We demonstrate the capabilities of the coherent anti-Stokes Raman scattering (CARS) microscope and its multimodal operation to image a histological section of human intestinal tissue. The imaging of unstained and stained sections using various nonlinear optical contrasts was performed. The CARS configuration of our microscope allows probing which does not require a preliminary staining of tissue saving the treatment time and providing label-free investigation of original matter. Particular attention was paid to visualisation of unstained tissue. CARS images were recorded in the high wave number Raman spectroscopy region and the spectra of the most distinguished features of images are provided and discussed. Additionally, the two photon excitation fluorescence (TPEF) and second harmonic generation (SHG) contrast mechanisms were used for structural visualisation of both unstained and stained sections of human intestinal tissue. Visualisation of a histological section using all contrast mechanisms mentioned above is analysed and discussed. The research is aimed to draw attention to a potential of CARS/nonlinear microscopy in routine healthcare.
Wide-field Polarization-resolved Second-Harmonic Generation microscopy is a label-free imaging technique which highlights molecular organization of collagenous tissues, enabling high-throughput quantitative biomedical imaging and cancer diagnostics.
Intralobarinė sekvestracija – plaucio vystymosi anomalija,pasireiskianti nefunkcionuojancios parenchimosplotu, turinciu anomalią kraujotaką. Klinikiniai simptomai yra nespecifiniai ir neretai si anomalija gali imituoti bronchektazes. Tyrimo tikslas: įvertintiintralobarinės plaucių sekvestracijos diagnostikos ir gydymo rezultatus VU MF Krūtinės chirurgijos centre. Tyrimo metodai ir tiriamieji. Į tyrimą įtraukti 44 asmenys, operuoti dėl bronchektazėms būdingų simptomų (is jų 10-ciai diagnozuota intralobarinė sekvestracija). Tyrimo rezultatai: pagrindiniai ligoniųsu intralobarine sekvestracija simptomai buvo kosulys (80%), skrepliavimas (30%), karsciavimas (30%). Diagnostikai naudoti tyrimai: krūtinės ląstosrentgenograma (100%), kompiuterinė tomografija(100%), aortografija (30%), kompiuterinės tomografijosangiografija (20%), fibrobronchoskopija (80%). Klaidinga pradinė diagnozė nustatyta pusei ligonių. Gydymas – chirurginis. Visiems ligoniams pasalinta plaucio apatinė skiltis (7 - kairiojo, 3 – desiniojo).Isvados: intralobarinė plaucių sekvestracija dažnai sukelia simptomus, identiskus sergant bronchektazėmis.Tiriant tokius ligonius būtina nustatyti,ar nėra anomalinių arterijų, patenkancių į vieną ar kitą plaucių dalį. Kompiuterinė angiografija įgalinatiksliai diagnozuoti anomalines plaucių arterijas. Jas (ją) nustacius pries operaciją, ligonis apsaugomasnuo galimo grėsmingo kraujavimo operuojant.
Silicosis remains a common occupational respiratory disease. Even in this era of highly sophisticated hygiene in European countries, new occupational cases of silicosis continue to be reported. Four cases of silicosis which developed after a relatively short occupational exposure to respirable silica among the members of one family are described. Four young men worked illegally abroad in mining in one of European countries. All of them were employed together in the same working conditions. One of the brothers died due to the acute form of the disease (lipoproteinosis). Two of the brothers suffered from simple nodular silicosis, and the fourth brother developed very early nodular silicosis and small airway dust disease. A one year follow-up revealed moderate/severe worsening of the disease in all surviving brothers.
Extracellular matrix (ECM) has important functions in cell proliferation, differentiation, and migration, which influence the development and progression of cancer. ECM in tumor microenvironment experiences changes in composition and structure that can appear early in tumor development and could serve as a biomarker for cancer diagnostics. In addition, some changes in ECM may correlate with the rate of tumor progression or its tendency to form metastases and would allow to predict future tumor development [1]. Collagen is an important structural protein found in ECM. It has a non-centrosymmetric structure, and, thus, can be easily visualized using second harmonic generation (SHG) microscopy. SHG microscopy employs certain polarimetric techniques to gain detailed information about the organization of collagen in various tissues [2]. In this work, polarimetric SHG microscopy is used to acquire collagen images from normal and cancerous regions of human colon and pancreas histological samples. Texture analysis is performed on SHG intensity and polarization images to characterize the distribution of ultrastructure parameters in the tissue. Significant differences are observed in collagen ultrastructure between normal and tumor areas. Further, collagen structures of colon and pancreas tumor microenvironments are compared to investigate relative differences in ECM organization between the tissues. Finally, a machine learning classifier is used to group the acquired images in tumor and normal groups. The results show potential for development of novel cancer diagnostic technique using polarimetric second harmonic generation microscopy and texture analysis. [1] Winkler, J. et al., "Concepts of extracellular matrix remodelling in tumour progression and metastasis", Nat Commun 11, 5120 (2020). [2] Golaraei, A. et al., "Polarimetric second-harmonic generation microscopy of the hierarchical structure of collagen in stage I-III non-small cell lung carcinoma," Biomed. Opt. Express 11, 1851-1863 (2020).
The mechanisms driving the transition from inflammation to fibrosis in sarcoidosis patients are poorly understood; prognostic features are lacking. Immune cell profiling may provide insights into pathogenesis and prognostic factors of the disease. This study aimed to establish associations in simultaneous of lymphocyte subset profiles in the blood, bronchoalveolar lavage fluid (BALF), and lung biopsy tissue in the patients with newly diagnosed sarcoidosis.A total of 71 sarcoid patients (SPs) and 20 healthy controls (HCs) were enrolled into the study. CD31, CD38, CD44, CD103 positive T lymphocytes in blood and BALF were analysed. Additionally, the densities of CD4, CD8, CD38, CD44, CD103 positive cells in lung tissue biopsies were estimated by digital image analysis.Main findings: (I) increase of percentage of CD3+CD4+CD38+ in BALF and blood, and increase of percentage of CD3+CD4+CD44+ in BALF in Löfgren syndrome patients comparing with patients without Löfgren syndrome, (II) increase of percentage of CD3+CD4+103+ in BALF and in blood in patients without Löfgren syndrome (comparing with Löfgren syndrome patients) and increase of percentage of CD3+CD4+103+ in BALF and in blood in more advanced sarcoidosis stage. (III) Increasing percentage of BALF CD3+CD4+CD31+ in sarcoidosis patients when comparing with controls independently of presence of Löfgren syndrome, smoking status or stage of sarcoidosis. Several significant correlations were found.Lymphocyte subpopulations in blood, BALF, and lung tissue were substantially different in SPs at the time of diagnosis compared to HCs. CD3+CD4+CD31+ in BALF might be a potential supporting marker for the diagnosis of sarcoidosis. CD3+CD4+CD38+ in BALF and blood and CD3+CD4+CD44+ in BALF may be markers of the acute immune response in sarcoidosis patients. CD4+CD103+ T-cells in BALF and in blood are markers of the persistent immune response in sarcoidosis patients and are potential prognostic features of the chronic course of this disease.
The distribution of nanoparticles (NP) in an organism is an important issue for developing NP-based drug delivery systems and for general nanotoxicology. The knowledge of NP localisation in the skin is crucial for the optimisation of NP behaviour in vivo. Therefore, we have used semiconductor quantum dots (QD) to investigate their biodistribution in the skin by means of confocal fluorescence microscopy after subcutaneous injection. The results obtained showed that the diffusion of QD in the dermis is limited by basement membrane and dense connective tissue fibres, which resulted in negligible QD penetration into the epidermis, hair follicles, sebaceous and sweat glands, nerves and blood vessels. Low permeation of QD through the tissues results in slow clearance and raises the risks of potential immune, inflammatory and cytotoxic responses. The study reveals the significance of the tissue architecture for the interstitial and intracellular migration patterns of non-functionalised QD.
Multicontrast nonlinear microscopy with SHG and THG were used to image normal and cancerous human colon histology samples, and texture analysis was applied to investigate the changes in collagen structure occurring during carcinogenesis.
Abstract Chronic eosinophilic pneumonia is a rare interstitial lung disorder, which causes diagnostic difficulties. Often the disease is diagnosed correctly after several weeks or months following initial presentation. The aim of the study was to prospectively evaluate peculiarities of manifestation of idiopathic chronic eosinophilic pneumonia (ICEP), which may allow to improving early diagnosis. Twenty patients with ICEP were involved in this investigation. The cases of acute eosinophilic pneumonia and cases of chronic eosinophilic pneumonia of known origin were excluded. To define archetypal signs of the idiopathic chronic eosinophilic pneumonia, 3 comparable groups were selected. They were the group of 50 patients with community-acquired pneumonia (COP); the group of 21 asthmatic patients with COP, and the cluster of 10 patients with morphologically confirmed cryptogenic organizing pneumonia (OP). Clinical and radiological manifestation of ICEP was similar to COP and cryptogenic OP manifestation. We have found that chest pain; fine rales and pleurisy were unrepresentative for ICEP. However, blood eosinophilia was typical sign of ICEP and wheezing was a frequent observation. Usually ICEP patients had relative mild clinical symptoms and moderate increased C reactive protein (CRP) level even in cases of multiple pulmonary infiltrates. In conclusion, in cases of not typical pneumonia course, i.e. non-resolving or recurrent pulmonary infiltrates; relative mild clinical symptoms and moderate increased CRP level with multiple pulmonary infiltrates; blood eosinophilia and/or signs of airway obstruction eosinophilic pneumonia should be suspected and bronchoalveolar lavage and/or bronchoscopic lung biopsy performed.
Abstract PurposeMyocardial fibrosis in aortic stenosis (AS) is associated with worse survival following aortic valve replacement (AVR). We assessed myocardial fibrosis in severe AS patients, integrating echocardiographic, cardiovascular magnetic resonance (CMR) and histological data. MethodsA total of 83 severe AS patients (age 66.4 ± 8.3, 42% male) who were scheduled for surgical AVR underwent CMR with late gadolinium enhancement (LGE) and T1 mapping and global longitudinal strain (GLS) analysis. Collagen volume fraction (CVF) was measured in myocardial biopsies (71) that were sampled at the time of AVR. ResultsCVF correlated with imaging and serum biomarkers of LV systolic dysfunction and left side chamber enlargement and was higher in the sub-endocardium compared with midmyocardium (p<0.001). CVF median values were higher in LGE-positive versus LGE-negative patients [28.7% (19-33) vs 20.7% (15-30), respectively, p=0.040]. GLS was associated with invasively (CVF; r=-0.303, p=0.013) and non-invasively (native T1; r=-0.321, p<0.05) measured myocardial fibrosis. GLS and native T1 correlated with parameters of adverse LV remodelling, systolic and diastolic dysfunction and serum biomarkers of heart failure and myocardial injury. ConclusionOur data highlight the role of myocardial fibrosis in adverse cardiac remodelling in AS. GLS has potential as a surrogate marker of myocardial fibrosis, and high native T1 and low GLS values differentiated patients with more advanced cardiac remodelling.
