Olfactory disorders in childhood: A comparative study of olfaction in children with adenoid hyperplasia versus a control group and the postoperative effects of adenoidectomy with respect to olfactory ability.

PURPOSE: Hyposmia in childhood is poorly characterized. The "U-Sniff Test", validated for children with anosmia, can be used to objectify olfactory impairment but has not been used to distinguish between hyposmia and normosmia. Therefore, we investigated children with enlarged adenoids with respect to hyposmia, its correlation with adenoid size, and the sensitivity of questionnaires to predict olfactory impairment. METHODS: In a prospective comparison, olfaction was assessed by "U-Sniff Test" (score 0-12; <8 hyposmia) in 41 children (5-18 years) with adenoid hyperplasia and compared with 196 children without any respiratory affection (control) after exclusion of previous SARS-Cov2-infection from December 2020 to December 2021. ENT-related complaints were collected using a self-designed questionnaire. We were able to include 13 children in a follow-up examination to compare preoperative performance in the "U-Sniff Test" with postoperative outcome after adenoidectomy. STATISTICS: chi-square-test (p < 0.05), odds-ratio, Spearman's rho, ROC-, cluster analysis. RESULTS: Severe hyposmia was present in 36.6% of children with adenoid-hyperplasia compared to 3.1% of the control-group. Adenoid-children scored significantly more often between 8 and 10 points (58.5%) than the control (31.6%; p < 0.01). Adenoid size and olfactory performance correlate significantly (r: 0.83; CI -0.89 … -0.72). Hyposmia in the adenoid group is characterized predominately by loss of the odors banana, butter and rose. None of children with hyposmia or parents reported impaired olfactory performance. Postoperatively, olfactory function improved significantly in 85% of cases (p 0.01, SD ± 1.71, Δ3.54points). CONCLUSION: Questionnaires are insufficient to detect hyposmia in this cohort. In contrast, the "U-Sniff Test" detects even reduced olfactory performance without reaching the cut-off value, which represents the majority of test results in the adenoid group. Therefore, we recommend the classification of moderate hyposmia (8-10 points) to be included for our study population.

[Quantitative methylation-specific PCR for the diagnosis of lung cancer]. / Quantitative methylierungsspezifische PCR zur Lungenkarzinomdiagnostik.

Efficient, preferably early diagnosis of lung cancer represents a major challenge. Under this aspect the sensitivity of conventional histomorphology and cytomorphology procedures is unsatisfactory. This review highlights technical aspects, possibilities and drawbacks of the application of aberrant promoter methylation as a biomarker for lung cancer diagnostics using specimens of pulmonary exfoliative cytology.

[Aberrant promoter methylation as biomarker for molecular cytological diagnosis of lung cancer]. / Aberrante Promotor-Methylierungen als Tumormarker für die molekularzytologische Diagnostik des Lungenkarzinoms.

Aberrant promoter methylation represents a main mechanism of tumor suppressor gene inactivation and may serve as a new source for biomarker discovery. This study investigated its applicability as a molecular tool for lung cancer diagnostics on bronchial aspirates. A methylation assay was developed applying a quantitative methylation specific real-time PCR (QMSP). A total of 552 patients with the differential diagnosis of lung cancer were investigated. The QMSP findings on bronchial aspirates were compared with the methylation status of respective genes investigated in microdissected tumor tissues (QMSP, cloning and sequencing of promoter regions after bisulfite conversion). Among the genes tested a marker panel consisting of APC, p16(INK4a) and RASSF1A proved to be the best suited for lung cancer diagnostics. This panel allowed for a correct diagnosis of lung cancer in cases with an ambiguous or false negative conventional cytology. In a cohort study on 247 patients, the combination of histology (sensitivity 59 %), cytology (sensitivity 44 %) and QMSP-assay (sensitivity 53 %) raised the sensitivity of a single bronchoscopy for the diagnosis of lung cancer up to 81%. The methylation assay yielded its major diagnostic surplus with respect to peripheral tumors representing 59 % of all primaries detected. In patients without antecedent lung cancer its specificity considering malignancy was >99 %. Therefore, the QMSP-assay is a promising technique which could enhance the sensitivity and diagnostic impact of conventional cytology. The assay is applicable to residual material of regular diagnostic cytology even in retrospect.

Spontaneous malignant transformation of conventional giant cell tumor.

Spontaneous malignant transformation of conventional giant cell tumor (GCT) of bone is exceedingly rare. We report on a case of GCT of the iliac crest in a 35-year-old woman with malignant change into a high-grade osteosarcoma 10 years after the first appearance of GCT on a radiograph. Since the patient refused therapy for personal reasons the tumor remained untreated until sarcomatous transformation occurred. Image cytometry showed DNA aneuploidy and a suspiciously high 2c deviation index (2cDI) in the primary bone lesion. A thorough review of the world literature revealed only seven fully documented cases of secondary malignant GCT which matched the definition of a "sarcomatous growth that occurs at the site of a previously documented benign giant cell tumor" and not treated by radiotherapy. These cases as well as the current one suggest that a spontaneous secondary malignant GCT presents as a frankly sarcomatous tumor in the form of an osteosarcoma or malignant fibrous histiocytoma. It usually appears at sites of typical GCTs-often without any recurrent intermediate state-and is diagnosed 3 or more years after the primary bone lesion. The prognosis is poor.

[Molecular cytopathology of lung cancer--prevalence of genetic alterations and their role in the development of molecular biomarkers]. / Molekulare Zytopathologie bösartiger Lungentumoren--Prävalenz der genetischen Alterationen und ihre Bedeutung für die Entwicklung molekularer Biomarker.

Carcinogenesis of lung cancer proceeds via a complex process that involves multiple genetic abnormalities, which do not necessarily have a linear progression. Genetic alterations include aneuploidy, deletions and amplifications of chromosomal regions, loss of heterozygosity (LOH), microsatellite alterations, point mutations and aberrant promoter methylation. There is considerable effort to use these genetic alterations as molecular biomarkers for early cancer diagnosis applying different approaches. An ideal tumor marker should be highly sensitive, tumor specific, easily to handle and non-cost intensive. While previous studies used screening for mutations, LOH and microsatellite alterations, more recent strategies concentrate on multicolor fluorescence in situ hybridization (FISH) and aberrant promoter methylation. Since in general the genetic alterations are prone to be more extensive in tumor cells as compared to non-tumor cells, methods that provide quantitative data (e.g., methylation specific real-time PCR) are likely to improve specificity. Consequently, molecular biomarkers could constribute to a more accurate risk assessment in carcinogen exposed individuals and early molecular cytologic diagnosis of precancerous lesions and cancers of the lung.

Prognostic significance of DNA cytometry in carcinoma of the uterine cervix FIGO stage IB and II.

OBJECTIVE: To assess the prognostic value of DNA-image cytometry in cervical carcinoma of the uterus and its relation to other established prognostic factors. STUDY DESIGN: The study included 116 cases of cervical carcinoma FIGO stages IB and II which were treated with radical abdominal hysterectomy. The median follow-up was 55 months (range 1-162 months). DNA image cytometry was performed on cytologic specimens prepared by enzymatic cell separation from formalin-fixed, paraffin-embedded tissues. DNA stemline ploidy, DNA stemline aneuploidy, 5c exceeding rate, 9c exceeding rate, 2c deviation index, and DNA malignancy grade were computed. DNA-variables as well as various clinical and histological variables were related to survival rates. RESULTS: In multivariate statistical analysis DNA stemline ploidy using 2.2c as a cut-off value and FIGO stage showed to be statistically significant available presurgery predictors of survival, whereas the postsurgical parameters lymphonodal status, tumor size and parametrial involvement were significantly correlated with survival. The synopsis of all parameters in a multivariate Cox model indicated that - with declining relevance - the number of positive pelvic lymph nodes, DNA stemline ploidy using a cut-off level at a modal value of 2.2c, largest pelvic lymph node, 5c exceeding rate, and ratio of carcinoma area to cervix area, were of predictive value for survival. CONCLUSIONS: Our results suggest that prognostic information deducted from classical staging parameters is successfully complemented by DNA image cytometry which can be applied pretherapeutically.

Overexpression of cathepsin B and urokinase plasminogen activator is associated with increased risk of recurrence and metastasis in patients with chondrosarcoma.

BACKGROUND: Deregulation of the cellular protease network has been shown to be responsible for aggressive clinical behavior in several common human malignancies. In the current study, the authors evaluated the expression patterns of proteases in patients with chondrosarcoma of bone and correlated these patterns with clinical outcome. METHODS: The expression levels of urokinase plasminogen activator; matrix metalloproteinase types-1, -2, and -9; and cathepsins B and L were determined immunohistochemically in 114 cases of chondrosarcomas of bone and were correlated with their clinicopathologic parameters as well as with long term follow-up data. RESULTS: Overexpression of cathepsin B was associated with a high rate of local recurrence (P = 0.006) and a decreased recurrence free survival (P = 0.005). Overexpression of urokinase plasminogen activator was associated with an increased rate of metastasis (P = 0. 013), a decreased metastasis free survival (P = 0.016), and a decreased 5-year overall survival rate (P = 0.048). The univariate Cox model showed that tumor extension into soft tissue, high histologic grade, and overexpression of cathepsin B were predictors of adverse outcome. Multivariate analysis showed only overexpression of cathepsin B and tumor extension into soft tissue to be independent predictors of local recurrence. CONCLUSIONS: Overexpression of cathepsin B and urokinase plasminogen activator can be used to identify those patients with chondrosarcoma of bone who have an increased risk of local recurrence and distant metastases.

Mutation of p53 with loss of heterozygosity in the osteosarcomatous component of a dedifferentiated chondrosarcoma.

We investigated a dedifferentiated chondrosarcoma of a 61-year-old woman with an osteosarcomatous high-grade component for p53 alteration. The low-grade cartilaginous and the high-grade osteosarcomatous components of the tumor were macrodissected and evaluated separately by immunohistochemistry and molecular biology. We used PCR-SSCP analysis and direct sequencing to screen exons 4-8 for p53 mutations. The p53 intron 1-polymorphism was investigated for loss of heterozygosity. A functionally relevant p53 missense mutation in codon 193 of exon 6 (A-to-T transversion) with loss of wild-type allele was detected only in the dedifferentiated component. Using the monoclonal antibody DO-1, immunohistochemistry failed to show p53 overexpression. This evidence of p53 mutation may be regarded as at least a co-factor that "switched" the preexisting low-grade conventional chondrosarcoma to a highly malignant dedifferentiated tumor.

Rosai-Dorfman disease and generalized AA amyloidosis: a case report.

We report on a patient who, at 31 years of age, was found to suffer from sinus histiocytosis with massive lymphadenopathy (SHML; Rosai-Dorfman disease) with nodal and extranodal involvement as described previously. Five years later the patient presented with nephrotic syndrome caused by a generalized AA amyloidosis, and he subsequently died from pulmonary thromboembolism owing to renal vein thrombosis. Retrospective analysis of serum levels of C-reactive protein (CRP) showed that during the last 3 years before his death, he had a persistently elevated CRP level ranging from 73 to 161 mg/L, despite antiinflammatory treatment with prednisolone, methotrexate, or 6-mercaptopurine. These figures indicate that the patient was probably suffering from a permanent acute phase response which, in the absence of any other evidence of a chronic inflammatory disease which commonly causes AA amyloidosis, was most likely owing to SHML.

Osseous manifestation of Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy). A case report and review of the literature.

Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy) is an unusual form of histiocytic disorder. Bone lesions are infrequent. We describe a 33-year-old man with involvement of multiple bones but without lymphadenopathy at the time of presentation. The literature on osseous manifestation in this condition is reviewed.

[Pathology of Ewing sarcoma]. / Pathologie des Ewing-Sarkoms.

Ewing's sarcoma is a very rare tumor which has, however, attracted much oncological interest since the dramatic improvement of its prognosis under chemotherapy. Its histogenesis has been discussed controversially for a long time, including a possible origin in immature reticulum, myogenous, endothelial and undifferentiated mesenchymal cells. Repeated reports have also suggested a possible neuroectodermal genesis. Convincing arguments, however, have only been brought forward during recent years, since it was found that Ewing's sarcoma and malignant peripheral neuroectodermal tumor share a common chromosome translocation 11;22. In the meantime this hypothesis has been strengthened by numerous cell biological analyses. There seems to be no clear border between Ewing's sarcoma and malignant peripheral neuroectodermal tumors with definite neural differentiation. Histological differential diagnosis of Ewing's sarcoma has been improved by immunohistological methods. In most cases, they can be distinguished from lymphoma (leucocyte common antigen, B and T markers) and embryonal rhabdomyosarcoma (muscle specific actin, desmin) without problems. Apart from that, it is possible nowadays to obtain antibodies against the MIC 2-protein, which is preferably expressed in Ewing sarcoma. The diagnostics of Ewing's sarcoma and the malignant peripheral neuroectodermal tumor have considerably been enriched by the fact that the specific chromosome translocation t(11;22) can be proved molecular biologically. In contrast to the cytogenetic evidence, it is not necessary to establish cell cultures.

Polyostotic heterogeneity of the spine in osteoporosis. Quantitative analysis and three-dimensional morphology.

It was the aim of this study to record quantitatively and qualitatively the distribution of the three-dimensional microarchitecture throughout the human spine in osteoporosis. Bone biopsies of the iliac crest and the complete spine of 26 autopsy cases without skeletal disease and 11 female patients with proven osteoporosis were removed. Grindings of all vertebrae by a technique which we developed allowed two- and three-dimensional measurements simultaneously. The analysis included an evaluation of trabecular bone volume, trabecular interconnection, and trabecular thickness, as well as a qualitative investigation of the structure of cancellous bone. The bone loss in osteoporosis is a loss of structure. The relative loss of the trabecular microarchitecture is greater in the iliac crest than in the lumbar spine. It is a gradual change from normal bone to osteoporosis. Transformation from plates to rods and the loss of whole trabeculae are caused by perforations. The polyostotic heterogeneity in osteoporosis is remarkable. Adjacent vertebrae may show differences of up to 100% in bone structure and bone volume. This explains the difficulties in early diagnosis of osteoporosis. Due to the polyostotic heterogeneity it is impossible to define a threshold mineral content for osteoporotic fractures.

The microarchitecture of the axis as the predisposing factor for fracture of the base of the odontoid process. A histomorphometric analysis of twenty-two autopsy specimens.

The axis from twenty-two cadavera was removed at autopsy and was sectioned in the sagittal plane to a thickness of one millimeter with use of a surface-stained block-grinding technique. Combined two and three-dimensional analysis included an evaluation of the volume of the trabecular bone, the trabecular interconnection, and the cortical thickness as well as qualitative investigation of the structure of the cancellous bone. The body of the axis, the base of the odontoid process, and the odontoid process were analyzed separately. The base of the odontoid process is a region of least resistance for fractures because of its unique microarchitecture. The mean volume of trabecular bone of the base of the odontoid process is 55 per cent less than that of the axis and the odontoid process. The base also has a markedly poorer trabecular interconnection and a cortical thickness that is one-third that of the odontoid process. In all of the specimens, trabeculae that were disconnected from the trabecular lattice (trabeculae with free ends) were demonstrated in the base of the odontoid process. The formation of microcallus in six (27 per cent) of the specimens supports the hypothesis that microfractures occur as a result of stress peaks, mechanical fatigue, and the relative insufficiency of bone in the static condition. Therefore, the base of the odontoid process can be considered as a site of predilection for fractures.

[Micro-callus formation of spongiosa. An up to now underestimated repair mechanism of the skeletal system]. / Mikrokallusformationen der Spongiosa. Ein bisher unterschätzter reparativer Mechanismus des Skelettsystems.

Microcallus formations are demonstrable in nearly all cancellous bones by means of suitable preparation techniques. Histologically, these structures are immature fibrous bone formed in local overloaded parts of the trabeculae. Using a preparation technique that allows combined two- and three-dimensional analysis, 26 normal human spines and 11 osteoporotic spines were investigated for microcallus. Microcallus formations occur frequently in people over 45 years of age. They are mainly localized in the lower thoracic and lumbar spine and occur significantly more frequent in females than in males. The number of microcallus formations depends more on the microarchitecture of the cancellous bone than on individual trabecular parameters. In about 33% of cases microfractures are demonstrable in the centre of the microcallus formation. In non-invasive studies the bone mass could be misinterpreted due to microcallus. Although it indicates instability of the bone structure, microcallus formation is not a purely negative mechanism. It stabilizes and regenerates the bone tissue. Furthermore, complete new trabeculae can be formed due to bridges of microcallus between residual trabeculae. Osteoporosis is not the result of an inability to form microcallus.

Three-dimensional analysis of the spine in autopsy cases with renal osteodystrophy.

The spinal column was removed from nine autopsy cases with chronic maintenance dialysis and sectioned in the sagittal plane to a thickness of 1 millimeter using a surface-stained block grinding technique. A combination of two- and three-dimensional analysis included an evaluation of the spine deformity index (SDI), the bone volume (BV/TV), the trabecular interconnection (TBPf), the trabecular thickness (Tb.Th), the trabecular number (Tb.N) as well as a qualitative investigation of the structure of cancellous bone. The control group consisted of 26 autopsy cases with intact skeletons. An iliac crest biopsy made a direct comparison of the diagnostic biopsy location and the spinal column possible. The SDI showed vertebral fractures in renal osteodystrophy (ROD) of types I and II, in spite of a trabecular bone volume within normal limits. The trabecular bone volume showed mean values and a distribution throughout the spinal column familiar in the skeletally-intact control group. Those cases with a longer history of hemodialysis showed higher BV/TV values regardless of age and sex. A trabecular volume within normal limits did not mean physiological trabecular interconnection. Perforations were commonly observed with ROD. Classical, hidden and tunnelling perforations were distinguished. Microcallus formations were frequently seen in the periphery of vertebrae at rod nodes, rods and plate intersections. Three-dimensional analysis in ROD shows a greater alteration in architecture than can be assumed from the two-dimensional histological sections.

Enzyme-linked immunosorbent assay for detection of immunoglobulin A and G antibodies to Campylobacter pylori.

An enzyme-linked immunosorbent assay (ELISA) employing an acid-glycine extract was used to detect IgG and IgA antibodies to Campylobacter pylori in sera from 179 patients with upper gastrointestinal disease, 174 blood donors and 65 children. The incidence of positive ELISA results clearly increased with the severity of histopathologic findings in the antrum mucosa and was also high in patients with peptic ulcers and gastric cancer. The incidence in blood donors and children was much lower and increased with age. The results achieved with the ELISA were similar to those observed previously using the immunoblot method. Differences between whole cell preparation and acid-glycine extract with respect to their protein profiles and immunoblot reactivities were minor. IgM titres were very low and could not be related to histopathological findings, peptic lesions or culture findings. The ELISA may be particularly useful for monitoring the outcome of therapy aimed at eradication of Campylobacter pylori.

[A method for the gas chromatographic determination of long and medium chain free fatty acids in serum]. / Eine Methode zur gaschromatographischen Bestimmung der lang- und mittelkettigen freien Fettsäuren im Serum.

Free fatty acids in serum were determined by gas chromatography after esterification with boron tri-fluoride-methanol. Following prepurification of the serum lipid fractions by thin layer chromatography, the described analytical procedure selectively determines the medium and long chain free fatty acids. Accuracy and precision were tested. The method is suitable for the analysis of fatty acid kinetics in blood, following lipid infusions.