Zmiana ekspresji mRNA dla CTLA-4, CD28, VDR i CD45 w limfocytach T u osób z chorobą Hashimoto — badanie pilotowe

Introduction: CD28/T-cell receptor (TCR)/cytotoxic T-lymphocyte antigen 4 (CTLA4) complex controls T-cell tolerance and autoimmunity in Hashimoto’s thyroiditis (HT). In addition, CD45 protein tyrosine phosphatase (PTPase) and vitamin D receptor (VDR) cooperatively interact with the TCR complex to affect autoimmune processes central to the pathogenesis of HT. Nevertheless, their role in HT aetiology has been less well established. In this study, we aimed to explore mRNA expression levels of CTLA4, CD28, CD45, and VDR in T-cells, across different outcomes of HT. Material and methods: The study included 45 HT patients and 13 euthyroid, healthy controls. T-lymphocytes were isolated from peripheral blood mononuclear cells, total mRNA was extracted from T-cells, and gene expression was studied by reverse transcription-polymerase chain reaction (RT-PCR) and ImageQuant method relative to glyceraldehyde-3-phosphate dehydrogenase RT-PCR products. Results: Nominally higher expression levels of VDR, CTLA4, CD28, and CD45RAB mRNA were found in unsorted T-lymphocytes of healthy controls when compared to the HT patients. No difference was observed between hypothyroid/untreated, spontaneously euthyroid and LT4-treated HT patients. VDR mRNA expression was linked to both T3 levels and CTLA4 gene expression, whilst CD45RB mRNA expression coincided with CTLA4 and CD28 transcript levels. Conversely, older age and lower T3 levels were associated with increased abundance of CD45R0 isoform in HT patients. Conclusions: The results suggest a cross talk between endocrine and immune functions in HT pathology: an altered peripheral T cell mRNA profile with reduced VDR, CTLA4, CD28, and CD45RAB transcript levels is accompanied by age-related shift from naive to memory/late-differentiated T cell CD45R mRNA signature and associated with thyroid hormone status in the HT patients.Wstęp: Kompleks antygenu CD28/receptora limfocytów T (TCR)/antygenu 4 związan ego z limfocytem T cytotoksycznym (CTLA4) reguluje tolerancję limfocytów T oraz autoimmunogenność w chorobie Hashimoto (HT). Ponadto białkowa fosfataza tyrozynowa (PTPase) CD45 oraz receptor witaminy D (VDR) wchodzą w interakcję z kompleksem TCR, modyfikując procesy autoimmunologiczne mające podstawowe znaczenie w patogenezie HT. Jednak rola tych cząsteczek w etiologii HT nie została dokładnie ustalona. Celem autorów badania była ocean poziomów ekspresji mRNA dla CTLA4, CD28, CD45 i VDR w limfocytach T w zależności od różnego statusu HT. Materiał i metody: Do badania włączono 45 chorych na HT i 13 zdrowych osób z prawidłową czynnością tarczycy. Limfocyty T wyizolowano spośród komórek jędnojądrzastych krwi obwodowej, wyekstrahowano z nich całkowity mRNA i określono ekspresję genów za pomocą łańcuchowej reakcji polimerazowej z odwrotną transkryptazą (RT-PCR) i metody ImageQuant związanej produktami reakcji RT-PCR dehydrogenazy aldehydu 3-fosfoglicerynowego. Wyniki: U osób zdrowych stwierdzono nominalnie wyższy poziom ekspresji mRNA dla VDR, CTLA4, CD28 i CD45RAB w niesortowanych limfocytach T niż u chorych na HT. Nie zaobserwowano żadnych różnic między chorymi na HT z niedoczynnością tarczycy/nieleczonymi, u których samoistnie nastąpiło przywrócenie eutyreozy, i stosującymi leczenie LT4. Ekspresja VDR mRNA była powiązana zarówno ze stężeniem T3, jak i ekspresją genu CTLA4, natomiast ekspresja mRNA dla CD45RB wiązała się z poziomami transkryptu CTLA4 i CD28. Z kolei starszy wiek i niższe stężenia T3 wiązały się ze zwiększoną ilością izoformy CD45R0 u chorych na HT. Wnioski: Uzyskane wyniki sugerują interferencje między czynnością wewnątrzwydzielniczą i immunologiczną w patologii HT: zmiana profilu mRNA obwodowych limfocytów T z ograniczeniem poziomu transkryptu białek VDR, CTLA4, CD28 i CD45RAB współistnieje z zależnym od wieku przesunięciem sygnatury mRNA limfocytów CD45R od komórek naiwnych do komórek pamięci immunologicznej/ zróżnicowanych i jest powiązana ze stężeniami hormonów tarczycy u chorych na HT

PAX8-PPARg Oncogene in Follicular Thyroid Tumors: RT-PCR and Immunohistochemical Analyses

US-guided fine needle aspiration cytology is currently the best diagnostic tool for thyroid nodules. However, it is not sensitive and specific enough for differentiating between benign and malignant follicular tumors. A potentially useful marker for this differentiation is the PAX8-PPARg rearrangement, identified in follicular thyroid carcinomas, but not in follicular adenomas or other types of thyroid tumors. The aim of this research was to determine the clinical significance of the PAX8-PPARg oncogene in diagnostics follicular thyroid tumors. The study included 62 patients with follicular or Hürthle cell tumors. Gene expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) from paraffin embedded tissues, and PCR products were checked using the agarose gel electrophoresis. The immunohistochemical analysis was performed on archive paraffin embedded tissues with the monoclonal PPARã antibody. The statistical analysis has indicated that neither the expression of PAX8-PPARg mRNA, nor the immunohystochemical analysis with the PPARg antibody correlate with the patohystological diagnosis. The oncogene PAX8-PPARg has not met the expectations as a reliable tumor marker for differentiation between benign and malignant thyroid tumors, which makes the only reliable histological criteria – capsular and vascular invasion

PAX8-PPARg Oncogene in Follicular Thyroid Tumors: RT-PCR and Immunohistochemical Analyses

US-guided fine needle aspiration cytology is currently the best diagnostic tool for thyroid nodules. However, it is not sensitive and specific enough for differentiating between benign and malignant follicular tumors. A potentially useful marker for this differentiation is the PAX8-PPARg rearrangement, identified in follicular thyroid carcinomas, but not in follicular adenomas or other types of thyroid tumors. The aim of this research was to determine the clinical significance of the PAX8-PPARg oncogene in diagnostics follicular thyroid tumors. The study included 62 patients with follicular or Hürthle cell tumors. Gene expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) from paraffin embedded tissues, and PCR products were checked using the agarose gel electrophoresis. The immunohistochemical analysis was performed on archive paraffin embedded tissues with the monoclonal PPARã antibody. The statistical analysis has indicated that neither the expression of PAX8-PPARg mRNA, nor the immunohystochemical analysis with the PPARg antibody correlate with the patohystological diagnosis. The oncogene PAX8-PPARg has not met the expectations as a reliable tumor marker for differentiation between benign and malignant thyroid tumors, which makes the only reliable histological criteria – capsular and vascular invasion

Importance of Interleukin 6 in Pathogenesis of Inflammatory Bowel Disease

Inflammatory bowel disease (IBD), encompassing ulcerative colitis (UC) and Crohn’s disease (CD), is an uncontrolled chronic inflammation of the gastrointestinal tract caused by an interaction of diverse genes and environmental factors. There is growing evidence that cytokine production plays an important role in IBD. One of the key roles in signaling pathway in development of IBD is performed by interleukin 6 (IL-6), although molecular mechanism of this pathway is not yet fully understood. In order to assess the clinical relevance of IL-6 serum concentration in patients with CD and UC we performed cross-sectional, case-control study of IL-6 levels in patients’ and healthy blood donors’ sera. A total of 100 CD and UC patients and 71 healthy blood donors were investigated. Clinical activity of CD and UC was evaluated using the Crohn\u27s disease activity index and Truelove-Witt\u27s criteria, respectively. Quantitative assessment of serum IL-6 was performed with solid-phase, enzyme-labeled, chemiluminescent sequential immunometric assay. Our results indicate that serum IL-6 is a clinically relevant parameter for CD and UC that strongly correlates with inflammatory activity of disease. We confirmed and extended the role of cytokine production patterns for IBD presentation in Croatian population

Usporedba 18F-FDG pozitronske emisijske tomografije i kompjutorizirane tomografije u bolesnika s kolorektalnim karcinomom i limfomom: naša početna klinička iskustva

Findings obtained by fluorine-18-fluorodeoxyglucose positron emission tomography (18F-FDG PET) and computed tomography (CT) were compared in patients with malignant lymphoma and colorectal carcinoma. In 14 malignant lymphoma patients, 16 18F-FDG PET procedures were performed to assess chemotherapy and/or radiotherapy outcome (remission). One patient with clinically overt relapse of non-Hodgkin.s lymphoma underwent PET to assess disease dissemination prior to prescribing second-line chemotherapy. Two patients were submitted to PET on two occasions. PET pointed to residual disease in six of 14 patients and was inconclusive in one patient. These patients underwent computed tomography (CT), some of them before and others after PET examination. Then PET and CT findings were compared and therapeutic response, i.e. disease remission was assessed. The signs of residual disease were present in four and absent in nine patients, whereas inconclusive findings in terms of residual disease were recorded in one patient. Although our initial clinical experience was acquired in quite a small number of patients, CT modified clinical evaluation of residual disease in two patients and should be included along with PET in diagnostic work-up of these patients.Usporedili smo nalaze fluoro-18-fluorodeoksiglukoza (FDG) pozitronske emisijske tomografije (18F-FDG PET) i kompjutorizirane tomografije (CT) u bolesnika s malignim limfomom i kolorektalnim karcinom. U 14 bolesnika 18F-FDG PET je učinjen 16 puta radi procjene ishoda kemoterapije i/ili radioterapije, odnosno remisije. U jednog bolesnika s klinički jasnim recidivom ne-Hodgkinova limfoma PET je proveden radi procjene proširenosti bolesti prije ordiniranja druge linije kemoterapije. U dvoje bolesnika PET je učinjena dva puta. U šestoro od 14 bolesnika nalaz PET ukazivao je na rezidualnu bolest, dok je u jednog bolesnika bio dvojben. Stoga je u tih bolesnika učinjena i CT; u nekih bolesnika CT je izvedena prije PET, a u drugih nakon PET. Tada smo usporedili nalaze PET i CT te procijenili terapijski odgovor, tj . remisiju bolesti. Znaci rezidualne bolesti bili su prisutni u četvoro bolesnika, odsutni u devetoro bolesnika, dok je kod jednog bolesnika i dalje bilo nejasno je li rezidualna bolest prisutna ili nije. Iako se ovo naše početno kliničko iskustvo odnosi na mali broj bolesnika, CT je promijenio kliničku procjenu rezidualne bolesti u dvoje bolesnika i smatramo da bi uz PET i CT trebao biti sastavni dio dijagnostičke obrade takvih bolesnika

Usporedba 18F-FDG pozitronske emisijske tomografije i kompjutorizirane tomografije u bolesnika s kolorektalnim karcinomom i limfomom: naša početna klinička iskustva

Findings obtained by fluorine-18-fluorodeoxyglucose positron emission tomography (18F-FDG PET) and computed tomography (CT) were compared in patients with malignant lymphoma and colorectal carcinoma. In 14 malignant lymphoma patients, 16 18F-FDG PET procedures were performed to assess chemotherapy and/or radiotherapy outcome (remission). One patient with clinically overt relapse of non-Hodgkin.s lymphoma underwent PET to assess disease dissemination prior to prescribing second-line chemotherapy. Two patients were submitted to PET on two occasions. PET pointed to residual disease in six of 14 patients and was inconclusive in one patient. These patients underwent computed tomography (CT), some of them before and others after PET examination. Then PET and CT findings were compared and therapeutic response, i.e. disease remission was assessed. The signs of residual disease were present in four and absent in nine patients, whereas inconclusive findings in terms of residual disease were recorded in one patient. Although our initial clinical experience was acquired in quite a small number of patients, CT modified clinical evaluation of residual disease in two patients and should be included along with PET in diagnostic work-up of these patients.Usporedili smo nalaze fluoro-18-fluorodeoksiglukoza (FDG) pozitronske emisijske tomografije (18F-FDG PET) i kompjutorizirane tomografije (CT) u bolesnika s malignim limfomom i kolorektalnim karcinom. U 14 bolesnika 18F-FDG PET je učinjen 16 puta radi procjene ishoda kemoterapije i/ili radioterapije, odnosno remisije. U jednog bolesnika s klinički jasnim recidivom ne-Hodgkinova limfoma PET je proveden radi procjene proširenosti bolesti prije ordiniranja druge linije kemoterapije. U dvoje bolesnika PET je učinjena dva puta. U šestoro od 14 bolesnika nalaz PET ukazivao je na rezidualnu bolest, dok je u jednog bolesnika bio dvojben. Stoga je u tih bolesnika učinjena i CT; u nekih bolesnika CT je izvedena prije PET, a u drugih nakon PET. Tada smo usporedili nalaze PET i CT te procijenili terapijski odgovor, tj . remisiju bolesti. Znaci rezidualne bolesti bili su prisutni u četvoro bolesnika, odsutni u devetoro bolesnika, dok je kod jednog bolesnika i dalje bilo nejasno je li rezidualna bolest prisutna ili nije. Iako se ovo naše početno kliničko iskustvo odnosi na mali broj bolesnika, CT je promijenio kliničku procjenu rezidualne bolesti u dvoje bolesnika i smatramo da bi uz PET i CT trebao biti sastavni dio dijagnostičke obrade takvih bolesnika

Vrijednost mjerljivih i nemjerljivih značajka ultrazvučnog prikaza limfnih čvorova u otkrivanju zloćudnog zauzeća

The aim of the study was to assess diagnostic value and utility of selected morphological features in predicting lymph node (LN) malignancy using B-mode, Doppler ultrasonography and multivariate settings in a tertiary radiological referral center. The study included 123 patients having undergone ultrasound-guided fine-needle aspiration and cytologic analysis (FNAC) of cervical, axillary and inguinal LNs. Each LN was characterized by long/L and short/T-axis, shape, margins, echogenicity, cortical thickness, vascularization, and examiner’s subjective impression. Within the limitations of FNAC, altered shape and vascularization had relatively high specificity and positive predictive value (>80%), whereas subjective impression had high sensitivity and negative predictive value (100%) for malignancy. The cut-off levels for different features of LN by ROC analysis were as follows: long-axis 23 mm, short-axis 11 mm, L/T ratio 2.19, and maximal cortical thickness 5.1 mm. On multivariate analysis (adaptive regression splines, n=108), the addition of long-axis, L/T ratio, age and sex considerably improved diagnostic accuracy (88%), sensitivity (margins + vascularization) and specificity (subjective impression) of the diagnostic model. The combination of morphological and demographic features could improve diagnostic accuracy, usually with a trade-off between the sensitivity and specificity of the predictive model. The performance may depend on the level of expertise and institutional settings.Cilj istraživanja bio je odrediti dijagnostičku vrijednost i korisnost odabranih morfoloških značajka u predviđanju malignosti limfnog čvora (LČ) upotrebom B-moda, Dopplerova ultrazvuka i multivarijatnih postavka u tercijarnom radiološkom referentnom centru. U 123 bolesnika je učinjena ultrazvučno vođena aspiracijska punkcija i citološka analiza LČ vrata, pazuha i prepona. Svaki LČ je opisan uzdužnim/L i poprečnim/T promjerom, oblikom, rubom, ehogenošću, debljinom kore, vaskularizacijom i subjektivnim dojmom pregledavatelja. Unutar ograničenja citološke analize, izmijenjen oblik i vaskularizacija su imali visoku specifičnost i pozitivnu prediktivnu vrijednost (>80%), dok je subjektivni dojam imao visoku osjetljivost i negativnu prediktivnu vrijednost (100%) za malignost. Optimalne granične vrijednosti za različite značajke LČ dobivene analizom ROC su bile 23 mm za uzdužni promjer, 11 mm za poprečni promjer, 2,19 za omjer L/T i 5,1 mm za maksimalnu debljinu korteksa. U multivarijatnoj analizi (adaptive regression splines, n=108) dodatak uzdužnog promjera, omjera L/T, dobi i spola semikvalitativnim obilježjima LČ značajno je povećao dijagnostičku točnost (88%), osjetljivost (rubovi + vaskularizacija, 87%) i specifičnost (subjektivni dojam, 83%) konačnog dijagnostičkog modela. U zaključku, kombinacija morfoloških i demografskih značajka može poboljšati dijagnostičku točnost, obično uz kompromis između osjetljivosti i specifičnosti prediktivnog modela. Učinkovitost može ovisiti o razini stručnosti i institucionalnim postavkama

The Influence of the Different Morphological Changes on Gastric Mucosa on Somatostatin Cell Number in Antrum Mucosa and Serum Somatostatin

The aim of our paper was to investigate the influence of the different morphological changes on gastric mucosa on somatostatin D-cell number in antral mucosa and serum Somatostatin. We analyzed according to Sydney classification to what extent the severity of gastritis affect the observed hormonal values. somatostatin D-cell number in antral mucosa and serum Somatostatin values were compared between three grups of patients; mild, moderate and severe cronic gastritis. The average number of somatostatin cell in biopsy sample of antrum mucosa was 30.41±35.38 (N=17) in the case of middle form, 18.69±26.65 (N=56) in moderate and in severe case of chronic gastritis 5.23±5.93 (N=7) cells in mm² of mucosa. The level of somatostatin in the serum of middle form gastritis were 26.43±28.76, moderate 19.95±35.93 and severe 17.88±17.66 pg/mL. In order to determine the number of somatostatin cells in antrum mucosa and serum somatostatin with present morphological changes of mucosa, it might helpful to exclude the patients with non-ulcer dyspepsia, but with the higher risk of premalignant and malignant changes

Polymorphisms of Interleukin-23 Receptor in Patients with Inflammatory Bowel Disease in a Croatian Tertiary Center

The Interleukin-23 signalling pathway is important for the differentiation of TH17 lymphocytes and is involved in the pathogenesis of Inflammatory bowel disease. Polymorphisms in the IL-23 receptor gene were previously found to be associated with Inflammatory bowel disease in various populations. The aim of this study was to determine whether the specific rs11209026 and rs7530511 single-nucleotide polymorphisms in the Interleukin-23 receptor gene are associated with Crohn’s disease and ulcerative colitis in a Croatian patient population. A total of 50 patients with Crohn’s disease and 93 patients with ulcerative colitis, as well as 99 healthy control subjects were included in the study. The results deter- mined a significantly higher occurrence of rs11209026 in control group compared to patients with inflammatory bowel disease, suggesting a protective effect of this polymorphism. The rs11209026 variant was strongly associated with Crohn’s disease, but it was absent in ulcerative colitis. However, there was no significant association between the rs7530511 poly- morphism with either ulcerative colitis or Crohn’s disease. Associations presented in this study give potentially impor- tant insight into the roles of specific Interleukin-23 receptor polymorphisms in Crohn’s disease pathogenesis in the Cro- atian population