Early serial Q-switched ruby laser therapy for medium-sized to giant congenital melanocytic naevi.

BACKGROUND: Medium-sized to giant congenital melanocytic naevi (CMN) are difficult to treat, especially if the lesions appear on the face or extremities where treated areas are visible and cosmesis is important. OBJECTIVES: In infants, nests of pigmented naevus reside more superficially and the skin is more transparent than in adults, so we treated medium-sized to giant CMN with early serial Q-switched ruby laser therapy from infancy. PATIENTS AND METHODS: We treated nine patients with medium-sized to giant CMN on the face or upper limbs from 1 month of age with early serial Q-switched ruby laser therapy. The laser power was initially 5 J cm(-2) and increased in 0.5 J cm(-2) steps to a maximum of 10 J cm(-2). There were three treatment sites on the forehead, one on the temple, one on the cheek and four on the upper arm. RESULTS: It took 8-15 treatments for the CMN to become a colour similar to the surrounding skin. The mean number of treatments was 9.6. The colour was reduced to 0-20% of the colour of the baseline lesion in all nine patients. Partial slight repigmentation occurred in eight of these patients. These naevi were treated with an additional one or two Q-switched ruby laser irradiations and successfully lightened for at least 1 year. In the remaining patient, pigmentation returned to a level similar to the original lesion within 1 month of the last treatment. Therefore, the lesion was excised for cosmetic reasons. After the treatment series, the skin texture was fine and no patients had hypertrophic scarring. CONCLUSIONS: Although treatment of one patient with the Q-switched ruby laser therapy failed, the remaining patients responded well and had good to excellent skin texture without hypertrophic scarring. Early serial Q-switched ruby laser treatment, starting from infancy, is a promising treatment method for this condition.

E-cadherin expression in the subepithelial nevus cells of the giant congenital nevocellular nevi (GCNN) correlates with their migration ability in vitro.

BACKGROUND: Giant congenital nevocellular nevi (GCNN) are histologically characterized by the broad distribution of nevus cells in the epidermis and dermis. OBJECTIVE: To characterize E-cadherin in GCNN and define its role in nevic cell migrations. METHODS: Twenty-four cases were immunohistochemically examined and in five cases cells were isolated for primary culture for migration assays. RESULTS: The nevus cells in the superficial region showed the immunoreactivity of E-cadherin in a membranous pattern, but those in the deep part of dermis had little immunoreactivity. Ultra-structural analysis of the superficial nevus cells revealed that E-cadherin immunodeposits in the fibrillar processes around the cell body in a spotted pattern. This distribution pattern is quite different from that in the adherens junction of skin squamous epithelial cells. Boyden chamber experiments were performed using primary cultures of intradermal nevus cells. EDTA pretreatment reduced cell migration to the E-cadherin positive side when the E-cadherin positive population was relatively large in the primary cultures. CONCLUSIONS: These results indicate that E-cadherin in the nevus cells may affect nevus cell motility rather than intercellular attachment.

Amelogenin positive cells scattered in the interstitial component of odontogenic fibromas.

BACKGROUND: Odontogenic tumours are often biphasic, consisting of epithelial and interstitial components, with an origin that is not well understood. Odontogenic fibromas are rich in mesenchymal component, but also have many epithelial nests. AIMS: To investigate the origin of this tumour by immunohistochemistry. METHODS: The expression of several odontogenic and epithelial markers, including amelogenin, was investigated by immunofluorescent studies. RESULTS: Immunohistochemical analysis showed that epithelial nests exhibited E-cadherin expression, but not amelogenin. Amelogenin positive cells were scattered in the fibrous tissue, which did not exhibit epithelial marker expression except for epithelial membrane antigen. In one case that had received a test biopsy before whole resection of tumour, amelogenin positive cells were distributed in the regenerating mucosal epithelium or subepithelial tissue. CONCLUSIONS: Results indicate that amelogenin positive cells of odontogenic fibromas have an epithelial origin and may have the potential for epithelial mesenchymal transition, which has not to date been investigated in benign tumours.

Rapid, severe repigmentation of congenital melanocytic naevi after curettage and dermabrasion: histological features.

BACKGROUND: Curettage and dermabrasion are effective in treating giant congenital melanocytic naevi (GCMN). We report two patients with rapid, severe postoperative repigmentation. To the best of our knowledge this is the first report on the histological features of such patients. OBJECTIVES: We wish to call attention to histological features that may cause rapid, severe repigmentation after curettage and dermabrasion of medium to giant CMN. PATIENTS/METHODS: From 1998 to 2002, we treated 23 patients with medium to giant CMN with curettage and dermabrasion. Patients being treated ranged in age from 1 month to 19 years. Histological samples were taken from the centre of naevi in all patients during surgery. Histological types were 12 intradermal and 11 compound. Follow-up after curettage lasted at least 3 years. RESULTS: Among our 23 patients only two showed repigmentation soon after surgery. Histological sections from these two patients indicated naevoid cells in the deep dermis along hair follicles or sebaceous glands. However, no such pigmented naevoid cells along hair follicles were observed in samples from patients successfully treated with curettage and dermabrasion with less repigmentation. CONCLUSIONS: Although we saw only two cases of repigmentation soon after curettage and dermabrasion, we suspect a correlation between pigmented naevoid cells around hair follicles and repigmentation. If histological sections of skin biopsies show pigmented cells along hair follicles in the deep dermis, other treatments such as total skin resection followed by skin grafting or tissue expansion may be better choices than curettage or dermabrasion.

Arthoscopic diagnosis of partial scapholunate ligament tears as a cause of radial sided wrist pain in patients with inconclusive x-ray and MRI findings.

To clarify the pathology of radial-sided wrist pain with inconclusive X-ray and MRI findings, we performed arthroscopic examinations of 11 wrists in 10 patients. Physical examination and various image investigations could not identify the cause of the pain. Arthroscopy revealed partial to complete tears of the scapho-lunate interosseous ligament and synovitis and/or chondral bone defects at the scaphotrapezio-trapezoidal joint in all 11 wrists. Surgical procedures consisted of eight simple synovectomies, two ligament reconstructions and one percutaneous pinning. Pain relief was achieved in 10 wrists. One wrist which had a simple synovectomy did not recover, so underwent secondary scaphotrapezio-trapezoidal fusion. In conclusion, we found that various degrees of scapholunate interosseous ligament tear and scaphotrapezio-trapezoidal joint osteoarthritis were the main causes of radial-sided wrist pain with inconclusive X-ray and simple MRI findings.

Proposal for a unified CCN nomenclature.

A proposal is put forth to unify the nomenclature of the CCN family of secreted, cysteine rich regulatory proteins. In the order of their description in the literature, CCN1 (CYR61), CCN2 (CTGF), CCN3 (NOV), CCN4 (WISP-1), CCN5 (WISP-2), and CCN6 (WISP-3) constitute a family of matricellular proteins that regulate cell adhesion, migration, proliferation, survival, and differentiation, at least in part through integrin mediated mechanisms. This proposal is endorsed by the International CCN Society and will serve to eliminate confusion from the multiple names that have been given to these molecules.

Effects of low-intensity pulsed ultrasound on proliferation and chondroitin sulfate synthesis of cultured chondrocytes embedded in Atelocollagen gel.

The effects of low-intensity pulsed ultrasound (US) on the proliferation and chondroitin sulfate synthesis of cultured chondrocytes embedded in Atelocollagen gel in vitro were examined. Articular cartilage was harvested from the hip, knee, and shoulder joints of 10-week-old Japanese white rabbits. Chondrocytes isolated by collagenase digestion were embedded in type I collagen gel, Atelocollagen gel, and were cultured in Dulbecco's modified eagle's medium for 3 weeks. The US apparatus, SAFHS, was used to deliver an ultrasound signal with spatial and temporal average intensities of 30 mW/cm(2) (US group). The frequency was 1.5 MHz with a 200-microsecond tone burst repeated at 1.0 kHz. US treatments were administered for 20 min per day under culture dishes, with the medium replaced twice a week. Another group of cells was exposed to sham ultrasound as a control. Cell number, histological findings, synthesis of isomers of chondroitin sulfate, and stiffness of the chondrocyte-collagen gel composites were analyzed. US exposure promoted synthesis of chondroitin sulfate, especially chondroitin 6-sulfate, although it did not significantly enhance cell number and stiffness. In this three-dimensional culture model, these results suggest that US exposure may be clinically useful in improving the quality of chondrocyte-Atelocollagen implants for transplantation into articular cartilage defects.

Loss of syndecan-1 and increased expression of heparanase in invasive esophageal carcinomas.

Heparan sulfate proteoglycans play important biological roles in cell-cell and cell-matrix adhesion, and are closely associated with growth factor actions. Loss of syndecan-1, a cell surface-bound heparan sulfate proteoglycan, has been reported for advanced head and neck carcinomas, and expression of endoglycosidic heparanase, which cleaves heparan sulfate glycosaminoglycans (HS-GAGs), is associated with invasion and metastatic potential of malignant tumors. Paraffin sections of 103 primary esophageal squamous cell carcinomas were immunohistochemically examined for the expression of syndecan-1 core protein, HS-GAGs and heparanase protein, and the results were compared with various clinicopathological parameters, such as invasion depth. For 16 cases, fresh tumor samples were quantitatively analyzed for heparanase and syndecan-1 mRNA expression by real-time RT-PCR in addition to the immunohistochemical studies. Syndecan-1 core protein and HS-GAGs expression was significantly decreased in pT2 and pT3 cases compared with their pTis and pT1 counterparts. Decreased expression of core protein and HS-GAGs was correlated with the incidence of lymphatic invasion, and venous involvement. Furthermore, decreased expression of HS-GAGs was correlated positively with the incidence of nodal metastasis and distant organ metastasis, and negatively with the grade of tumor cell differentiation. The percentage of cytoplasmic heparanase protein-positive cases increased significantly in pT2 and pT3 cases compared to that in pTis and pT1 cases, and this was associated with lymphatic invasion, and venous and lymph nodal involvement. The level of heparanase mRNA was inversely correlated with the degree of HS-GAGs expression rather than core protein. In conclusion, loss of syndecan-1 and heparanase overexpression in esophageal squamous cell carcinomas are closely associated with malignant potential. Regarding the mechanism of loss of HS-GAGs, heparanase upregulation appears to play an important role.

Perinuclear and cytoplasmic distribution of desmoglein in esophageal squamous cell carcinomas.

The desmosomal glycoproteins desmoglein (Dsg) and desmocollin (Dsc) are members of the cadherin family of cell adhesion molecules. They play an important role in epithelial adhesion. To observe the distribution pattern of Dsg in esophageal squamous cell carcinomas (SCC), immunohistochemical and immunoelectron microscopic analyses were performed. Immunohistochemically, normal esophageal squamous cells strongly expressed Dsg at the cell-cell boundaries, while moderately differentiated esophageal SCC cells showed a perinuclear distribution in addition to the cell boundary staining. At the ultrastructural level, the reaction product was concentrated at the desmosomes in the cell membrane region of normal epithelial cells, but was reduced at the membrane and found throughout the cytoplasm as well as in the surrounding outer nuclear envelope in SCC cells. These results demonstrate an aberrant distribution of Dsg in SCC cells. This may have important consequences for invasion and metastasis, as it may indicate loosened intercellular adhesion.

The expression of chicken NOV, a member of the CCN gene family, in early stage development.

The nephroblastoma overexpressed gene, NOV, is a member of the CCN gene family. We investigated the NOV gene expression pattern in the chicken during early stage embryogenesis. Several embryonic structures showed a distinct expression pattern. The initial expression was detected in Hensen's node (Hamburger and Hamilton stage (HH) 5). The expression was noted in the presumptive notochord and floor plate forming cells. The expression on the left side was more elongated posteriorly, a type of left-right asymmetry. Chicken NOV gene expression in the forming notochord and floor plate was observed until HH 18. The expression was also detected in the ventral area of the mesencephalon and isthmus at HH 14-16.

Proprioceptive improvement in knees with anterior cruciate ligament reconstruction.

The correlation between the prospective course of proprioceptive improvement and knee stability after anterior cruciate ligament reconstruction was investigated in 38 patients. Proprioception, on the basis of the patient's capacity to reposition the limb accurately, was evaluated at 3-month intervals for 24 months after hamstring graft anterior cruciate ligament surgery. Knee stability was evaluated concurrently with a KT-2000 knee arthrometer. Thirty patients experienced improvement in postoperative position sense in at least one of the examinations, although eight patients had no improvement at any time. Of the 30 patients who had improvement, 28 maintained improved position sense from 18 months to the final followup. Thirty patients maintained significantly better knee stability for a postoperative period of at least 24 months. These results indicated that a minimum of 18 months after anterior cruciate ligament reconstruction may be needed for complete restoration of the proprioceptive function in knees, although the mean position sense in all patients gradually improved from 9 months. Improvement in postoperative knee stability may have facilitated recovery of proprioception.

Repair of articular cartilage defects with cultured chondrocytes in Atelocollagen gel. Comparison with cultured chondrocytes in suspension.

We attempted to repair full-thickness articular cartilage defects in rabbit knee joints with allogeneic cultured chondrocytes embedded in Atelocollagen gel. An articular cartilage defect was created on the patellar groove of the femur. The defect was filled with chondrocytes cultured in the collagen gel and covered with periosteal flap (G group). In three other experimental groups, the same defects were transplanted with chondrocytes in monolayer culture with periosteal flap (M group), periosteal graft only (P group), or left empty (E group). At 4, 12, and 24 weeks after operation, the reparative tissue was analyzed macroscopically and histologically. At 4 weeks after operation, the surfaces of the reparative tissue were smooth, and the defects were filled with reparative tissues that resembled hyaline cartilage in all four groups. However, the reparative tissues degenerated gradually with time in the M, P, and E groups. In contrast, in the G group, the reparative tissue retained its thickness, and there was a steady integration of the grafted tissue into the adjacent normal cartilage at 24 weeks after operation. The results suggest that transplantation of allogeneic chondrocytes cultured in Atelocollagen gel is effective in repairing an articular cartilage defect.

Human chondrocyte proliferation and matrix synthesis cultured in Atelocollagen gel.

To evaluate the potential of Atelocollagen gel as a carrier for chondrocyte transplantation, histological and biochemical characteristics of the chondrocytes in gel culture were compared with those in conventional monolayer cultures. Articular chondrocytes from 20 patients were isolated by enzyme digestion, embedded in Atelocollagen gel, and cultured for up to 4 weeks. The effects on proliferation, morphological changes, and synthesis of proteoglycans were analyzed by cell counts, light and electron microscopy, and measurement of isomers of chondroitin sulfates. Chondrocytes embedded in the Atelocollagen gel gradually proliferated and produced chondroitin 6-sulfate, maintaining the chondrocyte phenotype for up to 4 weeks. In contrast, although monolayer chondrocytes increased in number, most could be characterized as being fibroblast-like cells with a reduced capability of producing chondroitin 6-sulfate. The results suggest that Atelocollagen gel permitted a gradual proliferation and matrix synthesis of chondrocytes and maintaining its phenotype. Atelocollagen gel represents an important carrier for the clinical application of cultured chondrocyte transplantation for repair of cartilage defects.

Successful restoration of the trapezius muscle using pedicle latissimus dorsi. A case report.

Reconstruction of the trapezius muscle using a pedicle latissimus dorsi flap was performed in a 27-year-old man with a large synovial sarcoma in his shoulder girdle. Size and location of the tumor required combined resection of surrounding muscles, including the trapezius, levator scapulae, and rhomboid major and minor. Thus, an extensive defect of the suspending muscles of the scapula was created after accomplishing an adequate resection of the tumor. The flap was performed to restore the trapezius functionally because there were no adjacent muscles available. The transferred muscle compensated for loss of the trapezius, thereby recovering excellent shoulder function. Although an opportunity of its application is thought to occur infrequently, the pedicle latissimus dorsi can activate scapular motion successfully in the absence of the levator scapulae. The technique may be extended to salvage failed conventional reconstruction after spinal accessory nerve palsy.

Identification of EPS8 as a Dvl1-associated molecule.

Dishevelled (Dsh) is involved in both Wingless (Wg) and Frizzled (Fz) signaling pathways. To further determine the function of Dsh, we have performed yeast two-hybrid screening and isolated several genes encoding the molecules associated with the PDZ domain of Dvl1, one of the murine Dsh homologs. During the screening, we found that EPS8, which is a substrate for activated EGF receptor (EGFR), specifically interacted with Dvl1. This interaction was also confirmed in vitro. Through transfection studies, we observed the mutual action between Dvl1 and EPS8. Dvl1 was hyperphosphorylated in the presence of EPS8, whereas the tyrosine phosphorylation of EPS8 by activated EGFR was inhibited in the presence of Dvl1. Immunohistochemistry showed that Dvl1 and EPS8 expression overlap in particular tissues during organogenesis. These results indicate that interaction between Dvl1 and receptor tyrosine kinase signal plays certain roles in developmental events.

Identification and characterization of a putative sevenless homologue in the malaria vector Anopheles gambiae.

Tyrosine kinase sequences were identified and characterized in Anopheles gambiae, the major vector of malaria in subsaharan Africa. One of these sequences has the characteristics expected for a homologue of the Drosophila sevenless gene, which is necessary for R7 photoreceptor cell fate determination in the developing compound eye. The putative Anopheles sevenless gene homologue is located in a telomeric region of the X chromosome and is expressed in the head of late larval and pupal stage mosquitoes. Identification of the Anopheles homologue of the sevenless gene is a first step towards the development of a dominant phenotypic marker that could be used for detecting transformed Anopheles mosquitoes in a wide variety of genetic backgrounds and, as such, could be used in the development of transgenic mosquitoes for the control of parasite transmission. Preliminary evidence for sevenless sequences were also found in DNA from blackfly, Mediterranean fruit fly and the honeybee.

Successful nerve regeneration and persistence of donor cells after a limited course of immunosuppression in rat peripheral nerve allografts.

BACKGROUND: The origin of Schwann cells and effect of a limited course of immunosuppression using cyclosporine (CsA) were examined in rat peripheral nerve allotransplants. METHODS: Phenotypes of Schwann cells in groups without, with continuing, and with limited (12 weeks) CsA treatment were examined immunohistochemically in allogeneically and syngeneically transplanted animals from 4 to 36 weeks after transplantation. RESULTS: In the group receiving no CsA, little nerve regeneration was obtained; donor Schwann cells were rejected and replaced by recipient cells. In continuing and limited-course CsA groups, successful nerve regeneration was achieved at postoperative week 36, as was also observed in the syngeneic group. Schwann cells in the continuing CsA group remained donor-derived. In the limited-course CsA group, graft rejection and loss of function occurred after the withdrawal of CsA, and donor Schwann cells were replaced by recipient cells in the part of the graft where rejection had been complete. However, many donor Schwann cells remained at week 36, when the rejection response subsided. CONCLUSION: Possible clinical use of a limited course of immunosuppression was supported by this demonstration of long term persistence of donor Schwann cells.

Ectopic expression of lunatic Fringe leads to downregulation of Serrate-1 in the developing chick neural tube; analysis using in ovo electroporation transfection technique.

Lunatic Fringe (l-Fng) is one of the vertebrate homologues of Drosophila Fringe, which interacts with the Notch signal pathway and regulates activation of the Notch ligands, Delta and Serrate. To elucidate the roles of l-Fng in vertebrate neurogenesis, we transfected chick l-Fng (C-l-Fng) to chick neural tube using the in ovo electroporation technique and examined the subsequent changes in expression of Notch-related genes. We observed downregulation of C-Serrate-1 by ectopic C-l-Fng expression which implied that C-l-Fng acts on the vertebrate Notch pathway to regulate the expression of its ligand.

Molecular cloning of the chick Nau gene and analysis of its expression patterns during neurogenesis.

We have previously identified a novel gene, Nau (Neuron-specific gene, which has a number of AU-rich RNA instability motifs), from a quail spinal cord cDNA library. Nau expression was observed mainly in post-mitotic neuroblasts, especially in the stage of axonogenesis. In this study, we cloned its chick orthologue (cNau, 5069 bp), which encodes 369 amino acid residues. cNau's open reading frame has three highly conserved regions with rat STOP (stable tubule-only polypeptide). cNau may be an orthologue of STOP gene considering its uniqueness in zoo blot analysis and its sequence similarity. Both the genetic structure and temporo-spatial expression patterns indicate that the cNau gene may be related to axonogenesis. In the hindbrain, cNau mRNA was accumulated in rhombomere boundary regions and in the rhombomere 2, 4 and 6. These observations suggest a specific role of cNau in the hindbrain or the difference in development between the hindbrain and spinal cord.

Identification of a chick homologue of Fringe and C-Fringe 1: involvement in the neurogenesis and the somitogenesis.

In Drosophila, Serrate plays an important role to the wing margin formation. A putative secretory protein, Fringe, is indispensable for the wing margin formation inducing Serrate and other genes. Recently, Xenopus homologues of Fringe were identified and one of them, lunatic Fringe (X-lFng), was demonstrated to be involved in mesoderm induction. We have identified two chick Fringe homologous genes by reverse transcription-polymerase chain reaction and cDNA library screening. One of them, C-Fringe 1, showed sequence similarity to X-lFng. In situ hybridization study of C-Fringe 1 has demonstrated its expression in the developing nervous system and in the presomitic mesoderm. The hindbrain and spinal cord showed the distinct stripe pattern expression which was complementary to that of C-Serrate, indicating the correlation between them in vertebrate.