Ultrastructural features resembling those of harlequin ichthyosis in patients with severe congenital ichthyosiform erythroderma.

Congenital ichthyoses are a group of heterogeneous disorders of cornification. Autosomal recessive congenital ichthyosis (ARCI) can be clinically subdivided into congenital ichthyosiform erythroderma and lamellar ichthyosis. Ultrastructurally, ARCI is classified into four groups: ichthyosis congenita (IC) types I-IV. The genetic background of the ARCI disorders is heterogeneous, but only one disease gene, transglutaminase 1, has been detected so far. We describe six patients with severe congenital ichthyosis from six different Scandinavian families. They could not be classified ultrastructurally into the four IC groups because of atypical findings of electron microscopy. These included abnormal lamellar bodies, alterations in keratohyalin, remnant organelles and lipid inclusions in the upper epidermal cells, which resembled the ultrastructural findings of harlequin ichthyosis (HI), although the HI phenotype was not present at birth. Some clinical features, such as thick scales, erythroderma, alopecia and ectropion were common to all patients. Ichthyosis was usually accentuated in the scalp and four patients had clumped fingers and toes. None of the patients carried the transglutaminase 1 mutation. We conclude that ultrastructural findings resembling those detected in previous HI cases (type 1 and 2) can also be found in patients who do not have classic clinical features of that rare ichthyosis. This may be due to lack of specificity of ultrastructural markers for HI or to its clinical heterogeneity.

Assignment of a novel locus for autosomal recessive congenital ichthyosis to chromosome 19p13.1-p13.2.

Autosomal recessive congenital ichthyosis (ARCI) is a rare, clinically and genetically heterogeneous genodermatosis. One gene (transglutaminase 1, on 14q11) and one additional locus (on 2q33-35, with an unidentified gene) have been shown to be associated with a lamellar, nonerythrodermic type of ARCI. We performed a genomewide scan, with 370 highly polymorphic microsatellite markers, on five affected individuals from one large Finnish family with nonerythrodermic, nonlamellar ARCI. The only evidence for linkage emerged from markers in a 6.0-cM region on chromosome 19p13.1-2. The maximum two-point LOD score of 7.33 was obtained with the locus D19S252, and multipoint likelihood calculations gave a maximum location score of 5.2. The affected individuals share two common core haplotypes, which makes compound heterozygosity possible. The novel disease locus is the third locus linked to ARCI, supporting previous evidence for genetic heterogeneity of ARCI. This is also the first locus for a nonlamellar, nonerythrodermic phenotype of ARCI.

A modified protocol for early treatment of osteomyelitis and osteoradionecrosis of the mandible.

BACKGROUND: The treatment of osteoradionecrosis (ORN) and early chronic osteomyelitis (COM) of the mandible and maxilla is controversial. Hyperbaric oxygen (HBO) at two to three times the atmospheric pressure at sea level can result in tissue oxygen tension of almost 400 mmHg. Herewith HBO increases oxygen supply in hypoxic tissue, thus inducing fibroblastic proliferation and capillary formation. METHODS: From 1981 to 1991, we used a monoplace chamber and since 1992, we have also had a multiplace chamber for HBO treatment. Hyperbaric oxygen was given at 2.5-2.8 atmosphere absolute pressure (ATA) for 90-120 minutes, once per day. The patients had five to 10 preoperative and five to seven postoperative sessions. Surgical therapy consisted of decortication of the affected bone, subsequently covered with a free periosteal transplant from the tibia. RESULTS: Thirty-six patients with ORN and 33 with COM of the mandible and maxilla was treated with this protocol. The median follow-up time in this material is 34 months, with a minimum of 10 months. Thirty-six ORN patients (92%) and 26 COM patients (79%) have remained symptom-free after the first treatment period. Three failed ORN patients were successfully treated with a free microvascular flap. The seven failed COM patients have been retreated, and five of them have occasional clinical symptoms. CONCLUSIONS: Hyperbaric oxygen is a promising adjunct to surgery in the treatment of mandibular and maxillary ORN and COM. Using this protocol, the necessary HBO treatment sessions have been reduced from earlier protocols, without adverse effect on the outcome.

The success rate of free flaps after preoperative irradiation in head and neck reconstruction.

BACKGROUND AND AIMS: The reconstructive treatment of large head and neck tumours involves serious problems. Preoperative irradiation increases the risk of postoperative complications. Microvascular free flaps offer definite advantages over conventional reconstruction techniques and permit immediate preoperative high-dose radiation therapy. MATERIAL AND METHODS: In 1985-1996, at the Turku University Central Hospital, 101 patients, 61 (60%) men and 40 (40%) women (age range 15-79 years, mean 61 years), with head and neck tumours, underwent microvascular reconstructions, and 90 of them were irradiated before surgery. All irradiated patients underwent primary reconstruction immediately after resection. The total irradiation dose was about 60 Gy, usual range 50 to 65 Gy. The patients were operated on 3 to 5 weeks after discontinuation of irradiation therapy. RESULTS: Of the whole material of 101 free flaps, 89 (88%) were transferred successfully. The success rate in irradiated patients was 88% and in nonirradiated patients 91%. Reoperation had to be performed within twelve hours in three cases, with good results. Only two hernias occurred as donor complications (2%). Two of the patients died postoperatively (2%). Partial flap necrosis occurred in five patients and five patients underwent another reconstruction using a new free flap or with conventional reconstruction techniques. CONCLUSIONS: Free flaps are often the only acceptable choice in extensive head and neck primary reconstructions after irradiation. Microvascular free tissue transfers to the head and neck after preoperative irradiation has become the accepted method of reconstruction as a result of increased success rates with superior aesthetic and functional results.

Microvascular free tissue transfers after preoperative irradiation in head and neck reconstructions.

Surgical reconstruction of large head and neck tumors involves great problems. Preoperative irradiation increases the risk of postoperative complications. The use of microvascular free flaps has definitive advantages over conventional reconstruction techniques and permits immediate preoperative high-dose radiation therapy. Eighty-eight patients, 55 (62%) men and 33 (38%) women, with head and neck tumors underwent microvascular reconstruction at the Turku University Central Hospital in 1985-1995; 77 of them were irradiated before surgery. All irradiated patients underwent primary reconstruction. The preoperative radiation dose was 50-65 Gy. The free tissue transfer was successful in 77 (88%) of the 88 patients. The success rate was 87% in irradiated patients and 91% in nonirradiated patients. Free tissue transfers are suitable for one-stage reconstruction in selected patients such as those receiving high-dose preoperative irradiation. The transfer of microvascular free flaps to the head and neck has become the most common reconstruction method owing to its increasing success rates as well as its excellent esthetic and functional results.

C1-tuning fork tests in school-aged children.

The Rinne and the Weber tests were carried out using a 256-Hz tuning fork on 687 6- to 15-year-old school children, the majority of whom were normally hearing subjects. The Rinne test was positive in 97% of ears with normal hearing or sensorineural hearing loss and negative in 72% of ears with conductive hearing loss. The Rinne test changed from positive to negative at 13 dB when performed using the normal loudness comparison method. In midline Weber results, the mean value of the air conduction difference at 0.25 kHz between the right and left ear was 1.4 dB. The lateralization in the Weber test occurred at a difference of 2.5 dB between the right and left ear. The Weber test was in midline in 96-98% of ears with bilateral normal hearing or sensorineural hearing loss. It lateralized to the poorer ear in 63% of ears with unilateral conductive hearing loss. The values of the Rinne and Weber tests were not good in the diagnosis of mild conductive hearing loss when the air-bone gap was 5-35 dB.

A modified protocol to treat early osteoradionecrosis of the mandible.

Hyperbaric oxygen (HBO) increases oxygen availability to hypoxic tissues thereby inducing fibroblastic proliferation and capillary formation in bone and soft tissue. From 1981 to 1991 we used a monoplace hyperbaric chamber, and since 1992 a multiplace chamber, for HBO treatments. HBO was given at 2.5-2.8 atm abs for 90-120 min, once per day. We used five to seven preoperative and five to seven postoperative sessions. Altogether, 17 patients with osteoradionecrosis (ORN) of the mandible or maxilla were treated with surgery and HBO. Surgical therapy consisted of decortication of the affected bone that was subsequently covered with a free periosteal transplant from the tibia. The shortest follow-up time is 18 mo. Sixteen patients have remained symptom free after the first treatment period. One case was a failure in the early protocol but was successfully treated using a free microvascular flap. It is concluded that HBO is a promising adjunct to surgery in treating mandibular or maxillary ORN.

Tympanic membrane changes in school-aged children.

A comprehensive otolaryngologic examination, including otomicroscopy, was performed for 687 school children for morphologically to study the distribution of normal and abnormal tympanic membrane findings and the association of different factors with the abnormal TM finding in 6 to 15-year-old children. Two-thirds of the children had completely healthy tympanic membranes without any visible pathological change. The findings were more often normal the older the child. In the total population, individual tympanic membrane findings were normal as follows: color 90.7%, position 93.8%, mobility 93.7%, structure 91.8% and position of pars flaccida 88.5%. Contrary to normal findings, tympanic membrane abnormalities were not in general associated with each other. The study provides new information on the occurrence of tympanic membrane changes among school-aged children.

Betahistine-induced vascular effects in the rat cochlea.

Betahistine (BH) has been used widely to treat cochlear disorders, such as tinnitus and Meniere's disease. The mechanism of action of BH in the cochlea is assumed to be based on its histamine-like effect on H1 receptors in the cochlear vasculature, leading to an increased cochlear blood flow (CBF). Recently it has been shown that BH can strongly affect H3 heteroreceptors (a novel histamine receptor subclass) in the periphery, via an autonomic ligand. This mechanism may also contribute to the BH effects on CBF. This study was to validate BH effects in the cochlear vasculature and to investigate the possible mechanisms of action of this drug in the inner ear vasculature. We assessed the effects of BH on CBF with the laser Doppler flowmeter in 23 rats and concluded that BH affects vascular conductivity in the cochlea in a dose-dependent fashion; betahistine diffuses through the round window, but does not have access to vascular receptors or ligands once in the labyrinthine fluids; and the H1 receptors mediate the systemic and peripheral vascular effects of BH, whereas the cochlear effect involves cholinergic receptors.

Healing of microvascular free skin flaps in irradiated recipient tissue beds.

This laboratory study compared the biomechanical and biochemical healing of microvascular free skin flaps in irradiated and nonirradiated recipient areas. In the adult rat, the skin of the right groin region was exposed to a single fraction dose of 20 Gy, using a linear megavoltage accelerator. The left groin was shielded. One week after radiotherapy, the left femoral artery and vein were divided, and an anastomosis was created in situ at the site of the epigastric pedicle. The epigastric skin flap was crossed over to replace the central area of the irradiated skin in the contralateral right groin. The healing of the interface between the irradiated recipient skin and the transferred free epigastric flap was studied at 4 weeks. A comparison was made with the microvascular free flaps of nonirradiated control animals. An additional group of control animals was used to verify the retarding effect of the 20-Gy radiation dose on wound healing. In control animals, the single dose of 20 Gy decreased the tensile strength of standardized surgical wounds by 23% (p = 0.03). Clinical healing of the transferred microvascular free flaps between the irradiated and control recipient areas was comparable, and the flaps showed no significant differences in the mechanical incorporation. The biochemical assays of the DNA, RNA-ribose, total nitrogen, and hydroxyproline contents of the flap showed an increased concentration of proteins (total nitrogen) expressed per cell count (DNA) (25%, p = 0.04), reflecting the excessive accumulation of organic matrix and a relatively decreased number of cells in irradiated recipient tissues. We conclude that there are no major differences in the healing capacity of microvascular free skin flaps between irradiated and nonirradiated recipient areas.

Experimental basis for lidocaine therapy in cochlear disorders.

In order to further our basic understanding of the effects of lidocaine hydrochloride in the inner ear, cochlear potentials and blood flow (CBF) were assessed after intravenous (i.v.), anterior inferior cerebellar artery (AICA), and local round window (RW) lidocaine administrations in guinea pigs and rats. Lidocaine RW applications produced a dose dependent decrease in compound action potentials (CAP) and cochlear microphonics (CM). The sensitivity changes were more pronounced at high frequencies. These findings suggest that lidocaine has specific pharmacological action in the inner ear other than simple anesthesia of the auditory nerve. The basal turn endocochlear potentials (EP) were not altered by topical lidocaine, implicating altered organ of Corti function following local application of lidocaine. RW applications of lidocaine had no effect on CBF or systemic blood pressure (BP). I.v. infusions caused substantial reductions in BP. In the case of systemic infusions the percent changes in CBF were equal to and accountable by the BP changes. The microinfusions (50 mg/ml, 100 nl/min) through AICA produced a 30%, long lasting increase in CBF. However, neither systemic lidocaine nor AICA infusions had an effect on CAP or CM. These findings indicate that systemically given lidocaine may not cross the blood-cochlear barrier and that the cochlear electrophysiological effects due to lidocaine when given locally are partly mediated by direct influence on cochlear hair cell function; they also suggest that lidocaine-induced interference with active ion transport in the lateral wall or an influence on CBF are not contributing factors.

Immediate reconstruction of mandibular defects with revascularized iliac bone grafts after radical surgery for osteosarcoma.

Excluding multiple myeloma, osteosarcoma is the most frequent primary malignant bone tumor. However, it is rare in the jaw bones, where it mainly occurs in young adult men. This report describes two mandibular osteosarcomas found in two males aged 32 and 30. In the former patient, osteosarcoma developed after bilateral mandibular sclerotic changes had been diagnosed radiologically as cementifying fibroma. This patient had received radiation therapy (56 Gy) because of a high grade astrocytoma of the brain three and a half years earlier. In the latter patient the tumor arose de novo in the mandibular molar area. The initial main symptom both patients was pain, which had been considered to be of dental origin because radiographs showed no evidence of malignancy. Radical surgery using immediate mandibular reconstruction with revascularized iliac bone grafts gave good results in both cases. The former patient died eight months after surgery because of recurrent astrocytoma and the latter is now free of disease five years after surgery. Immediate reconstruction of mandibular defects is possible with osteocutaneous groin flaps.

Clonogenic cell assay for anchorage-dependent squamous carcinoma cell lines using limiting dilution.

Clonogenic assays under either anchorage-dependent or -independent conditions are very useful for testing the sensitivity of tumor cells to cytotoxic drugs and radiation. These assays have not been widely used with squamous-cell carcinomas (SCC) because of poor tumor-cell viability and poor cloning efficiency, especially in semi-solid media. To find a clonogenic assay suitable for use with human squamous cancers we tested SCC lines, derived in our laboratory from patients with head and neck cancer, for the capacity to form colonies in soft agar and in 96-well plates. Of 13 UM-SCC lines tested for colony formation in agarose, only UM-SCC-11A was capable of growth in conventional semi-solid media. One other line, UM-SCC-14C, produced colonies in agarose only in the presence of epidermal growth factor. In contrast, all 17 of the SCC lines tested exhibited colony formation in adherent cell culture using limiting dilution in 96-well plates. The plating efficiencies of the SCC lines in the 96-well plate assay ranged from 0.02 to 0.52 colonies (wells)/cell whereas the PE values in soft agar were lower, ranging from 0.0055 to 0.0086 colonies/cell. The 96-well plate assay is not affected by cell migration, a problem encountered with some cell lines when clonogenic assays are performed in Petri dishes. UM-SCC-11A was tested for radiation sensitivity both in soft agar and in the 96-well plate assay. Comparable results were obtained. In summary, the majority of SCC cell lines did not form viable colonies in soft agar but the 96-well plate assay was applicable to a broad spectrum of anchorage-dependent human SCC cell lines and provides an efficient method for evaluating clonogenic cell survival.

Association of DNA aneuploidy with human papillomavirus-induced malignant transformation of sinonasal transitional papillomas.

The nuclear DNA content of 19 transitional papillomas of the sinonasal region and 9 maxillary squamous cell carcinomas was studied by flow cytometry; the presence of human papillomavirus (HPV) DNA types 11 and 16 was determined by the in situ hybridization technique from paraffin-embedded tissue. Thirteen (68%) of the papillomas and none of the carcinomas contained HPV genome. Six (32%) of the papillomas and 4 (44%) of the carcinomas had an aneuploid DNA content. The relative DNA content (DNA index) of the aneuploid maxillary carcinomas was larger than that of aneuploid papillomas (p = 0.004). Three of the papillomas underwent malignant transformation, all three of which contained HPV type 16 DNA; two were also aneuploid. Data indicate that papillomas containing HPV type 16 DNA have a tendency (p = 0.06) to undergo malignant transformation, and that this tendency is greater if DNA aneuploidy or HPV type 11 DNA is also present (p = 0.02).

Transitional papilloma of the nasal cavity and paranasal sinuses. Clinical course, viral etiology and malignant transformation.

Transitional papilloma is a rare but not uncommon tumor with an overall incidence varying between 0.5 and 4% of all primary nasal tumors. The probability of recurrence is high, between 20 and 62%, even after adequate therapy. Malignancies are associated with transitional papilloma in 2-13% of cases. A series of 21 patients with transitional papilloma of the nose and/or paranasal sinuses is presented. Human papillomavirus (HPV) structural proteins and DNA types were found in 13 cases (62%) of transitional papillomas studied. None of the 9 nasal polyps and none of the 9 squamous cell carcinomas of paranasal sinuses studied as control material contained HPV DNA. We found 4 recurrent papillomas (19%) during the 2-year observation time. All recurrences were HPV-positive. The incidence of malignant transformation was 14% in this series. Treatment of transitional papilloma is always surgical. In our opinion medial maxillectomy and ethmoidectomy via sublabial rhinotomy is the surgery of choice in the treatment of transitional papilloma.

Radiosensitivity of head and neck cancer cells in vitro. A 96-well plate clonogenic cell assay for squamous cell carcinoma.

Radiation sensitivity was determined for nine University of Michigan squamous cell carcinoma (UM-SCC) cell lines, MCF-7 and HeLa, using a 96-well plate clonogenic assay. Plating efficiencies (PE) of the UM-SCC cell lines were between 0.16 and 0.36. Higher PE values obtained were with MCF-7 (0.4) and HeLa (0.5). The UM-SCC cell lines were used at low passages (passage 13 to passage 20) to minimize artifacts attributable to long-term culture. Cells were irradiated in suspension using a cobalt 60 gamma source at a dose rate of 0.94 Gy/min (94 rad/min). Survival data were fitted well by either a linear quadratic function F = e-(alpha D + beta D2) or by a monoexponential function F = Ae-alpha D. Mean inactivation dose, equivalent to the area under the survival curve (AUC), was used as a measure of radiation sensitivity. The UM-SCC-1, 9, 11A, 11B, MCF-7, and HeLa were the most radiation resistant lines we tested (AUC greater than 2.1), while UM-SCC-14A was the most sensitive (AUC = 1.591). The assay was highly reproducible, and the difference in radiation sensitivity between cell lines were statistically significant.

Detection of human papillomavirus (HPV) structural antigens and DNA types in inverted papillomas and squamous cell carcinomas of the nasal cavities and paranasal sinuses.

To assess the suggested etiological role of human papillomavirus (HPV), biopsies from 14 patients operated on for an inverted papilloma (11 cases) and squamous cell carcinoma (3 cases) of the nasal cavity and paranasal sinuses were analysed for light microscopical evidence of HPV, by indirect immunoperoxidase (IP-PAP) to demonstrate HPV structural proteins, and using in situ DNA-hybridization to disclose the DNA of HPV types 6, 11 and 16. The majority of the inverted papillomas contained areas of metaplastic squamous cells, including koilocytes as well as dysplastic changes consistent with intra-epithelial neoplasia as described in uterine cervix. In 3 patients, frankly invasive squamous cell carcinomas were found, originating from dysplastic squamous epithelium. Of the 14 lesions, 7 (50%) expressed HPV antigens, usually confined to only a few cells close to the surface of the epithelium. None of the malignant lesions expressed HPV antigens. In situ DNA-hybridization disclosed HPV 11 DNA alone in 5 lesions, but none of the lesions contained HPV 6 DNA. HPV 16 DNA was found in 2 lesions as a single HPV type, and in 3 other lesions concomitant with HPV 11. All three carcinomas contained HPV 16 DNA. The HPV DNA distribution proved to be different from that found in the genital tract HPV lesions; HPV DNA was less abundant in the nasal papillomas, and it was also found in the basal and suprabasal cells.(ABSTRACT TRUNCATED AT 250 WORDS)