Comparative analysis of chemical profiles, antioxidant, antibacterial, and anticancer effects of essential oils of two Thymus species from Montenegro.

The essential oils of Thymus vulgaris (TVEO) and Thymus serpyllum (TSEO) show different biological activities. The aim of the study was to evaluate the biological activities of TVEO and TSEO from Montenegro. The main components of TVEO were p-cymene (29.52%), thymol (22.8%) and linalool (4.73%) while the main components of TSEO were p-cymene (19.04%), geraniol (11,09%), linalool (9.16%), geranyl acetate (6.49%) and borneol (5.24%). Antioxidant activity determined via DPPH for TVEO was 4.49 and FRAP 1130.27, while for TSEO it was estimated that DPPH was 4.88 µL/mL and FRAP was 701.25 µmol FRAP/L. Both essential oils were active against all tested bacteria, with the highest level of sensitivity of E. coli with MIC of 1.5625 µL/mL. Essential oils showed strong cytotoxic effects on human cancer cell lines, with IC50 values ranging from 0.20 to 0.24 µL/mL for TVEO and from 0.32 to 0.49 µL/mL for TSEO. TVEO caused apoptosis in cervical adenocarcinoma HeLa cells through activation of caspase-3 and caspase-8, while TSEO caused apoptosis through caspase-3. EOs decreased levels of oxidative stress in normal MRC-5 cells. HeLa cells treated with TVEO had reduced MMP2 expression levels, while cells treated with TSEO had lowered MMP2 and MMP9 levels. The treatment of HeLa cells with TVEO increased the levels of miR-16 and miR-34a, indicating potential tumor-suppressive properties. Our findings suggest that Thymus essential oils may be considered as good candidates for further investigation as cancer-chemopreventive and cancer-therapeutic agents.

Biowaste as a Potential Source of Bioactive Compounds-A Case Study of Raspberry Fruit Pomace.

Raspberry fruit pomace, a byproduct of juice production, was studied as a potential source of antioxidant compounds. Target high-performance liquid chromatography analysis of important polyphenolic compounds (gallic, p-coumaric, caffeic, quercitrin, chlorogenic, and ellagic acid) was performed together with analysis of total phenolic content (TPC), total flavonoid content (TFC), total anthocyanins content (TAC), and antioxidant capacity (via ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays). The differences in polyphenolic content of Rubus idaeus L. pomace were evaluated following ultrasound-assisted extraction and conventional maceration with different organic solvents. Additionally, the yield of free phenolics was measured in hydrolyzed pomace extracts. The results obtained show that the ultrasound method maximizes the quantity of antioxidant compounds in terms of TPC (27.79 mg/L gallic acid equivalent (GAE)), TFC (8.02 mg/g quercetin equivalent (QE)), TAC (7.13 mg/L cyanidine-3-glucoside equivalent (C3G Eq)), caffeic (19.17 µg/g), chlorogenic (3.56 µg/g), ellagic (105.52 µg/g), and gallic acids (8.75 µg/g), as well as FRAP (1002.72 µmol/L) and DPPH assays (969.71 µmol/mL vitamin C equivalent (vit C Eq); 567.00 µmol/100 g Trolox equivalent (TE)). On the other hand, conventional maceration maximizes the yield of quercetin and p-coumaric acid. In terms of biowaste valorization, raspberry fruit pomace has a promising industrial potential and may prove to be useful in the development of antioxidant dietary supplements.

Long-term stability of the urogenital microbiota of asymptomatic European women.

BACKGROUND: To date, information on healthy female urinary microbiota is available mostly at genus level and at one time point. However, profound species-level characterization of healthy urinary microbiome and its stability over time are essential for further correct interpretation of its role in healthy urogenital tract. In this study, we investigated female urogenital microbiome (FUM) at two timepoints (within 2.5-year interval) in young asymptomatic European women. We used culturomics with accurate isolates' identification (MALDI-TOF MS and gene markers sequencing) to understand species stability within healthy FUM. RESULTS: Extended culturomics of voided midstream urine sample pairs revealed a mean Shannon diversity index of 1.25 and mean of 19 species/sample (range 5-39 species; total of 115 species; 1830 isolates). High overall species variability between individuals was captured by beta diversity and a variety of community structure types, with the largest cluster characterized by Lactobacillus crispatus, often in combination with Gardnerella vaginalis or Gardnerella genomospecies 3. Significant FUM composition differences, related to Finegoldia magna and Streptococcus anginosus, according to smoking status were found. A high species variability within individuals (Shannon index SD > 0.5 in 7 out of 10 sample pairs) with a mean of 29% of shared species (range 9.1-41.7%) was observed. Moreover, 4 out of 10 sample pairs clustered in the same community structure type. The stable FUM sample pairs presented high abundance of Lactobacillus crispatus, Streptococcus agalactiae or Lactobacillus paragasseri and Bifidobacterium spp.. Moreover, Gardnerella vaginalis, Gardnerella genomospecies 3 or Gardnerella swidsinskii were often maintained within individuals in high abundance. CONCLUSIONS: Shift in species composition at two distant timepoints was frequently observed among urogenital microbiome of European asymptomatic women. This suggests possible interchange of particular species in healthy FUM and the existence of multiple health-associated FUM compositions in certain individuals. Additionally, we provided additional evidence on resilience of particular bacterial communities and identified certain species more prone to persist in urogenital tract. This study revealed important details on the FUM composition complexity relevant for studies aiming to understand microbiota role in the urogenital tract health and for identification of eubiotic and dysbiotic FUM.

The association of hs-CRP and fibrinogen with anthropometric and lipid parameters in non-obese adolescent girls with polycystic ovary syndrome.

Background The aim of the study was to evaluate high-sensitivity C-reactive protein (hs-CRP) and fibrinogen in non-obese normoinsulinemic adolescent girls with polycystic ovary syndrome (PCOS) and their relationship with anthropometric and lipid parameters. Methods The study comprised a total of 26 adolescent girls newly diagnosed with PCOS and 12 healthy controls with regular ovulatory menstrual cycles. The concentration of hs-CRP, fibrinogen, anthropometric measurements, and biochemical and hormonal testing were assessed. Results PCOS adolescent girls had significantly higher levels of hs-CRP and fibrinogen compared to healthy controls. In univariate regression analysis, statistically significant associations of hs-CRP and fibrinogen levels of PCOS patients have been shown with body mass index (BMI), waist circumference (WC), hip circumference (HC) and low-density lipoprotein (LDL)/high-density lipoprotein (HDL) ratio, while hs-CRP levels were also associated with cholesterol and LDL. In the multivariate regression model, we found that hs-CRP levels were predicted by BMI (ß=0.541, p<0.001) and LDL (ß=0.507, p=0.014), while fibrinogen levels were predicted by BMI (ß=0.449, p=0.004). Conclusions We have shown an association of proinflammatory indices hs-CRP and fibrinogen with anthropometric and lipid parameters of adolescent women with PCOS. The inflammatory markers might be useful in monitoring normal-weight adolescent women with PCOS in an effort to timely prevent unfavorable changes in body mass and lipid profile.

Ovarian Preservation from Tumors and Torsions in Girls: Prospective Diagnostic Study.

STUDY OBJECTIVE: To develop a new decision tree system (DTS) for the management of adnexal masses in prepubertal and adolescent girls, aimed to improve the distinction between benign and malignant masses, help preserve affected ovaries during surgery, and reduce the rate of surgical management of uncomplicated functional ovarian cysts. DESIGN: A prospective diagnostic study using clinical and ultrasound data collected for all patients younger than 19 years of age with adnexal masses managed between 2006 and 2015. SETTING: Mother and Child Health Institute of Serbia 'Dr Vukan Cupic' (Belgrade, Serbia). PARTICIPANTS: Patients (N = 1499) with adnexal masses, of whom 318 were surgically treated. INTERVENTIONS AND MAIN OUTCOME MEASURES: Ultrasonographic characteristics (Ueland's Morphology Index [MI] and the ovarian crescent sign [OCS]). Patients were managed expectantly or surgically, in line with 3 rules of the DTS. Rule 1: asymptomatic patients having a mass with MI of 4 or less and OCS present, were managed expectantly. Rule 2 (emergency): malignancy was suspected if the MI was 7 or more and no edema of the OCS was present. Rule 3 (nonemergency): malignancy was suspected if the OCS was absent and MI was 5 or more. The diagnostic accuracy was assessed using sensitivity and specificity (P < .05). RESULTS: No malignancy was found in the group of 1236 patients selected according to the DTS rule 1. Torsion was confirmed in 36% of surgically treated masses (n = 115). The OCS was present in 96% of benign masses in the non-emergency group (n = 149) and in three with microscopic malignancy. In predicting malignancy, the DTS (rules 2 and 3) showed a sensitivity of 93 (95% confidence interval [C.I.]: 82-98); and a specificity of 97 (C.I.: 95-99). Ovarian tissue was preserved from benign (n = 254, 93%) and malignant tumors (n = 3, 7%). Only five (2%) uncomplicated ovarian cysts were surgically treated. CONCLUSION: The DTS with 3 rules is a very accurate diagnostic tool in the differentiation between benign and malignant ovarian masses. The DTS rule 1 reduces the number of surgical procedures on functional cysts, rules 2 and 3 are very useful in choosing the optimal treatment of adnexal masses, whether or not they are twisted.