HOST AND BACTERIAL FACTORS IN PATHOGENESIS OF LISTERIOSIS

SAŽETAK Listerioza jest zoonoza do koje u većem broju slučajeva dolazi nakon konzumacije zagađenih namirnica. U zdravih odraslih osoba infekcija bakterijom Listeria monocytogenes posljeduje prolaznom, asimptomatskom kolonizacijom crijeva, ili slikom neinvazivna febrilnoga gastroenteritisa. Infekcija fetusa, novorođenčeta, ili imunokompromitirane osobe, može posljedovati razvojem invazivne infekcije praćene visokim postotkom smrtnosti. Zbog svojstva unutarstaničnoga parazitiranja, bakterija Listeria monocytogenes dugi se niz godina koristi kao oruđe za proučavanje stanične biologije i mehanizama stanične imunosti.ABSTRACT Listeriosis is a zoonosis usually transmitted through contaminated food or vertically from mother to foetus. In an immunocompetent, adult person the infection results in transitory asymptomatic gastrointestinal colonization or febrile, non-invasive gastroenteritis. The infection of the foetus, newborn or immunocompromised person can result in a serious invasive disease with high mortality rate. As a facultative intracellular pathogen, Listeria monocytogenes is widely used as model for studying cellular biology and mechanisms of cellular immunity

Evaluation of the HPV L1 Capsid Protein in Prognosis of Mild and Moderate Dysplasia of the Cervix Uteri

Cervical intraepithelial neoplasia (CIN) can be detected in the cytologic smears years before invasive squamous cancer arises, but no reproducible morphologic criteria exist to predict behavior of cervical lesions. The possibility of predicting the clinical course of cervical lesions could be of high value in clinical practice and some women will spare of unnecessary treatment. HPV L1 capsid protein represents about 90% of the total protein on the surface of the virus and can be detected in mild to moderate dysplasia and rarely in severe dysplasia. The purpose of the study was to evaluate the use of immunodetection of HPV L1 protein on archival Pap smears with findings of mild and moderate dysplasia in predicting its clinical course. Immunochemical analyses with L1 antibody revealed positively stained nuclei of squamous epithelial cells in 56 of 114 smears (49.1%). The staining results were correlated with follow-up smears or with histologic verification. Regression (negativisation of the Pap smear for 24 months or longer) was noticed in 31 of 56 (55.4%) L1-positive cases and in 20 of 58 (34.5%) L1-negative cases. Persistent disease occured in 13 (23.2%) L1-positive cases and in 14 (24.1%) L1-negative cases. Progressive disease occured in 12 (21.4%) L1-positive cases and in 24 (41.4%) L1-negative cases. The difference in the clinical course between the L1-positive and L1-negative patients was statistically significant (p=0.025). Also, the difference in the clinical course of the L1-negative staining in the under-30 and over-30 years age group was statistically significant (p=0.04). For conclusion, our data confirm that immunostaining for HPV L1 capsid protein could offer prognostic information about mild and moderate intraepithelial cervical squamous lesions

Evaluation of the HPV L1 Capsid Protein in Prognosis of Mild and Moderate Dysplasia of the Cervix Uteri

Cervical intraepithelial neoplasia (CIN) can be detected in the cytologic smears years before invasive squamous cancer arises, but no reproducible morphologic criteria exist to predict behavior of cervical lesions. The possibility of predicting the clinical course of cervical lesions could be of high value in clinical practice and some women will spare of unnecessary treatment. HPV L1 capsid protein represents about 90% of the total protein on the surface of the virus and can be detected in mild to moderate dysplasia and rarely in severe dysplasia. The purpose of the study was to evaluate the use of immunodetection of HPV L1 protein on archival Pap smears with findings of mild and moderate dysplasia in predicting its clinical course. Immunochemical analyses with L1 antibody revealed positively stained nuclei of squamous epithelial cells in 56 of 114 smears (49.1%). The staining results were correlated with follow-up smears or with histologic verification. Regression (negativisation of the Pap smear for 24 months or longer) was noticed in 31 of 56 (55.4%) L1-positive cases and in 20 of 58 (34.5%) L1-negative cases. Persistent disease occured in 13 (23.2%) L1-positive cases and in 14 (24.1%) L1-negative cases. Progressive disease occured in 12 (21.4%) L1-positive cases and in 24 (41.4%) L1-negative cases. The difference in the clinical course between the L1-positive and L1-negative patients was statistically significant (p=0.025). Also, the difference in the clinical course of the L1-negative staining in the under-30 and over-30 years age group was statistically significant (p=0.04). For conclusion, our data confirm that immunostaining for HPV L1 capsid protein could offer prognostic information about mild and moderate intraepithelial cervical squamous lesions

Villoglandular Papillary Adenocarcinoma of the Uterine Cervix with Aggressive Clinical Course – A Case Report

Villoglandular papillary adenocarcinoma (VGA) of cervix is an uncommon but well recognized histologic subtype of cervical adenocarcinoma which usually affects young women. Based on the favorable outcomes reported in most previous cases the tumor is generally considered to have an indolent clinical course with excellent prognosis. We present a case of a 22-year-old woman admitted at our Department for glandular abnormality on cervical smear and episodes of vaginal discharge. In the Pap smear, the cytological features were suspicious but not diagnostic of adenocarcinoma, therefore reported as atypical glandular cells (AGC). Histological examination confirmed VGA associated with lymphovascular space invasion. The patient underwent radical operative procedure. Intraoperative cytologic examination detected pelvic lymph nodes metastasis. The patient was confirmed to be in an advanced stage – III B (FIGO). During a two years follow-up period a rapid dissemination of the tumor occurred and resulted with a fatal outcome. Although VGA has been reported to have a favorable prognosis, several cases with lymph node involvement have already been described. Cervical smears examination would be helpful for an early diagnosis of VGA, however the cytologic recognition is often difficult. Further investigation of the pathogenesis, diagnosis and therapy of the tumor is needed

Villoglandular Papillary Adenocarcinoma of the Uterine Cervix with Aggressive Clinical Course – A Case Report

Villoglandular papillary adenocarcinoma (VGA) of cervix is an uncommon but well recognized histologic subtype of cervical adenocarcinoma which usually affects young women. Based on the favorable outcomes reported in most previous cases the tumor is generally considered to have an indolent clinical course with excellent prognosis. We present a case of a 22-year-old woman admitted at our Department for glandular abnormality on cervical smear and episodes of vaginal discharge. In the Pap smear, the cytological features were suspicious but not diagnostic of adenocarcinoma, therefore reported as atypical glandular cells (AGC). Histological examination confirmed VGA associated with lymphovascular space invasion. The patient underwent radical operative procedure. Intraoperative cytologic examination detected pelvic lymph nodes metastasis. The patient was confirmed to be in an advanced stage – III B (FIGO). During a two years follow-up period a rapid dissemination of the tumor occurred and resulted with a fatal outcome. Although VGA has been reported to have a favorable prognosis, several cases with lymph node involvement have already been described. Cervical smears examination would be helpful for an early diagnosis of VGA, however the cytologic recognition is often difficult. Further investigation of the pathogenesis, diagnosis and therapy of the tumor is needed

Cervical cytology (Pap test) – terminology and importance in screening for cervical cancer

. Papa-test je najbolji morfološki test probira za rak koji pomoću analize citologije vrata maternice otkriva predstadije i rane stadije raka vrata maternice. U Hrvatskoj se koristi u okviru oportunističkog probira, a od 1. 12. 2012. u organiziranom nacionalnom probiru. U svrhu klasifikacije i standardizacije citoloških nalaza vrata maternice u cijeloj Hrvatskoj koristi se modifikacija Bethesda klasifikacije. Obrazac sadrži kliničke podatke o pacijentici koje ispunjava ginekolog i citološki nalaz koji na osnovi analize Papa-testa pod svjetlosnim mikroskopom izdaje klinički citolog. U ovom preglednom radu opisuju se citomorfološke osobine abnormalnih pločastih i cilindričnih stanica vrata maternice te njihov klinički značaj. Zbog dokazane povezanosti humanog papiloma-virusa (HPV) s rakom vrata maternice sve češće se detekcija visokorizičnog HPV-a uvodi u probir za rak vrata maternice. U novije vrijeme metode temeljene na biološkim biljezima primjenjuju se na citološkim uzorcima u svrhu poboljšanja osjetljivosti i specifičnosti citologije u otkrivanju lezija visokog stupnjaThe Pap test is considered to be the best morphological screening test for cancer which reveals precancerous lesions and early stages of cervical cancer by analyzing cervical cytology. In Croatia, it is used in the context of opportunistic screening and since December 1, 2012 it has been used within an organized national screening programme. In order to classify and unify the terminology in cervical cytology reporting throughout Croatia a modification of the Bethesda classification is used. The form contains clinical data on the patient which is fulfilled by the gynecologist and the cytological report based on light microscope analysis of Pap smears which is made by the cytopathologist. In this article we will describe the cytological characteristics of abnormal squamous and glandular cells as well as their clinical significance. Since the connection between human papillomavirus (HPV) and cervical cancer has been clearly confirmed the detection of high-risk HPV is increasingly implemented in cervical cancer screening. More recently, in order to improve the sensitivity and specificity of cytology methods in detecting high-grade lesions, biological markers are applied on cytological specimens of the cervi

The role of cytology in preoperative assessment od primary fallopian tube carcinoma: case report

Cilj: Prikazati slučaj pacijentice s primarnim seroznim karcinomom jajovoda, opisati dijagnostički postupak te analizirati ulogu citološke dijagnostike u detekciji ovog tipa tumora. Prikaz slučaja: Pedesetogodišnja pacijentica prethodno je obrađivana u drugoj ustanovi zbog bolova u trbuhu i radiološki dokazanih uvećanih supraklavikularnih, aortokavalnih i ilijačnih limfnih čvorova. Nakon učinjene laparoskopske ekstirpacije limfnih čvorova zbog sumnje na limfoproliferativnu bolest, patohistološkom i imunohistokemijskom analizom utvrđeno je da se radi o metastatskom adenokarcinomu, vjerojatno podrijetla iz ginekološkog sustava ili dojke. Radiološkom obradom dojki nisu pronađene promjene suspektne na malignitet. PET/CT analizom (pozitronska emisijska tomografija/kompjutorizirana tomografija) uočeno je pojačano nakupljanje radiofarmaka u predjelu vrata i tijela maternice. U preoperativnoj obradi učinjena je citološka punkcija uvećanog supraklavikularnog limfnog čvora te je imunocitokemijskom analizom utvrđeno da se radi o metastatskom seroznom adenokarcinomu. U Papa-testu pronađene su maligne stanice jednake morfologije, bez tumorske dijateze, što je upućivalo na ekstrauterini adenokarcinom, a sugerirano je podrijetlo jajnika ili jajovoda. Učinjena je eksplorativna kiretaža, ali je patohistološki nalaz bio uredan. Pacijentica je predviđena za histerektomiju s obostranom adnekstektomijom. Tijekom operativnog zahvata uočeno je proširenje desnog jajovoda tumorskim tkivom koje je probijalo stijenku jajovoda. Učinjena je lavaža peritonealne šupljine u kojoj su pronađene maligne stanice adenokarcinoma. Patohistološka analiza potvrdila je da se radi o primarnom seroznom karcinomu jajovoda s metastazama u jajnicima. Zaključak: Zahvaljujući brzini i preciznosti primijenjenih citodijagnostičkih pretraga dijagnostičko-terapijski postupak pravilno je usmjeren i pravovremeno proveden. Prikazani slučaj potvrđuje da citološka analiza može pridonijeti dijagnostici tumora s neuobičajenom kliničkom prezentacijom ili tumora nejasnog podrijetla.Aim: To report a case of a patient with primary serous carcinoma of the fallopian tube, describe the diagnostic pathway and analyze the role of cytology in the detection of this tumor type. Case report: A 50-year-old patient was previously treated in another hospital for abdominal pain and radiologically confirmed enlargement of supraclavicular, aortocaval and iliac lymph nodes. A lymphoproliferative disorder was suspected and laparoscopic lymph node excision was performed. Histopathological and immunohistochemical analysis revealed a metastatic adenocarcinoma probably of gynecologic or breast cancer origin. Radiographic examination of the breast did not confirm any lesion suspicious for malignancy. Positron emission tomography/computed tomography observed increased radiotracer uptake involving the uterine cervix and corpus. Preoperative fine-needle aspiration cytology and immunocytochemical analysis of an enlarged supraclavicular lymph node was done and metastatic serous adenocarcinoma was confirmed. In the Pap smears malignant cells of equal morphology were found without tumor diathesis indicating an extra-uterine adenocarcinoma. It was suggested to consider a tumor of ovarian or fallopian tube origin. A total hysterectomy with bilateral salpingo-oophorectomy was performed. During surgery an enlargement of the was observed due to tumor growth extending through the tubal wall. The presence of malignant adenocarcinoma cells was found in the peritoneal washing. Histopathological analysis confirmed a primary serous carcinoma of the fallopian tubes with metastatic spread to the ovaries. Conclusion: Applying fast and precise cytology and immunocytochemistry techniques a correct diagnostic and therapeutic approach can be performed and implemented on time. The present case confirms that cytology is a wellestablished and widely accepted method for diagnosing tumors with unusual clinical presentation or unknown origin

EXPRESSION OF E-CADHERIN AND β-CATENIN IN EPITHELIAL-MESENCHYMAL TRANSITION AND THEIR PROGNOSTIC VALUE IN ENDOMETRIAL CARCINOMA

Cilj istraživanja: Ključni mehanizam odgovoran za progresiju karcinoma endometrija (KE) je aktivacija epitelno-mezenhimalne tranzicije (EMT), procesa kojim tumorske stanice stječu mezenhimalni fenotip. Glavnim obilježjem EMT smatra se gubitak adhezijske molekule E-kaderina, nuklearna akumulacija β-katenina i stjecanje N-kaderina. Takve tumorske stanice ulaze u kaskadu invazija-metastaziranje, a u metastazama prolaze obrnuti proces mezenhimalno-epitelne tranzicije (MET). Na histološkom nivou malo je podataka o značaju aberantnog izražaja ovih proteina u KE. Ispitanici i metode: U istraživanje je uključeno 86 bolesnica. Za imunohistokemijsku analizu napravljeni su tkivni rezovi primarnih tumora KE i njihovih metastaza te određen postotak i jačina obojanosti tumorskih stanica na E-kaderin, β-katenin i N-kaderin u centru tumora (CT), na invazivnoj fronti (IF), u limfovaskularnom prostoru (LVP) te u pripadajućim metastazama. Rezultati: Utvrđen je značajan pad izražaja E-kaderina od CT prema IF (p< 0.0001) uz istovremeno pojačan izražaj N-kaderina (p=0.005) što govori u prilog nastupu procesa EMT. Uočeno je smanjenje membranskog β-katenina od CT prema IF i LVP, dok nuklearni izražaj β-katenina nije potvrđen kao značajni pokazatelj EMT. Usporedbom u odnosu na IF u metastazama je dokazan ponovni izražaj E-kaderina (p=0.049) i gubitak N-kaderina (p=0.008) što govori u prilog aktivaciji MET. Negativni membranski β-katenin na IF povezan je s dubokom invazijom miometrija (p=0.004) i značajno kraćim preživljenjem bolesnica (p=0.0439). Zaključak: Rezultati ovog istraživanja doprinijeli su boljem razumijevanju E-kaderina, N-kaderina i β-katenina u procesu EMT i MET u KE. Imunohistokemijska analiza ispitivanih biljega dokazala je da postoji heterogenost u njihovom izražaju unutar tumora i u metastazama te poduprla hipotezu da je IF ključna tumorska regija u kojoj nastupa EMT.Objectives: The key mechanism responsible for the progression of endometrial carcinoma (EC) is the activation of epithelial-mesenchymal transition (EMT) a process which helps epithelial cells to acquire mesenchymal phenotype. One of the hallmarks of EMT is the loss of the adhesion molecule E-cadherin, nuclear accumulation of β-catenin and the acquisition of N-cadherin. Such tumor cells enter the invasion-metastasis cascade; while through the process of mesenchymal-epithelial transition (MET) re-establish their epithelial phenotype at distant sites. Little is known about aberrant expression of these proteins at histological level in EC. Patients and Methods: The study included 86 patients. For the immunohistochemical analysis tissue samples of primary tumors and their metastases were selected and the expression of E-cadherin, N-cadherin and β-catenin on the basis of percentage of positive tumor cells and staining intensity assessed in the central area (CA) of the tumor, at the invasive front (IF), in lymphovascular spaces (LVS) and metastases. Results: We found a significantly reduced expression of E-cadherin at the IF when compared to CA (p <0.0001) and a simultaneous higher expression of N-cadherin (p=0.005) indicating the appearance of the EMT process. A less significant reduction of positive membrane β-catenin from CA to IF was observed, while nuclear β-catenin expression was not correlated to EMT. We confirmed a re-expression of E-cadherin in metastases when compared to the IF (p=0.049) and a simultaneous significant loss of N-cadherin (p=0.008) suggesting the activation of MET. Negative membrane β-catenin at the IF was associated with deep myometrial invasion (p=0.004) and significantly shorter patient survival (p=0.0439). Conclusion: The results of our research have contributed to a better understanding of the role of E-cadherin, N-cadherin and β-catenin in the EMT and MET processes in EC. The immunohistochemical analysis of these markers confirmed that there is great heterogeneity within tumor areas in their expression as well as in metastases and support the hypothesis that the IF is the main tumor region in which EMT occurs

EXPRESSION OF E-CADHERIN AND β-CATENIN IN EPITHELIAL-MESENCHYMAL TRANSITION AND THEIR PROGNOSTIC VALUE IN ENDOMETRIAL CARCINOMA

Cilj istraživanja: Ključni mehanizam odgovoran za progresiju karcinoma endometrija (KE) je aktivacija epitelno-mezenhimalne tranzicije (EMT), procesa kojim tumorske stanice stječu mezenhimalni fenotip. Glavnim obilježjem EMT smatra se gubitak adhezijske molekule E-kaderina, nuklearna akumulacija β-katenina i stjecanje N-kaderina. Takve tumorske stanice ulaze u kaskadu invazija-metastaziranje, a u metastazama prolaze obrnuti proces mezenhimalno-epitelne tranzicije (MET). Na histološkom nivou malo je podataka o značaju aberantnog izražaja ovih proteina u KE. Ispitanici i metode: U istraživanje je uključeno 86 bolesnica. Za imunohistokemijsku analizu napravljeni su tkivni rezovi primarnih tumora KE i njihovih metastaza te određen postotak i jačina obojanosti tumorskih stanica na E-kaderin, β-katenin i N-kaderin u centru tumora (CT), na invazivnoj fronti (IF), u limfovaskularnom prostoru (LVP) te u pripadajućim metastazama. Rezultati: Utvrđen je značajan pad izražaja E-kaderina od CT prema IF (p< 0.0001) uz istovremeno pojačan izražaj N-kaderina (p=0.005) što govori u prilog nastupu procesa EMT. Uočeno je smanjenje membranskog β-katenina od CT prema IF i LVP, dok nuklearni izražaj β-katenina nije potvrđen kao značajni pokazatelj EMT. Usporedbom u odnosu na IF u metastazama je dokazan ponovni izražaj E-kaderina (p=0.049) i gubitak N-kaderina (p=0.008) što govori u prilog aktivaciji MET. Negativni membranski β-katenin na IF povezan je s dubokom invazijom miometrija (p=0.004) i značajno kraćim preživljenjem bolesnica (p=0.0439). Zaključak: Rezultati ovog istraživanja doprinijeli su boljem razumijevanju E-kaderina, N-kaderina i β-katenina u procesu EMT i MET u KE. Imunohistokemijska analiza ispitivanih biljega dokazala je da postoji heterogenost u njihovom izražaju unutar tumora i u metastazama te poduprla hipotezu da je IF ključna tumorska regija u kojoj nastupa EMT.Objectives: The key mechanism responsible for the progression of endometrial carcinoma (EC) is the activation of epithelial-mesenchymal transition (EMT) a process which helps epithelial cells to acquire mesenchymal phenotype. One of the hallmarks of EMT is the loss of the adhesion molecule E-cadherin, nuclear accumulation of β-catenin and the acquisition of N-cadherin. Such tumor cells enter the invasion-metastasis cascade; while through the process of mesenchymal-epithelial transition (MET) re-establish their epithelial phenotype at distant sites. Little is known about aberrant expression of these proteins at histological level in EC. Patients and Methods: The study included 86 patients. For the immunohistochemical analysis tissue samples of primary tumors and their metastases were selected and the expression of E-cadherin, N-cadherin and β-catenin on the basis of percentage of positive tumor cells and staining intensity assessed in the central area (CA) of the tumor, at the invasive front (IF), in lymphovascular spaces (LVS) and metastases. Results: We found a significantly reduced expression of E-cadherin at the IF when compared to CA (p <0.0001) and a simultaneous higher expression of N-cadherin (p=0.005) indicating the appearance of the EMT process. A less significant reduction of positive membrane β-catenin from CA to IF was observed, while nuclear β-catenin expression was not correlated to EMT. We confirmed a re-expression of E-cadherin in metastases when compared to the IF (p=0.049) and a simultaneous significant loss of N-cadherin (p=0.008) suggesting the activation of MET. Negative membrane β-catenin at the IF was associated with deep myometrial invasion (p=0.004) and significantly shorter patient survival (p=0.0439). Conclusion: The results of our research have contributed to a better understanding of the role of E-cadherin, N-cadherin and β-catenin in the EMT and MET processes in EC. The immunohistochemical analysis of these markers confirmed that there is great heterogeneity within tumor areas in their expression as well as in metastases and support the hypothesis that the IF is the main tumor region in which EMT occurs

Činitelji Bakterije I Domaćina U Patogenezi Listerioze

Listeriosis is a zoonosis usually transmitted through contaminated food or vertically from mother to foetus. In an immunocompetent, adult person the infection results in transitory asymptomatic gastrointestinal colonization or febrile, non-invasive gastroenteritis. The infection of the foetus, newborn or immunocompromised person can result in a serious invasive disease with high mortality rate. As a facultative intracellular pathogen, Listeria monocytogenes is widely used as model for studying cellular biology and mechanisms of cellular immunity.Listerioza jest zoonoza do koje u većem broju slučajeva dolazi nakon konzumacije zagađenih namirnica. U zdravih odraslih osoba infekcija bakterijom Listeria monocytogenes posljeduje prolaznom, asimptomatskom kolonizacijom crijeva, ili slikom neinvazivna febrilnoga gastroenteritisa. Infekcija fetusa, novorođenčeta, ili imunokompromitirane osobe, može posljedovati razvojem invazivne infekcije praćene visokim postotkom smrtnosti. Zbog svojstva unutarstaničnoga parazitiranja, bakterija Listeria monocytogenes dugi se niz godina koristi kao oruđe za proučavanje stanične biologije i mehanizama stanične imunosti