Marjeta Tercelj

In vivo assessment and evaluation of lung tissue morphologic and physiological changes from non-contact endoscopic reflectance spectroscopy for improving lung cancer detection

We present a method for lung cancer detection exploiting reflectance spectra measured in vivo during endoscopic imaging of the lung. The measured reflectance spectra were analyzed using a specially developed light-transport model to obtain quantitative information about cancer-related, physiological, and morphologic changes in the superficial bronchial mucosa layers. The light-transport model allowed us to obtain the absorption coefficient (mua) and further to derive the micro-vascular blood volume fraction in tissue and the tissue blood oxygen saturation. The model also allowed us to obtain the scattering coefficient (mus) and the anisotropy coefficient (g) and further to derive the tissue scattering micro-particle volume fraction and size distribution. The specular component of the reflectance signal and the instrument response were accounted for during the analysis. The method was validated using 100 reflectance spectra measured in vivo in a noncontact fashion from 22 lung patients (50 normal tissue/benign lesion sites and 50 malignant lesion sites). The classification between normal tissue/benign lesions and malignant lesions was further investigated using the derived quantitative parameters and discriminant function analysis. The results demonstrated significant differences between the normal tissue/benign lesions and the malignant lesions in terms of tissue blood volume fraction, blood oxygen saturation, tissue scatterer volume fractions, and size distribution. The results also showed that the malignant lung lesions can be differentiated from normal tissue/benign lesions with both diagnostic sensitivity and specificity of better than 80%.

Fungal exposure in homes of patients with sarcoidosis - an environmental exposure study

BackgroundThere is increasing evidence that exposure to moulds (fungi) may influence the development of sarcoidosis. To assess the influence of the environmental exposure, a study was undertaken to determine the exposure to fungi in homes of subjects with sarcoidosis.MethodsSubjects were patients with clinically established sarcoidosis recruited during the period September 2007 till June 2010. Of these 55 were newly diagnosed and currently under treatment for less than one year, 25 had been treated and had no recurrence and 27 had been treated but had recurrence of the disease. Controls were healthy subjects without any respiratory symptoms (n = 30). Samples of air (about 2.5 m3) were taken in the bedroom of the subjects using a portable pump and cellulose ester filters. The filters were analysed for the content of the enzyme N-acetylhexosaminidase (NAHA) as a marker of fungal cell biomass, using a specific substrate and a fluorescent technique and expressed as NAHA units (U)/m3.ResultsCompared to controls, subjects undergoing treatment of the disease (newly diagnosed or with recurrence) had significantly higher activities of NAHA in their homes than controls (33.6 and 39.9 vs 10.0 U/m3, p < 0.001 and <0.001). Among controls only 5 out of 30 subjects had levels of NAHA above the second quartile value (14 U/m3). In homes of subjects with newly diagnosed disease with treatment less than one year, values above 14 NAHA U/m3 were found among 35 out of 55 and among those with recurrent disease among 18 out of 27.ConclusionsThe higher activities of NAHA enzyme found in homes of subjects with active and recurrent sarcoidosis suggest that exposure to fungi is related to the risk of sarcoidosis. Further environmental studies to assess the importance of this exposure for subjects with sarcoidosis are warranted. The results suggest that remedial actions in homes with high levels of fungi may be justified.

Chitotriosidase activity in sarcoidosis and some other pulmonary diseases

Abstract Background: Patients with sarcoidosis have elevated levels of several markers of inflammation. Particularly high levels have been reported for chitotriosidase. In this study, we evaluate whether determining chitotriosidase in serum would be useful in the diagnosis and clinical management of patients with sarcoidosis. Methods: Patients with newly diagnosed sarcoidosis and patients with asthma, fibrosis, asbestosis, lung cancer or chronic obstructive pulmonary disease (n=190) were recruited from an outpatient department. Individuals with no disease (n=26) served as controls. An X-ray was taken, diffusion capacity was measured and blood samples were taken for analysis of chitotriosidase, soluble receptor for interleukin-2, tumour necrosis factor alpha and angiotensin converting enzyme. In most patients with sarcoidosis, the analyses were done before and after regular treatment with corticosteroids over 6 months. Results: Some patients with sarcoidosis had markedly high activities of chitotriosidase, but activities above controls were also found among patients with asbestos, fibrosis and lung cancer. There were significant relationships between chitotriosidase and interleukin-2 receptor and angiotensin-converting enzyme. After treatment, chitotriosidase activity decreased in 52 of 69 patients. Conclusions: The results confirm that chitotriosidase activity is markedly increased in some cases of sarcoidosis. As increased activities are also found in other diseases, chitotriosidase cannot be considered a specific marker of sarcoidosis. In cases of sarcoidosis where high CTO activities are found, this enzyme could serve as a useful marker supporting the diagnosis of sarcoidosis when following the effects of treatment and in surveillance for recurrence of the disease.

Antifungal medication is efficient in the treatment of sarcoidosis.

Objectives: Fungi have been suspected of contributing to the pathogenesis of sarcoidosis. A previous intervention study demonstrated an improvement in the clinical condition in 15 out of 18 patients with a long-term history of sarcoidosis when antifungal medication was added to corticosteroids. The present study was performed to compare the effects of antifungal treatment with corticosteroid treatment in sarcoidosis. Methods: Patients with newly diagnosed sarcoidosis were recruited. Corticosteroids were given to 39 subjects, corticosteroid + antifungal to 31, and antifungal only to 22 subjects. The effects of the treatments were evaluated at 6 months. X-ray scores were measured before and after treatment together with pulmonary diffusion capacity and two markers of sarcoidosis activity, that is, angiotensin-converting enzyme in serum (sACE) and chitotriosidase (CTO). Results: X-ray scores as well as sACE and CTO decreased significantly in all groups. The X-ray score decreased slightly more among subjects in the groups that received antifungal medication compared with corticosteroids only (p < 0.001). Conclusion: The results suggest that antifungal treatment is as efficient as corticosteroid treatment against the granulomatous and inflammatory manifestations of sarcoidosis. This is probably because this treatment is directed towards the causative agent. Additional studies are required to define the phenotype, where the antifungal treatment was not efficient (4/22) and to perform long‐term follow up to determine the risk of recurrence.

Inflammatory markers and pulmonary granuloma infiltration in sarcoidosis.

Previous studies have demonstrated increases of inflammatory mediators in sarcoidosis while epidemiological studies have also demonstrated an association with increased fungi exposure. This study measured the level of β‐glucan in the lungs and of inflammatory mediators in serum, and correlated both with the extent of pulmonary granuloma infiltration.

Fungal cell wall agents suppress the innate inflammatory cytokine responses of human peripheral blood mononuclear cells challenged with lipopolysaccharide in vitro

Exposure to high levels of fungi might lead to diseases, such as airway inflammation, hypersensitivity pneumonitis and allergy. To comprehend the mechanisms behind the exposure to fungi and a disease, we examined the in vitro innate inflammatory cytokine response of human peripheral blood mononuclear cells (PBMC) challenged by fungal cell wall agents (FCWAs), i.e., soluble and particulate (1→3)-β-D-glucan-curdlan (BGS and BGP), zymosan (ZYM) and chitosan (CHT) in the absence or presence of lipopolysaccharide (LPS). We also studied FCWA effects on the mRNA expression of dectin-1, TLR2, TLR4 and mannose receptor (MR) by real-time RT-PCR. Our results demonstrated that BGP strongly induced the secretion of TNF-α, IL-6, IL-10 and IL-12; BGS, ZYM and CHT were weaker, but still significant cytokine inducers. We showed that BGS significantly augmented the LPS-induced in vitro secretion of TNF-α. On the other hand, BGP, ZYM and CHT suppressed the LPS-induced production of all cytokines. At the mRNA level, the dectin-1, TLR2 and TLR4 expressions were significantly reduced by all FCWAs in the absence of LPS and even more in the presence of LPS. While we demonstrated that the innate inflammatory cytokine response of PBMC induced by CHT was mediated by MR, the MR mRNA expression was significantly reduced by CHT. On the contrary, BGS significantly enhanced the MR mRNA expression. In conclusion, a long-term and massive exposure to LPS and FCWA (e.g., organic dust) may cause an important disruption of normal immune response and allow development and/or persistence of various immunopathological events.

Nocturnal Asthma and Domestic Exposure to Fungi

Fungal growth indoors is a common problem both in older and modern buildings. Numerous studies demonstrate a deterioration of asthma in environments with fungal growth but precise estimates of exposure levels have not been reported. This study was carried out to investigate whether levels of airborne fungi were higher in homes of subjects with nocturnal asthma as compared to controls. Subjects with nocturnal asthma (n = 42) and non-nocturnal asthma (n = 14) were recruited from an outpatient department. Subjects without disease or respiratory symptoms (n = 24) served as controls. Among the asthmatics, determinations were made of pulmonary function before and after treatment with bronchodilatators. The subjects were equipped with a pump and a filter and instructed to sample air in their bedroom. The filters were analysed for the content of N-acetylhexosaminidase (NAHA) as a marker of fungal cell biomass. In some houses the number of colony forming units was also determined. Patients with asthma had significantly higher activities of NAHA in their bedrooms as compared to controls (31.2 vs. 12.5 U m−3, p = 0.004). When the material was divided according to the 2nd quartile of NAHA (14.0 U m−3), 29 out of 42 subjects with nocturnal asthma had high values compared to 5 out of 24 for the controls (p<0.0001). The results support the hypothesis that patients with nocturnal asthma have high levels of fungi in their homes. This provides a basis for treatment and exposure diminution. Whether that exposure is causative for their disease; or if it aggravates already existing airway symptoms remains to be elucidated.

The effect of age on the grip force control in lateral grip

In the paper we present the grip force tracking system for the evaluation of grip force control. We developed a grip measuring device which can be used for the computer assisted measurements of the grip force in real time. The device was used as an input to a force-tracking task where the subject applied the grip force according to the visual feedback from the computer screen. The performance of the task was evaluated by the tracking error between the target signal and the measured force. We assessed the grip force control in the groups of 10-year old children, 25- to 35-year old adults and 50- to 60-year old adults. The subjects performed a sinus tracking task which required periodic muscle activation to produce the desired output. The results of the average tracking errors show significant differences in grip force control among the three tested groups. The largest variability among subjects was observed in the group of children and older adults. No significant difference in force control was found between the dominant and non-dominant hand. The grip force tracking system presented is aimed to be used for the evaluation of grip force control in patients with different sensory-motor impairments.

Combining field imaging endoscopy with point analysis spectroscopy for improving early lung cancer detection

We propose to combine field imaging endoscopy with point spectral analysis for improving the overall diagnostic accuracy in clinical lung cancer detection. For this purpose, we developed an integrated endoscopy system that uses autofluorescence imaging and white light reflectance imaging to obtain high diagnostic sensitivity, while at the same time uses non-contact point reflectance/fluorescence spectroscopy to reduce false positive biopsies, thus, achieve high diagnostic specificity. A pilot clinical test on 22 lung patients demonstrated that using this system the malignant lung lesions can be differentiated from the benign lesions with both diagnostic sensitivity and specificity of better than 80%. To further reduce the number of false positive diagnosis and allow even higher diagnostic accuracies, we have also developed an endoscopic laser Raman probe for in vivo real-time biochemical analysis of the suspicious tissue areas identified by the field imaging modalities (white light imaging and autofluorescence imaging). Preliminary Raman spectroscopy results will be reported at the conference.

Fungal exposure and low levels of IL-10 in patients with sarcoidosis.

Background and Objectives. Sarcoidosis is an inflammatory disease with increased levels of inflammatory cytokines. Previous studies have shown a relation between the degree of granuloma infiltration and serum cytokine levels, except for interleukin- (IL-) 10. The aim of the study was to further investigate the serum levels of IL-10 in patients with sarcoidosis and relate them to fungal exposure in terms of the amount of fungi in the air of their homes and β-glucan in bronchoalveolar lavage (BAL) fluid. Methods. Patients with sarcoidosis (n = 71) and healthy controls (n = 27) were enrolled. IL-10 was determined in serum. BAL was performed and the amount of β-glucan was measured. Domestic exposure to fungi was determined by measuring airborne β-N-acetylhexosaminidase (NAHA) in the bedrooms. Results. At high levels of fungal exposure (domestic fungal exposure and β-glucan in BAL), serum IL-10 values were lower than at low and intermediate exposure levels. Conclusion. The low serum IL-10 values at high fungal exposure suggest that fungal cell wall agents play a role in granuloma formation in sarcoidosis by inhibiting the secretion of the anti-inflammatory cytokine IL-10.