Comparison of the 3-D patterns of the parasympathetic nervous system in the lung at late developmental stages between mouse and chicken.

Although the basic schema of the body plan is similar among different species of amniotes (mammals, birds, and reptiles), the lung is an exception. Here, anatomy and physiology are considerably different, particularly between mammals and birds. In mammals, inhaled and exhaled airs mix in the airways, whereas in birds the inspired air flows unidirectionally without mixing with the expired air. This bird-specific respiration system is enabled by the complex tubular structures called parabronchi where gas exchange takes place, and also by the bellow-like air sacs appended to the main part of the lung. That the lung is predominantly governed by the parasympathetic nervous system has been shown mostly by physiological studies in mammals. However, how the parasympathetic nervous system in the lung is established during late development has largely been unexplored both in mammals and birds. In this study, by combining immunocytochemistry, the tissue-clearing CUBIC method, and ink-injection to airways, we have visualized the 3-D distribution patterns of parasympathetic nerves and ganglia in the lung at late developmental stages of mice and chickens. These patterns were further compared between these species, and three prominent similarities emerged: (1) parasympathetic postganglionic fibers and ganglia are widely distributed in the lung covering the proximal and distal portions, (2) the gas exchange units, alveoli in mice and parabronchi in chickens, are devoid of parasympathetic nerves, (3) parasympathetic nerves are in close association with smooth muscle cells, particularly at the base of the gas exchange units. These observations suggest that despite gross differences in anatomy, the basic mechanisms underlying parasympathetic control of smooth muscles and gas exchange might be conserved between mammals and birds.

Newly raised anti-VAChT and anti-ChAT antibodies detect cholinergic cells in chicken embryos.

The autonomic nervous system consists of sympathetic and parasympathetic nerves, which functionally antagonize each other to control physiology and homeostasis of organs. However, it is largely unexplored how the autonomic nervous system is established during development. In particular, early formation of parasympathetic network remains elusive because of its complex anatomical structure. To distinguish between parasympathetic (cholinergic) and sympathetic (adrenergic) ganglia, vesicular acetylcholine transporter (VAChT) and choline O-acetyltransferase (ChAT), proteins associated with acetylcholine synthesis, are known to be useful markers. Whereas commercially available antibodies against these proteins are widely used for mammalian specimens including mice and rats, these antibodies do not work satisfactorily in chickens, although chicken is an excellent model for the study of autonomic nervous system. Here, we newly raised antibodies against chicken VAChT and ChAT proteins. One monoclonal and three polyclonal antibodies for VAChT, and one polyclonal antibody for ChAT were obtained, which were available for Western blotting analyses and immunohistochemistry. Using these verified antibodies, we detected cholinergic cells in Remak ganglia of autonomic nervous system, which form in the dorsal aspect of the digestive tract of chicken E13 embryos. The antibodies obtained in this study are useful for visualization of cholinergic neurons including parasympathetic ganglia.

Complications of Distal Radius Fractures Treated by Volar Locking Plate Fixation.

The current study investigated the incidence of complications after surgery for distal radial fractures. This multicenter retrospective study was conducted at 11 institutions. A total of 824 patients who had distal radius fractures that were treated surgically between January 2010 and August 2012 were identified. The study patients were older than 18 years and were observed for at least 12 weeks after surgery for distal radius fractures with a volar locking plate. Sex, age, fracture type according to AO classification, implants, wrist range of motion, grip strength, fracture consolidation rate, and complications were studied. Analysis included 694 patients, including 529 women and 165 men, with a mean age of 64 years. The mean follow-up period was 27 weeks. The fracture consolidation rate was 100%. There were 52 complications (7.5%), including 18 cases of carpal tunnel syndrome, 12 cases of peripheral nerve palsy, 8 cases of trigger digit, 4 cases of tendon rupture (none of the flexor pollicis longus), and 10 others. There was no rupture of the flexor pollicis longus tendon because careful attention was paid to the relationship between the implant and the tendon. Peripheral nerve palsy may have been caused by intraoperative traction in 7 cases, temporary fixation by percutaneous Kirschner wires in 3 cases, and axillary nerve block in 1 case; 1 case appeared to be idiopathic. Tendon ruptures were mainly caused by mechanical stress. [Orthopedics.2016; 39(5):e893-e896.].

[Roles and cooperation of medical professionals in natural disasters].

We have reconsidered the responsibility of occupational therapists who have been supporting the victims of the Great East Japan Earthquake. They can analyze problems and provide appropriate support for victims with rehabilitation and occupational therapy as well as for handicapped people. Support measures that can be provided by occupational therapists are as follows: 1) Maintenance and improvement of mind and body functions through occupational therapy. 2) Mental care. 3) Coordination of social circumstances for elderly and handicapped people. 4) Maintenance and improvement of ability to perform common activities of daily living. 5) Choice and adaptation of welfare equipment. Especially, occupational therapy provided with the aim to open victims' minds has an effect on mental care. Their mental wounds cannot be healed easily. However, networking and work activities play important roles in dealing with daily life. Occupational therapists will be expected to provide long-term treatment for victims through work activities with professional skills.

Radiographic features of Kirner's deformity.

We conducted a study of Kirner's deformity to investigate its radiologic features and consider its possible causes. Sixty-seven patients with Kirner's deformity, 41 in our series and 26 retrieved in a survey of the literature were investigated. We divided Kirner's deformity into three types according to the site of curvature: the epiphyseal line, diaphysis, and distal tip. Among our series, 12 hands were affected in males and 26 in females. The gender of the patients for the other three hands was unknown. Twelve cases occurred on the right side, five on the left side, and 24 on both sides. Radiographic data for 34 hands were available. The palmar surface of the distal phalanx was at a mean angle of 27.4 degrees to the long axis of the middle phalanx. Epiphyseal line curvature was seen in four hands (mean patient age, 11 years), diaphysis curvature in 10 (mean age, 12.4 years), and distal tip curvature in 20 (mean age, 26.5 years). Dorsal subluxation of the distal phalanx was noticed in six hands (18%). Radiographic data for 26 hands reported previously were analyzable. The differences between epiphyseal line or diaphysis curvature and distal tip curvature in terms of age were significant in both the present study and the literature.

Metacarpal hypoplasia associated with congenital constriction band syndrome.

PURPOSE: The amputation of digits that occurs in association with congenital constriction band syndrome (CCBS) is often described as similar to that resulting from trauma. Some previous studies have suggested that no bone hypoplasia is present in the affected limb proximal to the amputated part. The purpose of this study was to report our series of patients with metacarpal hypoplasia in amputated digits associated with CCBS. METHODS: We reviewed a total of 37 hands of 24 patients diagnosed as having amputated fingers owing to CCBS and evaluated the clinical features of the metacarpal bone hypoplasia. RESULTS: We found coexistent metacarpal hypoplasia in 11 hands in 9 patients. Three of the patients were male and 6 were female. The average age at the time of examination was 9 years (range, 4-20 y). We observed metacarpal hypoplasia in cases of amputation at the level of the proximal phalanx and not at the level of the middle phalanx. CONCLUSIONS: We found metacarpal hypoplasia in 38% of patients and in 30% of hands with amputated digits owing to CCBS. TYPE OF STUDY/LEVEL OF EVIDENCE: Prognostic IV.

Radial longitudinal deficiencies with hypoplastic/absent thumbs and cutaneous syndactyly of the most radial digits.

PURPOSE: To describe radial longitudinal deficiency with hypoplastic or absent thumb and cutaneous syndactyly between the most radial digits. In addition, to discuss the clinical relevance and unique treatment decisions involved in optimizing functional outcomes in these patients. METHODS: A total of 163 extremities of 122 patients with radial longitudinal deficiencies were reviewed. We reviewed radiographs and clinical images that were available, with most radial hypoplastic digit and cutaneous syndactyly to the adjacent finger. RESULTS: There were 7 hands with this type of deformity. Four cases had a hypoplastic thumb associated with cutaneous syndactyly between the thumb and index finger. Three cases had a deformity in which the thumb was absent and the hypoplastic index finger was syndactylized to the long finger. Proximal and distal radioulnar synostoses were associated with these deformities in 3 patients. CONCLUSIONS: Because the most radial digit is severely hypoplastic in this type of deformity, pollicization is usually indicated. However, the pollicization procedure must be modified due to associated syndactyly with different degrees of hypoplasia or absence of the intrinsic muscles. This type of deformity should be distinguished from hypoplastic thumb without syndactyly. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

Tissue morphogenesis coupled with cell shape changes.

When different tissues successively emerge during development, they need to be morphologically segregated from each other. Morphological segregation of tissues is often accompanied by robust changes in cell shape, and these two events are precisely coordinated. We overview recent progress in understanding how such coordination is regulated at the cellular and molecular levels using vertebrate somitogenesis asa unique model. In the formation of the somitic gap and its concomitant cell epithelialization, Ephrin-Eph intercellular signals play a central role, with Ephrin transducing intracellular signals via suppression of Cdc42. These signals lead to the activation of integrin where the segment border of somites forms, which in turn induces the assembly of fibronectin, the final player for the coordination. Intimate coupling of tissue-shape and cell-shape changes is also relevant to tumor suppression.

Snapping elbow with congenital radial head dislocation: case report.

An 11-year-old boy with congenital radial head dislocation experienced painful snapping of his left elbow upon movement. He had no previous history of trauma. A plain radiograph of his left elbow showed anterior dislocation of the radial head and flexion deformity of the hypoplastic radial neck. Arthroscopy showed that the snapping of the elbow occurred between the annular ligament and the dislocated radial head during elbow flexion and extension. After the annular ligament was released, the snapping immediately disappeared. Five years after the surgery, the patient has no pain or snapping upon elbow movement.

Long-term follow-up of radial shortening osteotomy for Kienbock disease. Surgical technique.

BACKGROUND: Three previous studies have investigated the long-term outcome of radial osteotomy in the treatment of Kienböck disease. However, none used patient-based assessment. The purpose of this study was to investigate the long-term clinical and radiographic outcomes of this osteotomy, including the subjective evaluation of the patient with use of the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire. METHODS: A DASH questionnaire was sent to nineteen patients with Kienböck disease who had undergone a radial shortening osteotomy, and thirteen replied. The mean age at the time of surgery was thirty-nine years. On the basis of the Lichtman classification, six patients had stage-II, four had stage-IIIA, and three had stage-IIIB disease. Prior to surgery, ulnar variance was positive in six patients, neutral in four, and negative in three. The mean duration of follow-up was twenty-one years. Clinical evaluation, including calculation of the modified Mayo wrist score, and radiographic evaluation were also performed on twelve of the thirteen patients. RESULTS: The mean DASH score was 8 points (range, 0 to 23 points), and patient satisfaction was high. Compared with the findings in the contralateral wrist, the mean range of motion was 81% in flexion and 82% in extension and mean grip strength was 88%. The mean modified Mayo wrist score was 83 points, and the clinical results were excellent in six patients, good in five, and moderate in one. The DASH scores tended to be worse in patients with Lichtman stage-IIIB disease. Follow-up radiographs revealed that the Lichtman stage had progressed in six of the twelve patients. CONCLUSIONS: Although most patients had mild wrist pain, patient satisfaction and the clinical results were satisfactory following a radial shortening osteotomy. This procedure is a reliable long-term treatment for Lichtman stage-II and IIIA disease and may be a reasonable option for patients with stage-IIIB disease.

De Quervain disease caused by abductor pollicis longus tenosynovitis: a report of three cases.

De Quervain disease is caused by a stenosing tenosynovitis in the first dorsal compartment, and the main aetiology is extensor pollicis brevis (EPB) tenosynovitis. We encountered three cases in which EPB tenosynovitis was absent and abductor pollicis longus (APL) tenosynovitis was confirmed during operation. In the treatment of de Quervain disease, APL tenosynovitis should be paid as much attention as EPB tenosynovitis.

EphrinB2 coordinates the formation of a morphological boundary and cell epithelialization during somite segmentation.

During early morphogenesis, tissue segregation is often accompanied by changes in cell shape. To understand how such coordination is regulated, somitogenesis was used as a model. When a somite forms in the anterior end of the presomitic mesoderm, an intersomitic boundary (gap) emerges, and it is rapidly followed by cell epithelialization at this border. It has been known that the gap formation is regulated by intercellular signals. We here demonstrate that cMeso-1, the chicken homolog of mouse Mesp2, up-regulates EphA4 in the cells located posteriorly to a forming boundary. This in turn activates EphrinB2-reverse signals in the anteriorly juxtaposed cells, where the EphrinB2 signal is sufficient to cause a gap formation and cell epithelialization cell-autonomously. During these processes, Cdc42 needs to be repressed via tyrosine phosphorylation of EphrinB2. This is the first demonstration that Ephrin-reverse signal acts as a platform that couples distinct morphogenetic changes in cell polarity and tissue shape.

Long-term follow-up of radial shortening osteotomy for Kienbock disease.

BACKGROUND: Three previous studies have investigated the long-term outcome of radial osteotomy in the treatment of Kienböck disease. However, none used patient-based assessment. The purpose of this study was to investigate the long-term clinical and radiographic outcomes of this osteotomy, including the subjective evaluation of the patient with use of the Disabilities of the Arm, Shoulder and Hand (DASH) Questionnaire. METHODS: A DASH questionnaire was sent to nineteen patients with Kienböck disease who had undergone a radial shortening osteotomy, and thirteen replied. The mean age at the time of surgery was thirty-nine years. On the basis of the Lichtman classification, six patients had stage-II, four had stage-IIIA, and three had stage-IIIB disease. Prior to surgery, ulnar variance was positive in six patients, neutral in four, and negative in three. The mean duration of follow-up was twenty-one years. Clinical evaluation, including calculation of the modified Mayo wrist score, and radiographic evaluation were also performed on twelve of the thirteen patients. RESULTS: The mean DASH score was 8 points (range, 0 to 23 points), and patient satisfaction was high. Compared with the findings in the contralateral wrist, the mean range of motion was 81% in flexion and 82% in extension and mean grip strength was 88%. The mean modified Mayo wrist score was 83 points, and the clinical results were excellent in six patients, good in five, and moderate in one. The DASH scores tended to be worse in patients with Lichtman stage-IIIB disease. Follow-up radiographs revealed that the Lichtman stage had progressed in six of the twelve patients. CONCLUSIONS: Although most patients had mild wrist pain, patient satisfaction and the clinical results were satisfactory following a radial shortening osteotomy. This procedure is a reliable long-term treatment for Lichtman stage-II and IIIA disease and may be a reasonable option for patients with stage-IIIB disease.

Notch mediates the segmental specification of angioblasts in somites and their directed migration toward the dorsal aorta in avian embryos.

We studied, using avian embryos, mechanisms underlying the three-dimensional assembly of the dorsal aorta, the first-forming embryonic vessel in amniotes. This vessel originates from two distinct cell populations, the splanchnic and somitic mesoderms. We have unveiled a role for Notch signaling in the somitic contribution. Upon activation of Notch signaling, a subpopulation of cells in the posterior half of individual somites migrates ventrally toward the primary dorsal aorta of splanchnic origin. After reaching the primary aorta, these somitic cells differentiate into the definitive aortic endothelial cells. This Notch-induced ventral migration is mediated by EphrinB2 and by an attractant action of the primary aorta. Furthermore, long-term chasing of cells by transposon-mediated gene transfer reveals that the segmentally provided endothelial cells of somitic origin in the dorsal aorta ultimately populate the entire region of the vessel. We demonstrate the molecular and cellular mechanisms underlying the formation of embryonic blood vessels from mesenchymal cells.

Stable integration and conditional expression of electroporated transgenes in chicken embryos.

The in ovo electroporation in chicken embryos has widely been used as a powerful tool to study roles of genes during embryogenesis. However, the conventional electroporation technique fails to retain the expression of transgenes for more than several days because transgenes are not integrated into the genome. To overcome this shortcoming, we have developed a transposon-mediated gene transfer, a novel technique in chicken manipulations. It was previously reported that the transposon Tol2, originally found in medaka fish, facilitates an integration of a transgene into the genome when co-acting with Tol2 transposase. In this study, we co-electroporated a plasmid containing a CAGGS-EGFP cassette cloned in the Tol2 construct along with a transposase-encoding plasmid into early presomitic mesoderm or optic vesicles of chicken embryos. This resulted in persistent expression of EGFP at least until embryonic day 8 (E8) and E12 in somite-derived tissues and developing retina, respectively. The integration of the transgene was confirmed by genomic Southern blotting using chicken cultured cells. We further combined this transposon-mediated gene transfer with the tetracycline-dependent conditional expression system that we also developed recently. With this combined method, expression of a stably integrated transgene could be experimentally induced upon tetracycline administration at relatively late stages such as E6, where a variety of organogenesis are underway. Thus, the techniques proposed in this study provide a novel approach to study the mechanisms of late organogenesis, for which chickens are most suitable model animals.

Tet-on inducible system combined with in ovo electroporation dissects multiple roles of genes in somitogenesis of chicken embryos.

The in ovo electroporation technique in chicken embryos has enabled investigators to uncover the functions of numerous developmental genes. In this technique, the ubiquitous promoter, CAGGS (CMV base), has often been used for overexpression experiments. However, if a given gene plays a role in multiple steps of development and if overexpression of this gene causes fatal consequences at the time of electroporation, its roles in later steps of development would be overlooked. Thus, a technique with which expression of an electroporated DNA can be controlled in a stage-specific manner needs to be formulated. Here we show for the first time that the tetracycline-controlled expression method, "tet-on" and "tet-off", works efficiently to regulate gene expression in electroporated chicken embryos. We demonstrate that the onset or termination of expression of an electroporated DNA can be precisely controlled by timing the administration of tetracycline into an egg. Furthermore, with this technique we have revealed previously unknown roles of RhoA, cMeso-1 and Pax2 in early somitogenesis. In particular, cMeso-1 appears to be involved in cell condensation of a newly forming somite by regulating Pax2 and NCAM expression. Thus, the novel molecular technique in chickens proposed in this study provides a useful tool to investigate stage-specific roles of developmental genes.

Immunologic effects of perinatal exposure to dioxins, PCBs and organochlorine pesticides in Japanese infants.

Effects of perinatal exposure to dioxins, PCBs and organochlorine pesticides on lymphocyte subsets were investigated in the peripheral blood from 101 Japanese infants with approximately 10 months of age. Perinatal exposure to these organochlorine compounds were estimated by their contamination levels in the breast milk of the mothers. Lymphocyte subsets such as CD16+, HLA-DR+, CD4+, CD4+8+, CD8+, CD3+ and CD20+ cells in peripheral venous blood were assessed in a subgroup of 92 infants. Greater exposures to HCE, chlordane and dioxins were significantly associated with the increase in the percentages of CD8+ and CD3+ T lymphocytes and CD4+/CD8+ T cell ratios, respectively. In addition, higher HCH exposure was also associated with a decrease in the percentage of HLA-DR+ T lymphocytes. Furthermore effects of dioxins, DDT and PCBs on the percentage of CD16+ T lymphocyte were more pronounced by the combined exposure of dioxins and PCBs or by the combined exposure of DDT and PCBs. Effects of HCE on the percentages of CD8+ T lymphocyte were also more pronounced by the combined exposure of HCE and chlordane. In conclusion, our study suggests that greater exposures to dioxins, PCBs and organochlorine pesticides determined in this study (except dieldrin) influence the immune system of Japanese infant, although the clinical significance of these changes is uncertain.

The short-term reliability of grip strength measurement and the effects of posture and grip span.

PURPOSE: Because of the difficulty in verifying the reliability and validity of grip strength there is still no consensus regarding its measurement, particularly short-term reliability. The present study was conducted to investigate the short-term reliability of grip strength measurement and the effects of posture and grip span. METHODS: One hundred healthy subjects (50 men, 50 women; mean age, 38.2 y; range, 22-58 y) were evaluated. Grip strength was measured twice as a single set by using a dynamometer and the mean value for each hand was recorded. First 3 sets of measurements were performed using 2 different approaches: (1) continuous measurement without rest and (2) interval measurement with a 1-minute rest after each set. Next 1 set of measurements was performed with 3 types of grip span: standard grip span (which was measured as one half the distance between the index finger tip and the metacarpophalangeal joint flexion crease at the base of the thumb), +10% of the standard grip span, and -10% of the standard grip span. Finally 1 set of measurements was performed in 3 postures: standing, sitting, and supine. RESULTS: During continuous measurement the grip strength decreased gradually as the number of sets increased. During interval measurement, however, there was no change among sets for both genders and each hand. On the basis of this result subsequent studies were performed using interval measurement. There was no significant difference in maximum grip strength between the standard and +10% of standard grip span measurements; however, the -10% of standard model gave the minimum grip strength in both genders. With regard to posture the minimum grip strength in both genders was obtained when the subject was supine, with no difference between standing and sitting. CONCLUSIONS: Our study showed that interval measurement with a 1-minute rest after each set yielded a constant value; therefore, we advocate this approach for rapid evaluation of grip strength under different conditions. In addition the influence of grip span and posture should be considered to maximize data accuracy.