Molecular analysis of non-syndromic preaxial polydactyly: preaxial polydactyly type-IV and preaxial polydactyly type-I.

Human GLI3 gene mutations have been identified in several phenotypes of digital abnormality such as Greig cephalopolysyndactyly syndrome, Pallister-Hall syndrome, preaxial polydactyly type-IV (PPD-IV) and postaxial polydactyly. However, the different phenotypes resulting from GLI3 mutations have not yet been properly defined. We have experienced two types of digital abnormality without other complicating developmental defects; a family with foot PPD-IV with syndactyly of the third and fourth fingers, and four sporadic cases with biphalangeal thumb polydactyly (PPD-I). The genes responsible for syndactyly of the third and fourth fingers (syndactyly type-I) and PPD-I have not yet been identified; we therefore examined the involvement of the GLI3 gene in these subtypes of digital abnormality. We found a non-sense mutation in the GLI3 gene in the family with foot PPD-IV accompanied with hand syndactyly of the third and fourth fingers, but no mutations were detected in the GLI3 gene in the four other cases with PPD-I alone. Thus, the phenotype of foot PPD-IV accompanied with hand syndactyly of the third and fourth fingers may result from a GLI3 mutation, whereas the PPD-I phenotype alone is not caused by GLI3 gene defect. These results will help to define the phenotypic spectrum of GLI3 morphopathies, which have been recently proposed.

Intraoperative identification of regenerated chorda tympani nerve and its relationship to recovered taste function.

This study demonstrated a simple method of repairing the severed chorda tympani nerve and a method of intraoperative identification of regenerated nerves, and evaluated taste function of regenerated nerves. Seven patients who underwent staged tympanoplasty and whose chorda tympani nerve was severed during primary surgery were evaluated. When the chorda tympani nerve was severed during primary surgery, proximal and distal stumps were anastomosed or approximated almost in the original position and fixed with fibrin glue on the temporal muscle fascia used to reconstruct the eardrum by the underlay method. During primary surgery, end-to-end anastomosis was possible in 3 patients but nerve gap defects remained in the other 4 patients. In all 7 patients, regenerated nerves were identified during secondary surgery not in the tympanic cavity but in the submucosal layer of the previously reconstructed eardrum. In all patients, complete or incomplete recovery of taste perception was observed by both the filter paper disk method and electrogustometry, suggesting that the regenerated nerves had actual taste function. From these results, it was concluded that the severed chorda tympani nerve could regenerate in the reconstructed eardrum even if nerve gap defects remained between the proximal and distal cut ends, when repair or approximation of the nerve was properly completed.

Hair follicle nevus occurring in frontonasal dysplasia: an electron microscopic observation.

We report a rare hair follicle nevus that occurred in a three-month-old Japanese boy with mild frontonasal dysplasia. It had been present since birth. Histologically, numerous tiny vellus hair follicles were found within the dermis. The constituent cells of these follicles showed the features of follicular germ cells under the electron microscope. The fibroblasts around the follicles were active and merged with the colloid substance. Many myofibroblasts were found in a collagenous stroma in the atrophic lesion of the frontonasal dysplasia.

Enhancement of cell killing by induction of apoptosis after treatment with mild hyperthermia at 42 degrees C and cisplatin.

Ohtsubo, T., Igawa, H., Saito, T., Matsumoto, H., Park, H. J., Song, C. W., Kano, E. and Saito, H. Enhancement of Cell Killing by Induction of Apoptosis after Treatment with Mild Hyperthermia at 42 degrees C and Cisplatin. Radiat. Res. 156, 103-109 (2001). We examined the interactive effects of cisplatin (1.0 microg/ml) combined with hyperthermia on cell killing and on the induction of apoptosis in IMC-3 human maxillary carcinoma cells. The cytotoxic effects of hyperthermia on IMC-3 cells at 44 degrees C were greater than at 42 degrees C, as has been reported for many other cells. The induction of apoptosis, DNA fragmentation and poly(ADP-ribose) polymerase cleavage were greater after hyperthermia at 44 degrees C for 30 min compared with treatment at 42 degrees C for 105 min, even though both of these heat doses were isoeffective in reducing cell survival to 50%. Treatment with cisplatin at 37 degrees C for up to 120 min did not result in cytotoxicity or the induction of apoptosis. The enhancement ratio for treatment with cisplatin at 42 degrees C was greater than that at 44 degrees C. More apoptosis was induced after the treatment with cisplatin at 42 degrees C compared to treatment with cisplatin at 44 degrees C. Taking these findings together, the combination of cisplatin and hyperthermia at 42 degrees C appeared to be more effective than cisplatin with hyperthermia at 44 degrees C for the induction of apoptosis in IMC-3 cells.

IL-1 induced chemokine production through the association of Syk with TNF receptor-associated factor-6 in nasal fibroblast lines.

The fibroblasts stimulated by cytokines released the chemokine and recruited the infiltrating cells, including eosinophils, that play a key role in the pathogenesis of airway disease. We established the human fibroblast lines showing high Syk expression and the lines showing low Syk expression from pieces of nasal polyp. IL-1 induces the interaction of TNFR-associated factor (TRAF) 6 with IL-1R-associated kinase, which is rapidly recruited to the IL-1R after IL-1 induction, whereas TRAF2 participates in TNF-alpha-signaling. In the present study, we found that Syk played a different role in IL-1- and TNF-alpha-induced chemokine production through a signaling complex involving Syk and TRAF6. Overexpression of wild-type Syk by gene transfer enhanced RANTES production from nasal fibroblasts stimulated with IL-1. The decrease of Syk expression by the administration of Syk antisense inhibited RANTES production in response to IL-1. However, the change of Syk expression did not affect RANTES production by TNF-alpha stimulation. We concluded that Syk is required for the IL-1-induced chemokine production through the association with TRAF-6 in fibroblasts of nasal polyps.

Acidic environment modifies heat- or radiation-induced apoptosis in human maxillary cancer cells.

PURPOSE: The effects of hyperthermia or irradiation on cell killing and induction of apoptosis were evaluated using human maxillary carcinoma IMC-3 cells and low pH (pH 6.8) adapted cells (IMC-3-pH). METHODS AND MATERIALS: Cellular heat-sensitivity or radiosensitivity was determined using the clonogenic assay. Apoptosis was assessed on the basis of a flow cytometric determination of the DNA content, DNA fragmentation, and poly(ADP-ribose)polymerase cleavage. RESULTS: When IMC-3 cells or IMC-3-pH cells were exposed to heat at 44 degrees C in pH 6.8 medium, an increase in thermosensitivity was observed compared with when the IMC-3 cells were exposed to heat at 44 degrees C in pH 7.4 medium. However, the selective reduction in survival was not observed after irradiation. In IMC-3 cells, apoptosis after heating at 44 degrees C for 60 min in pH 7.4 medium occurred earlier than that after 8 Gy irradiation, although both thermal and irradiated doses decreased the cell count to 10%. The degree of apoptosis after heating at pH 6.8 in IMC-3 cells or IMC-3-pH cells was greater than that at pH 7.4 in IMC-3 cells. However, the degree of apoptosis after 8 Gy irradiation at pH 6.8 in IMC-3 cells or IMC-3-pH cells was smaller than that at pH 7.4 in IMC-3 cells. CONCLUSION: Hyperthermia treatment is more effective at inducing apoptosis than radiation is in tumors that contain a population of low pH adapted cells.

Long-term follow-up results of electrogustometry and subjective taste disorder after middle ear surgery.

OBJECTIVES: The present study compares the long-term follow-up results of electrogustometry with patient reports of taste dysfunction after middle ear surgery. STUDY DESIGN: Retrospective review of 371 patients who underwent middle ear surgery. METHODS: Patients were divided into the following groups depending on the degree of manipulation or surgical damage to the chorda tympani nerves: the no-touch group (group 1 [n = 109]); the touch group (group 2 [n = 149]); and the severed nerve group (group 3 [n = 113]). Electrogustometry was periodically performed over the course of several years. RESULTS: The incidences of postoperative subjective taste disorder in groups 1, 2, and 3 were 2.8%, 25.5%, and 38.9%, respectively. Although the subjective taste disorder usually recovered within 1 to 2 years after surgery in all groups, it persisted for more than 2 years in 2.7% of the touch group and 5.3% of the severed nerve group. Concerning postoperative electrogustometric results, in the no-touch group, 8.3% of patients showed threshold elevation on electrogustometry, but the elevated thresholds completely recovered in all cases. In the touch group, 45% of patients exhibited elevated electrogustometric thresholds on their first postoperative test, including 32.9% who subsequently had complete electrogustometric recovery, 10.1% who subsequently had incomplete recovery, and 2% who failed to recover during the follow-up period. In the severed nerve group, none of the patients was responsive to the electrical stimulus on the first postoperative test, including 8.8% of patients who subsequently exhibited complete electrogustometric recovery, 32.7% who later had incomplete electrogustometric recovery, and 58.4% who never recovered any electrogustometric responsiveness. Nerve repair in the severed nerve group produced better recovery, as measured electrically. CONCLUSIONS: The incidence of postoperative subjective taste disorder was low, although inconsistent with the high incidence of threshold elevation on electrogustometry, especially in the severed nerve group. Preservation or repair of the chorda tympani nerve is recommended in order to maintain or recover gustatory function.

Keloid-derived fibroblasts are refractory to Fas-mediated apoptosis and neutralization of autocrine transforming growth factor-beta1 can abrogate this resistance.

The pathogenesis of keloid remains poorly understood. As no effective therapy for keloid is as yet available, an insight into its pathogenesis may lead to novel approaches. Apoptosis has been found to mediate the decrease in cellularity during the transition between granulation tissue and scar. Here, we report that in contrast to hypertrophic scar-derived and normal skin-derived fibroblasts, keloid-derived fibroblasts are significantly resistant to both Fas-mediated and staurosporine-induced apoptosis. The caspases-3, -8, and -9 were not activated indicating that the block in the apoptotic pathway in keloid is upstream of the caspases. There were no significant differences in the level of expression of Fas, Bcl-2, and Bax between the three groups but addition of transforming growth factor (TGF)-beta1 significantly inhibited Fas-mediated apoptosis in hypertrophic scar-derived and normal skin-derived fibroblasts and neutralization of autocrine TGF-beta1 with anti-TGF-beta1 antibody abrogated the resistance of keloid-derived fibroblasts. Anti-apoptotic activity was not observed with TGF-beta2. This is the first study linking refractory Fas-mediated apoptosis to cellular phenotype in keloids and indicating a pivotal role for the anti-apoptotic effect of TGF-beta1 in this resistance. Hence, it becomes important to treat keloids as a separate entity different from hypertrophic scars and enhancement of Fas-sensitivity could be a promising therapeutic target.

Technique for selective microangiography of skin flaps with high contrast and fine detail.

The technical factors of microangiography were studied, and the authors found that the main limiting factors in resolution were the injection pressure of the contrast medium and the thickness of the materials. A gentle manual hand pressure injection, with the aid of visual monitoring under the operating microscope, were keys to obtaining beautiful microangiograms.

Synthetic oligodeoxynucleotides inhibit IgE induction in human lymphocytes.

Synthetic oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs have the capacity to stimulate T-helper (Th)1-type responses in mice. Th1 cytokines are known to act as downregulators of IgE production. In this study we investigated whether synthetic ODNs inhibited IgE production in human peripheral blood mononuclear cells (PBMC) from normal donors stimulated with interleukin (IL)-4 plus anti-CD40 monoclonal antibody (mAb) in vitro. Thirty-mer single-stranded ODNs were randomly selected from the complementary DNA encoding the MPB-70 of Mycobacterium bovis Bacillus Calmette-Guerin. Two ODNs, containing CGTACG or AACGTT inhibited IgE production by human PBMC. When other oligonucleotides were substituted in a portion of the sequence of the core or flanking oligonucleotides in the ODN containing CGTACG, ODNs containing NACGTTCG or A/CTCGTTCG sequences specifically inhibited IgE production by human PBMC in vitro. The inhibition of IgE production by certain ODNs was mediated by both interferon (IFN)-gamma and IL-12, since the ODN-induced suppression was blocked by the addition of anti-IFN-gamma or anti-IL-12 mAb. Also, the ODNs inhibited induction of epsilon germline transcripts by IL-4. Our findings indicate that synthetic ODNs appear to be candidates for the treatment of IgE-dependent allergic disease in humans.

Inner ear abnormalities in Kabuki make-up syndrome: report of three cases.

Three patients, a female and two males, 28, 15, and 14 years of age, with Kabuki make-up syndrome (KMS) were studied for middle and inner ear abnormalities by using CT scanning of the petrous bones. All three patients had bilateral dysplasia of the inner ear, i.e., hypodysplasia of the cochlea, vestibule, and semicircular canals (so-called Mondini dysplasia), whereas their middle ears had no abnormalities. Audiometry demonstrated a sharp decrease in hearing of the high tone range, bilateral in one and unilateral in another, while the third patient was noncooperative. In view of these findings, it would be advisable to study each individual with KMS and hearing impairment for possible inner ear abnormalities.

Gluteus maximus adipomuscular turnover or sliding flap in the surgical treatment of extensive sacral chordomas.

Two cases with extensive posterior peritoneal defects after high sacral amputation for sacral chordoma are presented. An adipomuscular flap as a modification of the conventional gluteus maximus muscle flap was designed to obliterate an extensive residual posterior peritoneal dead space. The deep adipose tissue beneath the superficial fascia left on the gluteus maximus muscle was effectively used to provide more volume to the flap. The adipomuscular flap was turned over into the posterior peritoneal defect in the first case, and the flap was slid into the cavity in the other case. The adipomuscular flap eventually enabled the successful reconstruction of the posterior peritoneal defect, and the volume of the flap was well maintained behind the rectum, according to the postoperative magnetic resonance imaging findings in both cases.

DNA from Mycobacterium bovis bacillus Calmette-Guérin (MY-1) inhibits immunoglobulin E production by human lymphocytes.

A DNA fraction purified from Mycobacterium bovis bacillus Calmette-Guérin (BCG) and designated MY-1 induced interferon (IFN)-gamma production by human peripheral blood mononuclear cells (PBMC). IFN-gamma is well known as a downregulator of IgE production. In this study we investigated whether MY-1 regulates IgE production by human PBMC in vitro. MY-1 inhibited IgE production in PBMC taken from normal donors and stimulated with interleukin (IL)-4 plus monoclonal anti-CD40 antibody, without affecting production of IgA. MY-1 enhanced production of IFN-gamma and IL-12 by PBMC. Inhibition by MY-1 of IgE production was mediated by both IFN-gamma and IL-12, since the MY-1-induced suppression was blocked by the addition of monoclonal anti-IFN-gamma antibody, monoclonal anti-IL-12 antibody or a monoclonal antibody (mAb) directed at the IL-12 receptor. MY-1 inhibited the induction of epsilon germ-line transcript by IL-4. Additionally, MY-1 inhibited spontaneous in vitro production of IgE by PBMC from atopic donors in the absence of IL-4 plus anti-CD40 mAb. These results suggest that exposure to MY-1 may be a novel strategy for the treatment of IgE-related allergic disease.

[Application of microvascular surgery in reconstructive digestive tract surgery].

In our institutes, microvascular surgery has been effectively used in reconstructive digestive tract surgery, including esophageal reconstruction and hepatic arterial reconstruction. Free jejunal transfer combined with a gastric pedicle or microvascularly augmented elongated gastric pedicle has been utilized for total esophageal reconstruction. A microvascularly augmented jejunal pedicle or colonic pedicle has been applied in thoracic esophageal reconstructive cases with gastrectomy. Moreover, microvascular surgery has been performed in the reconstruction of the hepatic arterial system in the surgical treatment of pancreatic or bile duct cancer and living related-donor liver transplantation. Some pitfalls in selection of the recipient vessels and handling the intraperitoneal vessels for microvascular anastomosis are also described. Although microvascular surgery has been carried out by plastic and reconstructive surgeons in a team surgical approach, revisions in the medical educational system to create a new-type of surgeon with practical skills and clinical experience in both digestive tract and microvascular surgery will be required in future.

Functional alveolar ridge reconstruction with prefabricated iliac crest free flap and osseointegrated implants after hemimaxillectomy.

We achieved functional alveolar ridge reconstruction after hemimaxillectomy using a prefabricated iliac crest flap. The iliac crest was vascularized secondarily by a long rectus abdominis muscle flap with its inferior epigastric vessels intact to obtain an ideal anatomic location between the maxillary defect and microvascular anastomosis site. The iliac crest was tightly resurfaced with a split-thickness skin graft as well. After a bony surgical delay, the prefabricated iliac crest flap was microsurgically transferred to the face. Three osseointegrated implants were placed in the prefabricated iliac crest, and a dental prosthesis was worn with immobilization and stability. Our procedure enabled recovery of a satisfactory facial appearance and excellent masticatory function.

Clinical application of expanded free flaps based on primary or secondary vascularization.

Prefabricated free flaps using an expansion technique were used for four reconstructive cases, including two leg reconstructions and two facial reconstructions. In this series, the prefabricated free flaps created by using the expander were classified into two types: the expanded flap based on the conventional vascular pedicle, which is called the expanded flap with primary vascularization; and the expanded flap based on the vascular pedicle in the carrier, which is called the expanded flap with secondary vascularization. The expanded flap with primary vascularization that is created in the trunk has a good indication for leg reconstruction, because it provides an wide and thin flap with minimal donor site morbidity. The expanded flap with secondary vascularization created in the pectoral region has a good indication for facial reconstruction, because it provides good color and texture matches. Although there are some disadvantages in the tissue expansion technique, the prefabricated free flaps using the expander are very effective in facial and leg reconstruction.

A fiberscopic analysis of velopharyngeal movement before and after primary palatoplasty in cleft palate infants.

There have been few studies done on the abnormal function of velopharyngeal muscles in unrepaired cleft palate infants. To examine and assess velopharyngeal movement before primary palatoplasty offers supposedly any valuable information for the successful operation and the restoration of excellent velopharyngeal function. We designed to investigate and analyze velopharyngeal movement before and after primary palatoplasty in 26 cleft palate infants with a fine nasopharyngeal fiberscope. We found three different patterns of velopharyngeal movement in unrepaired cleft palate infants when crying or strangulation reflex occurred: (1) posterior movement type (10 cases, 38.5 percent), where the soft palates moved only posteriorly and cephalically and did not move medially; (2) medial movement type (10 cases, 38.5 percent), where the soft palates moved only medially and did not move posteriorly or cephalically; and (3) posteromedial movement type (6 cases, 23.0 percent), where the soft palates moved both posteriorly and cephalically as well as medially. Postoperative velopharyngeal closure was classified into three patterns: (1) the soft palate type, in which the soft palate mainly operates; (2) the lateral wall type, in which compensational medial movement of the lateral pharyngeal wall is mainly observed; and (3) the mixed type, in which both the soft palate and the lateral pharyngeal wall operate. Also, we demonstrated a close relationship between velopharyngeal movement before and after primary palatoplasty in cleft palate infants. In total, 10 of 16 cleft palate infants with the posterior movement type or posteromedial movement type, in which posterior movement of the soft palates was observed before primary palatoplasty, postoperatively showed the soft palate type of velopharyngeal closure. On the other hand, only 2 of 10 cleft palate infants with the medial movement type, in which the soft palates did not move posteriorly but medially before primary palatoplasty, postoperatively showed the soft palate type of velopharyngeal closure. The Fisher's exact probability test clarified that cleft palate infants with the posterior movement type or posteromedial movement type were more likely to show postoperatively the soft palate type of the velopharyngeal closure compared with those with the medial movement type (p = 0.051). This is the first trial to examine velopharyngeal movement in unrepaired cleft palate infants. Our findings indicate the probability that velopharyngeal closure mechanism in repaired cleft palate infants is able to be predicted by velopharyngeal movement behavior before primary palatoplasty. Next, we must clarify a correlation between preoperative velopharyngeal movement and postoperative velopharyngeal function and speech outcome.

Use of intraperitoneal vessels in reconstructive microsurgery: an account of 117 cases.

The purpose of this study was to evaluate our experience of microvascular anastomosis of intraperitoneal vessels. Between 1985 and 1994, 117 microsurgical reconstructions were done using intraperitoneal vessels. These included oesophageal reconstruction (n = 106), reconstruction of the hepatic arterial, superior mesenteric arterial, or portal venous system (n = 8), and reconstruction of the chest or abdominal wall (n = 3). We used 129 intraperitoneal arteries and 117 intraperitoneal veins. Of a total of 246 intraperitoneal vessels, five hepatic arteries, seven splenic, 14 gastroepiploic, six superior mesenteric, 178 jejunal, 30 ileocolic, four middle colic arteries or veins, and two portal veins were used for microvascular anastomosis. The overall successful rate for these reconstructive cases was 97% (114/117). Microsurgical use of intraperitoneal vessels is a safe and developing procedure in plastic and reconstructive surgery.

Effects of lipo-prostaglandin E1 on random pattern flaps in rabbits.

The effects of lipo-prostaglandin (PG) E1, which are lipid microspheres containing PGE1, were examined using 2 x 12 cm random pattern skin flaps on the backs of five rabbits. Five rabbits acted as controls. Both surviving length (8.5 (3.1) compared with 4.9 (1.8) cm) and surviving area (22.1 (9.3) compared with 11.5 (6.1) cm2) of the skin flap were significantly increased (p < 0.03 in each case). In three additional rabbits the blood flow on the distal side of the skin flap, where it was poor, was measured using a laser image blood flowmeter. Blood flow was improved five to 15 minutes after injection of lipo-PGE1 and was maintained for at least 60 minutes. In conclusion, lipo-PGE1 increased the blood flow of the random skin flaps, resulting in significant enlargement of the surviving area of skin flaps.

The effects of lipo-prostaglandin E1 on axial pattern flaps in rabbits.

The effects of lipo-prostaglandin E1 (lipo-PGE1), lipid microspheres containing PGE1, on axial pattern flaps were examined using 12 x 20 cm axial pattern flaps based on the right superficial inferior epigastric vessels on the abdomens of rabbits. The surviving area of the skin flap in the lipo-PGE1 group (0.197 +/- 14.1 cm2; n = 10) was significantly increased (p < 0.02) compared with the saline control group (110.4 +/- 5.5 cm2; n = 10). Changes in pedicle blood flow after lipo-PGE1 administration were determined using an ultrasonic pulsed Doppler flowmeter (n = 5). Blood flow in the artery was increased immediately after administration, and it was maintained for 40 minutes. The surface blood flow at the caudal end of the flap on the nonpedicle side, where it was poor, was measured using laserflowgraphy. Blood flow in the skin flap surface was improved 5 to 15 minutes after lipo-PGE1 administration, and it was maintained for as long as 60 minutes. Venous occlusion of the skin flap was also improved (n = 3). In conclusion, lipo-PGE1 increased the blood flow in the axial pattern flaps, resulting in significant enlargement of the surviving skin flap area.