Cilj: Istražiti utjecaj angiogenih parametara, izražaja osteopontina (OPN) i vaskularnog endotelnog čimbenika rasta (VEGF; engl. vascular endothelial growth factor) u koštanoj srži na odgovor na prvu ...liniju terapije u pacijenata s multiplim mijelomom (MM). Ispitanici i metode: U istraživanje je uključeno 68 pacijenata s MM-om, liječenih na Odjelu hematologije Kliničkog bolničkog centra u Rijeci. Za kvantifikaciju angiogenih parametara uz klasičnu metodu utvrđivanja gustoće krvnih žila (MVD; engl. microvessel density) koristili smo i računalno potpomognutu analizu slike koja nam je omogućila izračunavanje broja krvnih žila po mm2, površine krvnih žila po mm2, srednjeg promjera krvnih žila i ukupne vaskularne površine (TVA; engl. total vascular surface) na bioptičkim uzorcima koštane srži obojanim s anti-CD34. Za određivanje izražaja OPN-a i VEGF-a u plazma stanicama korištena je dvostruka imunohistokemijska metoda CD138+OPN i CD138+VEGF. Rezultati: Utvrđeno je da su viši angiogeni parametri, MVD (P = 0,004) i broj krvnih žila po mm2 (P = 0,028) značajni negativni prognostički pokazatelji odgovora na prvu liniju terapije u pacijenata s MM-om. Zaključak: Objektivna metoda procjene angiogeneze u uzorcima koštane srži mogla bi postati dio rutinske patohistološke obrade pacijenata koja bi pomogla u prepoznavanju podskupina pacijenata s eventualnim agresivnijim tijekom i lošijim odgovorom na terapiju.
Internistička patologija relativno je nov pojam koji obuhvaća dio patologije koji se bavi patohistološkom dijagnostikom pretežito netumorskih stanja, a u manjoj mjeri tumora, u različitim granama ...interne medicine, poput pulmologije, hematologije, nefrologije, gastroenterologije, dermatologije i imunologije. Uvidom u morfološke promjene bolesnih tkiva patolog postavlja dijagnozu koja se uz dodatne kliničke podatke dorađuje, međutim, patolog često odmah postavlja i definitivnu dijagnozu koja određuje daljnji tijek liječenja pacijenta. Internistička patologija obuhvaća vrlo složen i sofisticiran sustav dijagnostike u patologiji koji implementira bazičnu patohistološku dijagnostiku upotpunjenu histokemijskim, imunohistokemijskim, imunofluorescentnim i molekularnim tehnikama te elektronskomikroskopskom analizom. Oskudnost tkiva dobivenog pretežito tijekom različitih endoskopskih procedura te putem biopsije iglom jedna je od osobitosti ove grane patologije. Interpretacija promjena u analiziranim stanicama i tkivima često je ograničena dostupnošću kliničkih podataka o pacijentovu stanju, a korelacija morfoloških promjena s kliničkim podacima neophodna za ispravno postavljanje konačne dijagnoze. Iz navedenog se može zaključiti kako je internistička patologija dijagnostički vrlo zahtjevna i izazovna grana patologije koja koristi najsuvremenije pomoćne alate u cilju spoznaje patološkog supstrata bolesti, kako poznatih tako i novih internističkih entiteta.
Cilj: Imunohistokemijski procijeniti ekspresiju ciklina D1 na mijelomskim stanicama u koštanoj srži pacijenata s multiplim mijelomom (MM), liječenih bortezomibom te dobivene podatke usporediti s ...kliničko-patološkim parametrima i odgovorom na terapiju. Ispitanici i metode: Ekspresija ciklina D1 analizirana je ubioptičkim uzorcima koštane srži 24 pacijenta s MM-om prije i nakon provedene terapije bortezomibom. Za određivanje ekspresije ciklina D1 na mijelomskim stanicama korištena je dvostruka imunohistokemijska metoda koja omogućava istovremeno bojanje uzorka s dva protutijela (u ovom slučaju s CD138 za identifikaciju plazma stanica i ciklinom D1). Pozitivnom reakcijom smatralo se nuklearno obojenje naciklin D1, bilo kojeg intenziteta, u ≥ 10 % plazma stanica. Rezultati su potom korelirani s kliničko-patološkim podacima pacijenata ispitivane skupine. Rezultati: Nuklearna ekspresija ciklina D1 u plazma stanicama oboljelim od MM-a prije terapije nađena u je 9/24 pacijenta, a ukupno negativnih pacijenata bez ikakvog nuklearnog bojanja 13/24 ili s bojanjem na ciklin D1 u < 10 % stanica 2/24 bilo je 15/24. Nakon terapije bortezomibom 5/24 pacijenta bilo je pozitivno na ciklin D1, dok je ukupno 19/24 pacijenata bilo negativno, tj. bez ikakvog nuklearnog bojanja 17/24 ili je imalo < 10 % tumorskih stanica 2/24 pozitivno na ciklin D1. U ovoj studiji nije nađena statistički značajna razlika u ekspresiji ciklina D1 prije i nakon provedene terapije bortezomibom (p = 0,084) niti s nekim od kliničko-patoloških parametara. Zaključak: Rezultati i razina ekspresije ciklina D1 uvelike variraju unutar različitih ispitivanih skupina, a također ovise i o terapiji koju su pacijenti dobivali, stoga je za utvrđivanje značaja ciklina D1 na prognozu i korelaciju s kliničko- -patološkim karakteristikama potrebna veća i homogenija skupina pacijenata.
Objective The objective of the study was to compare the performance of 3 different extraction instruments in conjunction with 4 different amplification and detection kits for detection and typing of ...human papillomavirus (HPV) deoxyribonucleic acid (DNA). Study Design A total of 42 cervical swabs were investigated. HPV DNA was extracted on the 3 different instruments. Each of the extracts was then amplified, and HPV DNA amplification products were detected with 4 different kits. Results In 31 samples, HPV DNA was detected by both the Amplicor HPV test and the LINEAR ARRAY HPV genotyping test in conjunction with DNA extraction on the easyMAG instrument. In another 6 samples, only low-risk types were detected with the LINEAR ARRAY HPV genotyping test. After extraction on the easyMAG instrument, 32 samples tested positive when the PapilloCheck with the HotStarTaq DNA polymerase was used. Conclusion Together with extraction on the easyMAG instrument, the Amplicor HPV test, the LINEAR ARRAY HPV genotyping test, and the new PapilloCheck with the HotStarTaq DNA polymerase provide comparable results allowing reliable and safe HPV diagnostics in the routine laboratory. Use of alternative assays may lead to an increase of invalid and divergent HPV typing results.
The goal of this research was to determine the existence of the significant time differences in the identification of the recurrences and neck metastases in the patients surgically treated for the ...oral cavity cancer by comparing three postoperative follow up methods. The study included 286 patients surgically treated for oral and pharyngeal cancer in period from 1991 to 2007 at three different institutions, divided into three groups based on the different postoperative follow up protocol. In this study we were able to show that the period of identification of recurrences and neck metastases was significantly shorter in the group of patients whose follow up included neck ultrasound, along with methods of inspection and palpation of the oral cavity and the neck. In conclusion, implementation of more contemporary methods such as the neck ultrasound is needed along with usual follow up methods, such as inspection and palpation of the oral cavity and the neck. Also, follow up of the patients surgically treated for the oral cavity cancer should be conducted systematically. Ultrasound examination of the neck should be recommended due to its low cost, harmlessness, possible frequent usage, high quality visual imaging and possibility of combination with the fine needle aspiration cytology (FNAC) of the suspicious lymph nodes.
Skin melanoma is by far the most lethal skin cancer, it is unpredictable by nature and presents a severe diagnostic problem. One of the major issues in melanoma diagnostics is to differentiate it ...with confidence from a dysplastic nevus. Thus, the aim of this study was to evaluate hTERT expression on a spectrum of dermal lesions (from normal skin toprimary melanoma) in order to examine its possible role as a diagnostic marker in melanoma diagnostics. In this study we analyzed the expression of hTERT by real-time PCR on 58 freshly obtained biopsy samples (4 samples of normal skin, 12 dermal nevi, 23 dysplastic nevi, 19 primary melanomas). Our results showed slightly greater hTERT expression in dysplastic nevi than melanomas with major data overlap. Considering the given results, hTERT does not seem to be a reliable diagnostic marker for melanoma.
The aim of this study is to investigate the differences of clinical and laboratory parameters between patients with JAK2-V617F positive myeloproliferative neoplasms (MPNs) and JAK2 wild type MPNs. ...DNA was isolated from peripheral blood granulocytes of 106 patients treated at Rijeka University Hospital Center: 41 with polycythemia vera (PV), 43 with essential thrombocythemia (ET), 9 with primary myelofibrosis (PMF) and 13 with myeloproliferative neoplasm--unclassifiable (MPN-u). The JAK2-V617F mutation was detected using allele specific PCR. Laboratory and clinical parameters were obtained from patient's medical records. The JAK2-V617F mutation was detected in 69% (73/106) patients with MPNs. The results revealed significantly different prevalence of JAK2-V617F mutation, between MPNs entities: 88% in PV 58% in ET, 56% in PMF and 54% in MPNs-unclassified disorders. The JAK2-V617F mutation significantly correlated with higher leukocyte count and alkaline phosphatase co re in ET group and with higher platelets count, leukocyte alkaline phosphatase score and serum lactate dehydrogenase in PV group. Vascular events were associated with elevated platelets count in whole MPNs group, with higher platelets and leukocyte count in ET and with splenomegaly in PVpatients. Clinical and laboratory data revealed significant contribution ofJAK2-V617F mutation to the development of clinical phenotype in patients with distinct subgroups of MPNs.
Infekcija humanim papiloma virusom (HPV) najčešća je spolno prenosiva bolest i pretpostavlja se da genitalnu infekciju ovim virusom ti jekom života stekne 75 – 80 % spolno aktivnih žena i muškaraca. ...Od ranih osamdeseti h kada je Harald zur Hausen (dobitnik Nobelove nagrade za fiziologiju ili medicinu 2008. godine) dokazao humani papiloma virus (HPV) genotipa 16 i 18 u karcinomu cerviksa, u velikom broju studija potvrđeno je i znanstvenim činjenicama dokazano da je infekcija HPV-om ključni čimbenik u razvoju karcinoma cerviksa. Maligne promjene cerviksa nastaju ako su ispunjeni višestruki uvjeti koji su defi nirani virusnim značajkama, staničnim protuvirusnim mehanizmima i imunim odgovorom domaćina. Virusnu komponentu čine genoti p virusa, perzistencija i intenzitet infekcije, a imunogenetička konstitucija pojedinca, stanično posredovani imuni odgovor i utjecaj vanjskih čimbenika kao što su
lijekovi i bolesti, definiraju imunopatogenetski doprinos domaćina. Ključni proteini uključeni u nastanak karcinoma cerviksa su virusni onkogeni, E6 i E7, koji interferiraju s nizom staničnih procesa te dovode do nekontrolirane proliferacije i stanične imortalizacije, stoga je značaj molekularnih studija koje se bave problematikom HPV-a upravo u boljem razumijevanju patogeneze HPV-a te primjeni stečenih spoznaja u prevenciji virusne infekcije, razvoju profilaktičkog cjepiva i uvođenju molekularne dijagnosti ke kao standarda u procjeni rizika za žene izložene kroničnoj infekciji HPV-om.
It is still controversial whether keratoacanthoma is to be considered as a well differentiated variant of squamous cell-carcinoma or a separate entity. As opposed to malignant potential of squamous ...cell-carcinoma, keratoacanthoma is characterized by a spontaneous regression. However, in some cases, otherwise typical keratoacanthoma can behave aggressively showing the signs of perineural and perivascular invasion and metastases in regional lymph nodes. The most important feature that separates these two closely related entities is a tendency of keratoacanthoma to regress. Causes and detailed mechanism of this regression are still not completely elucidated.
Within the past few years, it has become evident that the molecular events regulating cell survival and apoptosis are important contributors to the overall kinetics of benign and malignant cell growth. Immunological mechanisms have been implicated in a phenomenon of spontaneous tumor regression. Recent studies suggested that the tumor regression is dependent mainly on the immune response mediated by cytotoxic T lymphocytes (CD8+), together with helper T cells (CD4+). Cytotoxic T lymphocytes can kill tumor cells and mediate tumor regression in vivo through two distinct molecular mechanisms: Fas/Fas ligand and granzyme B/perforin mediated pathways.
Tumor cells are capable of developing different escape mechanisms in order to overcome their sensitivity to apoptotic signals. However, granzyme B, contained in cytolytic granules released upon target cell recognition, can also cause tumor cell death and consequently tumor regression by direct damage to non-nuclear structures through a caspase-independent pathway.
Therefore, we propose a key role of plasticity in the granzyme B mediated cell death pathway in the killing of changed tumor cells, resulting in keratoacanthoma regression through apoptosis or direct damage of tumor cells. On the other hand, insufficient activation of cytotoxic T lymphocytes and decreased release or activity of granzyme B could be responsible for squamous cell-carcinoma progression and occasional aggressive behavior in keratoacanthomas.
As a first step in confirming or refuting our hypothesis, we suggest a thorough immunohistochemical study of the presence of granzyme B and its activity in keratoacanthoma and squamous cell-carcinoma samples. To our knowledge, no such study has been performed so far.
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