P16/Ki67 Dual Staining in Glandular Cell Abnormalities of the Uterine Cervix.

Very limited information exists about the role of p16/Ki67 dual staining on glandular cells in detecting glandular precancerous lesions and cervical adenocarcinoma. In this study, we investigated the diagnostic accuracy of p16/Ki67 dual staining for the detection of glandular and squamous lesions on the uterine cervix and for cancer of the upper reproductive tract. We performed a retrospective analysis of prospectively collected data on 96 patients with glandular cell abnormalities. We analyzed the diagnostic accuracy of p16/Ki67 dual staining for atypical glandular cells, not otherwise specified (AGC-NOS); atypical glandular cells, favor neoplastic (AGC-FN); adenocarcinoma in situ (AIS); and A-CA (cervical adenocarcinoma). A separate analysis for the detection of squamous precancerous lesions and squamous-cell carcinoma (CIN3+) and for cancer of the upper reproductive tract (EC/OC) was performed. Among patients who had normal histology or a low-grade lesion on final analysis, only 8.5% had positive dual staining. On the other hand, 85.7% of patients with AIS+ on final histology had positive dual staining. The respective specificities of p16/Ki67 dual staining on AGC-NOS for the detection of AIS+ (adenocarcinoma in situ or cervical adenocarcinoma), CIN3+ and EC/OC were 91.5%, 88.7% and 86.4%. High specificity values of p16/Ki67 dual staining on cervical smears labelled as AGC-NOS for the detection of CIN3+ and AIS+ suggest that this method might be a useful addition in cervical cancer screening.

The Role of p16/Ki67 Dual Staining in Cervical Cancer Screening.

Cervical cancer screening has enabled a decrease in the incidence and mortality of cervical cancer. Various screening modalities have been studied to date. In many countries, screening is still based on cervical cytology, where cervical cells obtained either on glass or in a liquid medium are examined under a microscope. However, the fact that the vast majority of cervical cancers are a result of persistent infection with high-risk human papillomaviruses (hr-HPV) has led to the implementation of primary HPV screening in many countries. Taking into consideration the fact that the majority of HPV infections are transient and do not cause cervical precancer, effective triage methods are needed to prevent an increase in colposcopy referrals. Among these, the most extensively investigated are HPV genotyping, HPV methylation, and p16/Ki67 dual staining. In this manuscript, we briefly summarize the current knowledge regarding different screening strategies for the prevention of cervical cancer, with a focus on p16/Ki67 dual staining. In addition, we provide an explanation regarding the rationale for the use of various screening modalities based on the molecular biology of cervical cancer and cervical precancerous lesions.

The Role of Methylation of Host and/or Human Papillomavirus (HPV) DNA in Management of Cervical Intraepithelial Neoplasia Grade 2 (CIN2) Lesions.

Cervical intraepithelial neoplasia grade 2 (CIN2) is an intermediate stage between CIN 1, which is a low-grade lesion, and CIN3, which is the immediate precursor of cervical cancer (CC). Traditionally, CIN2 was regarded as a high-grade lesion and was treated with conization or ablative methods. In recent years, there has been a shift in the management of younger patients, who are now more often being managed conservatively due to frequent spontaneous CIN2 regression and possible adverse effects of treatment on future pregnancies. Because the risk of progression to CC still exists with conservative management, a personalized approach is needed to identify patients with a higher probability of progression. In this regard, research has focused on the role of host and human papillomavirus (HPV) gene methylation. This systematic review summarizes the current knowledge regarding conservative CIN2 management focusing on the main methylation markers and its implementation in conservative CIN2 management, and it describes major ongoing longitudinal studies on the subject. The review showed that DNA methylation is an accurate predictor of disease progression and a valid triage tool for HPV-positive women, with CIN2 performing better than triage cytology. Because virtually all CCs are methylation-positive, methylation-negative women at baseline have an extremely low risk of CC.

Comparison of Conservative Treatment of Cervical Intraepithelial Lesions with Imiquimod with Standard Excisional Technique Using LLETZ: A Randomized Controlled Trial.

(1) Background: There are limited data on the success of conservative treatment of high-grade cervical squamous intraepithelial lesions (HSIL) with imiquimod directly compared to standard of treatment with LLETZ. (2) Methods: Patients aged 18-40 with histological HSIL (with high-grade cervical intraepithelial neoplasia, CIN2p16+ and CIN3), were randomly assigned to treatment with imiquimod or LLETZ. The primary outcome was defined as the absence of HSIL after either treatment modality. The secondary outcomes were the occurrence of side effects. (3) Results: 52 patients were allocated in each group and were similar regarding baseline characteristics. In the imiquimod group, 82.7% of patients completed treatment, which was successful in 51.9%. All patients in the LLETZ group completed treatment, which was successful in 92.3% (p < 0.001). In the subgroup of CIN2p16+ patients, treatment with imiquimod was not inferior to LLETZ (73.9% vs. 84.2%, p = 0.477). During and after treatment, no cases of progression to cancer were observed. Side effects and severe side effects (local and systemic) were more prevalent in the imiquimod than in the LLETZ group (88.5% vs. 44.2% (p-value < 0.001) and 51.9% vs. 13.5% (p-value < 0.001), respectively). (4) Conclusion: Generally, in patients with HSIL, LLETZ remains the gold standard of treatment. However, in a subgroup analysis of patients with CIN2p16+, the success rate was comparable between the two treatment modalities. Due to the prevalence of side effects, the treatment compliance with imiquimod use may, however, present a clinically important issue.

Diagnostic performance of p16/Ki-67 dual immunostaining at different number of positive cells in cervical smears in women referred for colposcopy.

BACKGROUND: The aim of the study was to evaluate the diagnostic accuracy of p16/Ki-67 dual immunostaining (p16/ Ki-67 DS) in cervical cytology and the number of positive p16/Ki-67 cells to diagnose high grade cervical intraepithelial neoplasia (CIN2+) in colposcopy population. SUBJECTS AND METHODS: We performed an analysis on a subset cohort of 174 women enrolled within a large-scale randomised controlled human papillomavirus (HPV) self-sampling project organised as part of the population-based Cervical Cancer Screening Programme ZORA in Slovenia. This subset cohort of patients was invited to the colposcopy clinic, underwent p16/Ki-67 DS cervical cytology and had the number of p16/Ki-67 positive cells determined. RESULTS: Among analysed women, 42/174 (24.1%) had histologically confirmed CIN2+. The risk for CIN2+ was increasing with the number of positive cells (p < 0.001). The sensitivity of p16/Ki-67 DS for detection of CIN2+ was 88.1%, specificity was 65.2%, positive predictive value was 44.6% and negative predictive value was 94.5%. CONCLUSIONS: Dual p16/Ki-67 immunostaining for the detection of CIN2+ has shown high sensitivity and high negative predictive value in our study, which is comparable to available published data. The number of p16/Ki-67 positive cells was significantly associated with the probability of CIN2+ detection. We observed a statistically significant and clinically relevant increase in specificity if the cut-off for a positive test was shifted from one cell to three cells.

Identification of novel diagnostic biomarkers in endometrial cancer using targeted metabolomic profiling.

PURPOSE: Endometrial cancer (EC) is the most common gynecological malignancy with high disease burden especially in advanced stages of the disease. Our study investigated the metabolomic profile of EC patient's serum with the aim of identifying novel diagnostic biomarkers that could be used especially in early disease detection. MATERIAL AND METHODS: Using targeted metabolomic serum profiling based on HPLC-TQ/MS, women with EC (n â€‹= â€‹15) and controls (n â€‹= â€‹21) were examined for 232 endogenous metabolites. RESULTS: Top performing biomarkers included ceramides, acylcarnitines and 1-methyl adenosine. Top 4 biomarkers combined achieved 94% sensitivity with 75% specificity with AUC 92.5% (CI 90.5-94.5%). Individual markers also provided significant predictive values: C16-ceramide achieved sensitivity 73%, specificity 81%, AUC 0.83, C22-ceramide sensitivity 67%, specificity 81%, AUC 0.77, hydroxyhexadecenoylcarnitine sensitivity 60%, specificity 96%, AUC 0.76 and 1-methyladenosine sensitivity 67%, specificity 81%, AUC 0.75. The individual markers, however, did not reach the high sensitivity and specificity of the 4-biomarker combination. CONCLUSIONS: Using mass spectrometry targeted metabolomic profiling, ceramides, acylcarnitines and 1-methyladenosine were identified as potential diagnostic biomarkers for EC. Additionally, these identified metabolites may provide additional insight into cancer cell metabolism.

P16/Ki-67 Immunostaining in the Triage of Postmenopausal Women With Low-Grade Cytology Results.

OBJECTIVE: The interpretation of postmenopausal smears and the gynecological treatment of these patients can often be difficult. The objective of this study was to assess the performance of p16/Ki-67 dual-stained cytology as a triage of atypical squamous cells of undetermined significance and low-grade intraepithelial lesion cytology results in postmenopausal women. METHODS: All consecutive atypical squamous cells of undetermined significance and low-grade intraepithelial lesion smears in 1-year period were collected and p16/Ki-67 immunostaining was performed retrospectively. The results were compared with histology results or long-term cytology follow-up in cases with no biopsy. RESULTS: The sensitivity of p16/Ki-67 immunostaining for the detection of cervical intraepithelial neoplasia (CIN) 2 and CIN 3 was 57.1% and 85.0%, respectively. The specificity for the detection of CIN 2 was 94.3% and CIN 3 92.4%. Negative predictive values for the detection of CIN 2 and CIN 3 were 96.3% and 99.6%, respectively. CONCLUSIONS: Dual p16/Ki-67 immunostaining is a useful additional method in postmenopausal patients with low-grade cytology. Considering the high specificity and negative predictive value in our study, we believe that it could be helpful in avoiding unnecessary referrals to colposcopy and thus reduce the cost of the program.

Vitamin D and Ovarian Cancer: Systematic Review of the Literature with a Focus on Molecular Mechanisms.

Vitamin D is a lipid soluble vitamin involved primarily in calcium metabolism. Epidemiologic evidence indicates that lower circulating vitamin D levels are associated with a higher risk of ovarian cancer and that vitamin D supplementation is associated with decreased cancer mortality. A vast amount of research exists on the possible molecular mechanisms through which vitamin D affects cancer cell proliferation, cancer progression, angiogenesis, and inflammation. We conducted a systematic review of the literature on the effects of vitamin D on ovarian cancer cell.

The Association of Vitamin D Levels with Common Pregnancy Complications.

The association between vitamin D deficiency and various adverse pregnancy outcomes has been extensively investigated in recent years. The pregnant woman is the only source of vitamin D for the foetus. The main sources of vitamin D for pregnant women are sunlight, fortified dairy products, oily fish and dietary supplements. Vitamin D deficiency during pregnancy has been associated with some adverse neonatal outcomes as well as an increased risk of late pregnancy complications. The outcomes of the published studies investigating preeclampsia and gestational diabetes mellitus vary with some large trials suggesting a potential positive effect of vitamin D supplementation during pregnancy on the decreased risk of these complications. Research also suggests a possible connection between lower vitamin D concentrations and increased risk of preterm labour. In our manuscript, we aim to review the existing literature regarding the prevalence of vitamin D deficiency during pregnancy, the factors associated with vitamin D deficiency, and possible pregnancy complications arising from it.

Incidence of positive peritoneal cytology in patients with endometrial carcinoma after hysteroscopy vs. dilatation and curettage.

BACKGROUND: The aim of the study was to compare the frequency of positive peritoneal washings in endometrial cancer patients after either hysteroscopy (HSC) or dilatation and curettage (D&C). PATIENTS AND METHODS: We performed a retrospective analysis of 227 patients who underwent either HSC (N = 144) or D&C (N = 83) and were diagnosed with endometrial carcinoma at the University Medical Centre Maribor between January 2008 and December 2014. The incidence of positive peritoneal cytology was evaluated in each group. RESULTS: There was no overall difference in the incidence of positive peritoneal washings after HSC or D&C (HSC = 13.2%; D&C = 12.0%; p = 0.803). However, a detailed analysis of stage I disease revealed significantly higher rates of positive peritoneal washings in the HSC group (HSC = 12.8%; D&C = 3.4%; p = 0.046). Among these patients, there was no difference between both groups considering histologic type (chi-square = 0.059; p = 0.807), tumour differentiation (chi-square = 3.709; p = 0.156), the time between diagnosis and operation (t = 0.930; p = 0.357), and myometrial invasion (chi-square = 5.073; p = 0.079). CONCLUSIONS: Although the diagnostic procedure did not influence the overall incidence of positive peritoneal washings, HSC was associated with a significantly higher rate of positive peritoneal cytology in stage I endometrial carcinoma compared to D&C.

Determinants of maternal vitamin D concentrations in Slovenia : A prospective observational study.

OBJECTIVE: To analyze the vitamin D status of pregnant women in Slovenia and the factors influencing it. METHODS: The study was performed in Maribor University Medical Centre (location 46°N). Maternal serum 25-hydroxy-vitamin D (25(OH)D) in 4 groups of 100 pregnant women in 4 different seasons of the year was measured at the time of delivery. Data on life style, eating habits, sunbathing and intake of vitamin D supplements during pregnancy were obtained using a questionnaire. Information on maternal outcome was acquired from medical records. Duration of sunlight and average temperature in the month preceding delivery were calculated from meteorological data. RESULTS: The average maternal 25(OH)D concentration ranged from 28.5 ± 17.1 nmol/l in March to 54.8 ± 24.1 nmol/l in June (p < 0.001). Severe vitamin D deficiency, i.e. <25 nmol/l 25(OH)D was present in 23.6% of women overall. In multiple regression analysis supplements containing vitamin D (ß = 0.225, p < 0.001) and duration of intentional exposure to sunlight (ß = 0.192, p = 0.026) were associated with maternal 25(OH)D status. Significantly lower vitamin D levels were observed in women who had a cesarean section (t = 2.053, p = 0.041) and those with premature delivery (t = 2.296, p = 0.022). Vitamin D levels were not associated with the occurrence of gestational diabetes, gestational hypertension or infections during pregnancy. CONCLUSION: A high prevalence of vitamin D deficiency was detected at the end of pregnancy, especially in the months following autumn and winter and in women who did not take supplements containing vitamin D.

Prognostic Role of HER2 Status and Adjuvant Trastuzumab Treatment in Lymph Node-Negative Breast Cancer Patients - a Retrospective Single Center Analysis.

BACKGROUND: The natural course of traditionally prognostically unfavorable human epidermal growth factor receptor 2 (HER2)-positive breast cancer has been changed by anti-HER2 therapy. It is not clear whether the prognosis for HER2-positive patients treated with adjuvant trastuzumab differs from that of HER2-negative patients. METHODS: We performed a retrospective study including patients with lymph node-negative invasive breast cancer treated at our institution in the period 2000-2009. Disease-free (DFS) and overall survival (OS) were calculated using the Kaplan-Meier method. The Cox proportional hazards model was applied to control for other clinically important variables. RESULTS: Median follow-up was 90-109 months. The 5-year DFS rates for HER2-negative patients, HER2-positive patients without adjuvant trastuzumab and trastuzumab-treated HER2-positive patients were 88.1% (95% confidence interval (CI) 85.6-90.6%), 73.1% (95% CI 64.3-81.9%) and 90.7% (95% CI 83.1-98.3%), respectively. No significant difference in DFS was observed between trastuzumab-treated HER2-positive patients and HER2-negative patients in multivariate analysis (hazard ratio 1.15; 95% CI 0.53-2.46; p = 0.728). There were no differences in OS among the 3 groups. CONCLUSION: Based on our results, the negative prognostic effect of HER2 positivity seen before targeted anti-HER2 treatment has completely disappeared in the era of routine trastuzumab administration in the adjuvant setting.

Treatment of vulvovaginal candidiasis: a review of the literature.

Vulvovaginal candidiasis (VVC) affects around three-quarters of all women during their reproductive age, although the exact incidence of VVC is difficult to determine because many patients are self-treated. The infections are divided into complicated and uncomplicated. Uncomplicated VVC is most effectively treated with local azoles. Oral treatment with a single dose of fluconazole is also effective for treating uncomplicated VVC. Treatment of complicated VVC is prolonged and most commonly consists of multiple doses of oral fluconazole or at least 1 week of local azoles. The role of probiotics in treating VVC is still disputed. This article presents a review of the literature on the various treatment options for VVC. Treatment for the most common pathogens that cause complicated VVC is also discussed.

Seasonal variations of vitamin D concentrations in pregnant women and neonates in Slovenia.

OBJECTIVE: While foreign research shows a high prevalence of vitamin D deficiency in pregnant women and consequently in neonates, we do not have any data on vitamin D concentration in these risk groups for Slovenia. We performed a prospective study to evaluate vitamin D concentration in pregnant women and neonates in Maribor region. STUDY DESIGN: We determined 25-hydroxy-vitamin D concentration from blood samples taken before delivery from 100 pregnant women who gave birth in Maribor University Clinical Centre in September and December 2013, respectively, and from the cord blood of their neonates. We collected data on nutrition and sun exposure during pregnancy. We calculated the vitamin D concentrations in pregnant women and neonates according to season of birth and use of nutrition supplements, determined the vitamin D levels in some pregnancy complications and checked the correlation of maternal and neonatal vitamin D concentrations. RESULTS: The average vitamin D concentration in the September group was 54.3±25.2nmol/L, and in the December group 33.3±18.6nmol/L (p<0.001). Optimal vitamin D concentration (>80nmol/L) was reached by 12.0% of pregnant women in September and by only 2.0% in December. Women who took nutrition supplements containing vitamin D during pregnancy had significantly higher vitamin D levels than those who did not (September 68.9±27.0nmol/L vs. 46.5±20.3nmol/L, p<0.001; December 38.7±17.9nmol/L vs. 30.2±18.4nmol/L, p=0.028). Neonates had higher average levels of vitamin D than their mothers but there was a good correlation between maternal and neonatal vitamin D values. CONCLUSION: Vitamin D deficiency is very common in pregnant women in Slovenia as well, especially in winter and in those women who do not take nutrition supplements containing vitamin D.

The role of intraoperative ultrasound in breast-conserving surgery of nonpalpable breast cancer.

OBJECTIVE: This retrospective study was undertaken to evaluate the role of intraoperative ultrasound in breast-conserving surgery of nonpalpable breast cancer. METHODS: The data about 125 patients with preoperatively diagnosed breast cancer operated with the help of intraoperative ultrasound between January 2009 and December 2012 were retrospectively analyzed. RESULTS: The mean volume of the excised tissue was 42.1 cm(3) (± 22.0 cm(3)). Immediate re-excision after ultrasound examination of the excised tissue was performed in 27 patients (21.6 %). Secondary procedure owing to positive histological margins was necessary only in four patients (3.2 %). CONCLUSIONS: Intraoperative ultrasound offers some advantages in comparison with other localization techniques for nonpalpable breast cancer surgery.

Long term results of operative treatment of knee osteochondritis dissecans.

PURPOSE: This retrospective study was designed to evaluate the hypothesis that open reduction internal fixation (ORIF) of an unstable osteochondritis dissecans (OCD) lesion results in a high percentage of acceptable outcomes at long-term follow-up. METHODS: Eight patients who had been treated with ORIF for unstable knee OCD lesions were identified. Long-term outcomes were assessed with a Subjective Knee Evaluation Form and Knee Examination Form of the International Knee Documentation Committee (IKDC 2000) and an OCD specific Hughston rating scale. RESULTS: All lesions were healed and stable to arthroscopic probing at 3 months. At the average follow-up of 14.8 years (range 12-21 years), the mean IKDC subjective score was 80.9. In IKDC examination form, six patients reached group A (normal) and two patients reached groups B (near normal) and C (abnormal), respectively. According to Hughston criteria, three patients were rated excellent, three good, one fair, and one poor. CONCLUSIONS: We recommend aggressive attempts to preserve articular cartilage in OCD.

The role of genital pathogens in morbidity following diathermy loop excision of the transformation zone of the uterine cervix.

OBJECTIVE: To evaluate the role of genital pathogens in postoperative morbidity following diathermy loop excision of the transformation zone (LETZ) of the uterine cervix. METHODS: Patients with cervical intraepithelial neoplasia (CIN) who underwent diathermy LETZ were included in a prospective study. Cervical swabs for genital pathogens were collected before the diathermy procedure. After surgery, women were followed-up regarding the occurrence and severity of postoperative pain, bleeding, and discharge. RESULTS: Genital pathogens were present in 463 of 788 (58.8%) cases. The most frequently isolated groups of microorganisms were group B ß-hemolytic Streptococcus, α-hemolytic Streptococcus, Enterococcus species, and coliforms. In patients with genital pathogens, postoperative pain was present in 298 (37.8%), discharge in 262 (33.2%), and bleeding in 236 (29.9%) cases. In patients without genital pathogens, 199 (25.3%) experienced pain, 181 (23.0%) discharge, and 143 (18.1%) bleeding. The differences between the 2 groups were not statistically significant (χ(2) for pain=0.675; χ(2) for discharge=0.031; χ(2) for bleeding=3.444; P>0.05). CONCLUSION: Genital pathogens are very common among patients with CIN and do not affect the occurrence or severity of pain, discharge, or postoperative bleeding after diathermy LETZ.