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

In vitro potencijal antagonista Bacillus spp. za suzbijanje fitopatogena Xanthomonas euvesicatoria

Bacterial pathogen Xanthomonas euvesicatoria, principal causer of bacterial spot, represents a significant problem in agricultural practice due to high yield losses in the production of pepper and tomato. The development of resistance to copper pesticides has shifted research, in the field of its suppression, towards biopesticides. In this study, several Bacillus strains were tested against Xanthomonas euvesicatoria strains, isolated from pepper leaves with symptoms of bacterial spot, to select a sufficiently effective antagonist. When it comes to the testing of cultivation broth, containing biomass of tested antagonists, the best results were achieved using isolate Bacillus sp3. On the other hand, when biomass-free supernatants, containing produced antimicrobial compounds, were tested, Bacillus sp1 and Bacillus sp2 have shown the highest antimicrobial activity. The results of this study represent a basis for further development of bioprocess solutions for the production of biopesticides based on Bacillus spp. biomass or antimicrobial compounds, showing high efficiency in suppression of pepper bacterial spot.Bakterijski patogen Xanthomonas euvesicatoria predstavlja glavnog uzročnika bakteriozne pegavosti, stoga predstavlja značajan problem u poljoprivrednoj praksi usled izazivanja velikih gubitaka prinosa u proizvodnji paprike i paradajza. Uobičajena sredstva za suzbijanje i kontrolu ovog fitopatogena predstavljaju preparati na bazi bakra. Međutim, razvoj rezistentnosti prema pesticidima na bazi bakra doveo je do zaokreta u polju suzbijanja ovih patogena ka primeni biopesticida, odnosno bioloških kontrolnih agensa. Bakterije roda Bacillus i njihovi metaboliti sa izraženom antimikrobnom aktivnošću protiv ciljanih fitopatogena predstavljaju najperspektivnije aktivne komponente biokontrolnih preparata za zaštitu bilja. U ovom istraživanju nekoliko sojeva roda Bacillus ispitano je sa ciljem odabira antagonista dovoljno efikasnih u suzbijanju sojeva Xanthomonas euvesicatoria, koji su izolovani sa listova paprike sa simptomima bakteriozne pegavosti. Prilikom testiranja antimikrobne aktivnosti uzoraka kultivacionih tečnosti, koji sadrže i biomasu testiranih antagonista, najbolji rezultati u suzbijanju testiranih fitopatogena su postignuti primenom izolata Bacillus sp3. Sa druge strane, prilikom testiranja antimikrobne aktivnosti supernatanata oslobođenih biomase antagonista, koji sadrže samo produkovana antimikrobna jedinjenja, izolati Bacillus sp1 i Bacillus sp2 su pokazali najveću antimikrobnu aktivnost protiv fitopatogena Xanthomonas euvesicatoria. Rezultati ovog istraživanja predstavljaju osnovu za dalji razvoj bioprocesnih rešenja za proizvodnju biopesticida na bazi biomase ili antimikrobnih jedinjenja koja proizvode antagonisti roda Bacillus, a koji pokazuju visoku efikasnost u suzbijanju bakteriozne pegavosti paprike

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

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)

Selection of antagonists for biocontrol of xanthomonas euvesicatoria

Xanthomonas euvesicatoria is a worldwide causer of pepper bacterial spot, a bacterial plant disease responsible for massive losses of fresh pepper fruits. Considering the current problems in management of bacterial plant diseases, biological control using antagonistic microbial strains with high potential for plant pathogens suppression emerges as a possible solution. The aim of this study was to select suitable antagonists for suppression of X. euvesicatoria among the bacteria, yeast and fungi from the genera Pseudomonas, Lactobacillus, Saccharomyces and Trichoderma, based on in vitro antimicrobial activity testing using the diffusion disc method. The results of this study have revealed that cultivation broth samples of the antagonists Lactobacillus MK3 and Trichoderma reseii QM 9414, as well as supernatant samples of the antagonist Pseudomonas aeruginosa I128, have showed significant potential to be applied in biological control of X. euvesicatoria. Further research would be required to formulate suitable cultivation medium and optimize bioprocess conditions for production of the proposed pepper bacterial spot biocontrol agents

Prevalence of Human Papillomavirus Genotypes in Cervical Cancer and Precursor Lesions

There are no data obtained in biopsy material on the prevalence of human papillomavirus (HPV) and HPV genotypes in Croatian women with cervical carcinoma and precursor lesions. Therefore, the prevalence of HPV and HPV genotypes was investigated in archival material of cervical carcinoma and precursor lesions kept at Department of Pathology, School of Medicine, University of Rijeka. DNA was isolated from formalin fixed, paraffin embedded tissue, histologically classified as cervical intraepithelial neoplasia (CIN) III (n=43), squamous cell carcinoma (SCC) (n=54) and adenocarcinoma (ADC) (n=40). HPV testing was performed by polimerase chain reaction (PCR) using generic and genotype specific primers. The prevalence of HPV DNA was 93.02%, 92.59%, and 92.5% in CIN III, SCC and ADC, respectively. In CIN III and SCC, HPV-16 was the most common high-risk genotype, identified in 65% and 52%, followed by HPV-18 in 22.5% and 28% of cases, respectively. HPV-18 showed a statistically significant prevalence in ADC (67.6%) as compared with SCC (2=9.924; p 0.01). Study results revealed a high prevalence of HPV-DNA in examined cervical lesions (>90%). HPV-16 predominated in SCC and HPV-18 in ADC. Single infection was more frequently present than multiple infections in all three histological groups