GLCCI1 Polymorphism rs37973 and Response to Treatment of Asthma With Inhaled Corticosteroids.

BACKGROUND: The response to asthma treatment is highly variable, and having pharmacogenetic markers that predict response to treatment would bring us one step closer to personalized treatment. Genome-wide association studies have shown that polymorphisms in GLCCI1 could be associated with the response to inhaled corticosteroids (ICSs) in asthma patients. MATERIAL AND METHODS: We genotyped rs37973 in GLCCI1 in 208 adult asthma patients treated with ICSs. The percentage change in FEV1, % predicted was analyzed after short-term treatment (3 months) and long-term treatment (at least 3 years). Treatment was defined as successful when FEV1 decreased by <30 mL/year. RESULTS: After 3 months of treatment, FEV1, % predicted was higher in patients with the GG genotype than in patients with the AG+AA genotype, and this genotype-dependent difference was only evident in nonsmokers. Similar results were found in nonsmokers and patients with atopy after at least 3 years of treatment, when all patients were analyzed. Even though no differences were observed for success of treatment (good vs poor response) when the whole group of patients was analyzed, genotype-dependent treatment success was highly influenced by smoking and atopy. The GG genotype was overrepresented in nonsmokers and patients with atopy and a good response. CONCLUSIONS: rs37973 was associated with response to short- and long-term treatment; however, smoking and atopy had a considerable effect on pharmacogenetic association. Furthermore, in contrast with findings from genome-wide association studies, we found the GG genotype to be associated with better treatment response.

Cell count-based triaging of cytology samples for cell block preparation.

OBJECTIVE: Cytology samples can sometimes yield cell blocks of low cellularity that are inadequate for cytomorphological evaluation and ancillary studies. The purpose of this study was to analyse the value of cell count based-triaging of samples for cell block preparation. METHODS: Cell blocks were prepared from different cytology samples using HistoGel™ according to the manufacturer's instructions following cell count assessment. The adequacy of cell blocks was semi-quantitatively scored as empty, scanty, moderate or highly cellular by evaluating corresponding haematoxylin and eosin (H&E) sections. Cell blocks with moderate and highly cellular H&E sections were considered adequate whereas cell blocks with empty and scanty cells in H&E sections were inadequate for cytomorphological evaluation. The average, range and a median number of cells (×106 ) embedded in adequate and inadequate cell blocks were compared. RESULTS: Among 54 cell blocks prepared from fine needle aspiration biopsies (FNAs, 32), effusions (4), brushings (13) and cell lines (5), 20 cell blocks (37%) yielded moderate or highly cellular H&E sections and were considered adequate whereas 34 (63%) cell blocks were empty or scanty and thus considered inadequate. The average (± SD) number of cells (×106 ) embedded in adequate and inadequate cell blocks was 2.1 ± 4.3 and 0.5 ± 0.3, respectively. Samples containing at least 1 × 106 cells yielded adequate cell blocks in 12/16 cases (75%) whereas samples with less than 1 × 106 cells in 13/38 (34%) cases. CONCLUSION: The present study showed that cell count-based triaging of cytology samples could reduce the number of low cellular cell blocks.

Optimization and validation of immunocytochemical detection of oestrogen receptors on cytospins prepared from fine needle aspiration (FNA) samples of breast cancer.

OBJECTIVE: To optimize and validate immunocytochemical (ICC) assessment of oestrogen receptors (ERs) on cytospins prepared from fine needle aspiration (FNA) samples. METHODS: Optimal conditions and variability in ICC detection of ERs were established on cytospins prepared from the human breast cancer cell line MCF-7. Protocols that yielded adequate results were further validated on 52 FNA samples of resected breast cancer tumours using analysis of concordance with the ER status, determined by standard immunohistochemistry on corresponding formalin-fixed, paraffin-embedded tissue (FFPET). On 37 diagnostic FNA samples, manual immunostaining with antibody 1D5 was compared with automated immunostaining with antibody 6F11. RESULTS: The highest percentage of ER-positive MCF-7 cells with lowest variability was obtained on methanol-fixed cytospins with or without microwave pre-treatment: 72 ± 5% and 75 ± 7%, respectively. Microwave pre-treatment was mandatory for Papanicolaou-stained cytospins in order to achieve between 63 ± 14% and 67 ± 9% of ER-positive MCF-7 cells. The concordance between ICC assessment of ERs on FNA samples and corresponding FFPET sections was complete for methanol-fixed cytospins (100%, kappa = 1) and adequate for Papanicolaou-stained cytospins (94%, kappa = 0.84) and Papanicolaou-stained smears (92%, kappa = 0.75). Complete agreement in ICC detection of ERs was obtained for manual immunostaining with antibody 1D5 and automated immunostaining with antibody 6F11. CONCLUSIONS: Methanol-fixed cytospins prepared from FNA samples ensure highly reliable ICC assessment of ERs, whereas Papanicolaou-stained cytospins or smears are conditionally suitable because of the small risk of false negative results.

A polymorphism in ORMDL3 is associated not only with asthma without rhinitis but also with chronic obstructive pulmonary disease.

BACKGROUND: Asthma is a heterogeneous disease, and asthmatic patients without rhinitis more commonly have fixed airway obstruction, a feature that is also typical of chronic obstructive pulmonary disease (COPD). The Dutch hypothesis suggests that both COPD and asthma have common genetic risk factors. The purpose of this study was to assess the association between the polymorphism rs4795405 in the known asthma candidate gene ORMDL3 and asthma with and without rhinitis. We also analyzed COPD in order to investigate whether, in addition to a clinical overlap, there might also be a genetic overlap between COPD and asthma. METHODS: The population of this genetic association study comprised 493 Slovenian adults, distributed as follows: 131 patients with asthma (59 had asthma with rhinitis and 72 asthma without rhinitis), 59 patients with rhinitis only, 133 patients with COPD, and 170 controls. Genotypes for rs4795405 were determined using the TaqMan genotyping assay. RESULTS: rs4795405 was specifically associated with asthma without rhinitis. Assuming a recessive genetic model, we found the CC genotype in 26% of healthy controls, in 24% of patients with asthma with rhinitis (P = .862), and in 44% of patients with asthma without rhinitis (P = .006). Polymorphism rs4795405 was also associated with COPD, for which the CC genotype was found in 37% of cases (P = .045). CONCLUSIONS: rs4795405 was strongly associated with asthma without rhinitis, a subtype of asthma for which a higher degree of airway obstruction was found. These results show the importance of analyzing different asthma phenotypes in genetic association studies. We also observed a genetic overlap between COPD and asthma without rhinitis.

Airway angiogenesis in stable and exacerbated chronic obstructive pulmonary disease.

Angiogenesis is a prominent feature of structural tissue remodelling that occurs in chronic airway diseases, including chronic obstructive pulmonary disease (COPD). The aim of this study was to evaluate the airway levels of VEGF, angiogenin, IL-8 and TNF-α in patients with COPD during the stable phase and during acute exacerbation of the disease. We analysed induced sputum samples from 28 patients with COPD. Thirteen of these patients were followed up and second samples of sputum were obtained during acute exacerbation of the disease. The two control groups consisted of 12 healthy smokers and seven healthy non-smokers, all with normal lung function tests. Concentrations of VEGF, angiogenin, IL-8, TNF-α and bFGF were measured by cytometric bead array. In the induced sputum of patients with stable COPD, concentrations of VEGF (P < 0.001, P = 0.02), angiogenin (P < 0.0001, P < 0.0001), IL-8 (P < 0.0001, P = 0.0021) and TNF-α (P < 0.001, P = 0.03) were significantly elevated in comparison with healthy smokers and non-smokers. No additional elevation of angiogenic factors was demonstrated at the time of exacerbation. There was a significant negative correlation between FEV1 and VEGF (P < 0.05, r = -0.38), angiogenin (P < 0.0001, r = -0.68) and IL-8 (P < 0.001, r = -0.54) among smokers (smoking COPD patients and healthy smokers). No significant differences were observed between groups of healthy smokers and non-smokers. These results showed increased airway angiogenesis in patients with COPD. Moreover, VEGF, IL-8 and angiogenin negatively correlated with pulmonary function, which suggests their important role in COPD airway remodelling. However, no additional angiogenic activation was found during exacerbation of COPD.

Review of negative and low-grade cervical smears in women with invasive cervical cancer after the first 3 years of the national cervical screening programme in Slovenia.

OBJECTIVE: The purpose of the study was to perform a national review of negative, low-grade and inadequate smears reported during the latest screening period before cervical cancer diagnosis in 2006, after the first 3 years of the screening programme. METHODS: Among 162 new cervical cancer cases there were 47 (29%) without previous cytology, 47 (29%) with one high-grade smear prior to diagnosis and 68 (42.0%) with at least one previous negative, low-grade, atypical or inadequate smear 1-40 months before diagnosis. Of the latter 68 cases, 37 patients with 59 smears (together with 118 control slides) were included in the review as 31 had smears reported at laboratories no longer operating. Findings were related to the last cytology report before diagnosis as well as to histological type and stage of the cancer. RESULTS: In our study group, 19 (51%) of 37 patients had squamous cell carcinoma, 15 (41%) adenocarcinoma and 3 (8%) adenosquamous carcinoma, compared with 121 (75%), 26 (16%), 12 (7%), respectively, and 3 (2%) other types, for all carcinomas. Twenty-one of 37 women also had high-grade cytology prior to diagnosis of cancer. Women with previous cytology (with or without recent high-grade smears) were more likely to have stage I cancers than those without cytology (P < 0.0001). The expert group upgraded 17/33 smears in the patients with squamous carcinomas, which was more than in those with adeno- and adenosquamous carcinomas (5/24, P < 0.05). CONCLUSION: As expected, a higher proportion of smears preceding adenocarcinomas were true negative. Under-diagnosed smears were not related to cancer stage or last cytology report before diagnosis.

External quality control for immunocytochemistry on cytology samples: a review of UK NEQAS ICC (cytology module) results.

OBJECTIVE: To date, external quality control for immunocytochemistry on cytology samples is provided only by the United Kingdom national external quality assessment service for immunocytochemistry and in situ hybridisation (UK NEQAS ICC & ISH). For the purpose of this study a retrospective analysis of a comprehensive collection of quality-related data regarding immunocytochemistry on cytology samples collected through this service was analysed. METHODS: The quality of immunocytochemical reactions, using on-line collected data, was analysed for the last 23 UK NEQAS ICC cytology module external quality assessments carried out on cytology samples completed in the period from 2004 to 2010. RESULTS: Our study showed that the majority of participants in the cytology module (66%) sent formalin-fixed paraffin-embedded (FFPE) tissue sections for assessment as in-house control slides and only 34% sent cytology slides of various types. The highest UK NEQAS ICC score for the quality of immunocytochemical staining among in-house control slides was achieved on cell block sections, followed by cytospins, FFPE tissue sections, liquid-based cytology slides and smears. With regard to fixation, acetone-fixed slides achieved significantly lower scores than other reported fixatives. The strength of agreement in perception of immunocytochemical staining quality was good between in-house assessors (Kappa=0.64) but only fair between in-house and UK NEQAS ICC assessors (Kappa=0.22). CONCLUSIONS: Good quality of immunocytochemical staining can be achieved on cytology slides prepared and fixed in different ways as well as on cell blocks. Unified criteria for high-quality immunocytochemical staining and proper internal and external quality assurance could facilitate further improvement and standardization of immunocytochemistry on cytology samples.

Complement factor C5a in acute exacerbation of Chronic Obstructive Pulmonary Disease.

The complement component C5a is a potent inflammatory peptide, which may be involved in the pathogenesis of Chronic Obstructive Pulmonary Disease (COPD). We analysed the induced sputum and plasma of 28 patients with stable COPD, 12 healthy smokers and 7 non-smokers. In 13 of the patients with COPD, we also observed paired samples during acute exacerbation. The concentrations of C5a/C5a desArg and C3a/C3a desArg were measured using cytometric bead array. Both C5a and C3a concentrations in induced sputum of stable patients with COPD were significantly increased compared to the control groups of healthy smokers and non-smokers. In addition, there was a significant elevation in C5a values in exacerbation of COPD that was independent from the airway C3a levels. Airway C5a levels were negatively correlated with forced expiratory volume in first second (FEV1)% predicted and diffusing capacity of the lung (TLCO). Plasma C5a concentrations in patients with COPD were significantly higher than in healthy smokers, but no further significant systemic C5a elevation was detected with acute exacerbation of COPD. There was no important difference in local or systemic C5a concentrations between healthy smokers and non-smokers. These in vivo results clearly show that local and systemic C5a concentrations in COPD are elevated, and that the local, in contrast to systemic, C5a concentrations additionally increase in the acute exacerbation of COPD. It seems that the cigarette smoke is not related to C5a increase. The elevated local and systemic C5a levels, and additional individual local C5a increase during the exacerbation support the importance of C5a in COPD.

Airway angiogenesis in patients with rhinitis and controlled asthma.

BACKGROUND: Airway angiogenesis may be an important part of structural remodelling in the pathogenesis of asthma. The development of asthma is frequently preceded by rhinitis. OBJECTIVE: We sought to determine whether the levels of angiogenesis-related factors are elevated in airways of patients with rhinitis or controlled asthma. METHODS: We analysed the induced sputum of 18 rhinitis patients, 16 asthmatic patients, and 15 healthy controls. The concentrations of angiogenin, vascular endothelial growth factor (VEGF), IL-8, fibroblast growth factor (bFGF), and TNF-alpha were measured by cytometric bead arrays. RESULTS: We found significantly increased angiogenin and VEGF concentrations in the induced sputum supernatant of both rhinitis and asthma patients compared with that of the healthy control group (P< or =0.0005). With the exception of TNF-alpha, there was no difference in the other angiogenic factors; TNF-alpha levels were higher in the rhinitis group than in the control group (P=0.02). CONCLUSION: These in vivo results suggest increased airway angiogenesis in patients with rhinitis without asthma as well as in corticosteroid-treated and well-controlled asthma patients.

Identification of carcinoma origin by thyroid transcription factor-1 immunostaining of fine needle aspirates of metastases.

UNLABELLED: Identification of carcinoma origin by thyroid transcription factor-1 immunostaining of fine needle aspirates of metastases OBJECTIVE: To assess the diagnostic potential of thyroid transcription factor-1 (TTF-1) immunocytochemical staining for identifying the origin of unknown primary cancers from fine needle aspiration cytology (FNAC) of metastases. METHODS: Immunostaining with TTF-1 was conducted on FNACs suggesting metastatic carcinoma in 128 patients with unknown primary tumours as part of the initial diagnostic procedure. The majority of patients (78) presented with neck masses, four had lung lesions and 46 had masses at other locations. The patients' files were reviewed to obtain data on tumour origin. RESULTS: Thirty-seven of 128 (28.9%) cases stained positive with TTF-1. Pulmonary origin of metastases was confirmed in 27 cases, and four lung lesions were primary lung adenocarcinoma. Two patients had primary and one had mestastatic thyroid carcinomas. In two patients, no follow-up data was available. In one patient, the origin of the primary could not be determined at autopsy. TTF-1 was negative in 18 patients with subsequent diagnosis of metastatic pulmonary cancer and in seven patients diagnosed clinically and radiologically as most probably having a lung carcinoma. In 60 TTF-1 negative patients, the primary tumour originated in other organs (34) or remained unknown (26). CONCLUSION: TTF-1 immunostaining of FNAC in patients with metastatic carcinoma of unknown origin identified primary pulmonary or thyroid carcinomas in nearly a third of cases. Patients with TTF-1 positive metastases could avoid further extensive diagnostic investigations for the origin of the primary site.

Ultrasonographically and stereotactically guided fine-needle aspiration cytology of non-palpable breast lesions: cyto-histological correlation.

OBJECTIVE: To determine the diagnostic accuracy of ultrasonographically (US) and stereotactically guided fine needle aspiration cytology (FNAC) for the diagnosis of non-palpable breast lesions. PATIENTS AND METHODS: From January 2002 to December 2004, 470 women with 478 mammographically detected non-palpable breast lesions had US or stereotactically guided FNAC of the breast lesion. Subsequent histological evaluation of the same lesion was performed at the Institute of Oncology Ljubljana, Slovenia. The correlation between the original cytological and histological diagnosis was assessed and the sensitivity, specificity, positive predictive value and negative predictive value were calculated. RESULTS: Among US-guided FNACs, 144 (53.5%) were histologically verified benign lesions and 125 (46.5%) were carcinomas. Cytological diagnoses were: true positive (TP) in 63 cases (50.4%), suspicious in 35 (28%), false negative (FN) in eight (6.4%), and in 19 (15.2%) cases, the material was inadequate for diagnosis. In the stereotactically guided FNAC group, there were 209 women with 209 lesions, with 95 (45%) histologically proven carcinomas and 114 (55%) benign lesions. Cytological diagnoses were TP in 49 (51.6%) cases, true suspicious in 21 (22.1%), FN in nine (9.5%), and in 16 (16.8%) cases, the material was not adequate for the diagnosis. CONCLUSION: The sensitivity of 88.7% and specificity of 98.6% for US-guided FNAC and 84.5% and 100% for stereotactically guided FNAC, respectively, suggest that clinicians can rely upon cytological diagnosis for planning further management of women with non-palpable breast lesions.

Haemorrhagic cytology samples - how to get the best diagnostic results.

OBJECTIVE: To describe and evaluate the value of a simple filtration technique for use in processing haemorrhagic samples for cytomorphological evaluation and immunocytochemistry. METHODS: One hundred and sixty haemorrhagic cytological samples (133 FNAs, 27 effusions) received in our laboratory from August 2002 to September 2005 were included in this study. After preparing two smears for diagnostic evaluation, the residual sample was suspended in 2 ml of a cell medium prepared in our laboratory. These primary haemorrhagic suspensions were filtered through disposable nylon filter devices and the cells deposited on the upper membrane surface were transferred into 2 ml of fresh cell medium. From all three fractions - primary cell suspension, filter deposit and filtrate - cytospins were prepared and stained by Giemsa or Papanicolaou methods. Cytospins were examined under the microscope for the presence of diagnostic cells, red blood cells (RBCs) and debris. Additional cytospins for immunocytochemistry were prepared at the cytopathologist's request. RESULTS: RBCs and debris were successfully removed in 142 out of 160 haemorrhagic samples (88%) by using this new filtration technique. In all these cases the tumour cells were well presented and allowed substantially improved cytomorphological evaluation. Immunocytochemical staining was performed on 112 filtered samples with three different markers per case on average. Filtration did not improve the quality of cytospins in 18/160 haemorrhagic samples, mostly attributable to insufficient numbers of diagnostic cells in the original samples. CONCLUSION: The presented filtration technique is very simple and quick. It substantially improves the quality of haemorrhagic samples for cytomorphological evaluation; moreover, the samples are well suited for multiple immunocytochemical stainings.

Cutaneous squamous cell carcinoma metastatic to chronic lymphocytic leukaemia: diagnostic potential of fine needle aspiration cytology.

OBJECTIVE: The phenomenon of cancer-to-cancer metastasis of cutaneous squamous cell carcinoma (SCC) and chronic lymphocytic leukaemia (CLL)/small lymphocytic lymphoma (SLL) is a rare event and only occasionally documented in the medical literature. METHODS: Two patients with SCC of the skin that were previously treated for CLL are presented. Both had palpable lymph nodes in the neck and fine needle aspiration cytology (FNAC) was performed to evaluate the pathological process. In addition, the literature on cutaneous SCC metastatic to CLL/SLL with special emphasis on the role of FNAC in this specific clinical situation was reviewed. RESULTS: On examination of the FNAC smear, cancer-to-cancer metastasis of cutaneous SCC to lymph node replaced by CLL was found. In one of the patients, light microscopy examination of the smear was complemented by immunostaining of atypical cells with cytokeratin antibodies and immunophenotyping of lymphoid cells by flow cytometry. In addition to our two patients, nine cases of cutaneous SCC metastatic to CLL/SLL have been reported in the literature, and in only one was the diagnosis made by FNAC. CONCLUSION: FNAC supported by ancillary immunocytological techniques could also be used in diagnosis of specific clinical situations such as cancer-to-cancer metastasis of the tandem of SCC-CLL/SLL.

MIB-1 immunostaining on cytological samples: a protocol without antigen retrieval.

The aim of the study was to evaluate the effect of fixation procedures on MIB-1 immunostaining on microwave-treated Papanicolaou-stained slides and to establish protocol for MIB-1 immunostaining on cytological samples without microwave pre-treatment. Cytospins for immunostaining and nuclear suspension for DNA measurement were prepared from human breast cancer cell line (MCF-7). Following fixation, the cytospins were either stained by Papanicolaou, stored in methanol or air-dried. Antigen retrieval by microwave was used before MIB-1 immunostaining only for Papanicolaou-stained cytospins. Air-dried cytospins and cytospins stored in methanol were immunostained without pre-treatment. The percentage of MIB-1 positive cells was compared with the S phase fraction of MCF-7 cells calculated from DNA histograms. Variations in fixation procedures used before Papanicolaou staining had no influence on the percentage of MIB-1 positive cells. The difference between the percentage of the MIB-1-positive cells on microwave-treated Papanicolaou-stained cytospins and on methanol-fixed cytospins without microwave pre-treatment was not significant. There was a strong correlation between the percentage of the MIB-1-positive cells and S phase fraction. Monoclonal antibody MIB-1 recognized Ki-67 antigen in Papanicolaou-stained cytospins treated by microwave as well as in cytospins that were fixed and stored in methanol without microwave pre-treatment.

Acral myxoinflammatory fibroblastic sarcoma in FNAB samples: can we distinguish it from other myxoid lesions?

We analysed cytomorphological characteristics of three fine needle aspiration biopsy (FNAB) samples of acral myxoinflammatory fibroblastic sarcoma (AMIFS) as well as the features of a number of other benign and malignant myxoid lesions. The analysis showed that FNAB samples from two cases of AMIFS had similar cytomorphology, containing all the characteristic features described in surgical biopsies: myxoid material, spindle cells with bipolar cytoplasmic extensions, epithelioid cells with globules of extra-cellular material, ganglion-like and lipoblast-like giant cells. Only the inflammatory component was scarce. The third sample did not contain features typical of AMIFS. Samples from other myxoid tumours resembled AMIFS to some extent, however, none of them contained all three tumour components characteristic of AMIFS. Cytomorphology of AMIFS may be characteristic enough to enable a definitive diagnosis from FNAB, provided all the distinctive features are sampled.

Hydrolysis profiles of formalin fixed paraffin-embedded tumors based on IOD (integrated optical density) and nuclear texture feature measurements.

The aim of the study was to determine optimal hydrolysis time for the Feulgen DNA staining of archival formalin fixed paraffin-embedded surgical samples, prepared as single cell suspensions for image cytometric measurements. The nuclear texture features along with the IOD (integrated optical density) of the tumor nuclei were analysed by an automated high resolution image cytometer as a function of duration of hydrolysis treatment (in 5 N HCl at room temperature). Tissue blocks of breast carcinoma, ovarian serous carcinoma, ovarian serous tumor of borderline malignancy and leiomyosarcoma were included in the study. IOD hydrolysis profiles showed plateau between 30 and 60 min in the breast carcinoma and leiomyosarcoma, and between 40 and 60 min in the ovarian serous carcinoma and ovarian serous tumor of borderline malignancy. Most of the nuclear texture features remained stable after 20 min of hydrolysis treatment. Our results indicate that the optimal hydrolysis time for IOD and for nuclear texture feature measurements, was between 40 and 60 min in the cell preparations from tissue blocks of three epithelial and one soft tissue tumor.

BMI and Heath-Carter somatotypes of female students in Ljubljana.

Measurements of 22 anthropometric and two spirometric variables were taken from 86 healthy female students of the University of Ljubljana who were not actively engaged in sports. The BMI and Heath-Carter somatotypes were calculated according to the anthropometric method. The average value of FEVC (Forced Expiratory Vital Capacity) amounted to 4.2 liters, and that of FEV 1 (Forced Expiratory Volume 1) to 3.7 liters and they exceeded the European standards for the corresponding age and height. Central and mesomorph-endomorph somatotypes predominated, and the majority of students were classified into the second group of BMI with a normal body weight. The values of FEVC, FEV 1 and the Tiffeneau index increased in line with the ectomorphic component. No differences in the average FEVC and FEV 1 were found between BMI groups. The Tiffeneau index increased from the third to the first group. It was concluded that the ventilation measures were higher in ectomorphic students, who actually exhaled values that exceeded the requirements of standards the least of all groups. Due to rare statistically significant differences in FEVC and FEV 1 between somatotypes and BMI groups it would be necessary to repeat the analysis on a larger sample. There also appeared to be a need for new Slovenian standards.

Construction of anthropometric measuring chair (AMC) for lung function prediction.

An anthropometric measuring chair (AMC) was constructed in order to predict lung function parameters (vital capacity--FVC and forced expiratory flow in the first second-FEV1) in normal Caucasian subjects. Design of a chair was aimed toward the fast and reliable recording of body dimensions, particularly of the human thorax. Static and dynamic measurements of thorax dimensions, arm span and sitting height were used to predict FVC and FEV1 with an accuracy better than standard prediction equations based only on body height and age. Computer program for automatic calculation of FVC and FEV1 according to prediction equations was constructed in VisualBasic 4.0 software. The prediction values for FVC and FEV1 based on anthropometric variables could be used in paraplegic, disproportional and handicapped individuals, where standard body height is unobtainable.

The effect of primary fixation with standard postfixation and the duration of hydrolysis on nuclear features in image cytometry.

The effect of three primary fixation procedures, used in the preparation of routine cytological samples: air-drying, Delaunay, and Saccomanno fixation, with postfixation in modified Böhm-Sprenger fixative, on nuclear features as a function of hydrolysis time is reported. Three different cell types: lymphatic cells (tonsil), epithelial cells (buccal mucosa) and mesenchymal cells (uterine myometrium) were used for the study. Our findings show, that generally not all features have the same plateau times as the IOD (integrated optical density), and that many features show different values depending on cell type and fixation method. It is therefore recommended that for any primary fixative used in routine clinical work and for each cell type, the hydrolysis curve for all nuclear features to be used in sample analysis should be established.