Morphological features of the deep component of the posterior inferior tibiofibular ligament.

PURPOSE: The deep component of the posterior inferior tibiofibular ligament (PITFL) was classified by type in this large-scale cadaveric study to provide basic information that will help elucidate the mechanisms underlying ankle joint posterior impingement syndrome. METHODS: This investigation examined 100 legs from 49 Japanese cadavers (mean age at death, 79 ± 11 years; 58 sides from men, 42 from women). In classification, absence of an independent fiber of the deep component of the PITFL were classified as Type I, an independent fiber of the deep component of the PITFL was classified as Type II-a, an independent fiber of the deep component of the PITFL with bundles of fibers connected to the posterior intermalleolar ligament (PIML) was classified as Type II-b, and an independent fiber of the deep component of PITFL with a band shape connected to the PIML was classified as Type III. RESULTS: A deep component of the PITFL was present in all specimens. An independent fiber of the deep component of the PITFL was present in 37 legs (37%), connecting to the deep component of the PITFL and PIML in 26 (70.3%). Several types of deep component of the PITFL were identified: Type I in 63 legs (63%); Type II-a in 11 (11%); Type IIb in 12 (12%); and Type III in 14 (14%). No significant differences were seen between the right and left legs. Type I male were significantly more than Type I female (p < 0.05). Type III female were significantly more than Type III male (p < 0.05). CONCLUSIONS: Strong relationships exist between the PITFL and PIML.

Morphological features of the cervical ligament.

PURPOSE: The purpose of this study was to clarify the morphological characteristics of the cervical ligament (CL). METHODS: This study examined 80 legs from 40 Japanese cadavers. The CL was classified by the number of fiber bundles. The morphological features measured were fiber bundle length, width, thickness, and angle with the sagittal plane. RESULTS: The CL was classified as follows: Type I, the CL is a single fiber; Type II, the CL consists of a superficial fiber and an inferior fiber; and Type III, the CL consists of a superficial fiber, intermediate fiber, and inferior fiber. Type I was seen in 15 feet, Type II in 57 feet, and Type III in 8 feet. In comparisons of morphological features within each type, significant differences were seen in fiber bundle length, width, and angle between superior fiber bundles and inferior fiber bundles of Type II and Type III. In comparison among types, the total fiber bundle width was significantly wider in Type II and Type III than in Type I, and the angle was significantly smaller in Type III than in Type I. CONCLUSION: The results of this study suggested that each type may have different sub-talar joint control functions.

Morphological features of the inferior fascicle of the anterior inferior tibiofibular ligament.

In this study, the inferior fascicle of the anterior inferior tibiofibular ligament (AITFL) was classified to provide basic information to help elucidate the mechanism of ankle joint anterolateral impingement, and the morphological features of each type were compared for the purpose of clarification. This investigation examined 100 feet from 52 cadavers. The AITFL was classified into four types according to the presence or absence of the inferior fascicle and the positional relationship between the AITFL and the inferior fascicle of the AITFL. The morphological features of the AITFL that were measured included the fibre bundle length, fibre bundle width, fibre bundle angle, and the distance between the joint levels. A distinct, independent inferior fascicle of the AITFL was identified in 15 feet (15%). There were no significant differences in the morphological features based on differences in the AITFL classification. Therefore, these findings suggest that the presence or absence of the inferior fascicle and the difference in the positional relationship between the AITFL and the inferior fascicle of the AITFL are less likely to be involved in impingement during ankle dorsiflexion.

Morphological features of the posterior intermalleolar ligament.

PURPOSE: In the present study, the posterior intermalleolar ligament (PIML) was classified by type using large-scale cadavers to provide basic information to help elucidate the mechanism of ankle joint posterior impingement syndrome. METHODS: This investigation examined 100 legs from 49 Japanese cadavers (mean age at death, 79 ± 11 years; 58 sides from men, 42 from women). In the classification method, an absent PIML was classified as Type I, a PIML with one fiber bundle (attachment to one place) was Type II, a PIML with two fiber bundles (attachment to two places) was Type III, and a PIML with three fiber bundles (attachment to three or more places) was Type IV. Furthermore, according to other adhering tissues, they were further subdivided and classified by type. RESULTS: There were various types of PIML: 19 (19%) Type I; 24 (24%) Type II; 23 (23%) Type III; and 34 (34%) Type IV. A PIML was present in 81 legs (81%). There were no significant differences between men and women and between left and right sides. CONCLUSIONS: The complex relationships of the PIML with the surrounding ligaments and tissues are considered to be among the factors that make interpretation of imaging findings difficult.

Morphological features of the bifurcated ligament.

INTRODUCTION: Damage to the bifurcate ligament is one of the most difficult injuries to diagnose from imaging techniques. A probable reason for this is that the morphological characteristics of this structure have yet to be sufficiently elucidated. We, therefore, endeavored to elucidate the morphological characteristics of the bifurcate ligament through a large-scale study involving numerous specimens. MATERIALS AND METHODS: This study included 100 feet from 52 formalin-fixed cadavers. The bifurcate ligament was classified into three types: presence of both calcaneonavicular ligament and calcaneocuboid ligament (Type I); absence of calcaneocuboid ligament (Type II); and absence of calcaneonavicular ligament (Type III). Morphological characteristics of the bifurcate ligament were determined by measuring fiber bundle length, width, and thickness at the center of each ligament. RESULTS: This classification resulted in 68 Type I feet (68%), 32 Type II feet (32%), and 0 Type III feet (0%). The calcaneonavicular ligament was 20.8 ± 2.9 mm long, 4.9 ± 1.2 mm wide, and 3.8 ± 1.1 mm thick. The calcaneocuboid ligament was approximately 10.5 ± 2.7 mm long, 4.7 ± 2.4 mm wide, and 1.5 ± 0.6 mm thick. The bifurcate ligament was located deep under the extensor hallucis brevis and extensor digitorum brevis muscles in all specimens. The calcaneal origin of the calcaneonavicular ligament was situated deep under the interosseous talocalcaneal ligament in all specimens. Two sides were identified in which the calcaneocuboid ligament was located deep under the dorsal calcaneocuboid ligament. CONCLUSION: Such variations and positional relationships were suggested to be factors complicating the diagnostic imaging of bifurcate ligament injuries. The present study results will likely form useful basic data for diagnostic imaging.

Anatomical study of the inferior patellar pole and patellar tendon.

In this study, detailed investigations into the shape of the inferior patellar pole, the site of the patellar tendon attachment, and the length and course of the patellar tendon were performed with the aim of examining the anatomical factors involved in the developmental mechanism of patellar tendinitis. The investigation examined 100 legs from 50 cadavers. The inferior patellar pole was classified into three types: pointed, intermediate, and blunt. The attachment of the patellar tendon to the inferior patellar pole was classified into two types: an anterior and a posterior. The length of the patellar tendon was measured from the tibial tuberosity to the inferior patellar pole. The pointed type was seen in 57% of legs, the intermediate type in 21%, and the blunt type in 22%. Twenty-one legs were the pointed type, as well as the anterior type. The patellar tendon was significantly shorter with the posterior type than with the anterior type. The blunt type also had a significantly shorter patellar tendon than the pointed type. In legs that were both the pointed type and the anterior type, the inferior patellar pole and the proximal posterior surface of the patellar tendon impinged during knee flexion due to the posterior tilt of the patella, suggesting the possibility that this may induce damage. With the posterior type and blunt type, on the other hand, the possibility of strong tensile stress on the tendon fibers of the posterior facet of the inferior patellar pole was suggested.

Gender differences of muscle and crural fascia origins in relation to the occurrence of medial tibial stress syndrome.

Although women reportedly have a higher prevalence of medial tibial stress syndrome (MTSS) than men, the possible role of gender-based anatomical differences has not been investigated. The aim of the present study was to investigate the presence of gender-based differences in the range of muscle attachments along the entire medial tibia, the proportion of muscle attachment at the middle and distal thirds of the medial margin of the tibia, the structure of the crural fascia, and chiasm position. The specimens were 100 legs of 55 Japanese cadavers. Statistical analysis was carried out using a chi-square test to compare anatomical features between the sexes. The flexor digitorum longus (FDL) had a higher proportion of attachment to the middle and distal thirds of the medial margin of the tibia than the soleus (SOL; P < 0.001). The proportion of the SOL attachment to the middle and distal thirds of the medial margin of the tibia was 33.3% in men and 72.5% in women (P < 0.001). The soleal aponeurosis was not observed in any specimen. In all specimens the FDL formed the top layer of both chiasms. These results suggest that the higher prevalence of MTSS reported among women may be the result of gender-based anatomical differences.

The twisted structure of the human Achilles tendon.

The Achilles tendon (AT) consists of fascicles that originate from the medial head of the gastrocnemius (MG), lateral head of the gastrocnemius (LG), and soleus muscle (Sol). These fascicles are reported to have a twisted structure. However, there is no consensus as to the degree of torsion. The purpose of this study was to investigate the twisted structure of the AT at the level of fascicles that originate from the MG, LG, and Sol, and elucidate the morphological characteristics. Gross anatomical study of 60 Japanese cadavers (111 legs) was used. The AT fascicles originated from the MG, LG, and Sol were fused while twisting among themselves. There were three classification types depending on the degree of torsion. Further fine separation of each fascicle revealed MG ran fairly parallel in all types, whereas LG and Sol, particularly of the extreme type, were inserted onto the calcaneal tuberosity with strong torsion. In addition, the sites of Sol torsion were 3-5 cm proximal to the calcaneal insertion of the AT. These findings provide promising basic data to elucidate the functional role of the twisted structure and mechanisms for the occurrence of AT injury and other conditions.

Dynamic changes in amniotic tight junctions during pregnancy.

The amniotic membrane encloses and retains amniotic fluid during pregnancy. In general, fluid flux is regulated by epithelial tissues, which have tight junctions (TJs). However, TJs have not yet been identified in the amniotic epithelium. In this study, we have determined whether the mouse amniotic epithelium contains TJs. Freeze-fracture electron microscopy revealed the presence of strand-like TJs in the amniotic epithelium. Amniotic TJs were composed of occludin; zona occludens (ZO)-1; and claudins 1, 3, 4, and 7. These claudins underwent developmental changes during pregnancy. The localization patterns of the claudins and their detergent solubility drastically changed between embryonic day (E) 16 and E17; the volume of the amniotic fluid also decreased sharply. Furthermore, in vitro assessment of amniotic membrane permeability showed that the amniotic membrane was more permeable on E17 than on E16. On E17, TJ components were sparsely distributed in parts of the amniotic epithelium. The results of Annexin V-fluorescein staining and Terminal dUTP nick-end labeling (TUNEL) assay revealed ongoing apoptosis in all the cells in such regions. The above findings suggest that TJs in the amniotic epithelium maintain amniotic fluid volume during pregnancy, while apoptosis of amniotic epithelial cells between E16 and E17 causes disruption of the TJs.

Effects of defect configuration, size, and remaining teeth on masticatory function in post-maxillectomy patients.

The aim of this study was to investigate the correlation between the masticatory function and the maxillary defect configuration, size, and remaining teeth in post-maxillectomy patients restored with an obturator prosthesis. Fifty consecutive post-maxillectomy patients (mean age 67.0) participated in this study. The subjects consisted of 26 dentate and 24 edentulous patients. Data were collected from the patients' clinical records, diagnostic casts, and a questionnaire on masticatory function to evaluate the defect configuration, size, and the masticatory function scores associated with wearing obturator prostheses. The defect size was evaluated by the ratio of defect area to the horizontal impression area. The defect configuration was assessed according to Aramany's classification and separated into unilateral and bilateral defects. The multiple regression analysis and the Mann-Whitney U-test (P<0.05) were used to assess statistical significance. The Spearman's correlation coefficient by rank test was also used to detect correlation. The following conclusions were made: (i) The presence of teeth in the maxillary dentition and the different type of the defect configuration had significant correlation with the masticatory function score (r=0.616). (ii) The masticatory function scores of the subjects differed significantly with the presence of teeth in the maxillary dentition and the different types of defect configurations (P=0.005, P=0.002, respectively). (iii) There was significant correlation between the masticatory function score and the size of the defect area in the edentulous group (r=-0.648, P=0.001). The presence of teeth, the size and configuration of the defect influenced the masticatory function of post-maxillectomy patients with obturator prostheses.

Differential expression of major gap junction proteins, connexins 26 and 32, in rat mammary glands during pregnancy and lactation.

We examined the expression pattern of two major gap junction proteins, connexin 26 (Cx26) and connexin 32 (Cx32), in rat mammary glands during pregnancy and lactation. Immunohistochemically the two different Cxs were coexpressed in acinar cells and were independently modulated according to the physiological cell activity. Western blot analysis demonstrated that Cx26 gradually increased from early pregnancy, while Cx32 rapidly and dramatically increased at 16 h after parturition, and that both Cxs reached a maximum early in lactation. Increased expression of both Cxs was confirmed by Northern blot analysis showing that their mRNA transcripts were significantly induced on the day of parturition. We also analyzed double-immunofluorescent staining for Cx26 and Cx32 on a confocal laser scanning microscope, in order to examine colocalization of these Cxs in situ. Cx26 immunoreactivity mostly overlapped with Cx32-positive sites in acinar cells of lactating mammary glands, indicating that both Cxs were colocalized together in the same gap junctional plaques in lactation. These results suggest that upregulation of Cx26 and Cx32 in acinar cells at lactating stages, with colocalization in the same gap junctional plaques, may be important for control of secretion by acinar cells in rat mammary glands.

Dental rehabilitation using endosseous implants and orthognathic surgery in patients with cleft lip and palate: report of two cases.

We describe the use of endosseous implants in the autogenous particulate cancellous bone and marrow grafted alveoli after orthognathic surgery for dental rehabilitation of patients with cleft lip and palate. This procedure has been applied to two patients and produced good results functionally and aesthetically. The results are encouraging and indicate that implant placement after orthognathic surgery is useful for patients both with congenital missing teeth and retrognathic maxillae.

Distribution of gap junction protein connexin 37 in smooth muscle cells of the rat trachea and pulmonary artery.

Connexin 37, one of the gap junction protein families, has been detected by Northern blotting in various organs and tissues, and found to be especially abundant in the lung. However, detailed information on the precise types of cells which express connexin 37 has not been previously published. We therefore prepared site-specific connexin 37 antibodies and examined the distribution of connexin 37 immunohistochemically. Connexin 37 was detected in endothelial cells, in the tunica media of both the pulmonary artery and the aorta, and in the smooth muscle layer of the trachea and bronchioles. In the tracheal smooth muscle layer, connexin 37 overlapped with desmin-positive areas, but was clearly segregated from vimentin- and von Willebrand factor-positive areas. These results suggest that connexin 37 is expressed in smooth muscle cells in the trachea, but not in fibroblastic cells or endothelial cells. Connexin 37 was partially colocalized with connexin 43 in tracheal smooth muscle cells, and showed a gradual increase in expression during postnatal development. To our knowledge, this is the first report to be published regarding the expression of connexin 37 in smooth muscle cells.

[Postoperative prophylactic intravesical instillation of tetrahydropyranyl-adriamycin (THP) for superficial bladder cancer].

Intravesical instillation of tetrahydropyranyl-adriamycin (THP) was performed on 51 patients with superficial bladder cancer after transurethral resection (TUR) for prophylaxis of recurrence. The instillation was carried out with 20 mg of THP dissolved in 40 ml of distilled normal saline. Instillation was performed once 24 hours postoperatively, 9 times every 2 weeks, and 8 times every 4 weeks. These drugs were instilled for 30 to 60 minutes. The recurrence-free survival at 1, 2 and 3 years was 74.5%, 64.6% and 58.0%, respectively. Side effects of THP instillation were observed in only 4 cases (7.8%) as slight urinary frequency or micturition pain. Cases involving 5 or more tumors, or tumors measuring 3 cm or larger, more frequently demonstrated recurrence. The cases that did not respond to preoperative intravesical instillation of THP demonstrated a high frequency of recurrence. Intravesical instillation of THP as a prophylaxis against recurrence of superficial bladder cancer was effective in selected patients.

Possible involvement of histamine in muscular fatigue in temporomandibular disorders: animal and human studies.

As an approach to clarifying the molecular basis of pain and fatigue in muscles involved in temporomandibular disorders, we examined the activity of histidine decarboxylase (HDC), the enzyme which forms histamine, in the masseter muscles of mice. In the resting muscle, HDC activity was very low. Direct electrical stimulation of the muscle markedly elevated HDC activity. HDC activity rose within 3 hrs of the electrical stimulation, peaked at 6 to 8 hrs, and then gradually declined. Intraperitoneal injection of a small amount of interleukin-1 (IL-1) (from 1 to 10 microg/kg) produced a similar elevation of HDC activity in the masseter muscle. We also examined the effect of an antihistamine, chlorphenylamine (CP), on temporomandibular disorders in humans and compared it with that of an anti-inflammatory analgesic, flurbiprofen (FB). Two groups received one or the other of the drugs daily for 7 days, and they were asked about their signs and symptoms before and after the treatment. A positive evaluation of their treatment was made by 74% of the CP group, but by only 48% of the FB group. Although the effects of CP on the limitation of mouth-opening and on joint noise were negligible, about 50% of the CP group answered positively concerning the drug's effect on spontaneous pain or pain induced by chewing or mouth-opening. The positive evaluation for CP (50%) in relieving associated symptoms (headache or shoulder stiffness) was significantly greater than for FB (13%). FB showed effectiveness similar to but sometimes weaker than that of CP on several symptoms. On the basis of these and previous results and the known actions of histamine, we propose that the histamine newly formed following the induction of HDC activity, which is itself mediated by IL-1, may be involved in inducing pain and, possibly, stiffness in muscles in temporomandibular disorders.

Claudin-1 contributes to the epithelial barrier function in MDCK cells.

Tight junctions (TJs) create a paracellular permeability barrier and also act as a fence preventing intermixing of proteins and lipids between the apical and basolateral plasma membranes. Recently, claudin-1 has been identified as an integral membrane protein localizing at TJs, and introduced claudin-1 can form TJ-like networks in fibroblasts. To investigate the function of claudin-1, MDCK cells were transfected with a mammalian expression vector containing myc-tagged mouse claudin-1, and four stable clones were obtained. The myc-tagged claudin-1 precisely colocalized with both occludin and ZO-1 at cell-cell contact sites, indicating that exogenous claudin-1 was properly targeted to the TJs. Immunoblot analysis revealed that overexpression of claudin-1 increased expression of ZO-1 but not of occludin or ZO-2. The barrier functions of these cells were evaluated by transepithelial electrical resistance (TER) and paracellular flux. Claudin-1-expressing cells exhibited about four times higher TER than wild-type MDCK cells. Consistent with the increase of TER, the cells overexpressing claudin-1 showed reduced paracellular flux, estimated at 4 and 40 kD FITC-dextrans. These results suggest that claudin-1 is involved in the barrier function at TJs.

Immunohistochemical localization of connexin43 in the enamel organ of the rat upper incisor during ameloblast development.

We examined the localization of connexin (Cx)43 in the enamel organ during ameloblast development. The specificity of monoclonal anti-Cx43 antibody was elucidated by immunoblot analysis and immunoelectron microscopy. Gold particles for Cx43 were detected by immunoelectron microscopy on gap junctions but not on other structures such as desmosomes. Punctate and intense immunofluorescence for Cx43 was detected in all cell types of the enamel organ. Cx43 expression in ameloblasts showed a transient decrease and then increase during ameloblast development. Double staining of Cx43 and amelogenin, one of the enamel proteins, revealed that immunofluorescence for Cx43 markedly decreased in some late presecretory ameloblasts just prior to enamel formation. Moreover, the localization of Cx43 changed during enamel formation. Cx43 was distributed randomly on the lateral plasma membranes of presecretory ameloblasts, but tended to gather on those corresponding to the supranuclear regions of secretory ameloblasts. Immunofluorescence for Cx43 in maturation ameloblasts appeared linear rather than punctate. These results suggest that Cx43 in the late presecretory ameloblasts is degraded just before enamel formation and then newly synthesized Cx43 is redistributed during the secretory stage. These changes in Cx43 expression may be related to the cellular differentiation of ameloblasts.

Use of an osseointegrated implant for dental rehabilitation after cleft repair by periosteoplasty: a case report.

OBJECTIVE: We report here on a patient with unilateral cleft lip and alveolus who underwent dental rehabilitation of cleft alveolus using an osseointegrated implant after cleft repair by periosteoplasty. The patient, whose lateral incisor was congenitally missing, had periosteoplasty, followed by excellent bone formation at the cleft alveolus within 2 years. After the completion of orthodontic alignment of the maxillary dental arch, a Brånemark single-tooth implant was placed into the bone-formed alveolus. CONCLUSION: This treatment procedure offers an option of dental rehabilitation for the alveolar clefts of patients with cleft lip and/or palate.

[Mutagenicity studies of (+/-)-4-diethylamino-1,1,-dimethylbut-2-yn-1-yl 2-cyclohexyl-2-hydroxy-2-phenylacetate monohydrochloride monohydrate (NS-21), a novel drug for urinary frequency and incontinence].

The mutagenicity of (+/-)-4-diethylamino-1,1-dimethylbut-2-yn-1-yl 2-cyclohexyl-2-hydroxy-2-phenylacetate monohydrochloride monohydrate (NS-21), a new drug for the treatment of urinary frequency and incontinence, was investigated by the reverse mutation test in bacteria, the chromosome aberration test in vitro, and the micronucleus test in mice. The reverse mutation test was performed at a dose from 31.3 to 4000 micrograms/plate, at which dose cell killing was observed, using Salmonella typhimurium TA100, TA1535, TA98, and TA1537, and Escherichia coli WP2uvrA. NS-21 did not increase revertant colonies significantly in any of the test strains with or without metabolic activation system (S9 mix). The chromosome aberration test was carried out at a dose from 3.75 to 140 micrograms/ml, at which dose more than 50% cell proliferation was inhibited, using cultured Chinese hamster lung cells (CHL/IU). No significant increases of the frequencies of cells with chromosome aberrations were observed with or without S9 mix. The micronucleus test was conducted in the bone marrow cells of Slc : ddY male mice. Mice were given NS-21 by a single oral administration at doses of 0, 43.8, 87.5, 175, and 350 mg/kg, the geometric mean dose between the maximum tolerated dose and the minimum lethal dose. There were no significant increases in the frequencies of micronucleated polychromatic erythrocytes at any dose levels. These results show that NS-21 has no mutagenic activity in vitro or in vivo.

Changes in the phosphorylation states of connexin43 in myoepithelial cells of lactating rat mammary glands.

Using specific antibodies and cDNA probe, we examined the expression pattern of a major gap junction protein, connexin43 (Cx43), in rat mammary glands during pregnancy and lactation. Double immuno-fluorescence revealed that the labeling of Cx43 was superimposed in the alpha-smooth muscle actin-positive cells, suggesting that myoepithelial cell were interconnected by gap junctions formed of Cx43. Just after delivery, the Cx43-labeled plaques were enlarged and increased in intensity. Northern and Western blot analyses confirmed the dramatic induction of Cx43 at both mRNA and protein levels on the day of parturition. Cx43 mRNA transcript immediately declined, while the increase of Cx43 protein continued for a few days. During pregnancy, immunoblots showed two bands of almost equal amounts at 43 and 45 kDa. Following delivery, the 45-kDa band gradually increased in intensity with a concomitant decrease of the 43-kDa band. From the sixth day of lactation, Cx43 was always detected as a single band at 45 kDa. Alkaline phosphatase treatment of immunoprecipitated Cx43 revealed that both bands represented phosphorylated forms, thus indicating that Cx43 was naturally phosphorylated and that it altered its phosphorylation states during lactation stages. These results suggest that the induction of Cx43 with the changes in the phosphorylation states plays an important role in the lactating function of myoepithelial cells in rat mammary glands. This is the first report on the changes of Cx43 phosphorylation states during physiological stages in vivo.