Prevalence of Human Papillomavirus (HPV) among Females in the General Population of the Split and Dalmatia County and Its Association with Genital Microbiota and Infections: A Prospective Study.

We conducted a prospective study with the aim of determining HPV prevalence and type distribution in the general female population of Southern Croatia (SDC), and to detect the presence of other microorganisms in the lower part of the female reproductive system and their possible influence on the frequency of HPV infection. Data were collected during routine check-up exams. All participants were examined by a gynecologist, and cervico-vaginal scrapings/swabs were collected, for cytological (Pap smear) and microbiological (for bacterial growth, genital mycoplasmas, chlamydia, and HPV) analysis. Informed consent was obtained from all participants with accompanying questionnaire. A total of 1050 asymptomatic women living in SDC participated in the study during a one-year period, and 107 of them (10.2%) had HR-HPV infection. We found that the presence of some bacteria (Ureaplasma, Chlamydia, and Gardnerella) in the lower part of the female genital system has a positive correlation with the frequency of HPV infection and, consequently, a possible influence on faster progression to cervical dysplasia caused by HPV. We consider that inclusion of screening for sexually transmitted infections as monitoring in women with HPV infection could help to find women at risk of cervical cancer progression.

SARS-CoV-2 Seroprevalence and Neutralizing Antibody Response after the First and Second COVID-19 Pandemic Wave in Croatia.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel coronavirus with a pandemic spread. So far, a total of 349,910 SARS-CoV-2 cases and 7687 deaths were reported in Croatia. We analyzed the seroprevalence and neutralizing (NT) antibody response in the Croatian general population after the first (May-July 2020) and second (December 2020-February 2021) pandemic wave. Initial serological testing was performed using a commercial ELISA, with confirmation of reactive samples by a virus neutralization test (VNT). A significant difference in the overall seroprevalence rate was found after the first (ELISA 2.2%, VNT 0.2%) and second waves (ELISA 25.1%, VNT 18.7%). Seropositive individuals were detected in all age groups, with significant differences according to age. The lowest prevalence of NT antibodies was documented in the youngest (<10 years; 16.1%) and the oldest (60-69/70+ years; 16.0% and 12.8%, respectively) age groups. However, these age groups showed the highest median NT titers (32-64). In other groups, seropositivity varied from 19.3% to 21.5%. A significant weak positive correlation between binding antibody level as detected by ELISA and VNT titer (rho = 0.439, p < 0.001) was observed. SARS-CoV-2 NT antibody titers seem to be age-related, with the highest NT activity in children under 10 years and individuals above 50 years.

Genital human papillomavirus infections.

Human papillomavirus (HPV) infections can have clinical presentations from self-limited benign growth in the skin and mucosal epithelia to malignant growth. HPV infects basal epithelial cells (undifferentiated keratinocytes) of the squamous-columnar junction, especially of the cervix. Although today we understand HPV oncogenesis very well, we have very powerful methods of diagnosis, treatment and prevention of HPV related precancerous lesions, however, more than 270,000 women annually die from cervical cancer worldwide. Integrating HPV vaccination with new, more sensitive, cervical screening assays as part of routine preventive care will improve healthcare for all women. The availability of prophylactic HPV vaccines has provided powerful tools for primary prevention of cervical cancer and other HPV-associated diseases. Secondary prevention through primary high-risk HPV (hr-HPV) testing has the potential to further reduce morbidity and mortality of cervical cancer. However, to achieve the maximum benefit of screening, there is need to continue to identify women who are either unscreened or under-screened. Synergies between HPV vaccination and HPV screening is recommended to improve the effectiveness and cost-effectiveness of prevention HPV-related disease.

Acinetobacter baumannii in Southern Croatia: clonal lineages, biofilm formation, and resistance patterns.

BACKGROUND: Acinetobacter baumannii is one of the most prevalent causes of severe hospital-acquired infections and is responsible for the dramatic increase in carbapenem resistance in Croatia in the last 5 years. Such data have encouraged multicenter research focused on the organism's ability to form biofilm, susceptibility to antibiotics, and particular genotype lineage. METHODS: Biofilm formation in 109 unrelated clinical isolates of A. baumannii recovered in six cities of Southern Croatia was investigated. Genotyping was performed by pulsed-field gel electrophoresis and antibiotic profile was tested by applying the disc diffusion method and confirmed by determining the minimum inhibitory concentrations. The ability to form biofilm in vitro was determined from overnight cultures of the collected isolates on microtiter plates, after staining with crystal violet, and quantified at 570 nm after solubilization with ethanol. The statistical relevance was calculated in an appropriate program with level of statistical confidence. RESULTS: There was no significant difference in biofilm formation due to the genotype lineage. Isolates collected from intensive care units (ICUs) and isolated from respiratory samples were more likely to create a biofilm compared with isolates from other departments and other samples. There was a significant difference in the ability to produce biofilm in relation to antibiotic resistance pattern. A large proportion of A. baumannii isolates that were resistant to ampicillin/sulbactam, carbapenems, and amikacin were found to be biofilm-negative. In contrast, isolates susceptible and intermediately susceptible to ampicillin/sulbactam, carbapenems, and amikacin were biofilm producers. CONCLUSION: Clinical isolates of A. baumannii from respiratory samples in ICUs with a particular susceptibility pattern are more prone to form biofilm.

Prevalence and genotyping of the human papillomavirus in the cervical specimens among women of Southern Croatia (Dalmatia County).

AIM: The aim of this study was to evaluate the prevalence of HPV types in the female population with either normal or abnormal Pap smears in Southern Croatia (Dalmatia County) and to evaluate the distribution of HPV types and their association with different cytological findings. METHODS: During one year period, we routinely screened 1,160 women for HR HPV at the Educational Public Health Institute of Split and Dalmatia County, by using Hybrid Capture 2 HPV DNA test. Only HR HPV positive specimens determined by HC2 were further genotyped by polymerase chain reaction (PCR) based assay at the Department of Pathology and Forensic Medicine, Split. RESULTS: Out of the total number of tested women, 406 (35%) of them were positive to HR HPV. The following frequency was observed: HPV 16 in 10.8%, HPV 18 in 7.8%, HPV 31 in 2.3%, HPV 33 in 1.1%, HPV 52 in 0.9%, HPV 45 in 0.6%, and HPV 59 in 0.4% specimens. CONCLUSION: The data obtained from our study indicate that HPV 16 is the most common HPV type found in the cervical specimens among women of Southern Croatia, as in most studies worldwide, followed by HPV 18 and HPV 31. The positivity rates in our study slightly differed for HPV types 18 and 31 from those found in the other studies conducted in Northern Croatia. These suggesting geographic and environmental factors are related to the prevalence of HPV types. We consider it very important to gather information on regional variations in the HPV type spectrum.

An intrauterine device as a possible cause of change in the microbial flora of the female genital system.

AIM: An intrauterine device (IUD), used by millions of women worldwide, is one of the most efficient methods of contraception. The goal of our study was to compare a group of women using the IUD to a control group. MATERIAL & METHODS: The survey included 236 women of fertile age from gynaecological practices in the area of Split and Dalmatia County, Croatia. The subjects were divided into two groups: IUD users and a control group (women not using any contraception methods). Sampling, transportation, sample processing in the laboratory and interpretation of results were conducted using standard microbiological procedures and methods. RESULTS: Opportunistic bacteria were statistically more frequently isolated among IUD users (P < 0.001). The most frequently isolated bacteria in both groups were Escherichia coli and Ureaplasma urealyticum with significantly higher rates in IUD users (P < 0.001). Both colonization and infection had higher rates in IUD users (P < 0.001). There were no significant differences either in the frequency of bacteria isolation regarding different IUD types (P = 0.93), or in relation to duration of IUD use (P = 0.67). CONCLUSIONS: Based on the data in our study IUD users have an increased chance of developing a cervical infection caused by the bacteria Escherichia coli and Ureaplasma urealyticum. Therefore, before IUD insertion women should be screened and treated for asymptomatic vaginal or cervical infections to prevent possible serious IUD-associated infections.

[Can introduction of guidelines for use of an intrauterine device reduce changes in microbial flora of female genital system in IUD-users?]. / Moze li uvodenje postupnika za postavljanje intrauterinoga uloska smanjiti promjenu mikrobioloske flore spolnoga sustava u zena koje ga rabe?

The intrauterine device (IUD), one of the most effective and most common methods of reversible contraception, is used by millions of women worldwide. Although various studies indicate the benefits of its use as a contraceptive method, the greatest concern regarding IUDs is the potential risk for infection. The presence of the IUD gives a solid surface for bacterial attachment and biofilm formation. Infections due to biofilm formation are chronic and difficult to resolve. However, women should be screened and treated for asymptomatic vaginal or cervical infections before IUD insertion. Based on the data in the literature and the data in study conducted in Croatia, it can be concluded that IUD users are at increased risk for development of infection. Treatment strategies in Croatia should take into consideration that Escherichia coli and Ureaplasma urealyticum are dominant causative agents. We conclude that guidelines for the use of IUD in Croatia are a necessity.

Ki-67 quantitative evaluation as a marker of cervical intraepithelial neoplasia and human papillomavirus infection.

OBJECTIVE: To assess the value of Ki-67 quantitative analysis in cervical intraepithelial neoplasia (CIN) in relation to CIN grading and human papillomavirus (HPV) group typing. METHODS: Cervical samples selected retrospectively from 106 cases were analyzed immunohistochemically for Ki-67-positive nuclei in 3 epithelial layers and by polymerase chain reaction for HPV typing. RESULTS: The proportion of high-risk HPV positivity was 0% in normal controls and 30% in CIN 1, 57% in CIN 2, and 90% in CIN 3 groups, and there was no low-risk HPV finding in CIN 2 and CIN 3 cases (P < 0.001). High-risk HPV-positive cases exhibited significantly more Ki-67-positive nuclei per 100-mum basal membrane, which were more frequent in the middle and upper third layers of the epithelium compared with low-risk HPV and HPV-negative cases (P < 0.001). The differences among the CIN groups in the total number and in the percentages of Ki-67-positive nuclei in the lower, middle, and upper third layers of the epithelium were significant (P < 0.001). With the cutoff value of more than 33% Ki-67-positive nuclei in the middle and the upper third layers of the epithelium, Ki-67 staining demonstrated 98.4% sensitivity (60/61 cases) and 97.8% specificity (44/45 cases) for the detection of CIN 2/CIN 3 in our study group. CONCLUSIONS: The Ki-67 immunostaining proved to be predictive for high-risk HPV infection, and it can differentiate reactive lesions from cervical dysplasias. Ki-67 quantitative analysis in 3 epithelial layers is a sensitive and specific method of differentiation between CIN 1 and CIN 2/CIN 3 grades and can be a valuable adjunctive method for more accurate CIN grading.

HPV testing for cervical cancer screening in Croatia.

Opportunistic screening based on the Pap smear has been undertaken in Croatia since 1953. However, cervical cancer remains an important health problem in Croatia when compared to European countries with organised screening programmes. In Croatia, in addition to screening based on well established cytology, Human papillomavirus (HPV) testing is widely used as secondary test as a triage to borderline cytology and as a follow-up after treatment of severe cervical lesions. Many different approaches for HPV testing arose in Croatia over the last decade depending on the needs of each medical institution involved. Presently, there is an urgent need for better networking between the laboratories, the implementation of quality assessment and the adaptation of a uniform system of referring to and reporting of HPV testing. In conclusion, the best possible organisation for HPV testing would be essential for implementation of HPV testing as primary screening test in Croatia, an thus ultimately and hopefully, the more successful cervical cancer control.

Human papillomavirus DNA typing in the cervical specimens among women of Split and Dalmatian County.

Human papillomavirus (HPV) infection is the main cause of cervical cancer, the second most common cancer in women worldwide. More than 200 types of HPV have been described, and within this range more than 40 types attack epithelium of genital tract. The types that were most commonly related to the development of cervical cancer are called high-risk types (HR HPV). There are very few studies about HPV prevalence in Croatia and there is an absolute lack of data for Split and Dalmatian County. Therefore, during a 6 month period, we routinely screened 570 women for HPV DNA at the Educational Public Health Institute of Split and Dalmatian County. HR HPV was detected in cervical exfoliated cells, by using Hybrid Capture 2 HPV DNA test. Out of the total number of tested women, 200 (35%) of them were positive to HR HPV Polymerase chain reaction (PCR) based assays were employed for HR HPV genotyping in positive specimens. The following frequency was observed: HPV 16 in 10%, HPV 18 in 6.1%, HPV 31 in 2.6%, HPV 33 in 1.9%, HPV 52 in 1.4%, HPV 59 in 0.7%, HPV 45 in 0.4% specimens, while 11.9% of tested specimens currently remained untyped. It is necessary to expand this study to a larger number of women, in order to better evaluate genital HPV types distribution among women in this region.

Dermatophytoses in Split and Dalmatia, Croatia, 1996-2002.

The aim of the study was to investigate the epidemiology of dermatophyte infections in Split and Dalmatia County, Croatia, between 1996 and 2002. Of 5792 samples 1318 (23.8%) were positive. Tinea corporis and tinea cruris were the most common types of dermatophytoses. Microsporum canis (36.5%), Trichophyton mentagrophytes (24.8%), and T. rubrum (21.5%) were the main etiologic factors.

Urinary tract infections in South Croatia: aetiology and antimicrobial resistance.

The aim of this study was to determine the distribution of uropathogens isolated from outpatients living in South Croatia and the in vitro susceptibility of these organisms to antimicrobial agents. Of 5080 enrolled uropathogen isolates, 82.28% isolates were Gram-negative, the most frequent isolates being Escherichia coli (62.62%), enterococci (10.18%), Proteus mirabilis (5.31%), Streptococcus agalactiae (3.84%), Staphylococcus spp. (3.70%), Pseudomonas spp. (3.46%), Klebsiella spp. (2.38%). The E. coli resistance rate was 42.17% to amoxycillin, 20.59% to trimethoprim-sulphamethoxazole and 6.09% to norfloxacin. Almost all Klebsiella spp. isolates were resistant to amoxycillin and the resistance rate to trimethoprim-sulphamethoxazole was over 20%, and 14.15% to the fluoroquinolones. A high methicillin-resistance rate was found among S. aureus (61.22%) and coagulase negative staphylococci (41.48).

Tuberculous orchiepididymitis diagnosed by nucleic acid amplification test: a case report.

Symptoms of tuberculous orchiepididymitis in a 39-year-old male started with swelling of left scrotum, followed by fistula formation with suppurative discharge. There was no any improvement produced by antibiotics. Surgical extirpation of inflammatory destroyed testicle and epidydimis was performed. Presence of tubercle bacilli was not shown by bacteriological analysis of testicle tissue. Tuberculous etiology was suggested after histopathological examination of testis and epididymis. Exudate from surgical wound was examined on presence of Mycobacterium tuberculosis DNA. Etiology of orchiepididymitis was proved by positive assay and inflammatory process was completely cured by antituberculotics therapy. By this report it was clearly shown that sometimes only molecular methods could confirm etiology of inflammatory process.