: Of 313 patients, 182 (58%) had at least one underlying disease. Sixty-eight (22%) required additional treatment, and 24 (8%) experienced serious events. After adjusting for age, sex, and underlying diseases, the 60-69- and 70-74-year age groups had a lower risk of serious events than the ≤59-year age group (odds ratio [OR], 95% confidence interval [CI]: 0.24, 0.069-0.81; p = 0.022 and 0.045, 0.0051-0.47; p = 0.0055). Underlying disease was associated with serious events (OR, 95% CI: 3.2, 1.1-9.5; p = 0.036). CONCLUSIONS: Unexpected events can occur in patients on world cruises regardless of age. Preexisting diseases may confer higher risk of serious events.

False positive reactivity of a substance P-antibody in the ectodermal/epithelial plug of the nose, ear, eye and perineum of the human and mouse fetuses.

Epithelial/ectodermal plug formation in the developing nose, ear, and eye regions is followed by canalization/recanalization mediated by cell death. However, the mechanism is not well understood. Recently, substance P (SP)-mediated cell death, rather than cell apoptosis, has been reported in neuronal and non-neuronal cells. Horizontal paraffin sections of 5 human fetuses at 15-16 weeks of gestation were used to examine the entire area of the nose, ear, eye and perineum with immunohistochemistry for SP and its receptor neurokinin-1 (NK-1), and protein gene product (PGP) 9.5 and S100 protein to identify whether the positive cells had neural origins. The deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) method was also conducted to identify apoptosis. Four SP antibodies were commercially obtained and compared the results. In addition, using the same antibodies for SP, those results were compared with fetal mouse heads (E14-17). Substance P immunoreactivity of one of the 4 antibodies (sc9758) was clearly found in the nasal plug, the epithelium of the anterior nasal cavity, the entire excretory tear duct, the marginal palpebral conjunctiva, the auditory meatal plug, the parotid duct, the external urethral orifice and, the preputial lamella along the future prepuce. Immunoreactivity was usually seen in enlarged round cells in humans. In fetal mouse heads, in spite of negative reaction in all these sites, the midline epithelial seam at the palate fusion and the oral epithelium especially at and near the tooth germ specifically reacted with the sc9758. Nevertheless, the other 3 antibodies did not react at any of those sites both in human and mouse fetuses. NK-1 receptor-positive cells were seen in the nose and meatal plugs and preputial lamella, but not in the tear duct. S100 protein, PGP 9.5, and TUNEL method all demonstrated negative reactivity at any sc9758-positive sites. Consequently, the present immunoreactivity of the sc9758 antibody seemed to be false positive, but it was likely to react with a specific substance in the epithelial or ectodermal cell because of the clearly restricted staining. Which substance it crossed remains unknown.