Prognostic significance of immunohistohemical parameters and histomorphological score in assessment of degree of cervical dysplasia

Cilj:Karcinom grlića je treći po uĉestalosti u svetu meĊu malignim bolestima. Displazije epitela grlića predstavljaju niz prekursorskih lezija karcinoma grlića.Najznacajniji faktor rizika za displaziju epitela grlića jeste infekcija humanim papiloma virusima.Cilj ove disertacije je bio da se utvrdi dijagnostiĉki znaĉaj imunohistohemijskih metoda markerima p16, p63, ciklin d1 i survivin u epitelnim displazijama grlića,morfometrijske nalize i histomorfološkog skora. Metodologija:Retrospektivnom studijom je bilo obuhvaćeno 100 pacijentkinja pregledanih na Ginekološkom odeljenju Vojnomedicinske akademije u trogodišnjem periodu.Kriterijumi za ulazak u studiju su: primarna, ranije netretirana lezija, kolposkopski nalaz koji upućuje na prisustvo premaligne lezije grlića: uĉinjen citološki test epitela grlića po metodi Papanikolau koji je interpretiran po Bethesda kriterijumu sa nalazom displazije lakog stepena (low-grade squamous intraepithelial lesion,LSIL), displazije teškog stepena (high-grade sqamous intraepithelial lesion,HSIL), atipiĉnih ćelija neodreĊenog znaĉaja (atypical squamous cells of undeterminated significance,ASCUS);uĉinjena tipizacija humanih papiloma virusa reakcijom lanĉane polimerizacije sa dokaznim prisustvom virusa tipa 16,18,31,33 kod svih zena. Ovoj grupi pacijentkinja je uradjena biopsija, imunohistohemijske i morfometrijske analize bioptiranog tkiva. Primenjena su imunohistohemijska bojenja p16, p63, ciklin d1 i survivin. Analiza imunohistohemijskih reakcija je vršena svetlosnim mikroskopom u zonama displazije. Od 100 zena koje su ušle u studiju, kod kojih je prvom biopsijom dokazano prisustvo displazije (HSIL, LSIL), kod 48 zena je ucinjena ponovna biopsija u perodu do sedam meseci, na osnovu istih dijagnostickih kriterijuma kao za prvu biopsiju, gde su primenjivane sve metode istraživanja koje su korišćene u prvoj biopsiji. Kontrolnu grupu su ĉinile 12 žena,koje su se javile na kontrolni pregled bez displazije i bez verifikovane infekcije grlića materice visokoriziĉnim HPV virusima, zbog mioma, na materijalu dobijenom nakon histerektomije...Aim:Cervical cancer is the third most common cancer in the world.Cervical dysplasia, such as minor and severe dysplasia, is a group of precursor lesion of the squamous cell carcinoma. The most important etiologic risk factors for dysplasia are infection with the human papilloma virus (HPV).The aim of this study was to determine the relevance of correlation between epithelial p16,p63,ciklin d1 and survivin expression and the degree of human papilloma virus (HPV)- induced cervical epithelial dysplasia, as well as, to establish the significance of morphometric analysis of nuclear area and histomorphologic score in assessment of cervical dysplasia degrees Methodology: This retrospective study included 100 patients examined at the Department of Gynaecology of the Military Medical Academy in the period from three years.Criteria for inclusion into the study were primary, previously untreated lesions, colposcopic findings that suggest the presence of premalignant lesions of the cervix; findings from Papanikolaou cytological test, interpreted by the Bethesda criteria with the findings of LSIL, HSIL, ASCUS. HPV typisation was done by PCR with evidence of viruses type 16, 18, 31 and 33 in all women. After the bipsy of the cervical mucosa, we performed pathohistological analysis by hematoxilineosin staining, immunohistochemical analysis for p16,p63,ciklin d1 and survivin and morphometric analysis of cervical epithelium. After the first biopsy, during next seven months in group of 48 patients with dysplasias, according the same criteria for inclusion, we performed rebiopsy and uniform examination . The control group consisted of 12 women without dysplasia and without verified infection of cervical high-risk HPV, with pathological diagnosis of leiomyoma in material obtained after the hysterectomy.Criteria for exclusion from the study was previously diagnosed and treated dysplasia or squamous cell carcinoma..

Etiological Factors for Epithelial Atypia in Chronic Rhinosinusitis with Nasal Polyps

Aim: Chronic rhinosinusitis with nasal polyps (CRSwNP) or nasal polyposis (NP) is characterized by polypoid outgrowths of chronically inflamed respiratory mucosa. Cellular atypia of stromal fibroblasts as a result of increased intercellular fluid pressure is a well-known histological characteristic of CRSwNP. However, the presence of squamous metaplasia and dysplasia on the mucosal surface of nasal polyps (NPs) is not well investigated. The aim of this study was to evaluate the presence of epithelial atypia in ethmoidal NPs and to find possible etiological factors for this histological feature. Methods: This retrospective analysis of prospectively collected data involved 212 patients with NP undergoing endoscopic ethmoidectomy. To evaluate possible etiological factors for epithelial atypia, the patients in whom we histopathologically detected the presence of epithelial atypia were compared with patients with \u27normal\u27 NPs in accordance with the following characteristics as found in the patients\u27 medical data: gender, age, atopic status, sensitivity to non-steroid anti-inflammatory drugs (NSAIDs), cigarette smoking, and occupational exposure to different noxious factors. Results: Epithelial atypia were detected in 44 (20.7%) NP patients, whereas features of \u27true\u27 dysplasia were found in only one patient (0.5%). The presence of atypia was more frequent in males than in females (p=0.008). The association with non-steroid anti-inflammatory drug (NSAID)-exacerbated respiratory disease (N-ERD) and with long-term occupational exposure to different noxious chemicals, especially in workers exposed to salts of heavy metals, was more frequent in NP patients with epithelial atypia than in patients without atypia (p=0.023; p=0.006), respectively). Conclusion: Our results suggest epithelial atypia in NPs are associated with NSAID sensitivity and occupational exposure to different noxious chemicals. Although extremely rare, epithelial dysplasia may occasionally be noted in NPs, a fact potentially useful for both rhinologists and pathologists

Significance of survivin immunoreactivity and morphometric analysis of HPV-induced cervical dysplasia

Genomic integration of high-risk human papilloma virus in the nucleus of cervical epithelial mucosal cells leads to epithelial dysplasia. The aim of this study was to determine the relevance of correlation between epithelial survivin expression and the degree of human papilloma virus (HPV)-induced cervical epithelial dysplasia, and to establish the significance of morphometric analysis of the nuclear area in the assessment of the degree of cervical dysplasia. This retrospective study included 99 women with primary, previously untreated lesions, and colposcopic findings indicating dysplasia, in whom a cytological test by Papanicolaou method was interpreted according to the Bethesda criteria as lowgrade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL), and atypical squamous cells of undetermined significance (ASCUS). We performed human papilloma virus (HPV) typing by PCR for evidence of viruse types 16, 18, 31, 33. After biopsy of the cervical mucosa, we performed hematoxylin-eosin (H-E) and Periodic Acid Schiff (PAS) staining, and immunohistochemical and morphometric analysis of tissue samples. The control group consisted of 12 women without dysplasia and without a verified infection of cervical high-risk HPV. A high statistical correlation between the degree of dysplasia and expression of survivin was found in patients with different types of cervical dysplasia (p = 0.003). We observed a high statistical difference between the area of nuclei at different degrees of cervical dysplasias (p = 0.000). The high-grade cervical dysplasia had a more than 2-fold higher level of ranking in comparison to low-grade dysplasia, and a more than 10-fold higher ranking than the control group without cervical dysplasia

Koncentracije proupalnih citokina u nosnom sekretu kao indikatori težine nosne polipoze

The aim of this prospective study was to evaluate whether cytokine levels in nasal secretions may be clinical parameters of severity of nasal polyposis. Forty nasal polyp patients (20 asthmatic and 20 nonasthmatic) requiring surgical treatment were included. Nasal secretion samples were collected from nasal cavities of all 40 subjects. The levels of T-helper type 1 (Th1) proinflammatory cytokines IL -2, IL -12, IFN -g, IL -1β, TNF -α and TNF -β, Th2 cytokines IL -4, IL -5 and IL -6, antiinflammatory cytokine IL -10 and chemokine IL -8 were measured using flow cytometric method. Each of the 40 patients was staged clinically according to global nasal symptom score, endoscopic score, and Lund-Mackay computed tomography (CT) score. Eosinophils were counted in hematoxylin-eosin stained sections of all nasal polyp samples. The concentrations of Th2 proinflammatory cytokines IL -5 and IL -6 were significantly higher (p<0.05 and p<0.01, respectively) in patients with nasal polyposis and asthma compared with nasal polyp patients without asthma. Positive correlations were observed between IL -2 concentration in nasal secretions and nasal symptom score, endoscopic score, and Lund-Mackay score only in nasal polyp patients without asthma. We also found positive correlation between Lund-Mackay score and the levels of IL -8, IL -4, and IL -1β in nonasthmatic patients. A positive correlation between IL -5 levels in nasal fluid and endoscopic score was found only in asthmatic patients. Eosinophil counts were higher in asthmatic patients’ polyps compared with nonasthmatic ones, but without statistical significance. Nasal polyposis in asthmatic patients has different immunological patterns compared to those without asthma. The concentrations of cytokines measured in nasal fluid were not sensitive enough to be universal criteria to determine the severity of all forms of nasal polyposis.Cilj ove prospektivne studije bio je ispitati mogu li koncentracije citokina u nosnom sekretu poslužiti kao klinički parametri za procjenu težine nosne polipoze. U studiju je bilo uključeno 40 bolesnika s nosnom polipozom (20 astmatičnih i 20 neastmatičnih) u kojih je bilo potrebno kirurško liječenje. Uzorci nosnoga sekreta bili su sakupljeni iz nosnih šupljina svih 40 bolesnika. Koncentracije T pomoćničkih-1 (Th1) proupalnih citokina IL -2, IL -12, IFN -γ, IL -1β, TNF -α i TNF -β, Th2 citokina IL -4, IL -5 i IL -6, protuupalnog citokina IL -10 i hemokina IL -8 mjerene su primjenom metode protočne citometrije. Svaki od 40 bolesnika klinički je klasificiran prema zbiru nosnih simptoma, endoskopskom zbiru i Lund-Mackayevu zbiru. Eozinofili su se brojili u uzorcima nosnih polipa obojenim tehnikom hematoksilin-eozin. Koncentracije Th2 proupalnih citokina IL -5 i IL -6 bile su statistički značajno više u astmatičnih nego u neastmatičnih bolesnika s nosnom polipozom (P<0,05, P<0,01). Nađena je pozitivna korelacija između koncentracije IL -2 u nosnom sekretu i zbira nosnih simptoma, endoskopskog zbira i Lund-Mackayeva zbira samo u neastmatičnih bolesnika. Također su utvrđene pozitivne korelacije između Lund-Mackayeva zbira i razina IL -8, IL -4 i IL -1β u neastmatičnih bolesnika. Pozitivna koralacija između koncentracije IL -5 u nosnom sekretu i endoskopskog zbira nađena je samo kod astmatičnih bolesnika. Broj eozinofila bio je veći u tkivu nosnih polipa astmatičnih bolesnika nego u neastmatičnih, ali bez statističke značajnosti. Nosna polipoza u astmatičnih bolesnika ima drugačiji imunološki tijek u usporedbi s neastmatičnim bolesnicima. Koncentracije citokina mjerene u nosnom sekretu nisu dovoljno osjetljiv pokazatelj da bi bile univerzalni kriterij za procjenu težine svih oblika nosne polipoze

Impact of TLR9 and TLR7 gene polymorphisms on prognosis and survival of patients with oral squamous cell carcinoma

Despite significant efforts in developing new diagnostic and therapeutic modalities, oral squamous cell carcinomas (OSCCs) still exhibit a high recurrence rate, a low five-year survival rate, and an increasing prevalence. Toll-like receptors (TLRs), which initiate and perpetuate immune mechanisms upon activation, have been linked to immune surveillance and the antitumor immune response. The aim of this study was to investigate the association between the polymorphisms of the TLR7 rs3853839 and TLR9 rs187084 genes and OSCC risk, clinicopathological features, and survival. Genotyping was assessed by real-time polymerase chain reaction (PCR) in 95 HPV-negative OSCC patients and 107 age- and sex-matched healthy controls. Patients with lymph node metastases had higher frequencies of the TLR9 rs187084 CC variant genotype compared to the major TT genotype (P = 0.020) and to T-allele carriers (combined TT + CT genotypes, P = 0.015). A higher prevalence of advanced stage III was observed in patients with the TLR9 rs187084 variant CC genotype compared to TT (P = 0.047) and to T-allele carriers (combined TT + CT, P = 0.037). Kaplan-Meier analysis revealed a lower overall survival rate in patients with the TLR9 rs187084 variant CC genotype compared to TT genotype (P = 0.010, log-rank test) and to T-allele carriers (TT + CT genotypes, P = 0.002), though it was not an independent predictor of overall survival. Both TLR9 rs187084 and TLR7 rs3853839 polymorphisms were associated with high alcohol consumption (P = 0.027 and P = 0.001, respectively). The investigated genetic variations were not associated with OSCC susceptibility. The variant CC genotype of the TLR9 rs187084 polymorphism might be a marker of poor survival and tumor progression in OSCC

Eozinofilna upala u alergijskom rinitisu i nosnoj polipozi

On histopathological examination, nasal polyps and nasal mucosa in allergic rhinitis show different forms of pseudostratified respiratory epithelium, whereas the dominant characteristic of lamina propria is an eosinophilic infiltration. The aim of this study was to compare interleukin (IL)-5 and eosinophilic cationic protein (ECP) levels in the nasal fluid of 42 patients: 12 with allergic rhinitis and nasal septal deviation, 17 non-atopic patients with nasal polyposis, and 13 atopic nasal polyp patients were enrolled in this cross-sectional study. Nasal secretion samples were collected a few days before surgery. The levels of IL-5 were measured using fl ow cytometry and the ECP using a commercial ELISA kit. In addition, we counted eosinophils in hematoxylin-and-eosin-stained sections of all nasal polyp and all nasal mucosa samples taken from the inferior nasal turbinates during septoplasty. A significantly higher concentration of IL-5 was found in the nasal fluid of atopic patients with nasal polyposis than in non-atopic nasal polyp patients (p=0.025) and patients with allergic rhinitis (p=0.05). ECP was higher in atopic nasal polyp patients than in patients with allergic rhinitis (p<0.0001) and than in non-atopic nasal polyp patients (p<0.0001). Polyp eosinophils were higher in atopic’ than in non-atopic patients (p<0.0001) and higher than in the mucosa of patients with allergic rhinitis (p<0.0001). These however had signifi cantly more mucosal eosinophils than was found in the polyps of non-atopic patients’ (p=0.025). ECP levels in nasal fl uid and eosinophil counts in tissue specimens correlated well in all three groups of patients. Our study has shown that atopic nasal polyp patients have a higher level of eosinophilic inflammation than non-atopic patients with nasal polyps and patients with allergic rhinitis.Na histopatološkim pregledima nosni polipi, kao i nosna sluznica u alergijskom rinitisu pokazuju različite oblike pseudoslojevitog respiracijskog epitela, dok je osnovna karakteristika lamine proprije infiltracija eozinofilima. Cilj ove studije bio je usporediti koncentracije interleukina 5 (IL-5) i eozinofilnoga kationskog proteina (ECP) u nosnome sekretu pacijenata s alergijskim rinitisom, neatopičnih i atopičnih pacijenata s nosnom polipozom, kao i usporediti broj eozinofi la u tkivu nosnih polipa/nosne sluznice ovih pacijenata. Četrdeset dvoje (n=42) pacijenata, 12-ero s alergijskim rinitisom i devijacijom nosnog septuma, 17-ero neatopičnih pacijenata s nosnom polipozom i 13-ero atopičnih pacijenata s nosnom polipozom bilo je uključeno u ovu presječnu studiju. Uzorci nosnog sekreta bili su skupljeni iz nosnih šupljina svih ispitanika nekoliko dana prije kirurškog liječenja. Koncentracije IL-5 mjerene su metodom protočne citometrije, dok su koncentracije ECP mjerene komercijalnim ELISA-kitom. Eozinofi li su brojeni u svim uzorcima tkiva nosnih polipa, kao i u svim uzorcima tkiva sluznice uzetih s donje nosne školjke tijekom septoplastike. Značajno više koncentracije IL-5 izmjerene su u nosnom sekretu atopičnih pacijenata s nosnom polipozom u usporedbi s neatopičnim pacijentima s nosnom polipozom (p=0,025) i pacijentima s alergijskim rinitisom (p=0,05). Naši su rezultati pokazali više koncentracije ECP u atopičnih pacijenata s nosnom polipozom nego u pacijenata s alergijskim rinitisom (p<0,0001) i u usporedbi s nealergičnim pacijentima s nosnom polipozom (p<0,0001). Veći broj eozinofi la izbrojen je u tkivu polipa atopičnih pacijenata nego u tkivu polipa neatopičnih pacijenata (p<0,0001), kao i u sluznici pacijenata s alergijskim rinitisom (p<0,0001). U nosnoj sluznici pacijenata s alergijskim rinitisom našli smo značajno veći broj eozinofila nego u polipima neatopičnih pacijenata s nosnim polipima (p=0,025). Konačno, našli smo pozitivnu korelaciju između nivoa ECP u nosnom sekretu i broja eozinofila u uzorcima tkiva u sve tri skupine ispitanika. Zaključili smo da atopični pacijenti s nosnom polipozom imaju viši stupanj eozinofilne upale u usporedbi s neatopičnim pacijentima s nosnim polipima i s pacijentima s alergijskim rinitisom

Prognostic significance of immunohistohemical parameters and histomorphological score in assessment of degree of cervical dysplasia

Cilj:Karcinom grlića je treći po uĉestalosti u svetu meĊu malignim bolestima. Displazije epitela grlića predstavljaju niz prekursorskih lezija karcinoma grlića.Najznacajniji faktor rizika za displaziju epitela grlića jeste infekcija humanim papiloma virusima.Cilj ove disertacije je bio da se utvrdi dijagnostiĉki znaĉaj imunohistohemijskih metoda markerima p16, p63, ciklin d1 i survivin u epitelnim displazijama grlića,morfometrijske nalize i histomorfološkog skora. Metodologija:Retrospektivnom studijom je bilo obuhvaćeno 100 pacijentkinja pregledanih na Ginekološkom odeljenju Vojnomedicinske akademije u trogodišnjem periodu.Kriterijumi za ulazak u studiju su: primarna, ranije netretirana lezija, kolposkopski nalaz koji upućuje na prisustvo premaligne lezije grlića: uĉinjen citološki test epitela grlića po metodi Papanikolau koji je interpretiran po Bethesda kriterijumu sa nalazom displazije lakog stepena (low-grade squamous intraepithelial lesion,LSIL), displazije teškog stepena (high-grade sqamous intraepithelial lesion,HSIL), atipiĉnih ćelija neodreĊenog znaĉaja (atypical squamous cells of undeterminated significance,ASCUS);uĉinjena tipizacija humanih papiloma virusa reakcijom lanĉane polimerizacije sa dokaznim prisustvom virusa tipa 16,18,31,33 kod svih zena. Ovoj grupi pacijentkinja je uradjena biopsija, imunohistohemijske i morfometrijske analize bioptiranog tkiva. Primenjena su imunohistohemijska bojenja p16, p63, ciklin d1 i survivin. Analiza imunohistohemijskih reakcija je vršena svetlosnim mikroskopom u zonama displazije. Od 100 zena koje su ušle u studiju, kod kojih je prvom biopsijom dokazano prisustvo displazije (HSIL, LSIL), kod 48 zena je ucinjena ponovna biopsija u perodu do sedam meseci, na osnovu istih dijagnostickih kriterijuma kao za prvu biopsiju, gde su primenjivane sve metode istraživanja koje su korišćene u prvoj biopsiji. Kontrolnu grupu su ĉinile 12 žena,koje su se javile na kontrolni pregled bez displazije i bez verifikovane infekcije grlića materice visokoriziĉnim HPV virusima, zbog mioma, na materijalu dobijenom nakon histerektomije...Aim:Cervical cancer is the third most common cancer in the world.Cervical dysplasia, such as minor and severe dysplasia, is a group of precursor lesion of the squamous cell carcinoma. The most important etiologic risk factors for dysplasia are infection with the human papilloma virus (HPV).The aim of this study was to determine the relevance of correlation between epithelial p16,p63,ciklin d1 and survivin expression and the degree of human papilloma virus (HPV)- induced cervical epithelial dysplasia, as well as, to establish the significance of morphometric analysis of nuclear area and histomorphologic score in assessment of cervical dysplasia degrees Methodology: This retrospective study included 100 patients examined at the Department of Gynaecology of the Military Medical Academy in the period from three years.Criteria for inclusion into the study were primary, previously untreated lesions, colposcopic findings that suggest the presence of premalignant lesions of the cervix; findings from Papanikolaou cytological test, interpreted by the Bethesda criteria with the findings of LSIL, HSIL, ASCUS. HPV typisation was done by PCR with evidence of viruses type 16, 18, 31 and 33 in all women. After the bipsy of the cervical mucosa, we performed pathohistological analysis by hematoxilineosin staining, immunohistochemical analysis for p16,p63,ciklin d1 and survivin and morphometric analysis of cervical epithelium. After the first biopsy, during next seven months in group of 48 patients with dysplasias, according the same criteria for inclusion, we performed rebiopsy and uniform examination . The control group consisted of 12 women without dysplasia and without verified infection of cervical high-risk HPV, with pathological diagnosis of leiomyoma in material obtained after the hysterectomy.Criteria for exclusion from the study was previously diagnosed and treated dysplasia or squamous cell carcinoma..

Primary ovarian malignant melanoma

Background. Primary ovarian malignant melanoma is extremely rare. It usually appears in the wall of a dermoid cyst or is associated with another teratomatous component. Metastatic primary malignant melanoma to ovary from a primary melanoma elsewhere is well known and has been often reported especially in autopsy studies. Case report. We presented a case of primary ovarian malignant melanoma in a 45- year old woman, with no evidence of extraovarian primary melanoma nor teratomatous component. The tumor was unilateral, macroscopically on section presented as solid mass, dark brown to black color. Microscopically, tumor cells showed positive immunohistochemical reaction for HMB-45, melan-A and S-100 protein, and negative immunoreactivity for estrogen and progesteron receptors. Conclusion. Differentiate metastatic melanoma from rare primary ovarian malignant melanoma, in some of cases may be a histopathological diagnostic problem. Histopathological diagnosis of primary ovarian malignant melanoma should be confirmed by immunohistochemical analyses and detailed clinical search for an occult primary tumor

CLINICAL CASE REPORT OF A LARGE ANTROCHOANAL POLYP

Antrochoanal polyps are benign lesions originating from the mucosa of the maxillary sinus. Nasal obstruction and rhinorrhea are their main symptoms. Their endoscopical and radiological appearance makes them relatively easy to diagnose. These polyps are usually presented unilaterally, although bilateral presentation is also possible. We described two cases of atypically giant antrochoanal polyps: in a 15-year-old child and in a 38-year-old man. In both cases, the diagnosis was done by nasal endoscopy and computed tomography (CT) of the paranasal sinuses and supported by histopathological analysis. In the first patient, the excised polyp had the histological characteristic of an angiomatous antrochoanal polyp. Because of their unusual dimension, the combined transoral and endonasal endoscopic approach was performed for complete polyp excision. We discussed the clinical, histopathological and immunohistochemical characteristics of choanal polyps in comparison to inflammatory nasal polyps, and the applicable surgical techniques for treatment of these polyps