Human papillomavirus and cervical cancer: mechanisms of carcinogenesis, epidemiology, diagnostics and prophylaxis

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.Human papillomavirus (HPV) infecti on is esti mated to be the most common sexually transmitt ed disease and about 75-80% of sexually active men and women will be infected with HPV at some point in their lifeti me. Since the early 1980s when Harald zur Hausen (winner of the Nobel Prize in 2008 for Physiology or Medicine) detected the HPV genotypes 16 and 18 in cervical cancer, a large number of studies have provided scienti fi c evidence that HPV infection is a key factor in the development of cervical carcinoma. Malignant changes of the cervix occur if multiple conditions are met and they are defined by virus characteristics, anti viral mechanisms and by the cellular immune response of the host. Viral components important in carcinogenesis are viral genotype, intensity and persistence of the infecti on, while immunogenetic constitution of the individual, cell mediated immune responses and influence of the external factors such as drugs and diseases defi ne immunopathogenetic contributi on of the host. Key proteins involved in the development of cervical cancer are viral oncogenes, E6 and E7, which interfere with a variety of cellular processes and lead to uncontrolled cell proliferati on and immortalisation. Therefore, the importance of molecular studies dealing with HPV lies in the fact that they enable us to better understand the HPV pathogenesis and facilitate application of the acquired knowledge in the preventi on of HPV infections, development of prophylactic vaccines and introducti on of molecular diagnosti cs as a standard in the risk assessment for women exposed to the chronic HPV infection

Prevalence of Human Papillomavirus among Croatian Women Attending Regular Gynecological Visit

Human papillomavirus (HPV) infection has been identified as major risk factor for cervical intraepithelial neoplasia (CIN) and invasive cervical cancer. About 40 HPV viral types are commonly found in the genital tract. Most HPV infections resolve spontaneously, while persistent infection with oncogenic types, namely HPV 16 and 18 is necessary for CIN to occur and progress to cancer. Cervical screening is presently based on the Pap smear that is designed to diagnose precancerous lesions and cervical cancer. The aim of this study was to investigate the prevalence of HPV DNA and to determine HPV types distribution among 361 women attending regular gynecological visit. There were 205 women (29±8 years old) without determined abnormal cervical lesions and 156 women (34±15 years old) with abnormal Pap smear; low grade squamous intraepitehelial lesions (LSIL, n=69), high grade squamous intraepithelial lesions (HSIL, n=72) and atypical squamous cells of undetermined significance (ASCUS, n=15). HPV DNA detection and genotyping was performed by Hybrid Capture 2 assay and additionally by consensus and type-specific primers directed PCR. The overall prevalence of high-risk HPV (hrHPV) in women with abnormal Pap smears was 67.9% (106/156), of which in ASCUS 33.4% (5/15), LSIL 62.3% (43/69) and HSIL 80.6% (58/72). In HPV positive specimens, HPV 16 was found as predominant type in 60.4% cases, followed by HPV 31 (8.5%), HPV 33 (6.6%) and HPV 18 (3.7%). In the group of women without obvious cervical changes the overall hrHPV prevalence was 35.6% with HPV 16 found in 43.8% cases, followed by HPV 31 (17.8%), HPV 33 (9.5%) and HPV 18 (6.8%). In both study groups, women with and without cervical lesions, the prevalence of HPV of indeterminate type was 14.2% and 13.7%, respectively. Our results indicate that cervical intraepithelial lesions are largely associated with HPV type 16, followed by HPV types 31, 33, 18 and HPV of indeterminate type. Although there is a significant difference in hrHPV DNA prevalence among two groups, no significant differences between particular hrHPV types distribution were observed

HEPATITIS C IN TRANSFUSION MEDICINE

SAŽETAK Najnovije procjene upućuju na to da se svjetska prevalencija anti-HCV reaktivnosti kreće u široku rasponu, i to od 0,1% u razvijenim zemljama do 13,6% u sjevernoj Africi. U Europi prevalencija se kreće od 0,1% do najviše 4,9% u pojedinim zemljama istočne Europe. Epidemiološka istraživanja predviđaju da je približno 170 milijuna ljudi u svijetu kronično inficirano hepatitis C virusom (HCV). Na osnovi preliminarnih testiranja opće populacije i prema procjenama, Hrvatska je svrstana u zemlje s nižom HCV prevalencijom (1% – 2%), te incidencijom od 0,005%. Kronična infekcija HCV-om sve je važniji javnozdravstveni problem, poglavito kada se radi o dobrovoljnim davateljima krvi kao mogućim izvorima zaraze. Zadaća je suvremene transfuziologije liječiti bolesnika sigurnim i kvalitetnim krvnim pripravcima, a postojeći ostatni rizik za infekciju krvlju prenosivim bolestima, svesti na najmanju mjeru. U Hrvatskoj sva se darivanja krvi obvezno testiraju na HIV, HCV, HBV i sifilis. Poslijetransfuzijski hepatitis C (PTHC) jest krvlju prenosiva bolest, te je unatoč obveznu anti-HCV testiranju krvi, još uvijek moguća komplikacija transfuzijskoga liječenja u nas i u svijetu. U zemljama zapadne Europe rizik za prijenos HCV-a krvlju i krvnim pripravcima kreće se u prosjeku od 1:400.000 do 1:800.000. Razloga je tomu više: od nepostojanja simptoma i kliničkih znakova hepatitisa C u davatelja koji se nalazi u fazi inkubacije, nepostojanja idealnoga testa kojim bi se otkrili mnogobrojni podtipovi HCV virusa, te – iako u malomu postotku, postojanje seronegativnih trajnih nositelja virusa koji su doživotni kliconoše.SUMMARY Latest estimates indicate that world prevalence of anti-HCV reactivity spans widely from 0.1% in developed country to 13,6% in north Africa, while in Europe prevalence ranges from 0.1% to 4.9% in some eastern European countries. Epidemiological studies show, around 170 million people in world to be chronically infected with hepatitis C virus (HCV). Preliminary testing of general population and according to the estimates, Croatia has classified within countries with HCV prevalence of 1%–2%, and incidence rate of 0.005%. Chronic HCV infection represents increasingly significant public health problem, especially when we take in consideration blood donors as a possible source of infection. Modern transfusion medicine has an assignment to treat patients with safe and quality blood preparations while minimizing risk of infection with blood transmitted diseases. In Croatia all blood donations are mandatory tested for HIV, HCV, HBV and syphilis. Although posttransfusion hepatitis C (PTHC) is only one of the possible infections transmitted by blood, there is no reason to be more concern than in other countries of Western Europe, where risk of HCV transmission through blood and blood preparations averages from 1:400.000 to 1:800.000. Despite blood testing PTHC is still possible complication of transfusion treatments in Croatia and worldwide. Reasons for that are absence of symptoms and clinical signs of hepatitis C in incubation period of infected blood donors, absence of ideal test who would detect all cases of infected bloo

MOLECULAR DIAGNOSIS OF HEPATITIS C INFECTION

SAŽETAK Svjetska zdravstvena organizacija (WHO) prepoznala je hepatitis C kao svjetski problem. Na osnovi procjena iz godine 1999., 170 milijuna ljudi bilo je kronično inficirano hepatitis C virusom (HCV). HCV vodeći je čimbenik neizlječivih oboljenja jetre, uključujući karcinom jetre. Za otkriće HCV-a godine 1989., zaslužni su molekularno-biološki postupci na kojima se zasniva suvremena dijagnostika hepatitisa C. Dijagnostički testovi dijele se na serološke testove kojima se dokazuju anti-HCV protutijela, te na molekularne testove kojima se u inficirane osobe dokazuje, određuje i obilježuje HCV-RNA genom. Kvalitativni molekularni testovi primjenjuju se za potvrđivanje infekcije, te u kontroli darivane krvi. Praćenjem broja virusnih kopija, kvantitativni HCV-RNA testovi daju podatke o odgovoru na antivirusnu terapiju. HCV jest heterogeni virus koji se na osnovi genomske promjenljivosti svrstava u 6 osnovnih genotipova, te u više podtipova. HCV genotipizacija bitna je za kliničara, budući da se prema pojedinome genotipu može procijeniti odgovor na antivirusnu terapiju, a određivanjem HCV genotipa odabire se optimalni terapijski postupak vezan uz duljinu razdoblja terapije i dozu ribavirina.ABSTRACT The World Health Organization recognized hepatitis C (HCV) as a global health problem estimated that in 1999, over 170 million people were chronically infected with HCV. HCV is leading cause of end-stage liver disease and hepatocellular carcinoma. The discovery of hepatitis C virus (HCV) in 1989 using molecular biology methods has led to the rapid evolution of the field of HCV diagnostics. Diagnostic tests for HCV can be divided into serological assays that detect antibody to HCV and molecular assays that detect, quantify and characterize HCV-RNA genome within infected patient. Qualitative molecular nucleic acid tests are used for confirmation of HCV infection and for screening blood donation. Quantitative HCV-RNA tests provide prognostic information for monitoring the response to antiviral therapy. HCV is heterogeneous virus with six distinct genotypes and numerous subtypes. HCV genotype tests are important clinically because they predict most accurately the chance of antiviral response and are routinely used for selecting treatment regimens regarding the duration of interferon therapy and ribavirin dosage

Distribution of Human Papillomavirus Types in Different Histological Subtypes of Cervical Adenocarcinoma

Little information is available regarding distribution of HPV types in different histological subtypes of adenocarcinoma (AC). Thus, in this study we examined the frequency of high-risk (hr) HPV types in AC, adenocarcinoma in situ (AIS) and adenosquamous carcinoma (ADSQ). A total of 102 cases of primary cervical adenocarcinoma (26 AIS and 76 invasive AC) obtained from pathology files from 1995–2006 were histologically subtyped. Our results demonstrated that endocervical type occupied the major subtype of AC (22/66) followed by ADSQ (17/66) where as in the group of AIS endocervical type (12/23) was followed by intestinal type of AIS (7/23). Successful DNA extraction was obtained in 89 samples; 81 out of 89 (91.0%) tested positive for HPV DNA. The prevalence of HPV DNA in AIS, AC and ADSQ was 91.3% (21/23), 90.9% (60/66) and 94.1% (16/17), respectively. We found HPV 18 type to be the most predominant type in AIS (11/21) and AC (17/60) followed by HPV of undeternmined type in AIS (3/21) and HPV 16 in AC (9/60) as the sole viral type. HPV 18 was most frequently detected type in all histological subtypes of AIS and AC. We have detected HPV DNA in all 5 samples of clear cell carcinoma (CCC), although other studies have reported a highly variable prevalence of HPV DNA in CCC. The most prevalent HPV type in ADSQ was HPV-16 followed by HPV 33 as single type. The observed overall predominance of HPV 18 in AIS ( 2= 6.109, p£ 0.025) and AC ( 2 = 8.927, p£0.01) as well as of HPV 16 in ADSQ ( 2 = 10.164, p £ 0.01) was statistically significant. Our data revealed statistically significant predominance of single hrHPV infections in AIS (16/21; 2 = 11.523, p £ 0.001) and AC (37/60; 2 = 6.533, p £ 0.025) whereas multiple hrHPV infections were more abundant in AC comparing to AIS (23/81and 5/81, respectively; 2 = 13.989, p £ 0.001)

Hypoxia inducible factor-1α correlates with vascular endothelial growth factor A and C indicating worse prognosis in clear cell renal cell carcinoma

<p>Abstract</p> <p>Background</p> <p>The role of angiogenesis in the pathogenesis of renal cell carcinoma is well recognized, however, the influence of tumor cells in this activity has not yet been fully clarified. The aim of this study was to analyze the expression of hypoxia inducible factor-1α (HIF-1α), a regulatory factor of angiogenic switch, in comparison to vascular endothelial growth factor A and C (VEGF-A and VEGF-C), recognized to be involved in blood and lymph vessel neoangiogenesis, with potential association in the prognosis of patients with renal cell carcinoma.</p> <p>Methods</p> <p>Ninety-four patients with diagnosis of clear cell renal cell carcinomas (CCRCC), all clinicopathological characteristics and overall survival were unrolled in this study. Immunohistochemicaly VEGF-A, VEGF-C, HIF-1α and Ki67 were detected on tumor cells and the staining was performed on tissue microarrays (TMA). The staining was evaluated as a percentage of cytoplasmic or nuclear positive tumor cells.</p> <p>Results</p> <p>Variable expression of all three proteins was confirmed. Both angiogenic factors demonstrated perimembranous or diffuse cytoplasmic staining, with diffuse pattern positively associated (p < 0.001). Nuclear HIF-1α expression (nHIF-1α) showed inverse correlation with diffuse cytoplasmic VEGF-A (p = 0.002) and VEGF-C (p = 0.053), while cytoplasmic HIF-1α expression (cHIF-1α) showed positive correlation with diffuse staining of both angiogenic factors (p < 0.001; p < 0.001, respectively). In comparison to clinicopathological characteristics, a higher nuclear grade (p = 0.006; p < 0.001, respectively), larger tumor size (p = 0.009; p = 0.015, respectively), higher stage (p = 0.023; p = 0.027, respectively) and shorter survival (p = 0.018; p = 0.024, respectively) were associated with overexpression of cHIF-1α and diffuse cytoplasmic VEGF-A expression. In contrary, overexpression of nHIF-1α was associated with better diagnostic parameters i.e. lower nuclear grade (p = 0.006), smaller tumor size (p = 0.057), and longer survival (p = 0.005).</p> <p>Conclusion</p> <p>Overexpression of VEGF-A and cHIF-1α in tumor cells highlights a more aggressive subtype of CCRCC that might have some clinical implications. The significance of nHIF-1α expression associated with better differentiated tumors should be further elucidated.</p

Fifty Years of the Department of Forensic Medicine and Criminalistics, Faculty of Medicine, University of Rijeka

Prvog veljače 2016. godine Zavod za sudsku medicinu i kriminalistiku Medicinskog fakulteta Sveučilišta u Rijeci proslavio je svoj 50. rođendan. U pedeset godina aktivnosti Zavod za sudsku medicinu i kriminalistiku razvijao se i napredovao u stručnom i znanstvenom smislu, suočavajući se s brojnim profesionalnim izazovima. Djelatnosti Zavoda obuhvaćaju nastavne, stručne i znanstvene aktivnosti. Nastavnici Zavoda uključeni su u programe obavezne i izborne izobrazbe studenata Medicine, Dentalne medicine i Fakulteta zdravstvenih studija. Stručne aktivnosti Zavoda su provođenje sudsko-medicinskih obdukcija, mrtvozornička služba, sudska vještačenja, procjene težine tjelesnih ozljeda, toksikološke i molekularne analize DNK-a. Stručna djelatnost provodi se u suradnji sa sudovima, državnim odvjetništvom, policijom i Uredom za javno zdravstvo Primorsko-goranske županije. Znanstvenom aktivnošću i razvojem novih metodoloških pristupa Zavod nastoji pridonijeti unapređenju sudskomedicinske prakse u Hrvatskoj.On the first day of February 2016, the Department of Forensic Medicine and Criminalistics, School of Medicine, University of Rijeka celebrated its 50th birthday. At fifty years of activity, the Department evolved and progressed in both professional and scientific senses, confronting the many professional challenges. Activities of the Institute include teaching as well as many professional and scientific obligations. Department’s teachers are included in mandatory and elective programs of training medical and dental medicine students and they cover the classes in Faculty of Health Studies. Professional activities comprise of the performing forensic autopsies, conducting a medical examiner’s office, expert witnessing, assessing the severity of injuries, performing toxicological and molecular DNA analyses. In performing professional projects, collaboration with the State’s judiciary (i.e. courts, state’s attorney, and the police) or the County’s Office of Public Health is the most crucial. With its scientific activities and the development of new methodological approaches, the Institute seeks to contribute to the advancement of forensic practice in Croatia

Mitochondrial DNA Heritage of Cres Islanders – Example of Croatian Genetic Outliers

Diversity of mitochondrial DNA (mtDNA) lineages of the Island of Cres was determined by high-resolution phylogenetic analysis on a sample of 119 adult unrelated individuals from eight settlements. The composition of mtDNA pool of this Island population is in contrast with other Croatian and European populations. The analysis revealed the highest frequency of haplogroup U (29.4%) with the predominance of one single lineage of subhaplogroup U2e (20.2%). Haplogroup H is the second most prevalent one with only 27.7%. Other very interesting features of contemporary Island population are extremely low frequency of haplogroup J (only 0.84%), and much higher frequency of haplogroup W (12.6%) comparing to other Croatian and European populations. Especially interesting finding is a strikingly higher frequency of haplogroup N1a (9.24%) presented with African/south Asian branch almost absent in Europeans, while its European sister-branch, proved to be highly prevalent among Neolithic farmers, is present in contemporary Europeans with only 0.2%. Haplotype analysis revealed that only five mtDNA lineages account for almost 50% of maternal genetic heritage of this island and they present supposed founder lineages. All presented findings confirm that genetic drift, especially founder effect, has played significant role in shaping genetic composition of the isolated population of the Island of Cres. Due to presented data contemporary population of Cres Island can be considered as genetic »outlier« among Croatian populations