A model for the study of autoimmune diseases applied to pemphigus: transplants of human oral mucosa to athymic nude mice binds pemphigus antibodies in vivo.

The present paper describes a new in vivo method to study the action of pemphigus antibodies against human tissue. Oral mucosal biopsies from healthy donors were transplanted to athymic nude mice, which, a week later, were injected with serum from pemphigus patients. From 1 to 5 days after the injection the epithelial transplants were removed and preparations were studied by immunofluorescence microscopy. Pemphigus antibodies were demonstrated in preparations from each of 23 mice which had received pemphigus serum, but in none of 6 which had received control serum. Transplants from about 2/3 of the experimental mice showed intercellular edema of the basal layers of the epithelium and in transplants from 3 mice supra-basilar splitting of the epithelium was found. None of these changes was seen in the control mice. Passive transfer of human serum or lymphocytes to nude mice transplanted with human tissue may be use in future studies of autoimmune diseases, including pemphigus.

Volume-referent bone turnover estimated from the interlabel area fraction after sequential labeling.

In the compact bony otic capsule remodeling is low, and bone remodeling units are distributed centrifugally in relation to inner ear tissues and spaces. Fluorochrome-labeled bone remodeling units are scarce, abortive, and tortuous with no uniform direction of movement. This study presents a method for the estimation of volume-referent bone turnover based on measurements of the fractional area between labels after sequential labeling with osteofluorochromes. The applicability of this method is tested against a classical quantification method in undecalcified cortical specimens from the canine humerus, where both methods can be used. The estimate of bone turnover derived from the new sequential labeling in eight dogs is 7.4% (SEM 2.1%) per year and the classic estimate derived from calculations of the formative osteonal area and the formative period yields 6.9% (SEM 2.1%) per year. Agreement is sufficient to justify future measurements of absolute bone turnover in sequentially labeled perilabyrinthine bone.

Experience in qualitative and quantitative FDG PET in follow-up of patients with suspected recurrence from head and neck cancer.

We evaluated positron emission tomography (PET) with 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG) in the detection of recurrent head and neck cancer, and compared visual and quantitative interpretation of PET images for their accuracy in the identification of tumour recurrence. Sixty-two FDG PET studies were performed in 56 patients having a total of 81 lesions, which were clinically suspected for recurrent carcinoma of the head and neck. The PET images were interpreted visually, and tracer uptake was quantitated as the standardised uptake value adjusted to body weight (SUV). Sensitivity of visual interpretation of the PET images for the presence of malignancy ranged from 84 to 95%, and specificity from 84 to 93%, respectively, depending on the selected scheme for grading of the lesions. Malignant lesions accumulated significantly more FDG than the benign ones (the median SUVs were 6.8 and 3.3, respectively, P<0.001). However, there was a wide overlap of the FDG uptake values between these two groups. Hence, the highest accuracy of quantitative analysis in correct identification of tumour recurrence (75% at Receiver Operating Curve analysis) was inferior to that of visual analysis (89%). FDG PET is feasible for the detection of recurrent head and neck cancer. Although quantitation of FDG uptake using SUV shows significantly higher tracer concentrations for malignant than benign lesions, the wide overlap of individual SUVs between these two groups is a serious concern in diagnostic evaluation. Therefore, in clinical practice it may be preferable to identify the presence of tumour recurrence within this patient group by qualitative interpretation of the PET images.

Melanotic schwannoma of the orbit. Immunohistochemical and ultrastructural study of a case and survey of the literature.

Light microscopical, immunohistochemical and ultrastructural study of a case of melanotic (pigmented) schwannoma in the left orbit of a 22-year-old man is presented. The tumour caused exophthalmus for three months and was located under the eyeball. Partial removal resulted in recurrence after three years. The recurrent tumour with extension to the nasal cavity and maxillary sinus was extirpated without loss of vision, and a year later the patient was without symptoms. The tumour tissue was Fontana positive, could be bleached, and showed immunoreactivity for S-100, vimentin and HMB 45. Ultrastructurally, the tumour cells, particularly the pigment loaded cells, showed a varying amount of basal lamina at the surface, melanosomes in varying stages of melanization, fine cytoplasmic filaments and micropinocytotic vesicles. Survey of the literature and discussion of the melanogenesis in Schwann's cells are presented.

Ménière's disease: endolymphatic sac decompression compared with sham (placebo) decompression.

The placebo effect in surgery for Ménière's disease was investigated in a double-blind, controlled surgery by comparing the effect of a regular endolymphatic shunt with the effect of a purely placebo operation (regular mastoidectomy). Thirty patients with typical Ménière's disease participated in the study. They were selected for surgery because of unsuccessful medical treatment and were chosen randomly for each treatment group. The patients filled in daily dizziness questionnaires for 3 months before and 12 months after surgery, registering nausea, vomiting, vertigo, tinnitus, hearing impairment, and pressure in the ears. The patients were operated on in two university ENT departments. Those operated on in one department were controlled each month at the other department, and vice versa. At the termination of the trial, the investigators as well as the patients gave their overall opinion of the efficacy of the operation. Minor differences could be demonstrated between the active and the placebo group, but the greatest difference in symptoms was found when comparing pre- and postoperative scores, in which both groups improved significantly.

Human pleomorphic adenomas transplanted to nude mice.

Tissue from 13 human pleomorphic adenomas was transplanted to a total of 64 nude mice. Eight of the tumors were transplanted into a second passage of mice, 17 in all. In 39 mice of first passage, there was a definite increase in graft size. Microscopic examination showed no change in the histologic pattern from the donor tumor to the transplanted tissue. The heterochromatin pattern after staining with the DNA-specific fluorochrome D287/170 allowed distinction between human and murine cells and showed that both epithelial and mesenchymal cells were of human derivation. Autoradiographic studies with tritiated thymidine showed that both epithelial and mesenchymal tumor cells were labeled. Our results thus show that cell proliferation in the human pleomorphic adenoma takes place in epithelial areas as well as in mesenchymal areas.

Placebo effect in surgery for Meniere's disease: three-year follow-up.

In 1981 we published the results of a double-blind, placebo-controlled study in which the efficacy of a regular endolymphatic sac mastoid shunt was compared with a purely placebo operation (mastoidectomy) in controlling the symptoms of 30 patients with typical Meniere's disease. Minor differences could be demonstrated between the shunt and the sham operation, but the greatest difference was between the preoperative and postoperative scores, and both groups improved significantly. It was concluded that the impact of the various endolymphatic sac shunts on the symptoms of patients with Meniere's disease is nonspecific, and that the 70% improvement in both groups was most likely caused by a placebo effect. At the time of the presentation, the results were based on a 1-year follow-up of all patients. As of January 1982 the patients had been followed for a minimum of 3 years. The 3-year results are the same as our results from the first year: no significant difference could be found between the two groups.

Otospongiosis: morphologic and microchemical investigation after NaF-treatment.

Element analysis of microareas of otospongiotic specimens is described. A total of 36 otospongiotic stapes are ultrasectioned without decalcification and examined using a transmission and a scanning electron microscopy (STEM-mode). The latter was equipped with an energy dispersive x-ray analyzer. Twenty of the stapes came from patients who have had sodium fluoride (NaF) treatment (15 to 45 mg/day) for a minimum of 12 months. The otospongiotic stapes are classified as spongiotic and sclerotic according to their pathologic characteristics and state of mineralization. Using the Ca/P ratio as criterion--measured by the characteristic x-ray fluorescence--it was shown in a blind study that the NaF-treated otospongiotic stapes had a statistically higher Ca/P ratio, indicating that the fluoride may stabilize otospongiotic lesions, particularly the spongiotic type with unstable mineralization.

Surface morphology of the endolymphatic duct in the rat. A scanning electron microscopy study.

Following intracardiac vascular perfusion fixation of 8 rats with glutaraldehyde in a buffered and oxygenated blood substitute, the vestibular aqueduct and endolymphatic duct were opened by microsurgery of the resulting 16 temporal bones. Optimum preservation of the epithelium for scanning electron microscopy was attained by coating of the specimens with osmium tetroxide and thiocarbohydrazide followed by a continuous dehydration procedure. This technique permitted, for the first time, an investigation of the surface morphology of the epithelial cells in the endolymphatic duct. Three types of cells were identified with the scanning electron microscope. A polygonal and oblong epithelial cell was observed in the largest number throughout the duct, and in the juxtasaccular half of the duct, two additional types of epithelial cells were observed. The scanning electron microscopic observations are compared and discussed with reference to previous transmission electron microscopic studies of the endolymphatic duct.

Remodeling patterns in the bony otic capsule of the dog.

Quantum-type bone remodeling was studied in undecalcified temporal bones and long bone specimens from adult mongrel dogs by means of combined microradiography, osteofluorochrome labeling, and osteoid staining. The overall turnover rate of capsular bone was estimated to be low compared to that of extracapsular compact bone. Evolving secondary osteons were distributed centrifugally relative to the perilymphatic space. Moreover, a zone of bone tissue enclosing perilymphatic spaces was found completely devoid of evolving secondary osteons. Throughout the otic capsule, segmentary perivascular foci of resorption and formation were present. These findings indicate that remodeling of capsular bone is subject to sustained local inhibition. The significance of this conclusion is considered with special reference to otosclerosis.

Demonstration of intravital microfissures in undecalcified plastic-embedded temporal bones with the prestaining technique.

Microfissures in the human otic capsule have been observed since the start of the century, but it was Otto Mayer, in 1930, who first realized that some of them were of intravital origin and not just processing artifacts. Since then, a small number of publications, based on decalcified temporal bones, have mostly confirmed his findings. With the introduction by Frost in the late 1950s of the undecalcified bone technique and the bulk staining technique for peripheral bones, a method was developed and refined for identifying even very small intravital microfissures (microdamage). Bulk staining of undecalcified otic capsules has not yet been used to verify the findings from the previous decalcified specimens. The present report presents our experience with the pertinent techniques, and suggests modifications and shortcuts pertinent to temporal bone research. Both large and tiny microfissures of intravital genesis are demonstrable within bulk-stained undecalcified human otic capsules. The importance of microfissures in the petrous bone in the causation of otosclerosis and perilymphatic leakage has long been discussed, and the present techniques may advance our understanding of these pathological conditions

Endolymphatic sac-mastoid shunt surgery. A nonspecific treatment modality?

The reason for the effectiveness of endolymphatic sac-mastoid shunt surgery in the treatment of patients with Meniere's disease is still open for debate. In a double-blind study, published in 1981, we could not demonstrate any difference between the effect of a simple mastoidectomy and a regular endolymphatic sac-mastoid Silastic sheet shunt. However, a significant reduction in symptoms could be demonstrated in both groups, and 70% of patients in both groups could be classified as successes. The patients were reexamined 3 years after surgery, and it was still not possible to demonstrate any differences between the sham and the active surgery. In this study, no significant differences between the two groups have been found at follow-up averaging 84 months, and success has been maintained in about 70% of patients. The only three failures, who have consistent vertiginous attacks, have been confined to the actual shunt group. Two patients in the active group have lost their hearing as compared with none in the sham group, and 35% of the patients have now developed bilateral disease. We believe that endolymphatic sac-mastoid shunt surgery is a nonspecific treatment modality, and we find no need for sac shunt surgery. The vast majority of the patients can be successfully treated by nonsurgical means, but we emphasize that above all the patient must be assured that in the event of persistent debilitating symptoms, a surgical solution to the problem is available.

Estimation of volume referent bone turnover in the otic capsule after sequential point labeling.

Using fluorochrome labeling and a newly validated method for bone turnover estimation, we determined absolute values for canine perilabyrinthine bone remodeling. The overall capsular bone turnover was found to be 2.1% per year, compared to 13.9% per year for the neighboring cranial bones and 7.4% per year for the humerus compacta. This gross 2.1% per year conceals a vast range, from 0.13% per year for the innermost perilymphatic zone, through a centrifugal increment toward 8% to 10% per year in the periphery. The underlying individual bone remodeling units exhibit a similar centrifugal pattern in numerical density and size. These findings indicate an inhibition of remodeling, seemingly emanating from the perilymphatic spaces, and affecting both the activation of osteoclasts and the extent of resorption by the osteoclasts. These values satisfactorily explain the preservation of such fetal remnants as the globuli ossei, the interglobular spaces, and the skein bone. In humans, local ineffective inhibition of bone resorption may play a role in the initiation of otosclerosis.

Audiological and temporal bone findings in myxedema.

Fifteen patients with confirmed myxedema at a median age of 48 years (range 32 to 60 years) were referred for audiological evaluation before and after treatment with levothyroxine. The median interval between the pretreatment and posttreatment investigations was 18 months (range 9 to 27 months). In addition, 13 patients at a median age of 78 years (range 64 to 95 years) were audiologically reexamined after long-standing levothyroxine treatment. The observation period upon treatment with levothyroxine was 40 months (range 32 to 46 months). No improvement in hearing sensitivity could be demonstrated either in the younger patients or in the elderly. When compared to an age- and sex-matched unscreened population, the myxedematous patients did not demonstrate any different degree of hearing loss. Histological investigation of the temporal bones from an 83-year-old woman with myxedema, however, showed no morphological changes or deposition of glycosaminoglycans, changes which are compatible with true age-related hearing loss. It is concluded that no association exists between myxedema and hearing impairment and that no morphological or structural changes due to myxedema can be demonstrated in the temporal bones.

Otospongiosis and sodium fluoride. A blind experimental and clinical evaluation of the effect of sodium fluoride treatment in patients with otospongiosis.

The effect of sodium fluoride treatment in patients with otospongiosis has been evaluated blindly in a morphological and microchemical element analysis of otospongiotic specimens together with a prospective clinical double-blind, placebo-controlled study. The results show that using the calcium/phosphorus ratio as an indication for bone maturity, the sodium fluoride treatment can stabilize otospongiotic lesions in retaining calcium relative to phosphorus. The clinical double-blind, placebo-controlled study of 95 patients showed a statistically significant worse deterioration of the hearing loss in the placebo group than in the active treated (40 mg sodium fluoride daily) group, supporting the view that sodium fluoride can change otospongiotic, active lesions to more dense, inactive otosclerotic lesions. We have postulated in the past that the actual mechanism of the cochlear loss is toxic enzymes produced by histiocytes at the periphery of the microfoci, and it may be that sodium fluoride has some effect on these enzymes.

Heterotransplantation of human pleomorphic adenomas to nude mice.

The purpose of the present study was to establish a model for in vivo studies of human pleomorphic adenomas by heterotransplantation of tumour tissue to nude mice. Tissue from 7 tumours was transplanted to a total of 34 mice. Take with obvious growth occurred in 12 mice, and survival of the tissue was seen in an additional 8 mice. The overall histological picture was unchanged from the donor tumours to the transplanted tissue. The possibilities of the model are discussed.

Prevalence of otitis media in a survey of 591 unselected Greenlandic children.

In an unselected survey in two Greenlandic towns, 591 children were examined to study the prevalence of otitis media (OM). The attendance rate in Nuuk was 80%, while 93% participated in Sisimiut. The children were three-, four-, five- and eight-years-old and represented 18% of children in these age groups in all Greenland. In total, 51.7% in Nuuk and 54.1% in Sisimiut presented pathologic middle ear affections ranging from slight to severe. The prevalence rates of chronic OM (COM) were 6.8% in Nuuk and 11.7% in Sisimiut (P = 0.055) but without significant age or sex difference. The acute OM point prevalence rate was between 1.5% and 0.4%. The prevalence rate of middle ear effusion (MEE) was between 23.0% and 28.2%. Secretory OM was significantly more prevalent in the younger age groups. The odds ratio of having COM was significantly higher in children with two Greenlandic born parents (3.07) than in children with only one Greenlandic born parent. A follow-up study after one year in Sisimiut revealed unchanged or aggravated middle ear disease in 56.8% of 82 children with middle ear pathology at the primary survey. Thus, OM persists as a major health problem among Greenlandic children, although the general socio-economic and medical conditions have improved during the last decades. Proposals are provided for increased otologic efforts.

Acute otitis media and sociomedical risk factors among unselected children in Greenland.

OBJECTIVE: To describe the sociomedical risk factors associated with episodes of acute otitis media (AOM), recurrent AOM (rAOM), and chronic otitis media (COM) in Greenlandic children and especially to point out children at high risk of rAOM (defined as > 5 AOM episodes since birth) and COM which are prevalent among Inuit children all over the Arctic. METHODS: The study design was cross-sectional and included 740 unselected children, 3, 4, 5, and 8-years-old, living in two major Greenlandic towns, Nuuk and Sisimiut. All children were otologically examined and the parents answered a questionnaire containing sociomedical variables including ethnicity, family history of OM, housing, insulation, crowding, daycare, passive cigarette smoking, breast feeding, type of diet, allergy, and chronic diseases. Historical data were cross-checked in medical records which also formed the basis for the drop-out analyses. Statistical analyses included frequency tests, calculation of odds ratio (OR), and multiple logistic regression. RESULTS: The attendance rate was 86%. Former episode of AOM was reported by 2/3 of the children, rAOM by 20%, and COM by 9%. The following variables were found significantly more often in children with AOM by simple frequency testing: Parental (OR = 1.83), sibling (OR = 1.62), and parental plus sibling (OR = 2.56) history of OM, crowding (OR = 5.55), long period of exclusive breast feeding ( > 4 months) (OR = 2.47), and recent acute disease (P = 0.034). The following variables were found significantly more often in children with rAOM or COM by simple frequency testing: Parental history of OM (OR = 1.60; OR = 2.11, respectively) and no recall of breast feeding (P = 0.005; P = 0.003, respectively). Also, COM was found significantly more often in children with two Greenlandic parents (OR = 3.07). A multiple logistic regression test denoted only parental history of OM (OR = 1.82) and long period of exclusive breast feeding (OR = 1.14) as significant predictors of AOM. CONCLUSIONS: Many of the risk factors usually associated with AOM could not be confirmed as risk factors in this survey. Parental history of OM and long period of exclusive breast feeding were the strongest factors associated with AOM in Greenlandic children and ethnicity was associated with COM. However, the study confirms that AOM is a multifactorial disease determined by a number of genetic and environmental factors.

Fine structure of the endolymphatic duct in the rat. A scanning and transmission electron microscopy study.

To investigate the surface morphology of the endolymphatic duct epithelium, 8 rats were vascularly perfused with glutaraldehyde in a buffered and oxygenated blood substitute. Optimal preservation of the epithelium for scanning electron microscopy was attained by coating of the specimens with OsO4 and thiocarbohydrazide followed by a continuous dehydration procedure. Three types of cells were identified with the scanning electron microscope: A polygonal and oblong epithelial cell was observed in the largest number throughout the duct, whereas in the juxta-saccular half of the duct two additional types of epithelial cells were observed. The scanning electron microscopical observations are compared and discussed with reference to transmission electron microscopical observations of the endolymphatic duct.

The suspensory ligament of the endolymphatic duct in the rat. An ultrastructural study.

Following optimized fixation and specimen handling, the endolymphatic duct was investigated by transmission and scanning electron microscopy. Tubular microfibrils, 10-12 nm thick and of indefinite length, occupied the subepithelial compartment abundantly and seemed to insert into the basal lamina of the epithelial cells of the duct and to merge with the bony wall of vestibular aqueduct thus forming a suspensory ligament of the endolymphatic duct. This ligament is suggested to protect the endolymphtaic duct from collapsing and to play a role in the mechanical protection of the vestibular apparatus from sudden increases in intracranial pressure. The capillaries and the aqueductal vein were located in intimate relationship to the epithelial cells. Both types of vessels were provided with fenestrations of the endothelial cells, supporting the proposal that the endolymphatic duct may be involved in the resorption of endolymph.