Cytometria przepływowa w diagnostyce różnicowej choroby Hashimoto i chłoniaka MALT tarczycy

 Introduction: A combination of traditional cytology methods with fluorescence activated cell sorting (FACS) analysis of fine-needle aspiration biopsy (FNAB) material is considered a powerful diagnostic tool in the differential diagnosis of thyroid lesions suspected of mucosa-associated lymphoid tissue lymphoma (MALT-L). The aim of this study was to demonstrate the FACS-based diagnostic process of thyroid lesions in a clinical situation where ultrasound and cytological examinations did not allow differentiation between Hashimoto’s thyroiditis (HT) and MALT-L. Material and methods: The patients analysed in this study presented significantly different clinical courses of thyroid disease: quickly enlarging painless tumour of the thyroid right lobe in the first case, and chronic HT with palpable tumour in the thyroid isthmus in the second patient. Due to the suspicion of MALT-L resulting from indeterminate ultrasound and FNAB-cytology results, FNAB material was obtained from all the previously examined thyroid lesions and directly subjected to FACS assessment, encompassing κ/λ light chain restriction analysis, as well as measurements of B and T cell surface antigens. Results: The FACS analysis of FNAB material obtained from our patients did not show any definite signs of light chain restriction. Although one of the samples showed a borderline value of κ/λ ratio (κ/λ = 0.31), further immunophenotyping confirmed clonal expansion in none of the examined thyroid regions. Histopathological findings documented the diagnosis of HT in both clinical cases. Conclusion: We believe that FACS represents a useful and reliable complementary diagnostic measure in FNAB-based differential diagnosis of lymphoproliferative thyroid disorders. Wstęp: Skojarzenie oceny cytologicznej oraz cytometrii przepływowej (FACS, fluorescence activated cell sorting) materiału uzyskanego podczas biopsji aspiracyjnej cienkoigłowej (BAC) jest uważane za niezwykle skuteczną metodę w diagnostyce różnicowej zmian tarczycy podejrzanych o obecność pozawęzłowego chłoniaka strefy brzeżnej systemu MALT (MALT-L). Celem pracy było zaprezentowanie opartego na FACS procesu diagnostycznego zmian ogniskowych tarczycy u chorych, u których badanie ultrasonograficzne i cytologiczne nie umożliwiło zróżnicowania przewlekłego zapalenia tarczycy (HT, Hashimoto's thyroiditis) od MALT-L. Materiał i metody: Chorzy opisani w pracy charakteryzowali się całkowicie odmiennym przebiegiem klinicznym choroby tarczycy — ujawniającej się w pierwszym przypadku jako szybko powiększający się guz płata prawego, w drugim jako zmiana ogniskowa w cieśni u chorej z przewlekłym wywiadem HT. Ze względu na podejrzenie MALT-L postawione na podstawie badania cytologicznego oraz podejrzany wzorzec ultrasonograficzny, przeprowadzono ponownie BAC wszystkich wcześniej ocenianych zmian tarczycy, a uzyskany materiał poddano bezpośrednio ocenie za pomocą FACS, obejmującej analizę restrykcji łańcuchów lekkich immunoglobulin κ/λ oraz antygenów powierzchniowych limfocytów T i B. Wyniki: Analiza FACS materiału uzyskanego za pomocą BAC nie ujawniła definitywnych cech restrykcji łańcuchów lekkich. Pomimo granicznych wartości współczynnika κ/λ (κ/λ = 0,31) w jednej z próbek, dalsza analiza fenotypowa nie potwierdziła klonalnej ekspansji w żadnym z badanych obszarów tarczycy. Wyniki histopatologiczne potwierdziły diagnozę przewlekłego zapalenia tarczycy w obu przypadkach klinicznych. Wnioski: Cytometria przepływowa jest badaniem wiarygodnie uzupełniającym ocenę cytologiczną w diagnostyce różnicowej przewlekłego zapalenia tarczycy i limfoproliferacyjnych chorób tarczycy

Patogeneza stwardnienia rozsianego

Stwardnienie rozsiane (SM, sclerosis multiplex) jest postępującym schorzeniem ośrodkowego układu nerwowego (OUN). Mechanizmy leżące u podstaw rozsianych zmian demielinizacyjnych charakterystycznych dla tej choroby nie zostały jak dotąd w pełni poznane. Uważa się, że w podatności na występienie SM duże znaczenie ma skomplikowane podłoże genetyczne, najprawdopodobniej różniące się w poszczególnych populacjach. Za jeden z kluczowych składników procesu patologicznego uznaje się powszechnie układ immunologiczny, a zwłaszcza takie jego składowe, jak autoreaktywne limfocyty T, komórki o charakterze regulatorowym oraz cytokiny i chemokiny prozapalne. Rozważa się także zaangażowanie czynników środowiskowych w tym wirusów i bakterii, jednak dotąd nie udało się zidentyfikować patogenu związanego bezspornie z występowaniem choroby. Poznanie złożonych mechanizmów chorobowych w SM wymaga dalszych szczegółowych badań, jednak uzyskana dotychczas wiedza pozwala na znacznie lepsze zrozumienie patogenezy tego schorzenia

Renin-angiotensin system – new link between brain and thyroid?

Renin-angiotensin system (RAS) belongs to the most effective regulatory circuits in human biology. Research-data generated in the last years considerably improved the understanding of RAS structure and function. Alternative enzymatic pathways have been described as well as new angiotensin-metabolites reacting with their specific receptors. An existence of local – tissue and organ specific RAS has been suggested in opposition to the classical hormonal system. This review has been focused on the local RAS of the central nervous system and its role in the development of cognitive dysfunction and in the pathogenesis of dementia. A potential interaction of local RAS and thyroid hormones was also discussed.Układ renina-angiotensyna (ang. renin-angiotensin system, RAS) jest jednym z najbardziej efektywnych systemów regulacyjnych ludzkiego organizmu. Badania ostatnich lat znacznie poszerzyły wiedzę na temat struktury i funkcji RAS. Opisane zostały alternatywne szlaki enzymatyczne i nowe aktywne metabolity angiotensyny oraz specyficzne dla nich receptory. Powstała również koncepcja swoistych dla poszczególnych tkanek i narządów lokalnych układów RAS. Niniejsza praca skoncentrowana została na przedstawieniu lokalnego układu RAS ośrodkowego układu nerwowego i jego roli w rozwoju zaburzeń poznawczych oraz patogenezie demencji. Przedyskutowano również potencjalne znaczenie interakcji lokalnych układów RAS z hormonami tarczycy

Optical Coherence Tomography in the Differential Diagnosis of Patients with Multiple Sclerosis and Patients with MRI Nonspecific White Matter Lesions

In the differential diagnosis of nonspecific white matter lesions (NSWMLs) detected on magnetic resonance imaging (MRI), multiple sclerosis (MS) should be taken into consideration. Optical coherence tomography (OCT) is a promising tool applied in the differential diagnostic process of MS. We tested whether OCT may be useful in distinguishing between MS and NSWMLs patients. In patients with MS (n = 41) and NSWMLs (n = 19), the following OCT parameters were measured: thickness of the peripapillary Retinal Nerve Fibre Layer (pRNFL) in superior, inferior, nasal, and temporal segments; thickness of the ganglion cell-inner plexiform layer (GCIPL); thickness of macular RNFL (mRNFL); and macular volume (MV). In MS patients, GCIPL was significantly lower than in NSWMLs patients (p = 0.024). Additionally, in MS patients, mRNFL was significantly lower than in NSWMLs patients (p = 0.030). The average segmental pRNFL and MV did not differ between MS and NSWMLs patients (p > 0.05). GCIPL and macular RNFL thinning significantly influenced the risk of MS (18.6% [95% CI 2.7%, 25.3%]; 27.4% [95% CI 4.5%, 62.3%]), and reduced GCIPL thickness appeared to be the best predictor of MS. We conclude that OCT may be helpful in the differential diagnosis of MS and NSWMLs patients in real-world settings

A Loop That Matters—An Unusual Case of Bow Hunter’s Syndrome

Bow Hunter’s syndrome (BHS), also known as rotational vertebral artery occlusion (VAO), is a rare entity in which vertebral artery is reversibly compressed due to rotation or extension of the head, causing vertebrobasilar insufficiency. Because of VAO, BHS should be considered as a possible life-threatening condition. Diverse aetiologies of BHS may trigger a broad spectrum of non-specific symptoms and may result in frequent misdiagnosis of this disorder in daily clinical practice. Herein, we present a case of BHS caused by previously non-described vascular aetiology

The Role of Optical Coherence Tomography in Differential Diagnosis of Multiple Sclerosis and Autoimmune Connective Tissue Diseases with CNS Involvement

The purpose of this study was to examine whether application of optical coherence tomography (OCT) measurements can provide a useful biomarker for distinguishing central nervous system (CNS) involvement in autoimmune connective tissue diseases (CTD) from multiple sclerosis (MS). An observational study included non-optic neuritis eyes of 121 individuals: 59 patients with MS, 30 patients with CNS involvement in CTD, and 32 healthy controls. OCT examination was performed in all subjects to measure retinal nerve fiber layer (RNFL) thickness, ganglion cell complex (GCC) thickness, ganglion cell layer-inner plexiform layer (GCIPL) thickness, and volume of the macula. There was a significant group effect with regard to superior optic disc RNFL, macular RNFL, GCC, and GCIPL thickness, and macular volume. Post-hoc analysis revealed that MS patients have significantly smaller macular volume and thinner superior optic disc RNFL, macular RNFL, GCC, and GCIPL compared to healthy controls. CTD patients have significantly smaller superior optic disc RNFL, GCIPL, and GCC thickness compared to healthy controls. However, no significant group differences were observed between the patient groups (MS vs. CTD) on any outcome. Although a prominent retinal thinning may be a useful biomarker in MS patients, in a general population of individuals with a confirmed CNS involvement the use of OCT is not specific enough to discriminate between MS and autoimmune CTD

Cerebrospinal Fluid Biomarkers in Differential Diagnosis of Multiple Sclerosis and Systemic Inflammatory Diseases with Central Nervous System Involvement

Background: Diagnosis of multiple sclerosis (MS) is established on criteria according to clinical and radiological manifestation. Cerebrospinal fluid (CSF) analysis is an important part of differential diagnosis of MS and other inflammatory processes in the central nervous system (CNS). Methods: In total, 242 CSF samples were collected from patients undergoing differential MS diagnosis because of the presence of T2-hyperintensive lesions on brain MRI. The non-MS patients were subdivided into systemic inflammatory diseases with CNS involvement (SID) or cerebrovascular diseases (CVD) or other non-inflammatory diseases (NID). All samples were analyzed for the presence of oligoclonal bands and ELISA was performed for detection of: INF gamma, IL-6, neurofilaments light chain (NF-L), GFAP, CHI3L1, CXCL13, and osteopontin. Results: The level of IL-6 (p = 0.024), osteopontin (p = 0.0002), and NF-L (p = 0.002) was significantly different among groups. IL-6 (p = 0.0350) and NF-L (p = 0.0015) level was significantly higher in SID compared to NID patients. A significantly higher level of osteopontin (p = 0.00026) and NF-L (p = 0.002) in MS compared to NID population was noted. ROC analysis found weak diagnostic power for osteopontin and NFL-L. Conclusions: The classical and non-standard markers of inflammatory process and neurodegeneration do not allow for sufficient differentiation between MS and non-MS inflammatory CNS disorders. Weak diagnostic power observed for the osteopontin and NF-L needs to be further investigated

The Molecular Effect of Diagnostic Absorbed Doses from 131I on Papillary Thyroid Cancer Cells In Vitro

Diagnostic whole-body scan is a standard procedure in patients with thyroid cancer prior to the application of a therapeutic dose of 131I. Unfortunately, administration of the radioisotope in a diagnostic dose may decrease further radioiodine uptake—the phenomenon called “thyroid stunning”. We estimated radiation absorbed dose-dependent changes in genetic material, in particular in the sodium iodide symporter (NIS) gene promoter, and the NIS protein level in a K1 cell line derived from the metastasis of a human papillary thyroid carcinoma exposed to 131I in culture. The different activities applied were calculated to result in absorbed doses of 5, 10 and 20 Gy. Radioiodine did not affect the expression of the NIS gene at the mRNA level, however, we observed significant changes in the NIS protein level in K1 cells. The decrease of the NIS protein level observed in the cells subjected to the lowest absorbed dose was paralleled by a significant increase in 8-oxo-dG concentrations (p < 0.01) and followed by late activation of the DNA repair pathways. Our findings suggest that the impact of 131I radiation on thyroid cells, in the range compared to doses absorbed during diagnostic procedures, is not linear and depends on various factors including the cellular components of thyroid pathology

Naturally Occurring Nervonic Acid Ester Improves Myelin Synthesis by Human Oligodendrocytes

The dysfunction of oligodendrocytes (OLs) is regarded as one of the major causes of inefficient remyelination in multiple sclerosis, resulting gradually in disease progression. Oligodendrocytes are derived from oligodendrocyte progenitor cells (OPCs), which populate the adult central nervous system, but their physiological capability to myelin synthesis is limited. The low intake of essential lipids for sphingomyelin synthesis in the human diet may account for increased demyelination and the reduced efficiency of the remyelination process. In our study on lipid profiling in an experimental autoimmune encephalomyelitis brain, we revealed that during acute inflammation, nervonic acid synthesis is silenced, which is the effect of shifting the lipid metabolism pathway of common substrates into proinflammatory arachidonic acid production. In the experiments on the human model of maturating oligodendrocyte precursor cells (hOPCs) in vitro, we demonstrated that fish oil mixture (FOM) affected the function of hOPCs, resulting in the improved synthesis of myelin basic protein, myelin oligodendrocyte glycoprotein, and proteolipid protein, as well as sphingomyelin. Additionally, FOM reduces proinflammatory cytokines and chemokines, and enhances fibroblast growth factor 2 (FGF2) and vascular endothelial growth factor (VEGF) synthesis by hOPCs was also demonstrated. Based on these observations, we propose that the intake of FOM rich in the nervonic acid ester may improve OL function, affecting OPC maturation and limiting inflammation

Distribution of subpopulations of dendritic cells in peripheral blood of patients treated with exogenous thyrotropin

Abstract Background Dendritic cells (DCs) play a major role as regulators of inflammatory events associated with thyroid pathology. The immunoregulatory function of DCs depends strongly on their subtype, as well as maturation and activation status. Numerous hormonal factors modulate the immune properties of DCs, however, little is known about effects exerted by the hypothalamus-pituitary-thyroid-axis. Recently, we have shown a direct regulatory influence of thyroid hormones (TH) on human DCs function. The aim of the present study was to analyze the effect of systemically administered thyrotropin (TSH) on human blood DCs ex vivo. Methods Blood samples for the cytometric analysis of peripheral blood plasmacytoid and myeloid DCs subtypes were collected from patients subjected to total thyroidectomy because of differentiated thyroid carcinoma at 2 time points: (i) directly before the commencement of TSH administration and (ii) 5 days after first TSH injection. The whole blood quantitative and phenotypic analysis of plasmacytoid and myeloid DCs subtypes was performed by flow cytometry. Results Administration of TSH did not influence the percentage of plasmacytoid DCs in peripheral blood of study participants. Also the percentage of the two main myeloid DCs subpopulations – CD1c/BDCA1+ DCs and CD141/BDCA3+ DCs did not change significantly. TSH administration had no effect on the surface expression of CD86 – one of the major costimulatory molecules – neither in the whole peripheral blood mononuclear cell (PBMC) fraction nor in particular DCs subtypes. Conclusions In the present study, we demonstrated no influence of systemic TSH administration on human peripheral blood DCs subtypes. These results are in accordance with our previous work suggesting the direct effect of TH on human DCs ex vivo.</p